ISSN 1443-0193 Australian Biochemist The Magazine of the Australian Society for Biochemistry and Molecular Biology Inc. Volume 48 APRIL 2017 No.1

SHOWCASE ON RESEARCH Figure 1 Lactation

Stem cell

Common progenitor

Luminal Myoepithelial progenitor progenitor

Myoepithelial Alveolar Ductal

THIS ISSUE INCLUDES

Showcase on Research Other Features  Appetite Control  Great Expectations  Science Talent Search Hormones in Human Milk  SDS (Students) Page  Honours for Members  Cells in Human Milk –  Competition  Special Interest Groups What Do They Tell Us  Travel Report  Our Sustaining Members  Human Milk Bioactivity  FAOBMB  Forthcoming Meetings  Pesticides in Human Milk  Intellectual Property  Directory

INSIDE Profiles of ASBMB Medallists and Awardees Vol 48 No 1 April 2017 AUSTRALIAN BIOCHEMIST Page 1 ‘Chemical Cryptogram’ Competition

We have another competition for the readers of the Australian Biochemist. All correct entries received by the Editor (email [email protected]) before 14 May 2017 will enter the draw to receive a gift voucher. With thanks to Rebecca Lew.

Solve the coded words below to reveal a special list, described in the underlined title. Each letter codes for another letter – to get you started, the coded letters RPAUMS = BLONDE. The dates are hints.

DAYSU RBAFQSYBJEJ DQA DAU UARSP KCBLSJ 1. VSCET FACB (1947) 2. MACAEQT QAMVGBU (1964) 3. VSCECXMS SPBAU (1988) 4. SPBLIRSEQ RPIFGRXCU (2009) 5. IMI TAUIEQ (2009)

Showcase on Research Great Expectations Technical Features SDS (Students) Page Coverage of all issues from 2000 to the present

Page 2 AUSTRALIAN BIOCHEMIST Vol 48 No 1 April 2017 From the Editor’s Desk

As we head towards Autumn, it isn’t just the leaves and the weather that are changing; change is also coming to the Australian Biochemist, in the form of a new editorial team. Beginning with the next issue, Suresh Mathivanan and Tatiana Soares da Costa will be taking over the reins from me. As you well know, the Australian Biochemist has already undergone some changes, most notably the transition to an electronic only format. Under the guidance of Suresh and Tatiana, I have no doubt that the magazine will improve by leaps and bounds, as they take the magazine further forward into the digital era. Suresh’s expertise with technology (evident from his leadership in revamping the ASBMB website) combined with Tatiana’s fresh perspective (having written our Short Discussions for Students Page for the last few years) will ensure that the magazine remains a relevant and an effective tool to meet the communication needs of our membership. As part of these changes, this issue will feature our very last Showcase on Research, a feature that has been running in the magazine for at least 19 years. Thankfully, we will be finishing on a high note, with a fascinating feature on human breastmilk. Put together by Donna Geddes and her team of authors, the Showcase will explore several topics surrounding this powerhouse solution of nutrients, cells and bioactive Chu Kong Liew molecules, which has effects not only on the developing infant but also on its life Editor Australian Biochemist further down the track. One feature that isn’t going anywhere (at least for now) is Great Expectations, with this issue’s feature written by Rebecca Lew. As a former Editor of the Australian Biochemist, Rebecca is certainly used to organising and editing Great Expectations but in this issue, she proves herself equally adept at writing an interesting piece too! In her feature, Becky takes us on her journey from biology-loving child in the US to senior lecturer at Monash University and now a senior medical writer. Following on from her article on the considerations behind filing a patent, Sarah Hennebry describes the sequence of events that take place once a patent been filed. Tatiana Soares da Costa shares some valuable advice on writing and publishing a manuscript. Rounding off this issue are the profiles of the ASBMB award winners, travel award and Special Interest Group reports, and Australia Day Honours for ASBMB members. As I step down from my role, I’d like to acknowledge all the wonderful help that I’ve received from the Editorial Committee during my time as editor. First and foremost, I’d like to thank Liana Friedman, our Editorial Officer, for her skill, patience and dedication to putting out the highest quality publication with every issue. It is difficult to overstate her contribution to the success of the magazine but I know that with Liana committed to staying on, the Australian Biochemist is in great hands! Thanks go to Rebecca Lew, who introduced me to the job and has provided valuable guidance over the years. I’d also like to thank the other members of the Editorial Committee, both past and present for kindly giving up their time to the magazine. Many thanks also to Sally and Chris Jay, particularly to Sally for her tireless efforts in serving as the nexus between the magazine and our Sustaining Members. Thanks also to the ASBMB Executive Council for their continual help and advice. Last but not least, a big thank you to all our contributors, who generously provide the wonderful content that fills these pages. Chu Kong Liew

Would you like to help out with the Australian Biochemist?

Then please join our Editorial Committee! If you have an interest in being part of the magazine, we warmly welcome your involvement. To volunteer or to find out more about what is involved, please contact the Editor at [email protected].

Australian Biochemist - Editor Chu Kong Liew, Editorial Officer Liana Friedman © 2017 Australian Society for Biochemistry and Molecular Biology Inc. All rights reserved.

Vol 48 No 1 April 2017 AUSTRALIAN BIOCHEMIST Page 3 SHOWCASE ON RESEARCH

EDITORIAL Lactation in the Limelight Recently, there has been widespread recognition infants self-regulate their milk intake and that human among the scientific community of the programming of milk contains bioactive appetite control factors, it is long-term health outcomes during pregnancy and the compelling to deduce that both the composition of milk early years of life. Indeed, it is believed that the first and the volume consumed may be related to infant 1000 days of life is the most sensitive window of human growth and body composition. Gridneva and Geddes development. Epidemiologic and interventional animal explore this scenario with a complex analysis of human models provide compelling evidence that nutrition milk composition, carried out with respect to gastric profoundly affects organ development and metabolism, emptying (a key appetite regulator), infant growth and with consequences extending into adult life. body composition over the first year of life. In a landmark series recently published in The Lancet, Often overlooked is the fact that human milk is an extensive meta analysis confirms the significant a ‘live potion’ that contains amongst its bioactive advantages of breastfeeding, which include decreased components, live cells. Most recently, immune cells in childhood infections and malocclusion, likely milk have been shown to increase in response to both reductions in overweight and diabetes later in life infant and maternal infection, highlighting the intimate along, as well as increased intelligence (1). However, relationship between the mother and infant. Further, the mechanisms by which breastfeeding confer a full cellular hierarchy has also been identified, with advantages remain to be elucidated. human milk stem cells being shown to be pluripotent Further, whilst the benefits to the infant are often cited and able to develop into numerous cells types when to encourage breastfeeding, the lactating woman also placed in stimulating environments. Twigger and reaps substantial health benefits, such as decreased Hartmann discuss the potential of these cells in helping risk of breast and ovarian cancer and type 2 diabetes to elucidate the development of the human mammary (1). It has been predicted that if breastfeeding were to gland during lactation as well as the possibile discovery be scaled up to near universal levels, 823,000 deaths of of biomarkers for dysfunction and disease. children under five years of age would be prevented, Sharp et al. discuss the fascinating evolution of the along with 20,000 deaths from breast cancer per year (1). mammary gland and development of the young with Finally, human milk and breastfeeding are moving into respect to the bioactive components of milk. Further, the spotlight as researchers try to uncover aspects of they explore gene expression in the milk of women breastfeeding that confer benefits; these aspects include with mastitis, which is one of the major causes of early milk components and the very act of feeding and weaning in breastfeeding women. whether these are affected by maternal characteristics. Unfortunately, not all components of milk are Such knowledge may potentially provide windows of considered beneficial for the infant, in particular opportunity to improve nutrition and development of contaminants present in our environment. Gay et al. our infants. articulate the development of a highly sensitive assay This Showcase on Research will explore human milk to detect persistent organic pesticides in human milk. to a much greater extent than previously documented, Further, they calculate rather than estimate the actual due to recent research using cutting edge technology, dose ingested by the infant over the first year of life and allowing greater insight into the intriguing intricacies determine if this is related to infant growth. of this complex fluid. Enjoy this update of one of the most crucial Many of the benefits of breastfeeding are attributed physiological processes for the survival of the human in part to the reduced growth rate of breastfed infants race! compared to formula-fed infants. Given that breastfed 1. Victora, C.G., et al. (2016) Lancet 387, 475-490 Donna Geddes School of Chemistry and Biochemistry, University of Western Australia, Perth WA 6009 [email protected]

Lactation Guest Editor: Donna Geddes Cover Illustration Mammary stem and differentiated 5 Appetite Control Hormones in Human Milk – What is Their Role mammary epithelial cells isolated Zoya Gridneva and Donna Geddes from human milk. 9 Cells in Human Milk – What Do They Tell Us Image courtesy of Alecia-Jane Alecia-Jane Twigger and Peter Hartmann Twigger and Peter Hartmann, School 13 Human Milk Bioactivity: Lessons from the Evolution of Lactation of Chemistry and Biochemistry, Julie Sharp, Ashalyn Watt, Christophe Lefevre and Kevin Nicholas University of Western Australia. 19 Pesticides in Human Milk: Should We Be Concerned? Melvin Gay, Robert Trengove and Donna Geddes

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Appetite Control Hormones in Human Milk – What is Their Role Zoya Gridneva* and Donna Geddes School of Chemistry and Biochemistry, University of Western Australia, Perth, WA 6009 *Corresponding authors: [email protected]

Demand the Best synthesising mechanisms (9). Indeed, breastfed infants All parents seek optimal nutrition for their children and exhibit higher serum leptin levels compared to formula- whilst we believe most know that human milk (HM) meets fed infants, which is likely due to leptin being rendered all the needs of the infant (1), not everyone understands inactive during the processing of bovine milk for the form of delivery, that is breastfeeding on demand, is formula (10). Leptin in HM has been hypothesised to be also desirable. Breastfeeding mothers meet on average involved both in the short-term control of appetite and seven deadlines per day in the first months of baby’s life (2) in developmental programming of appetite and energy and these deadlines may happen anywhere and anytime. signalling pathways, promoting efficient energy control Modern society is still not keen on breastfeeding in public, and storage throughout life (8,11). In support of this, and any mother daring to address the demand may be leptin administered during the first 14 days of life acts as frowned upon, yet it is not the actions of a spoiled child a neurotrophic agent, promoting neural growth from the we are observing, but the amazing biochemistry behind the arcuate nucleus of the hypothalamus to multiple appetite appetite control in action. control centres located in the central nervous system (12). However the influence of leptin on GE, a key regulator of Human Milk Appetite Hormoness appetite, has not been well studied. Animal models show Breastfed infants eat less but more frequently than that central administration of leptin delays GE (13) and formula-fed infants and are leaner as a result of lower reduces food intake (14). In contrast, we have measured energy intake (3). They also display a variety of feeding infants’ stomachs (Fig. 1) sequentially to assess the effect of patterns and frequency (2) which can, in part, be explained an array of HM components on GE for a single breastfeed by HM composition. and we found that HM leptin does not appear to impact HM is a complex mixture consisting not only of nutrients GE in term infants (15,16), nor has it been linked to either but also bioactive molecules including hormones, growth the frequency of feeding or time between the feeds (15- factors, neuropeptides, immunomodulating and anti- 17). This suggests the action of HM leptin on short-term inflammatory agents (4). An increasing number of appetite appetite control may be mediated by upregulation of hormones have been recently identified in HM and a circulating melanocortins, potent anorexigenic agents that number of studies aimed to elucidate their effect on the promote satiety (18). Thus actions other than regulation of infant. These HM appetite hormones are mainly derived GE or downstream effects of HM leptin not yet measured from the maternal circulation, with a small contribution are likely responsible for the beneficial effects of HM in by the breast (5,6). As mediators between the adipose protecting against obesity later in life. tissue, gastrointestinal tract and infant brain, the potential functions of appetite hormones include regulation of satiety, development of appetite control pathways, and modulation of infant growth and development (7) . This review discusses the effect of the appetite hormones on infant gastric emptying (GE) and body composition (BC), as both have important roles in regulating appetite, food intake and energy balance.

Leptin Leptin is a polypeptide hormone synthesised by the white adipose tissue and is the most widely studied of all appetite hormones. It is involved in the regulation of adipose tissue, food intake and body weight. Leptin causes weight loss by suppressing appetite via signalling satiety and increasing metabolic rate. Leptin in HM is Fig. 1. Ultrasound technique for measuring infant’s derived mainly from maternal serum, following secretion stomach. The longitudinal and transverse planes for from white adipocytes and gastric chief cells into the measurement of stomach volume are indicated. bloodstream, with a small contribution from the mammary gland epithelium (5,8). With respect to regulation of BC, higher HM leptin levels In infants, HM is believed to be a major source of are associated with lower infant weight gain in the first leptin early in life, due to immature endogenous leptin- six months, lower infant BMI at two years of age (18),

Vol 48 No 1 April 2017 AUSTRALIAN BIOCHEMIST Page 5 SHOWCASE ON Appetite Control Hormones in Human Milk RESEARCH lower infant weight and adiposity (18,19) and greater lean mucosal gastric vagal afferent in an obesity-induced model body mass (total body water) in breastfed infants (20,21), (28). In humans, elevated serum levels of adiponectin are suggesting a pivotal role for leptin in regulating infant associated with more rapid GE in diabetic patients (29). growth and BC. Research in our laboratory confirms We are the first to have studied HM adiponectin in fully these results and also shows that both higher HM leptin breastfed term infants and have found that increased levels concentration and 24-hour intake of leptin are associated and doses are associated with longer time between feeds in with lower infant weight, lean body mass and fat mass this population (16). This indicates adiponectin’s potential (measured with bioimpedance spectroscopy, Fig. 2) action on the gastric vagal nerve via decreased sensitivity during first 12 months of lactation (Gridneva and Geddes, of tension receptors, potentially resulting in delayed satiety unpublished data). Given that many of the benefits of signalling, leading to higher food intake. breastfeeding are associated with the growth rate of the The modulating effects of GE in infants and potentially infant, these findings suggest that HM is indeed bioactive. on appetite and nutrient intake may partially explain the Further studies should incorporate exploration of the growth-regulating effect of adiponectin in infants in earlier regulation of leptin levels in HM, which have been shown months of life, as we and other researchers have shown to be influenced by maternal adiposity, but also display a (30). The relationship between changes in HM adiponectin circadian rhythm (17). and body composition and growth over the first two years of life has been studied. High levels of adiponectin are associated with lower weight, lean body mass and adiposity (30) (Gridneva and Geddes, unpublished data) in the first months of life, whereas in the second year of life, infants display more rapid growth and accretion of lean body mass and in a sense, ‘catch up’ to infants that received lower levels of adiponectin (31). Slower growth appears to be in part regulated by HM adiponectin and thus constitutes a potential mechanism for the lower risk of obesity in later life.

Ghrelin Ghrelin is a less studied orexigenic peptide produced primarily in the stomach, but also in the lactating breast, Fig. 2. Measurements of infant body composition resulting in HM having a higher ghrelin concentration with bioimpedance spectroscopy. than maternal serum (32). Ghrelin is an antagonist to leptin and stimulates appetite, gastric motility, acid secretion Adiponectin and food intake and is involved in long-term regulation of Adiponectin is the appetite hormone present in the growth, weight and energy metabolism; thus heightened highest concentrations in HM and its concentration is concentrations in HM could be vital for the infant drive more than 40 times higher than that of ghrelin and leptin to feed. (22,23). Adiponectin is secreted by adipose tissue and also Ghrelin stimulates appetite and food intake and increases synthesised by the breast (5). Adiponectin circulates as GE in rats (33) and humans (34) via the vagal nerve and oligomers of different sizes, from low-molecular-weight afferent activity. High infant serum ghrelin levels are trimers to high-molecular-weight octodecamers, the latter associated with increased age, length and weight in both being the most common in HM (24), in contrast to the formula-fed and breastfed infants (35,36), with formula-fed middle-molecular-weight form of bovine adiponectin (25). infants having higher ghrelin levels than breastfed infants It is present in a biologically active form and is resistant (due to higher levels in formula) (10,35). Further, longer to digestion (26). Interestingly, HM adiponectin levels are fasting times are associated with higher serum levels of only moderately related to both maternal circulating levels ghrelin in formula-fed infants in the first six months of life and adiposity, indicating adiponectin is regulated within (37). The inverse correlation between ghrelin and leptin the mammary gland (27). Amongst its various functions, it in infant serum (10) indicates that HM leptin may play a has anti-inflammatory properties, breaks down fatty acids role in lowering ghrelin levels. Since ghrelin and leptin and heightens sensitivity to insulin. Strong correlations actions are mediated by neuropeptide Y (NPY), reduced between adiponection levels in HM and infant serum leptin levels allow ghrelin NPY stimulation and increase (26) along with the presence of adiponectin receptors in in food intake (10). Indeed, more comprehensive studies the infant’s intestinal tract (26) emphasise its importance are required to clarify potential interactions between these for the infant. Indeed, higher levels of circulating plasma appetite hormones. adiponectin are related to lower BMI and healthier HM ghrelin levels increase progressively in first months metabolism. of lactation (38) and therefore, may influence infant feeding In animal models, adiponectin inhibits tension sensitive patterns and BC. Higher serum ghrelin levels in breastfed gastric vagal afferent mechanosensitivity, modulating infants are associated with lower weight gain (36) and in satiety signals in both lean and obese animals, while formula-fed infants, with lower BMI (10), indicating a role simultaneously increasing the mechanosensitivity of for ghrelin in the regulation of body weight in healthy

Page 6 AUSTRALIAN BIOCHEMIST Vol 48 No 1 April 2017 SHOWCASE ON Appetite Control Hormones in Human Milk RESEARCH infants. Higher ghrelin levels in HM are associated with metabolic development of infants and could be of interest higher infant weight gain (39,40) but lower weight (39). in further research. These somewhat contradictory results may be due to the ­­­ combined effects of other HM hormones that were not Conclusion measured in previous studies. Also it is not clear if ghrelin HM appetite hormones influence GE and BC in breastfed levels precede changes in appetite (37) and body weight infants and are associated with early infant growth and (36,41) or follow them. development and subsequently health later in life. So the Ghrelin influences novelty seeking behaviour in rodents next time you see an infant being breastfed think, of all the and humans (42), which may translate into an advantage delicate, complex mechanisms and pathways signalling to in terms of locating new sources of food. Levels of ghrelin ensure programming of appetite regulation, leading to a are blamed for impulsive shopping behaviour, as images reduced risk of obesity and chronic disease in our next of food become more desirable (43). These behaviour generation. modulations mean that raised ghrelin levels in the infant will translate into hunger cues to gain the attention of the References mother. 1. WHO (2007) Geneva, Switzerland 2. Kent, J., Hepworth, A., Sherriff, J., Cox, D., Mitoulas, L., Other Key Appetite Hormones and Hartmann, P. (2013) Breastfeed. Med. 8, 401-407 Insulin is a major hormone involved in glucose 3. Butte, N., Wong, W., Hopkinson, J., Smith, E., and Ellis, metabolism and is produced by the pancreas. Studies K. (2000) Pediatrics 16, 1355-1366 in rats show that dietary insulin is not degraded in the 4. Goldman, A.S. (2000) J Nutr 130, 426S-431S stomach and plays an important role in the maturation 5. Hassiotou, F., Savigni, D., Hartmann, P.E., and Geddes, and development of the small intestine (44); thus HM D.T. (2014) FASEB J. 28, 38.8 insulin could play a similar role in the development 6. Kugananthan, S., Lai, C.T., Gridneva, Z., Mark, P.J., of human intestinal epithelium. Insulin is actively Geddes, D.T., and Kakulas, F. (2016) Nutrients 8, 711 transported into HM in concentrations similar to maternal 7. Savino, F., Benetti, S., Liguori, S., Sorrenti, M., and circulating levels (45) and is significantly higher (almost Cordero Di Montezemolo, L. (2013) Cell. Mol. Biol. 59, four times) than in bovine milk and infant formula, where 89-98 it is barely detectable (46). While one study showed no 8. 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(2004) J. and leptin associated with beneficial microbial metabolic Neurosci. 24, 2797-2805 pathways that are able to reduce inflammation (48), 12. Proulx, K., Richard, D., and Walker, C. D. (2002) further highlighting the diverse effects of these hormones. Endocrinology 143, 4683-4692 Obestatin is an appetite-suppressing derivative of the 13. Smedh, U., Hakansson, M.L., Meister, B., and Uvnas- ghrelin peptide precursor, which further emphasises the Moberg, K. (1998) Neuroreport 9, 297-301 complexity of the appetite hormone system. Obestatin 14. Sanchez, J., Oliver, P., Miralles, O., Ceresi, E., Pico, C., is produced by the cells lining the stomach and small and Palou, A. (2005) Endocrinology 146, 2575-2582 intestine and salivary glands (49). Early studies reported 15. Cannon, A.M., Gridneva, Z., Hepworth, A., Lai, C.T., that obestatin opposes ghrelin’s effects on body weight, Tie, W.J., Khan, S., Hartmann, P.E., and Geddes, D.T. food intake and GE (50). However, obestatin was (2017) Pediatr. Res. in press later implicated in the inhibition of thirst and anxiety, 16. Gridneva, Z., Kugananthan, S., Hepworth, A.R., regulation of sleep, memory improvement, induction Tie, W.J., Lai, C.T., Ward, L.C., Hartmann, P.E., and of cell proliferation and increasing the secretion of the Geddes, D.T. (2017) Nutrients 9, E15 pancreatic juice enzymes (49). HM obestatin has been 17. Cannon, A., Kakulas, F., Hepworth, A., Lai, C., identified (51) with a concentration more than twice that Hartmann, P., and Geddes, D. (2015) Int. J. Environ. Res. of corresponding circulating maternal levels. Surprisingly Public. Health 12, 12340-12355 however, the source of obestatin has still not been 18. Miralles, O., Sanchez, J., Palou, A., and Pico, C. (2006) established. The relationship between obestatin and infant Obesity 14, 1371-1377 development and BC clearly requires further investigation. 19. Fields, D., and Demerath, E. (2012) Pediatr. 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Cells in Human Milk – What Do They Tell Us Alecia-Jane Twigger* and Peter Hartmann School of Chemistry and Biochemistry, University of Western Australia, Perth, WA 6009 *Corresponding author: [email protected] The physiological processes of human lactation are cells that can be used to non-invasively characterise not as foolproof as is often thought, as many women lactating mammary cell gene expression, morphology experience milk production difficulties, often leading to and regenerative capabilities (Fig. 1). early weaning. Most of these difficulties are attributed Early studies examining cells from human colostrum to pain during feeding and real or perceived low (HC) and HM cells were conducted by smearing undiluted milk supply. Some identified risk factors for impaired samples across slides to morphologically determine cell lactation include maternal obesity resulting in abnormal types and properties (7). Originally it was thought that development of the mammary gland (1) and maternal milk was dominated by leukocytes and ‘foam cells’ mammary ZnT2 transporter mutations being linked to (colostrum bodies) (7), however more modern techniques low production (2). Evidently, greater research into the of cell isolation and classification have advanced our mechanisms facilitating successful morphogenesis of understanding of HM cells. Studies from our laboratory the gland during pregnancy and lactation is desperately using freshly isolated cells extracted from either HC or required to develop strategies to assist at-risk women HM examined for CD45 with flow cytometry found that and optimise their milk production. In the past, studies whilst leukocytes dominate HC, they revert to <2.5% of into the development of the gland have been hindered by total mature HM cells when both the mother and infant the lack of readily accessible human lactating tissue (3). are healthy (8). On the other hand, infection of either As Figure 1a result, many previous studies have instead focused the infant or mother can affect the leukocyte content on animal models of pregnancy and lactation, although of HM, increasing slightly for infant only infections major differences exist in mammary gland development and more dramatically (>90% leukocyte content) in (4), gene expression (5) and milk composition (6) between cases of maternal mastitis (8,9) (Fig. 2). These findings, mammalian species. However, human milk (HM) together with animal studies illustrating immune cell contains immune, stem and differentiated mammary transfer through the milk to various infant organs (10), particularly the thymus (11,12), suggests a potential protective and programming role of these cells for the infant. Additionally, the heightened levels of immune cells in cases of maternal mastitis provides a unique opportunity to study pathways of mammary gland defence that may be optimised for rapid resolution of mammary gland infection. Unlike bovine milk, which contains low numbers of viable cells (primarily immune cells) (13,14), mature HM is a rich source of viable cells, representative of the lactating Stem cell mammary epithelium (3,15) (Fig. 3A). Numerous viable cells are available for isolation from mature HM, ranging Common from 1.16x104 – 3.9x106 cells per progenitor millilitre of milk, of which 76- 100% are viable (16-18). Lower Luminal numbers of viable cells can be Myoepithelial progenitor progenitor isolated from either HC or early HM samples (18). Variation in Fig. 1. Cells isolated from these values are likely due to human milk contain a differences in isolation protocols, hierarchy of stem and where storage and centrifugation differentiated mammary cells. Myoepithelial Alveolar Ductal speed are important in

Vol 48 No 1 April 2017 AUSTRALIAN BIOCHEMIST Page 9 SHOWCASE ON Cells in Human Milk RESEARCH determining the number and viability of extracted cells in obese mothers (16). In addition, higher expression of (19). Recently, polyploidal luminal cells have been estrogen related receptor beta (ESRRB) expression and identified in the human mammary gland and these play a lower expression of growth differentiation factor 3 (GDF3) vital role in lactation, as observed by low milk production expression were associated with earlier stages of lactation, in mice with inhibited binucleated cell formation (20). It implying ongoing development of the mammary gland appears polyploidal cells filter into the milk as seen in over the course of lactation. Indeed whole transcriptome Fig. 3A (Twigger, unpublished data), although more analysis between milk fat globules isolated from HC, detailed imaging should be carried out to confirm this. transitional HM and mature HM found differences in Examination of HM cells has confirmed the expression of expression of key genes such as milk protein genes alpha- many key mammary markers including integrin alpha-6 lactatalbumin and beta casein between these three stages (CD49f), membrane metalloendopeptidase (CD10), of lactation (24). Moreover, immune defence pathways epithelial adhesion marker (EPCAM) and cytokeratins were expressed specifically in HC, upregulation of milk 5, 14, 18 and 19 (CK5, CK14, CK18 and CK19) which are synthesis machinery was found in transitional HM and representative of mammary stem and differentiated basal lipid production pathways were dominant in mature HM and luminal cells (16,21,22). Furthermore, a previous (24). A subsequent study examined changes in the HM cell study found that mammary tissue and milk cells from transcriptome by comparing sequential samples collected lactating macaques had comparable gene expression, over the first 12 months of lactation with prepartum suggesting milk cells to be representative of mammary secretions from the same women and resting mammary tissue (23). Discrepancies in HM cell gene expression gland tissue (25). As well as confirming findings from the have been associated with different characteristics of the HM fat globule study, this study determined differences mother and infant, building our understanding of the in the expression of cell adhesion pathway genes between influencing factors that determine successful mammary prepartum secretion cells and resting tissue, as well as gland development and milk production. differences in the expression of cancer pathway genes The population of cells in HM vary in composition over between mature HM cells and resting mammary tissue the course of a breastfeed as well as within and between that were not identified in the HM fat globule study women, much like milk composition in general (3). HM (25). Studies such as these will elucidate the mechanisms cell content varies over the course of a breastfeed, similar to behind human milk synthesis and mammary metabolism the variation observed in fat content in HM, being greater that are trigged during secretory activation. It is evident towards the end of a feed and peaking approximately 30 that not only can HM cells be utilised to examine minutes post-feeding (17). In a recent study, variation mammary gene expression (16,26,27), they can also be of HM cell gene expression was established between 66 grown in culture to help further profile the developing participants for 17 different genes representative of the and functioning mammary gland (21,22,26,28,29). mammary cellular hierarchy (16). Gene expression was The first studies of HM cell culture were conducted investigated in light of maternal and infant demographics, in the late 1970s and early 1980s, where cells isolated which included maternal body mass index (BMI), breast from early milk (3 to 7 days post-partum) proliferated volume, gestational age of the infant and lactation stage, in 2D culture and survived multiple passages (30-33). in order to determine the presence of any associations Interestingly, it was noted that not all cells participated (16). Of note, lower expression of CK18, which is in cell division whilst many of the cells could proliferate representative of lactocytes, was associated with higher for 8–11 passages (30). Subsequent studies found cells maternal BMI, potentially suggesting undifferentiated isolated from more mature HM could also be cultured lactocytes as an underlying cause of low milk production and these displayed many of the features originally

A B i) B ii) Scatter Count Side i 70.5% CD45+ 55% CD45+ iii ii

Forward Scatter CD45 CD45 Fig. 2. Flow cytometric analysis of isolated milk cells collected from a woman with mastitis. A. Forward and side scatter plots reveal size and granularity of milk cells which were gated to examine subpopulations i), ii) and iii). B. Cells from subpopulations i) and ii) were examined for CD45, where subpopulation i) had 70.5% of cells positive for CD45, whereas subpopulation ii) had 55% of cells positive for CD45.

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Fig. 2. Human milk cells examined with an inverted microscope. A. Freshly isolated cells stained with Trypan Blue exclusion dye illustrate a large number of viable cells with some apparent bi-nucleated cells (red arrows), non-viable cells (black arrow) and few milk fat globules (yellow arrow). B. Cells placed in mammary media for two days begin to forming spheroids under non-adherent conditions. described with added immunofluorescence staining, can differentiate into two separate mammary cell lineages demonstrating the diversity in the resulting mammary and consequently undergo cell cycle arrest (28). Lineage- cell subtypes (21,22,26,28). Additionally, it was found specific progenitor markers such as CD105, CD29, CD90 that CK5 and CK14 gene expression was enriched after and Stro-1, which are representative of mesenchymal stem culture, providing an opportunity to select and expand cells, have been identified in HM, which are able to secrete the progenitor cell population from freshly isolated HM vascular endothelial growth factor (VEGF) in culture (36- cells (21). Another study using cultured monolayer HM 39). Whether these cells are true mesenchymal stem cells is cells was able to identify differences in the morphology debatable (40), however further studies suggest that HM and growth patterns of the cells after transfection of 14-3- cells exhibit lineage plasticity. An interesting study by 3s (Sigma) siRNA (28). Similar gene silencing techniques Hassiotou et al. demonstrated the presence of pluripotent could be utilised in the future to examine the importance transcription factors OCT4, SOX2 and NANOG in HM of specific genes for mammary gland development. cells and illustrated their plasticity by differentiating Using non-adherent culturing conditions, 3D structures them into neuronal, hepatocyte, pancreatic and bone–like can also be formed from HM cells (Fig. 3B) to create bi- cells (26). Similar studies have been conducted in resting layered spheroids containing CD49f+ positive basal cells mammary tissue, which also contain highly plastic cells and Prl-R+ (prolactin receptor) luminal cells, mimicking (41), albeit at lower densities (26). Evidently, stem cells mammary alveoli (22). These 3D structures were shown in the mammary gland are vital to achieving maturity in to synthesise and secrete the milk proteins alpha- pregnancy and to some degree lactation. However it is lactalbumin and lactoferrin into the culture media (26). not known why these cells are excreted into the milk. It is Enhancements of these protocols may utilise the latest possible that they exist for the benefit of the infant, where organoid culture methods, which are able to produce they have been found to survive the gastrointestinal tract organised tertiary alveolar and conn3cting ductal of the offspring and are transferred to distant tissues structures from resting mammary cells (34). Indeed, HM where they integrate to potentially contribute to tissue cell culture may hold the key to a greater understanding function (42). It has been speculated that HM cells have of human mammary gland development, which is driven a potential use in regenerative therapies; however a by progenitor and stem cells. great deal more work is needed to understand the cells’ Currently, human mammary stem cells are not well characteristics and proportions in milk. defined with respect to markers, location or differentiation Clearly HM contains a fascinating cellular composition capabilities due to the limited availability of tissue and that can be non-invasively isolated and used in a plethora heterogeneous methods of investigation (35). It is likely of experiments to examine lactating mammary cell gene that a spectrum of mammary stem cells exists with expression, growth and in vitro morphology. HM cells different differentiation capabilities. Investigations of will also assist in the understanding of the intricacies of lactating mammary cells isolated through HM have mammary gland development and function. Future milk found subpopulations that are positive for markers cell work investigating the characteristics of normal HM associated with bipotent, multipotent and pluripotent cells hold the promise of unravelling mammary gland markers (21,26,28). Nestin was the first multipotent stem dysfunction and pathologies such as low milk production cell marker identified in HM cells and co-stained with the and mastitis, both of which are major contributors to mammary stem cell marker CK5 (21). HM cells positive early weaning. for multipotent marker p63 have also been identified and

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References 22. Thomas, E., Lee-Pullen, T., Rigby, P., Hartmann, P., Xu, 1. Rasmussen, K.M. (2007) Ann. Rev. Nutr. 27, 103-121 J., and Zeps, N. (2012) Stem Cells 30, 1255-1264 2. Dempsey, C., McCormick, N.H., Croxford, T.P., Seo, 23. Lemay, D.G., Hovey, R.C., Hartono, S.R., Hinde, K., Y.A., Grider, A., and Kelleher, S.L. (2012) J. Nutr. 142, Smilowitz, J.T., Ventimiglia, F., Schmidt, K.A., Lee, J., 655-660 Islas-Trejo, P., Silva, P.I., Korf, I., Medrano, J., Barry, 3. Hassiotou, F., Geddes, D.T., and Hartmann, P.E. (2013) P.A., and German, J.B. (2013) BMC Genomics 14, 872 J. Hum. Lact. 29, 171-182 24. Lemay, D.G., Ballard, O.A., Hughes, M.A., Morrow, 4. Dontu, G., and Ince, T.A. (2015) J. Mammary Gland Biol. A.L., Horseman, N.D., and Nommsen-Rivers, L.A. Neoplasia 20, 51-62 (2013) PLoS One 8, e67531 5. Lemay, D.G., Lynn, D.J., Martin, W.F., Neville, M.C., 25. Twigger, A.J., Geddes, D.T., and Kakulas, F. 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(2013) Clin. Transl. Martin, T. (2011) Cell Cycle 10, 1-7 Immunology 2, e3 29. Hosseini, S.M., Talaei-Khozani, T., Sani, M., and 9. Riskin, A., Almog, M., Peri, R., Halasz, K., Srugo, I., and Owrangi, B. (2014) Neurol. Res. Int. 2014, 807896 Kessel, A. (2012) Pediatr. Res. 71, 220-225 30. Chang, S.E., Keen, J., Lane, E.B., and Taylor- 10. Jain, L., Vidyasagar, D., Xanthou, M., Ghai, V., Shimada, Papadimitriou, J. (1982) Cancer Res. 42, 2040-2053 S., and Blend, M. (1989) Arch. Dis. Child 64, 930-933 31. Stoker, M., Perryman, M., and Eeles, R. (1982) Proc. R. 11. Zhou, L., Yoshimura, Y., Huang, Y., Suzuki, R., Soc. Lond. B. Biol. Sci. 215, 231-240 Yokoyama, M., Okabe, M., and Shimamura, M. (2000) 32. Gaffney, E.V. (1982) Cell Tissue Res. 227, 563-568 Immunology 101, 570-581 33. Taylor-Papadimitriou, J., Shearer, M., and Tilly, R. 12. Ghosh, M.K., Nguyen, V., Muller, H.K., and Walker, (1977) J. Natl. Cancer Inst. 58, 1563 A.M. (2016) J. Immunol. 197, 2290-2296 34. 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Human Milk Bioactivity: Lessons from the Evolution of Lactation Julie Sharp1,2*, Ashalyn Watt1, Christophe Lefevre3,4,5 and Kevin Nicholas6,7 1Institute for Frontier Materials, Deakin University, Geelong, VIC 3216 2Cancer Program, Monash Biomedicine Discovery Institute and Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC 3800 3Division of Bioinformatics, Walter and Eliza Hall Medical Research Institute, Melbourne, VIC 3000 4Peter MacCallum Cancer Centre, Melbourne, VIC 3000 5Department of Medical Biology, University of Melbourne, Melbourne, VIC 3000 6Department of Anatomy and Developmental Biology, Monash University, Clayton, VIC 3800; 7School of Biosciences, University of Melbourne, Melbourne, VIC 3010 *Corresponding author: [email protected] Lactation has evolved over the past 200 million years Reproduction in the tammar is characterised by a short since the appearance of the aplacental, egg laying gestation period (26.5 days), birth of immature young and monotremes. Subsequently, there has been extensive a long lactation period (approximately 300 days), during adaptation to reproduction, particularly in lactational which the concentration of all the major milk constituents, strategies, when the Theria (subclass of mammals) and many minor milk bioactives progressively change split into the Metatheria (Marsupialia) and Eutheria (15) (Fig. 1A). There is increasing evidence that these (Placentalia) lineages over 140 million years ago (1). The changes in milk composition regulate growth of the role of milk in providing nutrition to the suckled young in tammar pouch young (10,16), particularly during the mammals is well established. However, over the past few first 100 days postpartum, when the development of the decades a focus on the functionality of milk has confirmed neonate is similar to a late stage eutherian fetus. Therefore that it also delivers a range of bioactives that are either the signalling factors involved in the development of the present in milk in the mammary gland or are released eutherian fetus are most likely delivered to marsupials in when the milk is processed in the oral cavity and the gut the milk of the mother (17) (Fig. 1B). Fostering experiments of the neonate. These bioactives may include proteins, demonstrated that transferring the early phase pouch peptides, complex oligosaccharides and miRNA and have young to a late phase lactating tammar can accelerate the the potential to stimulate development, regulate gut flora growth and physical development of pouch young (18,19), and provide protection for the young from infection (2-8). and also accelerate maturation of specific organs such as An interesting and often neglected group of bioactives is the stomach (20). More recent studies using in vitro models now being identified; they have a function locally within have shown that milk collected from marsupials during the mammary gland to remodel the mammary tissue early lactation (day 20–100), but not late lactation (day 100– (stimulate growth or signal cell death) and can provide 300), stimulated proliferation and differentiation of whole protection from infection and inflammation when the lung cultures from mouse embryos (21). Therefore the mammary gland is susceptible to these challenges. This temporal delivery of these bioactives is most likely crucial review will focus predominantly on the relationship for the development of the suckled young. between milk proteins and proteases, resulting in the The regulation of the lactation cycle in the tammar wallaby release of bioactive peptides. has fascinated and challenged scientists for many decades and the interesting interplay between the endocrine, Milk Bioactives – Impact on Development of the Young autocrine and paracrine mechanisms that are implicated in To better identify bioactives, scientists have increasingly this process are now beginning to be better understood. If used comparative genomics and bioinformatics to explore these animals are genuine biomedical research models that the lactational strategies of the Australian monotremes offer new insights into reproduction in eutherians, it must and marsupials. The lactation cycle in the only two extant be assumed that ‘evolution has not reinvented the wheel’ monotremes, the echidna (Tachyglossus and Zaglosus and that the basic mechanisms regulating cellular responses genera) and platypus (Ornithorhynchus anatinus), and an have been conserved over the past 140 million years. Australian marsupial, the tammar wallaby (Macropus This assumption appears to be reasonably sound. eugenii) have been studied extensively (9,10). The echidna Research over the past 50 years has shown that insulin, lays shelled eggs (11) and the hatchlings are altricial glucocorticoids and prolactin are required for lactation in (undeveloped) and not immune competent (12). Therefore, eutherians and a number of in vitro studies have shown milk is not only important for growth and development these hormones are a basic requirement for successful but particularly for protection of the young from disease lactation in the tammar and echidna (22-26). It is now while they are in a non-sterile environment (13,14). accepted that the mammary extracellular matrix (ECM) has

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Fig. 1. The lactation cycle of the tammar wallaby. A. The lactation cycle in the tammar has been divided into four phases, characterised by changes in milk composition and the sucking pattern of the pouch young. B. Development of the tammar pouch young from day 6 to day 220 postpartum (upper panels) compared to development of the human embryo at day 23 and preterm baby at 24 weeks of age (lower panels). an integral role and is essential for the development of the mammal. However, the reality is that the mammary gland mammary gland during gestation, the overt differentiation in these species is very sophisticated in terms of its capacity of mammary epithelial cells for galactopoesis and for temporal delivery of bioactives for multiple targets. The remodelling of the mammary gland during involution eutherian mammary gland is less sophisticated as many (27-29). More recent studies have shown that the ECM in of its previous functions have evolved to be delivered by the tammar has additional roles in regulating the cellular multiple tissues, particularly a well-developed placenta program for the lactation cycle in the tammar, particularly and the amniotic fluid. It is clear that the marsupial provides in controlling the progressive changes in milk composition a unique opportunity to more easily identify bioactives (30,31). Perhaps one of the most exciting and compelling that potentially play a role in the early development of studies on the regulation of lactation was reported recently the fetus. However, it may suggest that an increased focus by Jane Visvader’s laboratory (29). The experiments using on human colostrum would be appropriate to get a better a mouse model showed that binuclear mammary epithelial understanding of its potential role in regulating early cells appeared in the mammary gland at lactogenesis and development of the baby, particularly at a time when the disappeared at involution. The presence of these cells in gut is amenable to increased transfer of milk components. the mammary gland was essential for successful lactation. We have known for some time that significantly However it was fascinating to note that these binuclear premature and low birthweight babies have acute cells were also present in lactating tissue from the human challenges for survival, largely due to limited development and wallaby, indicating that this basic regulatory process of their lungs and gut, and these babies may show an was conserved during the evolution of lactation. increased frequency of mature onset disease (32). Studies It is ironic that marsupials may be considered a primitive using the tammar wallaby may lead to a new range of

Page 14 AUSTRALIAN BIOCHEMIST Vol 48 No 1 April 2017 SHOWCASE ON Human Milk Bioactivity RESEARCH human fortifiers that include bioactives with the potential wallaby mammary epithelial cells exhibiting increased to specifically target the growth and development of proliferation following inclusion of MaeuCath1b in the tissues in the human neonate to improve outcomes for media, confirming this hypothesis (38). premature and low birthweight babies (33,34). Another interesting tammar milk protein, WAP four- Therefore, developmental clocks are set in the neonate disulphide domain protein-2 (WFDC2) is part of a large and any potential disruption to this process may family of whey acidic protein (WAP) four disulfide core subsequently impact on mature onset disease. The tammar proteins (46-49). Tammar WFDC2 is comprised of two provides a new model to better understand this process four-disulfide core domains annotated as domain III on of developmental programming. It is not known if this the amino terminal end and domain II at the carboxyl developmental program is set during the short gestation terminal end (50). Expression of the WFDC2 gene is or whether the milk provided in early lactation includes elevated only in pregnancy, early lactation and during signals to the altricial neonate that have a role in this involution (51). Studies by Watt et al. (52) showed domain process. The option of cross fostering neonates to mothers II of the protein had antibacterial activity against a range at advanced stages of lactation to exclude the delivery of pathogenic bacteria but no antibacterial activity against of putative milk bioactives to the suckled young (33-35) Enterococcus faecalis. The elevated expression of WFDC2 may shed new light on the initiation of developmental during pregnancy and involution correlates with the programming. timing of increased risk of infection in the mammary gland (40,52,53), largely resulting from Staphylococcus aureus, Milk Bioactives That Impact on Mammary Function Streptococcus spp. and Escherichia coli in the mammary The early work of Li et al. (36) showed that ligating tissue (54-56). However, expression of this gene during the teats on one side of a lactating mouse led to these the first 100 days postpartum suggests an additional role mammary glands progressing only to involution, despite in protecting the pouch young when it is not immune all mammary glands being exposed to the same hormonal competent (52). Therefore, the antibacterial effect of milieu. This was clear evidence that milk bioactivity WFDC2 appears to be directed to pathogenic bacteria and could impact locally on mammary gland physiology. not commensal bacteria (40,53,57). The down regulation More recent work using the tammar has shown that the of WFDC2 around 100 days postpartum, when the young mammary gland has a sophisticated capacity to use milk detaches from the teat, correlates with the development of when under challenge from infection and that milk can an immune response in the young. stimulate growth, and program cell death of the tissue when appropriate (36,37). Mammary genes coding for Human Milk Bioactivity – Acute Response to Challenge milk proteins can be multifunctional and demonstrate The examples of the temporal regulation of either intact a temporal domain-specific bioactivity resulting from proteins or domain-specific expression of proteins to alternate splicing (38). For example, cathelicidin usually provide the kind of milk bioactivity required by the tammar exists in an inactive proform and is cleaved by specific mammary gland and the suckled young is dependent proteases to provide the two domains of cathelicidin on sophisticated programming of the mammary gland. that may have a variety of functions (39). The tammar However, as discussed previously, there is a reduced cathelicidin 1 gene (MaeuCath1) is differentially expressed need for this level of progressive change in the delivery of in the mammary gland throughout the lactation cycle specific bioactives in human milk, although it is evident as two splice variants (MaeuCathel 1a and 1b) (38). The that human milk composition can change during lactation level of MaeuCath1a transcripts are upregulated only and in response to specific challenges to the breast (58,59). during early lactation (2A) and late involution whereas A recent study by Sharp et al. (2016) using microarrays to MaeuCath1b transcripts are expressed throughout analyse the expression of genes in cells present in breast lactation and steadily increase from late lactation milk at days 24, 48 and 101 of lactation and days 7 and 14 until early involution. The protein corresponding to of involution identified a total of 206 protease genes (60). MaeuCath1a significantly inhibits a range of bacteria A large component of these genes were predicted to code (38). The expression of this transcript in the first 48 hours for proteins with a signal peptide. Predicted classes of postpartum to day 80 postpartum is consistent with a need secreted proteases included threonine, aspartate, cysteine, to provide protection from pathogens (40). MaeuCath1a serine and metallo proteases. Some of these genes were expression at day 10 of involution suggests an additional expressed throughout lactation whereas other genes were antibacterial role at a time when the mammary gland is expressed only at specific time points during lactation. more susceptible to pathogen-mediated mastitis (41). The Bioactive milk peptides resulting from proteolytic continued expression of the MaeuCath1b transcript when digestion had functions that included immunomodulatory the neonate has developed adaptive immunity suggests (61,62), antimicrobial (63-65), and antithrombotic (66,67) additional roles for the maintenance and proliferation of activity, and these peptides could act as opioid agonists mammary epithelia during increasing milk production (68-71), ACE inhibitors (72) and proliferative factors (73). (23,42). Indeed, several studies have suggested a role for In recent studies, skim breast milk was incubated at 37 cathelicidins in epithelial cell proliferation during wound degrees over a period of 5 days to allow digestion of milk healing, maintenance and re-establishment of the intestinal protein by endogenous proteases (Watt, Nicholas and barrier integrity and proliferation of lung epithelial Sharp, unpublished data). Peptides were predominantly cells (43-45). More recent in vitro studies have shown derived from casein proteins, and antibacterial assays

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Fig. 2. Mastitis-specific gene expression profile. Expression profiles of genes related to a defence response within cells collected from healthy milk and mastitic milk. Y axes represent expression values. COL: colostrum 24 hours prepartum; LACT: lactation at day 24, 48, 101; MAST; mastitis at day 23; INV: involution at day 7 and 14. using mastitis causing S. aureus showed that incubating this to explore with regard to the complexity of human milk. hydrolysate significantly increased antimicrobial activity. The established role of complex carbohydrate on gut flora Similarly, these peptides showed anti-inflammatory and the general health of the young and the emerging role activity. Importantly, the peptides did not show any of miRNA as legitimate signalling molecules for both the capacity to program apoptotic activity in human mammary breast and the young (74,75) emphasise the complexity of epithelial cells. Therefore these peptides may play a specific the multiple roles of milk, in addition to its role in nutrition. role in the breast to reduce infection and inflammation if However, it is increasingly evident that the interrogation there is an interruption to breastfeeding. of the evolutionary history of lactation has great promise for the application of comparative approaches to obtain a Acute Responses to Mastitic Challenge better understanding the role of milk in acute and chronic Determination of gene expression in normal lactating wellbeing of the baby. women demonstrated the utility of RNA obtained directly from human milk cells to detect mammary epithelial References cell (MEC)-specific gene expression. Therefore milk cell 1. Lefevre, C.M., Sharp, J.A., and Nicholas, K.R. (2010) RNA collected during mastitis was also analysed by Annu. Rev. Genomics Hum. Genet. 11, 219-238 human Affymetrix arrays and revealed regulation of a 2. FitzGerald, R.J., Murray, B.A., and Walsh, D.J. (2004) J. unique set of genes specific to this disease state, whilst Nutr. 134, 980S-988S maintaining regulation of milk synthesis genes (60). Genes 3. Hartmann, R. and Meisel, H. (2007) Curr. Opin. specific to infection and related to immune function, Biotechnol. 18, 163-169 antimicrobial response, and anti-inflammatory response 4. Allen-Blevins, C.R., Sela, D.A., and Hinde, K. (2015) Evol. were upregulated compared to healthy milk (Fig. 2). More Med. Public Health 2015, 106-121 recent studies (Watt, Nicholas and Sharp, unpublished 5. Zivkovic, A.M., German, J.B., Lebrilla, C.B., and Mills, data) have used human MECs cultured on a range of D.A. (2011) Proc. Natl. Acad. Sci. USA 108 Suppl 1, 4653- ECMs which allows the formation of three-dimensional 4658 acini (mammospheres) that respond to insulin, cortisol 6. Marcobal, A., Barboza, M., Froehlich, J.W., et al. (2010) J. and prolactin to express milk protein genes. These Agric. Food Chem. 58, 5334-5340 mammospheres were challenged with lipoteichoic acid 7. Zhou, Q., Li, M., Wang, X., et al. (2012) Int. J. Biol. Sci. 8, (LTA) and showed similar transcriptional responses 118-123 to milk-derived cells from women with mastitis (Fig. 8. Admyre, C., Johansson, S.M., Qazi, K.R., et al. (2007) J. 3). These in vitro acini therefore represent a robust and Immunol. 179, 1969-1978 relevant human mammary model to better understand 9. Sharp, J.A., Watt, A., Bisana, S., et al. In: Singh, H., the acute molecular responses to mastitic challenge, and Thompson, A., and Boland, M. (eds.), Milk Proteins: From may therefore provide new opportunities for improved Expression to Food (2014) Elsevier, Chapter 3, pp76-112 treatment of breast infection. 10. Sharp, J.A., Modepalli, V., Enjapoori, A.K., et al. (2015) J. This review has focused on the relationship between Mammary Gland Biol. Neoplasia 19, 289-302 milk proteins and peptide bioactivity. Although it is 11. Morrow, G.E., and Nicol, S.C. (2013) Aust. J. Zool. 60, 289- becoming evident that breastmilk bioactivity for the 298 development of the young has been diminishing with a 12. Griffiths, M. The Biology of Monotremes (1978) Acedemic greater contribution from the placenta, there is still much Press, USA

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Fig. 3. Mastitis-specific gene expression mimicked by in vitro mammospheres. Expression profiles of genes which were shown to be upregulated in mastitis were also induced when mammospheres were incubated with LTA for 4 and 24 hours. A. Haematoxylin and eosin stain of paraffin embedded milk cells collected from mature milk. Arrows indicate presence of intact cells showing stained nuclei and cytoplasm. Binucleated cells can be observed. B–D. Gene expression profile of milk cells collected from the different stages of lactation (24, 48 and 101 days) and mastitis (day 23). E. Bright-field microscopy of human mammospheres grown for 7 days. F–H. Gene expression profiles of 7 day mammospheres treated with and without LTA for 4 and 24 hours.

13. Bisana, S., Kumar, S., Rismiller, P., et al. (2013) PLoS One 22. Nicholas, K.R. and Tyndale-Biscoe, C.H. (1985) J. 8, e53686 Endocrinol. 106, 337-42 14. Enjapoori, A.K., Grant, T.R., Nicol, S.C., et al. (2014) 23. Bird, P.H., Hendry, K.A., Shaw, D.C., Wilde, C.J., and Genome Biol. Evol. 6, 2754-73 Nicholas, K.R. (1994) J. Mol. Endocrinol. 13, 117-25 15. Tyndale-Biscoe, C.H., and Janssens, P.A. The Developing 24. Ballard, F.J., Grbovac, S., Nicholas, K.R., Owens, P.C., Marsupial: Models for Biomedical Research (1988) Springer- and Read, L.C. (1995) Gen. Comp. Endocrinol. 98, 262-8 Verlag, Germany 25. Kwek, J.H., Wijesundera, C., Digby, M.R., and Nicholas, 16. Sharp, J.A., Lefevre, C., Kwek, J., et al. In: Deakin, J.E., K.R. (2007) Biochim. Biophys. Acta 1770, 48-54 Waters, P.D., and Marshall Graves, J.A. (eds.), Marsupial 26. Enjapoori, A.K., Lefevre, C.M., Nicholas, K.R., and Genetics and Genomics (2010) Springer, UK. pp317-334 Sharp, J.A. (2017) Gen. Comp. Endocrinol. 242, 38-48 17. Brennan, A.J., Sharp, J.A., Digby, M.R., and Nicholas, 27. Roskelley, C.D., Srebrow, A., and Bissell, M.J. (1995) K.R. (2007) IUBMB Life 59, 146-150 Curr. Opin. Cell Biol. 7, 736-47 18. Menzies, B.R., Shaw, G., Fletcher, T.P., and Renfree, M.B. 28. Schedin, P., Mitrenga, T., McDaniel, S., and Kaeck, M. (2007) Reprod. Fert. Develop. 19, 976-983 (2004) Mol. Carcinog. 41, 207-20 19. Trott, J.F., Simpson, K.J., Moyle, R.L., et al. (2003) Biol. 29. Rios, A.C., Fu, N.Y., Jamieson, P.R., et al. (2016) Nat. Reprod. 68, 929-36 Commun. 7, 11400 20. Kwek, J., De Iongh, R., Nicholas, K., and Familari, M. 30. Wanyonyi, S.S., Lefevre, C., Sharp, J.A., and Nicholas, (2009) J. Exp. Zool. B Mol. Dev. Evol. 312, 613-24 K.R. (2013) Matrix Biol. 32, 342-51 21. Modepalli, V., Hinds, L.A., Sharp, J.A., Lefevre, C., and 31. Wanyonyi, S.S., Lefevre, C., Sharp, J.A., and Nicholas, Nicholas, K.R. (2015) BMC Dev. Biol. 15, 16 K.R. (2013) Dev. Comp. Immunol. 40, 289-99

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32. Svedenkrans, J., Henckel, E., Kowalski, J., Norman, M., 55. Borm, A., Fox, L., Leslie, K., et al. (2006) J. Dairy Sci. 89, and Bohlin, K. (2013) PLoS One 8, e80869 2090-2098 33. Waite, R., Giraud, A., Old, J., et al. (2005) J. Exp. Zool. A 56. Bradley, A.J., and Green, M.J. (2001) J. Clin. Microbiol. 39, Comp. Exp. Biol. 303, 331-344 1845-1849 34. Kwek, J.H., Iongh, R.D., Digby, M.R., et al. (2009) Mech. 57. Old, J.M., and Deane, E.M. (1998) Comp. Immunol. Dev. 126, 449-463 Microbiol. Infect. Dis. 21, 237-245 35. Modepalli, V., Hinds, L.A., Sharp, J.A., Lefevre, C., and 58. Ballard, O., and Morrow, A.L. (2013) Pediatr. Clin. North Nicholas, K.R. (2016) Mech. Dev. 142, 22-29 Am. 60, 49-74 36. Khalil, E., Digby, M.R., O’Donnell, P., and Nicholas, K.R. 59. Hartmann, P.E., and Prosser, C.G. (1984) Fed. Proc. 43, (2008) Comp. Biochem. Physiol. B Biochem. Mol. Biol. 151, 2448-2453 64-69 60. Sharp, J.A., Lefevre, C., Watt, A., and Nicholas, K.R. 37. Brennan, A.J., Sharp, J.A., Lefevre, C.M., and Nicholas, (2016) Funct. Integr. Genomics 16, 297-321 K.R. (2008) J. Mol. Endocrinol. 41, 103-116 61. Adel-Patient, K., Nutten, S., Bernard, H., et al. (2012) J. 38. Wanyonyi, S.S., Sharp, J.A., Khalil, E., Lefevre, C., and Agri. Food Chem. 60, 10858-10866 Nicholas, K.R. (2011) Comp. Biochem. Physiol. A Mol. 62. Qian, B., Xing, M., Cui, L., et al. (2011) J. Dairy Res. 78, 72- Integr. Physiol. 160, 431-439 79 39. Yang, D., Biragyn, A., Hoover, D.M., Lubkowski, J., and 63. Dallas, D.C., Guerrero, A., Khaldi, N., et al. (2013) J. Oppenheim, J.J. (2004) Annu. Rev. Immunol. 22, 181-215 Proteome Res. 12, 2295-2304 40. Daly, K.A., Digby, M., Lefevre, C., et al. (2007) Vet. 64. Gifford, J., Hunter, H., and Vogel, H. (2005) Cell. Mol. Life Immunol. Immunopathol. 120, 187-200 Sci. 62, 2588-2598 41. Oliver, S.P., and Mitchell, B.A. (1983) J. Dairy Sci. 66, 65. Liepke, C., Zucht, H.-D., Forssmann, W.-G., and 1162-1166 Ständker, L. (2001) J. Chrom. B Biomed. Sci. App. 752, 369- 42. Dove, H., and Cork, S.J. (1989) J. Zool. 219, 385-397 377 43. Heilborn, J.D., Nilsson, M.F., Kratz, G., et al. (2003) J. 66. Fiat, A.-M., Migliore-Samour, D., Jollès, P., et al. (1993) J. Invest. Dermatol. 120, 379-389 Dairy Sci. 76, 301-310 44. Otte, J.M., Zdebik, A.E., Brand, S., et al. (2009) Regul. Pept. 67. Chabancea, B., Jollèsa, P., Izquierdoa, C., et al. (1995) Br. 156, 104-117 J. Nutr. 73, 583-590 45. Shaykhiev, R., Beisswenger, C., Kandler, K., et al. (2005) 68. Teschemacher, H., Koch, G., and Brantl, V. (1997) Peptide Am. J. Physiol. Lung Cell Mol. Physiol. 289, L842-L848 Sci. 43, 99-117 46. Bingle, L., Singleton, V., and Bingle, C.D. (2002) Oncogene 69. Meisel, H., and FitzGerald, R.J. (2000) Br. J. Nutr. 84, 27- 21, 2768-2773 31 47. Campbell, S.M., Rosen, J.M., Hennighausen, L.G., 70. Migliore-Samour, D., Floc’h, F., and Jollès, P. (1989) J. Strech-Jurk, U., and Sippel, A.E. (1984) Nucleic Acids Res. Dairy Res. 56, 357-362 12, 8685-8697 71. Gertrud, K., Klaus, W., and Hansjörg, T. (1985) Naunyn- 48. Topcic, D., Auguste, A., De Leo, A.A., et al. (2009) Evol. Schmiedeberg’s Arch. Pharmacol. 331, 351-354 Dev. 11, 363-375 72. Wu, Z., Pan, D., Zhen, X., and Cao, J. (2013) J. Sci. Food 49. Hennighausen, L.G., and Sippel, A.E. (1982) Nucleic Agri. 93, 1331-1337 Acids Res. 10, 2677-2684 73. Kanda, Y., Hisayasu, S., Abe, Y., Katsura, K., and 50. Sharp, J.A., Lefevre, C., and Nicholas, K.R. (2007) Evol. Mashimo, K. (2007) Life Sci. 81, 449-457 Dev. 9, 378-392 74. Kumar, A., Buscara, L., Kuruppath, S., Ngo, K.P., 51. Watt, A.P., Sharp, J.A., Lefevre, C., and Nicholas, K.R. Nicholas, K.R., and Lefèvre, C. In: Reyes Cruz, L.M., and (2012) Dev. Comp. Immunol. 36, 584-590 Ortiz Gutierrez, D.C. (eds) Lactation: Natural Processes, 52. Basden, K., Cooper, D.W., and Deane, E.M. (1997) Physiological Responses and Role in Maternity (2012) Nova Reprod. Fertil. Dev. 9, 243-254 Science Publishers, pp73-97 53. Old, J.M., and Deane, E.M. (2000) Dev. Comp. Immunol. 75. Modepalli, V., Kumar, A., Hinds, L.A., et al. (2014) BMC 24, 445-454 Genomics 15, 1012 54. Barkema, H., Green, M., Bradley, A., and Zadoks, R. (2009) J. Dairy Sci. 92, 4717-4729

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Pesticides in Human Milk: Should We Be Concerned? Melvin Gay1*, Robert Trengove2,3 and Donna Geddes1 1School of Chemistry and Biochemistry, University of Western Australia, Crawley, WA 6009 2Separation Science and Metabolomics Laboratory, Murdoch University, Murdoch, WA 6150 3Metabolomics Australia, Murdoch University Node, Murdoch, WA 6150 *Corresponding author: [email protected]

Breastfeeding has been the norm throughout primate via inhalation, ingestion and dermal contact, followed by evolution. Breastfeeding stimulates both the mother and bioaccumulation in organs with higher lipid contents, such the infant physiologically. Maternal effects include the as the liver and breast, as they are not easily metabolised release of oxytocin, which is essential for milk ejection in the body (12). but also induces a lower threshold to pain and stimulates Pesticides, such as organochlorine pesticides (OCPs), positive feelings, hence its nick name the ‘love’ hormone. organophosphate pesticides (OPPs) and carbamates, are The skin-to-skin contact and the act of breastfeeding also used to control pests in agriculture and in homes. Besides modulates infant temperament, heart rate, temperature their primary action as pesticides, many of these OCPs and and acid–base balance. In addition, there are a myriad of OPPs can interfere with the functions of normal endocrine positive effects on both the mother and infant that can in systems by mimicking, blocking, modulating or altering many cases be attributed to the unique species specific the synthesis, metabolism or transport of hormones composition of human milk (HM) (1,2). (12,13). Long term pesticide exposure has also been The short-term benefits of HM for the infant are extensive, associated with neuropsychiatric sequelae (14), chronic where meta-analyses show a lower incidence of infection diseases such as Parkinson’s disease (15), cancer (16) and including necrotising enterocolitis (NEC), gastrointestinal asthma (17), as well as congenital disorders (18,19). tract infections and a reduction in infant mortality While many of these pesticide groups, such as OCPs with increased doses of HM (3). Further, the lifetime and OPPs, have been banned since the 1980s/90s due programming effects of HM include a lower risk of to their persistence and long half-lives (over seven obesity and diabetes later in life (4,5), which are precursors years), many of these pesticides are still detected in the to metabolic disease as well as better neurological environment and in biofluids. A variety of pesticides, development and cognitive function compared to infants such as p,p’-dichlorodiphenyldichloroethylene (p,p’- that were formula-fed (6,7). Indeed mothers also reap DDE), p,p’-dichlorodiphenyltrichloroethane (p,p’-DDT) significant health benefits, particularly if they breastfeed and hexachlorocyclohexane (HCH), have been detected in their infants for more than 12 months, with a significant various biofluids of the inhabitants of different countries. reduction in risk of cardiovascular disease, hypertension, p,p’-DDE, which is a metabolite of p,p’-DDT, is one of diabetes and cancer (breast and ovarian) compared to the most commonly detected OCPs in humans as DDT those who have never breastfed (8-10). was widely used around the world in agriculture and The benefits of HM to infant are still therefore domestic households in the 1940s/50s. Since the 1980s, undeniable. However, the mechanisms associated many of these banned pesticides have been replaced by with the transfer and/or production of nutritional and more ‘human-friendly’ pesticides, such as pyrethroids, protective components in milk also act as pathways for which can be easily metabolised by mammals and have the passage of environmental contaminants into the milk. shorter half-lives of 2.5 to 12 hours in blood plasma (20). Indeed persistent organic pollutants (POPs) and heavy As humans are not only exposed to a single chemical, metals have been detected in HM. The question is: does but to a multitude of different chemicals simultaneously, this pose a risk to the infant? Further, is the monitoring of it is important to continuously measure the level of these contaminants worldwide sufficient to safeguard not environmental contaminants, both banned and legalised, only the infants, but also everyone that lives in the same to ensure that the total environmental pesticide exposure environment? is within the safe threshold limits.

Persistent Organic Pollutants Impact of Pesticide Exposure on Infant Health POPs, such as pesticides, polychlorinated biphenyls The fetus and infants are recognised as being more (PCBs) and polychlorinated dibenzo-p-dioxins (dioxins), susceptible to the harmful effects of pesticides due to their are synthetic chemicals that are ubiquitously present in small size and rapid growth as well as their immature the environment (11,12). These pollutants are usually by- immune and metabolic systems (21). Many of the studies products from industrial processes or agrichemicals used designed to investigate the adverse effects of pesticides on for pest and weed control on farms. As many of these infants have focused on maternal exposure to pesticides xenobiotics have lipophilic properties and are resistant by measuring pesticide levels in maternal biofluids, to environmental degradation, they can be transported such as blood (22,23), cord blood (24) and HM (25,26). over a long distances via the atmosphere and water. Associations are then drawn between these measures and Hence, these chemicals are then absorbed into mammals prenatal, infant and child development measures such as

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Table 1. Concentrations of DDTs (ng/g fat) in human milk from various countries. Adapted from (25).

Country/ Year of Mothers Human milk Fat content DDTs Reference region sampling (number) samples (g/L) (ng/g fat) Asia China 2006/07 60 60 31 583a, * (52) India 2011 53 53 32 1914b (47) Taiwan 2000/01 36 36 31 333c (53) Korean 2008 >50 50 22 225c (54) Japan 2008/09 90 9# 32 119c (55) Malaysia 2003 17 17 17 1600d (37) Philippine 2004 33 33 22 170b (56) Turkey 2009 47 47 36 338e (57) a Iran 2006 10 10 15 3560 (36) Australian and New Zealand

f New Zealand 2007/2010 39 37 39 385 (58) g Australia/WA 2013/15 40 40 33 63 (25) Europe

b Poland 2002/05 28 28 14 1621 (59) d Slovakia 2003 12 12 27 665 (60) g Norway 2002/06 377 377 36 53 (61) h Latvia 2002/04 15 15 na 267 (62) e Denmark 1997/01 43 43 na 82 (63) *, d Belgium 2009/10 84 84 44* 60 (64) d Germany 2007/08 516 516 36 125 (65) b, * Croatia/Zagreb 2009/10 20 29 na 278 (66) *, f UK 2001/03 54 54 na 157 (67) North and South Americas

a USA 2004 38 38 22 65 (68) Brazil 2001/02 69 69 na 493b (69) Africa

c South Africa 2001 28 28 na 6320 (70) b Ethiopia 2010 39 29 na 14460 (71)

na no data available * Values represented as median value # Pooled sample a Sum of o,p’-DDE + p,p’-DDE + p,p’-DDD + p,p’-DDT b Sum of p,p’-DDE + p,p’-DDD + p,p’-DDT c Sum of p,p’-DDE + p,p-DDD + o,p’-DDT + p,p’-DDT d Sum of p,p’-DDE + p,p’-DDT e Sum of o,p’-DDE + p,p’-DDE + o,p’-DDD + p,p’-DDD + o,p’-DDT + p,p’-DDT f Sum of o,p’-DDE + p,p’-DDE + p,p’-DDD + o,p’-DDT + p,p’-DDT g p,p’-DDE only h Sum p,p’-DDE + o,p’-DDT + p,p’-DDT

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Temporal Trends and Worldwide Comparisons of Pesticides in HM The ratio of p,p’-DDE to p,p’-DDT concentration indicates exposure history where a high DDE/DDT ratio (>5) represents historical exposure and a low DDE/DDT ratio (<5) suggests continuous exposure (35). This provides valuable insight into the maternal environment, lifestyle and dietary habits but also allows early intervention to minimiSe infant (and maternal) exposure to these pesticides if present at high concentrations. HM pesticide concentrations vary widely between countries (Table 1) and these are largely dependent on the extent of pesticide use and the timing of legal bans. Higher total Fig. 1. Trends of total DDT concentrations in human DDT concentrations (sum of DDTs and its metabolites, milk of mothers in Western Australia. DDE and DDD) are typically observed in malaria-prone countries such as Iran (36) and Malaysia (37), where DDTs growth, immune and metabolic status. were widely used to combat mosquitos and were only Studies have shown an association between higher recently banned (1990s to 2000s). Limited use of DDT is concentrations of the insecticide chlorpyrifos in maternal also still allowed indoors in malaria endemic countries, prenatal plasma and smaller infant birth weight and such as South Africa and India (38). We have shown a length (23). Similarly, smaller infant birth weight and declining trend in the total DDTs in HM from women in head circumference have been observed in mothers Western Australia since the 1970s (Fig. 1) (39-43). This is with high prenatal urinary DDE concentrations (27). in agreement with Muller et al., where an annual decrease Further, recent studies have associated high exposure of approximately 12–13% of DDTs was observed in earlier of OPPs with attention deficit/hyperactivity disorder in years (1960s to 1980s) slowing to around a 5% annual five-year-olds (28), and poor intellectual development decrease since the 1990s (40). On a national level, the HM and cognitive performance in seven-year-olds (29). Sex p,p’-DDE concentrations observed in WA are 2.4 to 7.5 differences have also been identified where male infants times lower than corresponding p,p’-DDE concentrations were more susceptible to OPP exposure with smaller in other states and territories such as South Australia, head circumference, delayed adaptive skills and social Victoria and New South Wales, and is reflective of the and motor development skills respectively (30). It is still inconsistent timing of legislation for pesticide control difficult however to decide whether the associations are prior to the 1990s. Since levels of pesticides are declining due to prenatal or postnatal pesticide exposure, therefore, in HM, it stands to reason infant dose is also declining. one should exercise caution in the interpretation of these studies. Infant Daily Dose of Pesticides Via HM While many studies have used maternal pesticide It is essential to monitor HM pesticide concentrations to levels and infant anthropometrical data at birth as a ensure that the daily infant pesticide intake is within the proxy measure of in utero development, our recent tolerable limit of 0.5–10 µg/kg body weight/day (44-46) epidemiological longitudinal and cross-sectional studies recommended by regulatory agencies (e.g. EPA, FAO, have shown no significant associations between p,p’-DDE RIVM, WHO) to decrease the risk of impaired infant concentrations in milk from mothers in Western Australia growth and development. Calculation of infant dose and infant growth outcomes such as weight, length, head requires infant weight, concentration of the pesticide, circumference and percentage fat mass (estimated using bioimpedance spectroscopy and ultrasound skinfolds) at 2, 5, 9 and 12 months postpartum (25). Furthermore, the concentration of p,p’-DDE significantly decreased by 68% from 70.9 ± 70.6 ng/g fat at 2 months to 22.4 ± 14.0 ng/g fat at 12 months (31), indicating that maternal bioburden is reduced via breastfeeding. Interestingly, expected relationships between maternal age (increased HM pesticide levels) and parity (reduced HM pesticide levels) are not always confirmed in the literature, however this is likely due to differences in study design and analysis. (22,25,27). One is tempted however, to speculate that the reduction of maternal bioburden during lactation could potentially contribute to the reported reduced incidence of breast and ovarian cancer in women that have breastfed (32-34). Whilst the Western Australia data is encouraging, Fig. 2. Comparison between the actual daily intake the question is whether the dramatic reductions in POPs (ADI) and estimated daily intake (EDI) of total in HM are paralleled globally? DDTs by infants during the first year of lactation.

Vol 48 No 1 April 2017 AUSTRALIAN BIOCHEMIST Page 21 SHOWCASE ON Pesticides in Human Milk RESEARCH milk fat and the volume of milk consumed. Many studies e827-841 have used estimated values of milk fat, infant milk intake 6. Newburg, D.S., and Walker, W.A. (2007) Pediatr. Res. and infant weight, to estimate daily intake (EDI) of the 61, 2-8 infant (36,47,48). In our recent cross sectional study we 7. Quigley, M. A., Hockley, C., Carson, C., et al. (2012) J. calculated daily intake (CDI) by using the average milk Pediatr. 160, 25-32 intake based on the infant age (49) and measuring milk 8. Ip, S., Chung, M., Raman, G., et al. (2009) Breastfeed. fat (25). This was further improved by calculating the Med. 4 Suppl 1, S17-30 actual daily intake (ADI) which is based on the actual 9. Schwarz, E.B., Ray, R.M., Stuebe, A.M., et al. (2009) measurements of milk fat, 24 hour milk intake and infant Obstet. Gynecol. 113, 974-982 weight in our longitudinal study (Fig. 2) (31). The EDI 10. Stuebe, A.M., Willett, W.C., Xue, F., et al. (2009) Arch. significantly overestimates the overall daily intake over Intern. Med. 169, 1364-1371 the first 12 months postpartum compared to both CDI 11. Loganathan, B.G., and Kannan, K. (1994) Ambio 23, and ADI by 53% and 44%. While Du et al. (31) observed 187-191 a significant decrease in pesticide intake by the infant 12. Tanabe, S. (2002) Mar. Pollut. Bull. 45, 69-77 throughout the 12 months, they also observed that the 13. Bergman, A., Heindel, J.J., Kasten, T., et al. (2013) ADI from several mothers exceeded the tolerable limit Environ. Health Perspect. 121, A104-106 (>0.5 µg/kg body weight/day) in the earlier months of 14. Freire, C., and Koifman, S. (2013) Int. J. Hyg. Environ. lactation (two and five months). This could be considered Health 216, 445-460 concerning, however this is counterbalanced by the fact 15. Costa, L.G., Giordano, G., Guizzetti, M., et al. (2008) that infant exposure to pesticides in HM via breastfeeding Front. Biosci. 13, 1240-1249 is a relatively short period as compared to a lifetime 16. Mostafalou, S., and Abdollahi, M. (2013) Toxicol. Appl. living in the same environment. Overall, the ADI we Pharmacol. 268, 157-177 calculated was approximately three times lower than 17. Hernandez, A.F., Parron, T., and Alarcon, R. (2011) the tolerable limit. Thus, the benefits of HM, such as the Curr. Opin. Allergy Clin. Immunol. 11, 90-96 immunological and nutritional benefits, far outweigh the 18. Cavieres, M.F., Jaeger, J., and Porter, W. (2002) potential negative effects that are yet to be adequately Environ. Health Perspect. 110, 1081-1085 demonstrated. Therefore breastfeeding should continue 19. Weselak, M., Arbuckle, T.E., and Foster, W. (2007) J. to be encouraged for up to two years and beyond, as Toxicol. Environ. Health B Crit. Rev. 10, 41-80 recommended by WHO (50). 20. Leng, G., Leng, A., Kuhn, K.H., et al. (1997) Xenobiotica 27, 1273-1283 Conclusions and Future Directions 21. Bruckner, J.V. (2000) Regul. Toxicol. Pharmacol. 31, 280- The maternal environment, diet and body composition 285 before and during pregnancy play a major role in 22. Reid, A., Callan, A., Stasinska, A., et al. (2013) Sci. Total fetal programming and the future health of the infant Environ. 449, 208-213 (51). As part of a continuum, lactation represents a 23. Perera, F.P., Rauh, V., Tsai, W.-Y., et al. (2003) Environ. period of further infant programming for optimum Health Perspect. 111, 201-205 health outcomes. Whilst the presence of pesticides in 24. Ribas-Fito, N., Julvez, J., Torrent, M., et al. (2007) Am. HM is of concern, the use of HM can provide a non- J. Epidemiol. 166, 1198-1202 invasive avenue to monitor pesticide exposure and to 25. Du, J., Gridneva, Z., Gay, M.C., et al. (2016) measure infant daily intake. Further, while new data Chemosphere 167, 247-254 does not show detrimental effects of POPs in HM on 26. Ribas-Fito, N., Grimalt, J.O., Marco, E., et al. (2005) early infant growth, longitudinal studies should be Environ. Res. 98, 8-13 conducted for longer periods (beyond three years of 27. Wolff, M.S., Engel, S., Berkowitz, G., et al. (2007) life) to elucidate relationships with infant growth and Pediatr. Res. 61, 243-250 developmental outcomes, including cognitive function. 28. Marks, A.R., Harley, K., Bradman, A., et al. (2010) As many countries now source commodities, such as food Environ. Health Perspect. 118, 1768-1774 produce, from various countries, it is essential that global 29. Bouchard, M.F., Chevrier, J., Harley, K.G., et al. (2011) restrictions are implemented and adhered to in order to Environ. Health Perspect. 119, 1189-1195 reduce population contaminant exposure via diet. 30. Liu, P., Wu, C., Chang, X., et al. (2016) Environ. Health Perspect. 124, 1637-1643 References 31. Du, J., Gridneva, Z., Gay, M. C., et al. (2016) Sci. Rep. 6, 1. Newburg, D.S. (2001) Adv. Exp. Med. Biol. 501, 3-10 38355 2. Hale, T.W., and Hartmann, P.E. (2007) Hale and 32. Danforth, K.N., Tworoger, S.S., Hecht, J.L., et al. (2007) Hartmann’s Textbook of Human Lactation, 1st Cancer Causes Control 18, 517-523 edition, Hale Publishing 33. Collaborative Group on Hormonal Factors in Breast 3. Meinzen-Derr, J., Poindexter, B., Wrage, L., et al. Cancer (2002) Lancet 360, 187-195 (2009) J. Perinatol. 29, 57-62 34. Shema, L., Ore, L., Ben-Shachar, M., et al. (2007) J. 4. Verduci, E., Banderali, G., Barberi, S., et al. (2014) Cancer Res. Clin. Oncol. 133, 539-546 Nutrients 6, 1711-1724 35. Mishra, K., and Sharma, R.C. (2011) Sci. Total Environ. 5. American Academy of Pediatrics. (2012) Pediatrics 129, 409, 4939-4949

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36. Behrooz, R.D., Sari, A.E., Bahramifar, N., et al. (2009) 54. Kim, D., Ryu, H.Y., Lee, J.H., et al. (2013) J. Environ. Chemosphere 74, 931-937 Sci. Health B 48, 243-250 37. Sudaryanto, A., Kunisue, T., Tanabe, S., et al. (2005) 55. Fujii, Y., Haraguchi, K., Harada, K.H., et al. (2011) Arch. Environ. Contam. Toxicol. 49, 429-437 Chemosphere 82, 25-31 38. Mandavilli, A. (2006) Nat. Med. 12, 870-871 56. Malarvannan, G., Kunisue, T., Isobe, T., et al. (2009) 39. Noren, K., and Meironyte, D. (2000) Chemosphere 40, Environ. Pollut. 157, 1924-1932 1111-1123 57. Çok, I., Mazmanci, B., Mazmanci, M.A., et al. (2012) 40. Mueller, J.F., Harden, F., Toms, L.-M., et al. (2008) Environ. Int 40, 63-69 Chemosphere 70, 712-720 58. Mannetje, A., Coakley, J., Bridgen, P., et al. (2013) Sci. 41. Stacey, C., Stanley, W., and Whitney, S. (1985) Arch. Total Environ. 458–460, 399-407 Environ. Health 40, 102-108 59. Hernik, A., Góralczyk, K., Struciński, P., et al. (2011) 42. Stacey, C.I., and Thomas, B.W. (1975) Pestic. Monit. J. Ann. Agric. Environ. Med. 18, 113-118 9, 64-66 60. Yu, Z., Palkovicova, L., Drobna, B., et al. (2007) 43. Stevens, M.F., Ebell, G.F., and Psaila-Savona, P. (1993) Chemosphere 66, 1012-1018 Med. J. Aust. 158, 238-241 61. Polder, A., Skaare, J.U., Skjerve, E., et al. (2009) Sci. 44. FAO/WHO (2000) Total Environ. 407, 4584-4590 45. RIVM (2001) Re-evaluation of human-toxicological 62. Bake, M.A., Linnika, Z., Sudmalis, P., et al. (2007) Int. J. maximum permissible risk levels. National Institute Hyg. Environ. Health 210, 483-489 of Public Health and the Environment, Bilthoven, The 63. Shen, H., Main, K.M., Virtanen, H.E., et al. (2007) Netherlands Chemosphere 67, S256-S262 46. USEPA (1996) Integrated Risk Information System 64. Croes, K., Colles, A., Koppen, G., et al. (2012) (IRIS). DDT: non-cancer assessment Chemosphere 89, 988-994 47. Bedi, J.S., Gill, J.P.S., Aulakh, R.S., et al. (2013) Sci. 65. Raab, U., Albrecht, M., Preiss, U., et al. (2013) Total Environ. 463-464, 720-726 Chemosphere 93, 461-467 48. Van Oostdam, J., Gilman, A., Dewailly, E., et al. (1999) 66. Klinčić, D., Romanić, S.H., Sarić, M.M., et al. (2014) Sci. Total Environ. 230, 1-82 Environ. Toxicol. Pharmacol. 37, 543-552 49. Kent, J.C., Mitoulas, L., Cox, D.B., et al. (1999) Exp. 67. Kalantzi, O.I., Martin, F.L., Thomas, G.O., et al. (2004) Physiol. 84, 435-447 Environ. Health Perspect. 112, 1085-1091 50. World Health Organization, and UNICEF. (2003) 68. Johnson-Restrepo, B., Addink, R., Wong, C., et al. Global strategy for infant and young child feeding, (2007) J. Environ. Monit. 9, 1205-1212 WHO, Geneva, Switzerland 69. Azeredo, A., Torres, J.P.M., de Freitas Fonseca, M., et 51. Godfrey, K.M., and Barker, D.J. (2001) Public Health al. (2008) Chemosphere 73, S246-S251 Nutr. 4, 611-624 70. Darnerud, P. O., Aune, M., Larsson, L., et al. (2011) Sci. 52. Leng, J.-H., Kayama, F., Wang, P.-Y., et al. (2009) Total Environ. 409, 4048-4053 Chemosphere 75, 634-639 71. Gebremichael, S., Birhanu, T., and Tessema, D.A. 53. Chao, H.-R., Wang, S.-L., Lin, T.-C., et al. (2006) (2013) Chemosphere 90, 1652-1657 Chemosphere 62, 1774-1785

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ultraconserved elements in the human genome and several The Lemberg Medal is awarded to a distinguished Australian biochemist or molecular biologist who will present the Lemberg classes of small RNAs, and, in conjunction with colleagues, Lecture at the ComBio meeting. The Medal is presented in developed new methods for high-resolution transcriptome memory of Emeritus Professor M.R. Lemberg, who was the analysis. Society’s first President and Honorary Member. Nominees must John returned to Sydney in 2012, where he established have been members of the Society for at least five years before the year in which the Medal nomination is to be considered. An the Garvan Institute as one of the world’s leading centres honorarium is provided by ASBMB. for human genome sequencing and oversaw the launch of Genome.One, a wholly owned subsidiary of the Garvan and one of the world’s first clinically accredited whole genome sequencing companies. John has published over 300 papers, which have been cited over 32,000 times (Scopus). Inter alia, he has been Chair of The Lemberg the Queensland Studies Authority and a member of the Medal: Australian Health Ethics Committee. John was recently named by the NHMRC as the one of the all-time high John Mattick achievers in Australian health and medical research.

The Merck Research Medal is awarded to an outstanding Australian biochemist or molecular biologist with less than 15 years postdoctoral experience. The successful candidate will present the Merck Medal Lecture at the ComBio meeting. Nominees must have been members of the Society for at least John Mattick is the Executive Director of the Garvan two years before the year in which the Medal nomination is Institute of Medical Research. He obtained his BSc with to be considered. An honorarium is provided through the generosity of Merck Pty Ltd. First Class Honours in Biochemistry from the University of Sydney, and his PhD in Biochemistry from Monash University, under the supervision of Professor Tony Linnane and Professor Phillip Nagley. He then undertook postdoctoral training with Professor Salih Wakil at Baylor The Merck College of Medicine in Houston, where his work on the Research architecture of the fatty acid synthase complex is now featured in biochemistry textbooks. Medal: John returned to Australia in 1982 to join the (then) CSIRO Lars Ittner Division of Molecular Biology, where he developed one of the world’s first genetically engineered vaccines (against ovine footrot). In 1988, he joined the University of Queensland as the Foundation Professor of Molecular Biology and Director of the Centre for Molecular Biology and Biotechnology, Professor Lars Ittner is a group leader in the Department of which later became the ARC Special Research Centre Anatomy of the Faculty of Medicine at the University of New for Molecular and Cellular Biology, and ultimately the South Wales (UNSW) and Senior Principal Research Fellow Institute for Molecular Bioscience (IMB). John was also the at Neuroscience Research Australia (NeuRA) in Sydney. He Foundation Director of the Australian Genome Research is a neuroscientist and heads the research laboratories of the Facility (1996–2002) and the ARC Special Research Centre Dementia Research Unit and the Transgenic Animal Unit of for Functional and Applied Genomics. the Mark Wainright Analytical Centre at UNSW. In 2006, John relinquished the IMB Directorship to take Lars received his medical degree from the University of up an ARC Federation Fellowship (and later an NHMRC Ulm in Germany in 2001, followed by an experimental Australia Fellowship) to work on his longstanding thesis in medical research at the University of Zurich conviction that the vast amounts of untranslated RNAs in Switzerland in 2002. Lars was then selected into the that are expressed from the genomes of complex organisms Postgraduate Course for Experimental Biology and are not junk, but rather an extensive regulatory system Medicine at the University of Zurich, Switzerland, from responsible for guiding the epigenetic trajectories of where he continued as a postdoctoral fellow with Professor development. Jan A. Fischer, working on G-protein-coupled receptors He had first published his hypothesis after a sabbatical and neuronal development. in Cambridge in 1994, and in the ensuing two decades, In 2005, Lars moved to Australia to work as a Postdoctoral showed the extensive transcription of intronic, intergenic Fellow with Professor Jürgen Götz at the University of and antisense RNAs, their highly dynamic and specific Sydney. With this move, Lars changed his research focus cellular and subcellular expression, their association with from early neuronal development to neurodegenerative chromatin-modifying complexes and their involvement diseases, which continues to be the major interest of his in development and disease. He also discovered research program. During his time at the University of Page 24 AUSTRALIAN BIOCHEMIST Vol 48 No 1 April 2017 Medallist and Awardee Profiles

Sydney, he made fundamental discoveries about the honours in genetics. She carried out a PhD at the Peter synaptic functions of one of the hallmark proteins in MacCallum Cancer Institute (back in East Melbourne) in Alzheimer’s disease, the tau protein. Furthermore, he yeast cell cycle genetics, but she then changed direction developed a range of novel transgenic mouse models of to study zebrafish developmental genetics at UCSF in San neurodegenerative diseases that continue to be used for Francisco and then at the Ludwig Institute. She has always understanding pathomechanisms and testing of novel loved to study complex systems, and to consider both the treatments. Lars became a Senior Lecturer and Associate detail of a reductionist approach at the same time as the Professor at the University of Sydney’s Brain and Mind effect on the whole organism. Heather gained a teaching Research Institute in 2009 and 2011, respectively. and research academic position at Monash University in Lars moved to UNSW in 2013, where he was appointed the School of Biological Sciences, but realised that she was as Professor in the School of Medical Sciences to head most fascinated with university education and the complex the Dementia Research Unit. His current work aims to processes by which students learn. In 2014, she was understand the earliest changes at the synapse of neurons fortunate to gain a Teaching Specialist position as a Senior during the onset of Alzheimer’s disease. Furthermore, Lecturer at the University of Melbourne in the Department Lars works together with industry partners to translate of Biochemistry and Molecular Biology. his findings into therapeutic avenues of the future. Lars Heather believes that student-centred activities are the is also part of a large chief investigator team, dedicated to key to quality teaching. She aims to teach in a way that understanding and treating rarer forms of dementia and results in students being actively engaged in thinking motor neuron disease. about (and discussing!) molecular biology problems in the Lars has published over 100 peer-reviewed articles in classroom, even though those classrooms are sometimes journals, including Science, Cell, Nature Medicine, Cell Stem very large. This approach is designed to make students Cell, Nature Reviews Neuroscience, Acta Neuropathologica and apply their understanding, even if at a basic level and PNAS. He has held prestigious fellowships throughout his without a laboratory, to enable students to fully internalise career, including a NHMRC Senior Research Fellowship, knowledge. Students in Heather’s classes are often trying to and he has been awarded multiple Project and Program interpret experimental data or design simple experiments. Grants from both the NHMRC and ARC as chief She trusts that students are ready to try challenging investigator. Amongst several awards he has received for activities with higher order thinking, as long as they are his research, he has also been awarded the Merck Young trained and supported and the expectations are clear and Achiever Award from the Australian Society for Medical consistent. Research (ASMR) in 2010. Heather has been researching ways to confront student misconceptions in the classroom in a way that allows The Shimadzu Education Award rewards outstanding achievement them to rethink their understanding. She has been in education of biochemistry or molecular biology, especially leading a research project funded by a University of innovation and creativity in education, with a view to fostering leadership in this important area of the Society’s objectives. The Melbourne Learning and Teaching Initiative grant, to Award will enable the recipient to participate in an international trial different ways to change student misconceptions conference with a significant focus on education, or to spend a in a range of different science, technology, engineering period of time at another institution (in Australia or overseas) for the and mathematics (STEM) fields, and a range of different purposes of undertaking developments in education of biochemistry and molecular biology. The recipient will present a lecture within educational situations. She also runs an Education the Education Symposium at the ComBio meeting. Applicants must Research Group within the School of Biomedical Sciences, have been members of the Society for at least two years before to support academics who wish to carry out Scholarship the year in which the Award application is to be considered. The of Teaching and Learning (SoTL) and be recognised for contribution to travel expenses is provided through the generosity of Shimadzu Excellence in Science Program. their achievements and innovations. One aim of this is to promote the roles of Teaching Specialist academics within the university. The Shimadzu Education Award: Heather Verkade

Heather Verkade’s interest in science began at the University of Melbourne, with a Bachelor of Science and

Vol 48 No 1 April 2017 AUSTRALIAN BIOCHEMIST Page 25 Medallist and Awardee Profiles

its receptor. This opportunity was undoubtedly the most The Beckman Coulter Discovery Science Award is awarded to an exciting in my career, not only because it offered me Australian biochemist or molecular biologist for distinguished significant independence but also because it gave me the contributions to the field of biochemistry and molecular biology. The Award is intended as a Travelling Lectureship to enable the opportunity to pursue a very important and still unsolved awardee to present his/her work at a number of centres within problem in structural biology, with direct application to Australia and New Zealand. Nominees must have been members of human health. The WEHI has proved to be an incredibly the Society for at least two years before the year in which the Award supportive and stimulating environment in which to nomination is to be considered. The award carries an honorarium to cover the travelling expenses, provided through the generosity work. In 2013, my lab finally obtained the first atomic of Beckman Coulter. picture of insulin bound to its receptor and published it in Nature. Since then my laboratory has sought to build on this finding, not only to fill in the many gaps remaining but to pursue active ways of translating it into better The Beckman insulins and into applications in cancer.

Coulter Discovery The Eppendorf Edman Award is awarded to a biochemist or molecular biologist with no more than seven years postdoctoral Science Award: experience, in recognition of their outstanding research work. The Award provides funds to assist the recipient to attend an Mike Lawrence overseas conference in a field associated with biochemistry or molecular biology or to briefly visit a research laboratory in Australia or elsewhere to access specialised equipment or to learn new research techniques. Applicants must have been members of the Society for at least 12 months before the year in which the Award application is to be considered. The contribution to travel expenses is provided through the I began my research career in 1978, undertaking a PhD generosity of Eppendorf South Pacific. degree in theoretical physics at the University of Cape Town (UCT) in South Africa. My thesis concerned the semi-classical modelling of the quantum mechanics of protein tunnelling in hydrogen-bonded compounds, and it was heavily based on my mathematics and physics background. However, I was always fascinated by the The Eppendorf application of these disciplines to the life sciences and in 1980, I was fortunate to secure funds for a two-year Edman Award: postdoc at the MRC Laboratory for Molecular Biology in Natalie Cambridge, UK. This was a turning point in my career and I found myself on a very steep learning curve! During Spillman the postdoc I determined the structure of the T-state of the allosteric enzyme phosphofructokinase from Bacillus stearothermophilus and developed techniques in electron micrograph image alignment, some of which are still used During my research-focussed Bachelor of Philosophy today in cryo-tomography. degree at the Australian National University, I developed an I then returned to South Africa for five years. interest in parasitology, and have spent my career studying Unfortunately, South Africa had no resources at the time the malaria parasite. Living inside a host red blood cell for structural biology, but nevertheless I secured the presents many challenges to the parasite, and the tricks and directorship of a small electron microscopy facility within unique biology it uses to thrive in this environment keep the science faculty at UCT and had quite a bit of fun. me fascinated every day. I completed my PhD at ANU in In 1988, I emigrated to Australia and joined the research 2012 under the supervision of Professor Kiaran Kirk and Dr group headed by Peter Colman, which proved to be a Richard Allen, investigating Na+ and pH regulation in the further turning point of my career. During my eighteen malaria parasite. Although I was interested in Na+-transport years at CSIRO, I worked on a number of particularly as an elementary and crucial aspect of cellular physiology, challenging projects in structural biology, including I demonstrated that a key transporter involved in parasite determination of the structure of phaseolin (a seed Na+-regulation, a P-type Na+-ATPase called PfATP4, was storage protein), the fusion protein of Newcastle disease involved in resistance to a suite of new antimalarial drugs. virus (NDV-F) and, ultimately, the extracellular domain My PhD work sparked a flurry of work in the malaria field of the human insulin receptor, in a project led by Dr concerning PfATP4 and new compounds targeting PfATP4. Colin Ward. The insulin receptor project terminated in Following this, for my postdoctoral work I switched 2006 and I decided to accept a position as a laboratory from cell biology techniques studying ion fluxes, to head at the Walter and Eliza Hall Institute of Medical using biochemical analyses and genetic manipulation to Research (WEHI) in Parkville, Victoria, to continue the understand how the parasite modifies and alters its red quest to determine how insulin bound to and activated blood cell host. I also switched continents, and moved Page 26 AUSTRALIAN BIOCHEMIST Vol 48 No 1 April 2017 Medallist and Awardee Profiles to Washington University School of Medicine, in Saint a component of the Raf/MEK/ERK signalling cascade that Louis, Missouri, USA. Under the mentorship of Professor is often deregulated in cancer. This experience not only Daniel Goldberg I investigated how the malaria parasite expanded her repertoire of cell biology and biochemistry alters erythrocyte signalling lipids, and hypothesised why techniques but also kick-started her interest in cell signalling. this is important for disease pathogenesis. My overseas In 2010, Yan started her PhD under the guidance of Dr postdoctoral studies were supported by two competitive Dominic Ng and Associate Professor Marie Bogoyevitch, fellowships, the Australia to USA Fellowship from the where she aimed to characterise signalling pathways that American Australian Association and an NHMRC CJ target the microtubule network, specifically in the context Martin Overseas Biomedical Early Career Fellowship. of cellular stress. She discovered that stress stimulation In 2016, I returned to Australia and moved to the activates both the c-Jun N-terminal kinase and cyclic-AMP Biochemistry and Molecular Biology department at the protein kinase pathways, which then act synergistically to University of Melbourne. Here I am part of the laboratory protect the microtubule network from depolymerisation by of Professor Leann Tilley, working on untangling the phosphorylation and subsequent inactivation of stathmin, a molecular mechanisms of drug action and drug resistance. In microtubule destabilising protein. This work, published in the future, I hope to combine my interests in cell physiology, the Journal of Biological Chemistry, highlights a mechanism by membrane signalling and drug resistance to continue to which the cell is able to adapt to deleterious stress stimuli in unravel the unique biology of the malaria parasite. order to maintain cell viability. I am honoured and excited to be the recipient of the 2017 Keen to continue working on aspects of microtubule Eppendorf Edman Award, which will allow me to present function, Yan joined the laboratory of Dr Mark Dodding my current research at the EMBO/EMBL Molecular and Cell at the Randall Division of Cell and Molecular Biophysics, Biology of Membranes meeting in Heidelberg, Germany. King’s College London for her postdoctoral work. The key research interest of the Dodding lab is to understand the The Boomerang Award is offered to an outstanding expatriate molecular basis of microtubule-based intracellular cargo Australian biochemist or molecular biologist to provide an transport, having solved the first crystal structure of the opportunity to return to Australia to present their work in a interface between the kinesin-1 microtubule motor protein symposium at the ComBio meeting and to give seminars at universities or research institutes. This will provide the awardee and its cargo. In collaboration with Dr Roberto Steiner, also with exposure in Australia and will facilitate interactions with at the Randall, Yan found that the light chains of kinesin-1 local researchers. The Award will include free registration undergo a cargo binding-induced conformational change at ComBio, as well as significant contributions to travel and that confers activity to the holoenzyme. Identification of accommodation expenses. Applicants must have been a member of a recognised Australian scientific society for at least this novel molecular switch extends our understanding of two years, and be no more than seven years since the award how cargo selectivity can impact kinesin-1 function and this of their PhD. The contribution to travel expenses is provided work was recently published in PNAS. by ASBMB. Yan is grateful to the ASBMB for the Boomerang Award. She will use this opportunity to present her work at ComBio2017 in Adelaide and to engage with research labs in Brisbane and Melbourne.

The Boomerang Award: Yan Yan Yip

Yan Yip’s first exposure to biomedical research was at the Department of Pathology, the University of Melbourne, as an Honours student under the supervision of Dr Wendy Cook. She studied the post-transcriptional regulation of a novel gene that is silenced in acute myeloid leukaemia, learning a variety of molecular biology techniques along the way. Yan then joined the laboratory of Dr Amardeep Dhillon at the Department of Biochemistry and Molecular Biology located within the Bio21 Molecular Science and Biotechnology Institute as a research assistant. Here, she helped identify key determinants of Raf-1 kinase activity,

Vol 48 No 1 April 2017 AUSTRALIAN BIOCHEMIST Page 27 ASBMB FELLOWSHIP PROFILES

The ASBMB Fellowships are awarded annually to biochemists or molecular biologists, in their early career and normally resident in Australia, in recognition of their outstanding work in an area of biochemistry and molecular biology. The Fellowships provide funds to assist the recipient to attend an overseas conference in a field associated with biochemistry or molecular biology or to briefly visit a research laboratory in Australia or elsewhere to access specialised equipment or to learn new research techniques. Applicants must be at least in the second year of PhD training and not more than 2 years subsequent to the award of the PhD degree. Applicants must have been members of the Society for least 1 year immediately prior to application.

SAMUEL ROBINSON - recipient of the Fred Collins Award for the most outstanding ASBMB Fellowship applicant Samuel Robinson completed his BSc and BSc (Honours) at the University of Auckland, New Zealand. In 2011, he received an Australian Postgraduate Award, to undertake a PhD at Monash University, supervised by Professor Ray Norton. Since the completion of his PhD in 2015, for which he was awarded the Mollie Holman medal, he has been based at the University of Utah, USA, working as a postdoctoral fellow in the lab of Professor Baldomero Olivera. This year he will return to Australia to join Professor Glenn King and Dr Eivind Undheim at the University of Queensland. His research interest is in understanding the composition and mechanism of action of animal venoms. A major research highlight has been his pioneering of the use of proteogenomics for the identification of new, previously hidden, classes of bioactive peptides. One outcome of this was the discovery that a class of specialised insulins form an integral part of the venoms of some species of marine cone snail. This provided ground-breaking insight into the structure-activity relationship of insulin, and has led to the preclinical development of a new anti-diabetes agent. This research was featured by numerous newspapers and science magazines around the world, including The Washington Post, The Guardian, New Scientist, Scientific Americanand National Geographic. This ASBMB Fellowship will allow him to spend time working in the lab of Dr Beatrix Ueberheide, a world expert in mass-spectrometry, at New York University, USA, where he will aim to expand his knowledge and skillset in the identification and characterisation of venom peptides.

GEORGIA ATKIN-SMITH Georgia Atkin-Smith completed a Bachelor of Biotechnology and Cell Biology at La Trobe University in 2013. She then completed her honours in Biochemistry under the supervision of Dr Ivan Poon and Dr Mark Hulett, finishing with first class honours. During this year, Georgia began to work on the molecular mechanisms and functions of apoptotic cell disassembly in monocytes, demonstrating a novel mechanism of disassembly which resulted in a first author Nature Communications paper. Now in the third year of her PhD, Georgia has continued her work on the disassembly of apoptotic monocytes with a first author Scientific Reports manuscript characterising a novel method to isolate apoptotic bodies. Georgia has also published a review in Trends in Cell Biology and contributed to publications in Nature Protocols and Apoptosis. Current unpublished findings include the identification of the first positive regulator of apoptotic cell disassembly and the elucidation of the role of disassembly in infection in both viral propagation and immune detection. Recently, Georgia was a finalist for the Research Impact Victorian Young Achievers Award 2016 and was awarded the overall People’s Choice. Additionally, after a two week research placement in a laboratory in Tokyo, Japan, Georgia was invited to present her current research findings at four universities throughout Tokyo and has also been selected for numerous domestic conference oral presentations. With the help of this ASBMB Fellowship, Georgia is planning to travel to Toronto, Canada, this year to present her research at the International Society of Extracellular Vesicles Annual Conference.

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STEPHANIE BEGG Stephanie completed a Bachelor of Science (Biomedical Science) at the University of Adelaide in 2012, majoring in Microbiology, Immunology, and Biochemistry. Following her undergraduate studies, she was accepted into the accelerated Masters of Philosophy program within the Research Centre for Infectious Diseases (RCID), under the supervision of Associate Professor Christopher McDevitt and Professor James Paton. She was later upgraded to a full PhD to further her research into the human bacterial pathogen Streptococcus pneumoniae. Her thesis investigated this bacterium’s cellular requirement for essential trace metals, such as manganese and zinc, and focussed on how S. pneumoniae steals these metals directly from the host environment. This scavenging by invading bacteria is crucial for both their survival and ability to cause disease within humans. Her research delivered both first author and co-authored publications in journals such asNature Chemical Biology, Nature Communications and ACS Chemical Biology. Stephanie was awarded the University Doctoral Research Medal upon completion of her PhD in 2016, and is now a postdoctoral researcher in the RCID. She is currently investigating the molecular details of how metal poisoning and starvation impact S. pneumoniae, as these processes are directly relevant to how the human immune system attempts to kill invading bacteria. This ASBMB Fellowship will allow Stephanie to attend the Cell Biology of Metals Gordon Research Conference in Vermont, USA, in July 2017, and present her recent research into the prokaryotic metalloproteome to an international audience.

YILIN KANG Yilin Kang obtained a Bachelor of Science with First Class Honours in 2014 from the University of Melbourne, majoring in Biochemistry and Molecular Biology. In the final year of her undergraduate degree, Yilin was exposed to mitochondrial protein biogenesis in the lectures of Dr Diana Stojanovski, a mitochondrial biologist from the Bio21 Institute. Mitochondria immediately caught Yilin’s interest and she was fortunate enough to undertake a mini project in the lab of Dr Stojanovski, where she developed a passion for mitochondrial research. Yilin stayed on with Diana for her Honours project in 2014 and is now in the third year of her PhD. Her doctoral studies have focused on the characterisation of a human mitochondrial protein import machinery – the Translocase of the Inner Membrane 22 (TIM22) complex. Although protein transport systems are well-studied in yeast, the human TIM22 complex were still poorly characterised when Yilin commenced her PhD. Yilin’s work has uncovered a novel, metazoan-specific subunit of the human TIM22 complex, Tim29, which has a role in mediating the assembly and stability of the mature TIM22 complex. Identification of Tim29 highlights the importance of analysing protein import systems across phylogenetic boundaries, which can reveal novel facilitators and mechanisms in protein trafficking. Yilin’s work on Tim29 was published in Elife in 2016 and she has presented her research at several conferences including AussieMit 2016, where she received a Young Investigator Award. This ASBMB Fellowship will allow Yilin to attend and present her studies on mitochondrial protein translocation at the 2017 EMBO Conference on Protein Translocation and Cellular Homeostasis in Dubrovnik, Croatia.

Short Discussions SDS Page for Students Page Publishing your First first-author paper You have worked hard in the lab and your supervisor thinks you have a nice story to tell. Job done, right? No, now it is time to convince your academic peers that your work is worth publishing in a peer-reviewed journal. Your research is only going to reach others if it is being communicated effectively and publishing in a journal is a good starting point. To be a successful researcher, not only do you need to have motivation, perseverance, time management and technical skills but you also need to be able to write well. Writing a scientific paper is something that comes naturally to some, but not to others. It may seem like a daunting task and a good paper may take a very long time to write, but hopefully the following tips can help you with the process (which I am still learning myself!).

Vol 48 No 1 April 2017 AUSTRALIAN BIOCHEMIST Page 29 SDS Page

1. Decide on your main message Writing a paper is merely a way to tell people what you have discovered. Decide on what story you want to tell and what the main point of your paper will be. Then you pick the results that best help you tell your story. List all your results and decide on whether you have enough ‘evidence’ to draw your conclusions. It can be a bit disheartening when you realise that only a small fraction of your results actually make it into a paper. You should resist the temptation to try and cram in as many results as possible in order to tell multiple stories in one paper. The key is to put yourself in the 100% Australian reader’s shoes to get your message across. owned company supplying scientific laboratories 2. Pick a journal throughout Australia and Once you have decided on your take home message, decide on which abroad since 1987. journal you want to submit your article to. Your supervisor and collaborators will be able to help you decide on the most appropriate journal. Finding the ‘right’ journal can be difficult and you may want to take into account the research area, reputation and target audience of the journal. Assessing the quality of a journal by its impact factor is a controversial issue as there are other factors that can influence the impact factor, not necessarily related to the quality of the papers being published. Nevertheless, the impact factor is still one of the main criteria to consider when choosing a journal. It is always a good idea to check which journals publish work similar to yours. You typically aim high (more prestigious journals) to begin with because you can resubmit to less prestigious journals if you get rejected. Be honest with yourself about where you realistically can see your work getting published, otherwise you may lose valuable time (and sleep) reformatting papers for submission. Chances are that your first paper will not be published in Nature or Science – many might go through their whole careers without ever getting one! So don’t be disheartened as plenty of influential, highly cited and high-quality papers are published outside of these journals. Consider open access policies for the journals and publication fees. Once you have picked the journal, read the guidelines for authors that you must adhere to.

3. Prepare your figures Whenever I get some exciting new data, I try to generate publication- www.rowe.com.au

quality figures as soon as possible, which can then be used in presentations and manuscripts. A well-constructed figure is worth a thousand words. Of course your figures do need to abide by the target journal’s guidelines, so make sure you can still edit them if need be. The figures and tables, including the legends, should be self-explanatory and the information conveyed in a figure should not be repeated in another figure. I also find it

is a good idea to have a mixture of different types of figures for a paper. Any at online Order figures that don’t add to the main story but are important in supporting the paper can be included in the supplementary material. Don’t forget to include appropriate axes titles and units. TARSONS 4. Write a cohesive story – introduction, methods, results and discussion It is unlikely that you will be writing your first paper alone. Remember that the reviewers are not necessarily in your niche area of research and you South Australia & NT Ph: (08) 8186 0523 should make the review experience as painless as possible for them. I find it [email protected] useful to find a good paper in the same journal that you can use as a guide. Queensland Ph: (07) 3376 9411 A good introduction should convince readers that your work is important [email protected] and will address an unresolved problem or gap in the literature. Add Victoria & Tasmania Ph: (03) 9701 7077 enough background information so that the reader can understand the [email protected] remainder of the paper. Western Australia Ph: (08) 9302 1911 [email protected] The methods you used in your paper should include enough detail that New South Wales & ACT Ph: (02) 9603 1205 a fellow researcher can reproduce the experiment. But if a method is well [email protected] established, use references to previously published procedures to keep this 415-Australian.Biochemist.10.10.16 section succinct.

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Once you have prepared your figures and tables, you will find the results section relatively straightforward to compile. I tend to group the results together into subheadings if this is permitted by the journal. Keep the data in a logical order that helps you tell a coherent story, which is not necessarily the order you performed your experiments in. After you have presented your results, it is in the discussion that you highlight the importance of your data and explain what it all means. You need to put your data into the context of your field. When writing your discussion, try not to overstate the conclusions. Try to come up with all plausible explanations for your results and let the results speak for themselves. Pledge to Chemists The order in which you write these sections is up to you. One of the key If chemists in Australia are points in writing a good paper is ensuring that one section flows into the next. experiencing difficulty in obtaining supply, please 5. Title and abstract send me an email; peter. Even though a title is only a few words long, it often takes time to come up [email protected] and with one that you are happy with. Keep the title succinct and descriptive of I promise to help you. the content but not so technical that it cannot be easily understood. I know a number of people who make the decision to read a paper or not based on This is not a ‘subtle’ attempt the title. Think of the title as the advertisement of your article. to obtain more business, The abstract should be succinctly written and avoid the use of jargon. It is but a sincere pledge to help a way of highlighting the aim of the paper and the main findings. In some journals, the editor may only look at the abstract to decide whether a paper fellow scientists source the should be further considered and sent for review. items they need to do their work, and thereby help 6. Feedback Australia grow. Try not to dwell on your first draft too much before sending it to your supervisor for revision. More often than not, the draft will come back to This is the raison d’etre for you covered in red writing. Once you have incorporated your supervisor’s Rowe Scientific Pty. Ltd. feedback, pass it on to your co-authors. It is a requirement that all authors on the papers have revised and approved the final version of the manuscript. Sincerely, I also find it useful to get someone outside of my discipline or even a non- Peter Sommers (FRACI) scientist to read the final draft of the paper. It is unbelievable the errors they CEO Rowe Scientific Pty. Ltd. can pick up even when they only understand the meaning of a few words.

7. Submission and revision The senior author on the paper will be in charge of submitting the revised draft of the paper alongside a cover letter. The review process may take 100% Australian a few days but more likely, a few months. If major/minor changes are owned company supplying required you will have a timeframe to resubmit the paper. Consider the reviewers’ comments carefully as you will often have to prepare a letter scientific laboratories to respond to their feedback. When preparing your response letter, make throughout Australia and sure you carefully address each comment in a calm and methodical way. abroad since 1987. I always find it a good idea not to address the comments immediately after reading it for the first time. Also, don’t think of the comments as a personal attack on your research but merely a way of improving the quality Order online at of your publication. A glass of wine may also ease the process! Sometimes, www.rowe.com.au reviewers may request changes to the manuscript or even more data. If you South Australia & NT Ph: (08) 8186 0523 decide not to follow the request, make sure you provide a good justification [email protected] for not doing so. Simply ignoring a request or criticism will not be looked Queensland Ph: (07) 3376 9411 upon favourably by the editor. Once you have addressed the reviewers’ [email protected] comments, with any luck, your paper will be accepted by the journal. Victoria & Tasmania Ph: (03) 9701 7077 When your work gets published in a journal, you tend to forget about the [email protected] Western Australia Ph: (08) 9302 1911 pain you had to endure during peer review and you start looking forward [email protected] to putting together your next publication! New South Wales & ACT Ph: (02) 9603 1205 [email protected] The Student’s Page is coordinated by Dr Tatiana Soares da Costa, who is an NHMRC Early Career Fellow at the La Trobe Institute for Molecular Science 415-Australian.Biochemist.10.10.16 ([email protected]).

Vol 48 No 1 April 2017 AUSTRALIAN BIOCHEMIST Page 31 Australia Day Honours for ASBMB Members

Professor David Vaux Professor Richard was awarded an Officer of Harvey was awarded the Order of Australia (AO) a Member of the the for ‘distinguished service Order of Australia (AM) to medicine in the field of for ‘significant service to biomedical cancer research, medicine in the field of cell to higher education as an biology and cardiovascular academic and mentor, and research, and through to professional integrity scientific leadership roles’. and ethics.’ Richard was born in Dave Vaux (or ‘Davo’) is Adelaide and grew up currently Deputy Director in Bute, a small one-pub of the Walter and Eliza country town in the upper Hall Institute (WEHI). He reaches of Yorke Peninsula trained in medicine at the in South Australia. He did University of Melbourne, a BSc at the University of spending a year in the middle of his degree to do research Adelaide, continuing with Honours and a PhD in the in Gus Nossal’s lab. He returned to WEHI after his intern Department of Biochemistry under the supervision of year at the Royal Melbourne Hospital to undertake a PhD Julian Wells. studying B cell malignancies supervised by Jerry Adams. After a short period with a biotech company in Working on the lymphoma associated Bcl2 gene, he Strasbourg, France, where he cloned and expressed (in showed that unlike other cancer genes known at the active form) the potent anti-thrombin anticoagulant time, it did not promote cell growth and proliferation, Hirudin from the medicinal leech, he undertook but stopped cells from being able to kill themselves. This postdoctoral studies at Harvard University with showed that cell division and cell death were separately Doug Melton, where he began his life-long interest in regulated independent processes. Bcl2 thus became the first embryonic development. component of the cell death mechanism to be recognised. He established an independent group at the Walter Because of the association of Bcl2 translocations with and Eliza Hall Institute in Melbourne, where he spent lymphoma, it also became clear that failure of cell death ten years. In 1998, he relocated to the Victor Chang could promote malignancies in humans, and avoidance of Cardiac Research Institute, where he is currently Co- apoptosis is now recognised as one of the ‘hallmarks of Deputy Director and Head of the Developmental and cancer’. Stem Cell Biology Division. His research focuses on the He showed that Bcl2 could synergise with the c-myc genetic basis of heart development and the pathological oncogene to transform cells in vitro, and by making Bcl-2 mechanisms underlying congenital heart disease, and transgenic mice and crossing them with myc transgenic cardiac regeneration. He holds the endowed Sir Peter mice, he showed they could also synergise to cause Finley Professorship of Heart Research at the University leukemia in vivo. of New South Wales, and is a Fellow of the Australian In 1989 he went to Stanford, and together with Stuart Academy of Science, Australian Academy of Health Kim made transgenic C. elegans worms expressing and Medical Science, EMBO and The Royal Society of Bcl2, and showed that the mechanism for apoptosis of London. He has been a member of the ASBMB since 1998 mammalian cells and the mechanism for programmed and in 2010 was awarded the Lemberg Medal. cell death in the worm were evolutionarily conserved. Expression of human Bcl2 in the worms prevented most of the cell deaths that were normally programmed to occur during development, and showed that the mechanism for apoptosis of mammalian cells and the mechanism for programmed cell death in the worm were evolutionarily conserved. His association with biotech companies IDUN and TetraLogic helped in the translation of discoveries of the basic biology of cell death into drugs that inhibit the Bcl2 and IAP families of proteins. He has an interest in promoting integrity in research, and has long advocated establishment of an office for research integrity in Australia.

Page 32 AUSTRALIAN BIOCHEMIST Vol 48 No 1 April 2017 Communicating the Message

Rebecca Lew recounts her journey from budding young biologist to endocrinology researcher and medical writer.

experience on the role of the pineal gland in non-shivering thermogenesis in woodchucks, which was intense Rebecca Lew. (especially when I killed one by accidently giving it ten times the intended dose of adrenalin – oops). Then, the Catching the Biology Bug next summer, I worked at the Monell Chemical Senses Most scientists seem to express their scientific nature Center in Philadelphia investigating how alpha female early in life, and I was no exception. I was the youngest marmosets (a type of little monkey) use pheromones of five children growing up in a suburb of Philadelphia. and social cues like grooming to suppress reproduction My father was a chemist at DuPont, developing specialist in subordinate females. In my final year at UD, I did a spray paints for General Motors cars and the lining of thesis project (essentially the same as Australian Honours Coors beer cans, among other things, while my mother projects) on the role of the sympathetic nervous system in was a self-taught librarian and local historian. I spent the vasopressor effect of angiotensin in bullfrogs (yes, frogs most of my childhood days catching frogs in our ponds, again!). That thesis project resulted in my first published collecting all manner of natural ‘treasures’, playing with paper in General and Comparative Endocrinology. Although my stuffed animals (I never really played with dolls), these projects were disparate in scope and species, they and roaming the neighbourhood with my friends, as you all had one thing in common – communication between could in those days. My scientific side was fostered by distant cells via chemical messengers, ie, endocrinology. my father, who was also a keen fisherman and keeper To this day, endocrinology is still my favourite discipline. of fish – I knew the Latin names of most of his tropical (Lesson 3: Pursue a range of topics in your research career; killifish, including my personal favourite, Aphyosemion you will likely find that the things that interest you have bivittatum (pronounced exactly as it’s written, folks). I a common underlying theme. Oh, and, always double- loved animals and always thought I would make a living check your calculations!) doing something involving animals. (Lesson 1: Your basic personality develops early in life, so revisiting your PhD Years childhood can help you remember who you are.) I did very well at UD and finished up with a 4.0 grade My childhood and adolescence were pleasant and point average, which meant I could pretty much pick and despite my early conviction of a career centred on choose where to go for a PhD (I never considered anything animals, I toyed with other options, including being a but more study and research). My short list came down to doctor and writing poetry. Mostly, I just drifted through Cornell University (where my dad did his PhD), with its school, although I was deemed smart enough to be in focus on veterinary science, and the University of Virginia the ‘advanced placement’ group and managed good enough grades. When the time came for university, I didn’t look far – just an hour down the road (albeit in a different state) at the University of Delaware (UD). I entered UD’s fledgling Honours Program in the autumn of 1980, which meant I was in a dorm with other Honours students and got to attend smaller freshman classes of about 25–30 students instead of ten times that number. I was a Biology major, and my freshman Honours Biology class was taught by Steven Skopik, who was fabulous! He was both enlightening and entertaining, and a perfect example of how a great lecturer can ‘turn you on’ to the subject. I definitely caught the bio bug, thanks to Professor Skopik. (Lesson 2: Never underestimate the value of good teaching.) My years at UD saw my first forays into research. Atop the now-gone World Trade Center in New During my second year, I spent a semester getting lab York with fellow PhD students, circa 1985.

Vol 48 No 1 April 2017 AUSTRALIAN BIOCHEMIST Page 33 (UVA), with a strong physiology department. I visited that didn’t entice me the first few times Michael asked me both campuses, but was swayed towards UVA mainly out. However, his persistence (and charm) eventually won because the PhD students there were great fun! Unlike out, and by the start of 1987, we were officially an item. in Australia, most PhD programs in the US (at least in After a year or so, we decided to get married, which those days) take 4 to 5 years, and include a coursework we did in April 1989. I was still about a year away from component as well as the independent research project. finishing my degree, so Michael had to scrounge to find UVA Physiology was particularly tough – two solid years money to extend his time in the States. His particular visa of coursework before you were allowed to start your PhD type required him to leave the US when his postdoctoral project, although you were encouraged to do lab rotations studies were complete, and he could not work in the US during your second year to help decide on your supervisor for two years after leaving – this despite being married and project. I personally think that this format provides to a US citizen and at the same time as a general amnesty you with an excellent, well-rounded scientific education, for illegal immigrants was declared (Mike, of course, was beyond just your focussed field of research. legal – he’d have been better off if he weren’t). So it was I didn’t appreciate it at the time, but the physiology always clear that I needed to find a postdoc in Australia, department at UVA in the ‘80s was among the very best which in those pre-internet, pre-email days, wasn’t so in the world, with big names like Bob Berne (department easy. I started looking through the journal Endocrinology, chair), Brian Duling, Gary Owens, and many more who and one name kept cropping up – John Funder. I wrote to Australian biochemists have probably never heard of – Funder, who replied that he was going to the Endocrine you’ll just have to trust me! (Lesson 4: Appreciate and Society meeting in Seattle and we could meet up there. make the most of the scientific talent around you.) In part It was a brief, but apparently successful, meeting, and because of my interest in endocrinology, I selected Alex eventually, I had a postdoc lined up with Ian Smith, who Baertschi as my supervisor. Alex was a Swiss import who worked with Funder at the old Prince Henry’s Institute had a number of research interests in the neuroendocrine of Medical Research (PHIMR) off St Kilda Road in field. I dabbled with a couple of ‘out there’ projects, Melbourne. Michael and I arrived in Melbourne on Anzac which in retrospect were really cutting-edge, including Day 1990 (after some harrowing days waiting for my visa investigating the neuroendocrine-immune axis, which to be approved), and I started work at PHIMR soon after. would have been really cool if I’d been able to make it (Lesson 5: Don’t be afraid to venture far from home, both work... In the end, I settled on investigating how hypoxia literally and figuratively.) stimulated the release of a newly discovered hormone from the heart, atrial natriuretic peptide (ANP), using Early Days with Enzymes in the Land of Oz both isolated rat hearts and cultured atrial cells. Funder had a project idea for me, which took advantage of my background in cardiac hormone research and Ian’s expertise in high-performance liquid chromatography, which I was quickly schooled in. The idea was based on the high expression of an amidating enzyme in the atria of the heart, the same tissue that synthesised ANP. This enzyme catalyses the C-terminal amidation of many hormones – but ANP was not one of them. The hypothesis was that there was another atrial peptide hormone that was amidated – so the search began. Ian and I developed methods to fractionate and assay heart extracts in such a way as to identify putative amidated peptides. The idea was sound in theory and the techniques worked on control samples, but we never did find anything. To the

With future husband Michael in the Everglades, circa 1988. Southern Charm When I decided to go to UVA, my grandmother, whose family was originally from the South, said “You’ll meet some nice Southern gentleman who you’ll marry.” Well, it turned out the gentleman was from further south than Nana intended! Towards the end of my second year, an Aussie postdoc joined Brian Duling’s lab, next door to Alex’s lab. This guy had red hair, a bushy beard, a funny accent, and was loud and brash – not anything like what I was looking for in a boyfriend. Crocodile Dundee had just Becky and son Brian with the Ian Smith lab on the occasion of Corie come out, so Aussies were all the rage (apparently), but Shrimpton’s PhD at the Baker Medical Research Institute in 1998.

Page 34 AUSTRALIAN BIOCHEMIST Vol 48 No 1 April 2017 best of my knowledge, no one else ever has either, and the accepted wisdom now is that the amidating enzyme plays a completely different role in the heart than in other endocrine tissues. I spent just a few months at PHIMR before moving to the Baker Medical Research Institute when Funder became director there. The Baker was, and still is, known for its focus on cardiovascular disease, and was where my husband did his PhD work. Michael had returned to the Baker after UVA, so we were reunited (for a while, anyway) with my move from PHIMR. The Smith lab settled in well, and what followed were some very enjoyable years with great colleagues. After the quest for the elusive amidated atrial peptide fizzled out, I branched out to other enzymes Rebecca with ProScribe colleagues at a rope-climbing involved in the metabolism of peptides regulating activity at Taronga Zoo, March 2017. cardiovascular function. The most famous such enzyme is angiotensin converting enzyme (ACE), the target for the To Monash and Beyond antihypertensive ACE inhibitor class of drugs that are in In 2004, the Smith lab moved to the biochemistry widespread therapeutic use. However, we focussed on department at Monash, soon after Chris Mitchell took more obscure enzymes, particularly one affectionately over as head. The lab had been collaborating with people known as ‘24.15’, after its Enzyme Commission number, there, such as Mibel Aguilar and Patrick Perlmutter, EC 3.4.24.15. This enzyme is involved in the degradation for several years, and both Ian and I had given lectures of several bioactive peptide hormones, including the there. Thus, the ties were strong, and the department was vasodilator bradykinin. In collaboration with labs in vibrant and more aligned with where our research was Australia and worldwide, the Smith lab characterised heading than the Baker. During this period, I started to 24.15 and its role in peptide metabolism. I spent most branch out on my own a bit more and come out from of the 1990s dedicated to this little enzyme (confession: under Ian’s shadow (Ian’s a big man, both physically and throughout the year 2015, I would often say “2415” because in personality and reputation). I had considered leaving I had said it so often) and its cousins, but came up short on Ian’s lab on several earlier occasions, but circumstances anything groundbreaking. However, I enjoyed the work, and inertia prevented that, undoubtedly to the detriment my labmates, and annual pilgrimages to the Lorne Protein of my research career. One area of interest was lecturing, Meeting, and Ian kept us well funded – what more could so I increased my involvement with the departmental I want? Although the publications came at a steady pace, education group. Another area was academic publishing I suppose my most important achievements during this – I was one of those freaks who actually enjoyed writing period were the creation of two sons, Brian in 1997 and papers – so when I read in the Australian Biochemist that the Daniel in 2000. (Lesson 6: As much as I enjoyed the Smith then-Editor, Phillip Nagley, was looking for people to join lab, I realise in retrospect that I was too comfortable and an editorial committee, I signed up. This little step turned spent too long there – be sure to change jobs a bit more out to have major consequences, in that it directly led to often than every 15 years…) me becoming Editor at the start of 2006, which, in turn, led to my big career shift later that year. (Lesson 7: You never know where things might lead, so if an opportunity comes up that interests you, take it.) During the early 2000s, I began to realise that I wasn’t a star researcher and was unlikely to ever become one. Worse still, I was enjoying research less and less – having to focus on one, relatively narrow topic was, quite frankly, becoming boring. After a couple of false starts with alternative careers, I stumbled upon an ad in Seek for a ‘medical writer’, a job which I had never known existed, with a company called ProScribe. I applied, completed the writing assessment and interview, and was offered the job! Now being the cautious person I am, I didn’t want to burn my bridges behind me, so for the first year or so, I worked half-time with ProScribe and half-time ASBMB Editorial Committee meeting held at ComBio2005 as a lecturer at Monash. I had great flexibility, thanks to (clockwise from top left): Sam Richardson, Nghia Le, Liana both employers, especially as ProScribe at that time was Friedman (Editorial Officer), Agnieszka Lichanska, Graham a ‘virtual company’ with only home office-based writers. Baldwin, Phillip Nagley (ASBMB President standing in for It didn’t take long before I knew I had found the perfect Editor Clem Robinson) and Rebecca Lew (Editor Elect). job for me.

Vol 48 No 1 April 2017 AUSTRALIAN BIOCHEMIST Page 35 Career Switch Despite having been a medical writer for ten years and Medical writing mainly includes assisting authors with rising through the ranks, I feel like I am still a beginner manuscripts, abstracts, posters, and slide presentations and am learning all the time. This is probably the most (usually for clinical trials), but can also include a range of rewarding aspect of my job – there is nothing quite like other document types, such as submissions to regulatory the feeling of starting a new project and knowing that, agencies like the Therapeutic Goods Administration. although you have done similar projects in the past, each Although the focus of a medical writing job is, of course, one will bring new challenges and new rewards. I believe writing, there are many other aspects that I discovered I that continually challenging yourself is a critical part of was good at. Analytical thinking, project management, fulfillment, whatever your career choice may be. attention to detail, diplomacy, efficiency, etc. The job is as deep as it is wide – everything from understanding the science behind a study and its place in the literature down to grammar and punctuation. Over the years, I have worked on a broad range of projects (including manuscripts, abstracts, posters, slides, grants, medical education programs, and regulatory documents) in a broad range of therapeutic areas (including oncology, diabetes, ophthalmology, psychiatry, dermatology, gastroenterology, neuroscience, osteoporosis, and more). I’ve also worked with countless clients and authors from around 25 countries worldwide and have had the opportunity to travel to several Asian countries as part of my job. Needless to say, boredom has never been an issue! I have also had the privilege to work alongside some incredibly smart, dedicated people, and have even helped train a few with ties to ASBMB… (Lesson 8: Understand Rebecca with husband Michael and your own interests, skills, and limitations, which will help sons, Daniel and Brian (right). you find a career that suits you best.) ELECTION OF COUNCIL 2018 Nominations are called for the following positions on the Council of the Australian Society for Biochemistry and Molecular Biology Inc for 2018: Secretary, Treasurer, Editor, Secretary for Sustaining Members and State Representatives for ACT, NSW, Qld, SA, Tas, Vic and WA.

The Council for the President L. Tilley period 1 January 2017 President Elect nominations sought to 31 December 2017 Secretary B. Forbes# is composed of the Treasurer T. Piva* following members: Editor S. Mathivanan# Education Representative S. Rowland# Secretary for Sustaining Members S. Jay#

Representatives for: ACT Y.P. Mabbitt* NSW K. Michie# * Retiring member, not Vic E. Lee# eligible for re-election Qld D. Ng* # Eligible for re-election SA S. Polyak# Tas K. Brettingham-Moore# WA N. Taylor*

Nomination forms are available on the ASBMB website. Nominations for all vacant positions must be signed and seconded by members of the Society. The nominations must be signed by the nominee to indicate his/her willingness to stand. If more than one nomination is received for any position, a ballot will be held to determine the successful candidate. All members may vote for all positions except those of State Representatives where election is by members in the State concerned.

NOMINATIONS MUST REACH THE SECRETARY BY 5PM 21 SEPTEMBER 2017 (14 DAYS BEFORE THE ANNUAL GENERAL MEETING TO BE HELD AT 1:15PM 5 OCTOBER 2017)

Page 36 AUSTRALIAN BIOCHEMIST Vol 48 No 1 April 2017 Sydney Protein Group: an ASBMB Special Interest Group

History The Sydney Protein Group (SPG) has a rich history of promoting protein science locally. The SPG is made up of protein scientists and students from academia, hospitals and industry that meet regularly to hear updates from local and international speakers on the latest developments in protein science.

SPG Meetings and Early Career Researcher Support The SPG organises and supports several annual events, with the help of funding from the ASBMB and dedicated trade partners. An important Sydney Protein Group ethos is to provide opportunities for young researchers working in the 2016 Thompson Prize finalists and judges. field of protein science to attend high quality scientific From left: Margie Sunde (USyd – Judge), Lisa Belfiore meetings, where they can hear about recent research, (UOW), Tony Ngo (VCCRI), Neil Robertson (USyd), Emma interact with guest speakers and other researchers, and Dawson (UTS), Samira Aili (UTS), Martina Sanderson-Smith present their own work. An initiative of the ASBMB in (UOW – Judge) and Mark Molloy (Macquarie – Judge). recent years is to offer a SIG-sponsored speaking slot at ComBio for a promising early career researcher. This year target. Next, Lisa Belfiore (UOW) spoke about her PhD the SPG-sponsored early career researcher who spoke at research on the development of drug-loaded liposomes ComBio2016 was Laura McCaughey from the University to target breast cancer cells. Her work included the use of Technology Sydney (UTS). Laura talked about of single molecule fluorescence to quantify how many characterising a new protein antibiotic from Pseudomonas targeting moieties there were per liposome — a first of its aeruginosa and its potential as a new target in the fight kind! Neil Robertson (University of Sydney; USyd) told us against bacterial drug resistance. about his innovative FRET-based techniques to elucidate the binding kinetics of the LIM-LID domains. These proteins are notoriously difficult to work with in isolation and hence were not amenable to standard biophysical analyses. Samira Aili (UTS) introduced us to the world of ant venom and its potential as a source of bioinsecticides. Laura McCaughey Her lead compound(s) looks promising and could one day (UTS) was the be applied in the field to help ensure our food security. SPG-sponsored As expected, the talks were of superb quality and the early career range of topics certainly impressed the judges [consisting researcher of Margie Sunde (USyd), Mark Molloy (Macquarie who spoke at University) and Martina Sanderson-Smith (UOW)]. In ComBio2016. the end, the judges agreed that Emma Dawson was to be awarded the 2016 Thompson Prize. The SPG also offers several prestigious prizes aimed at 2) The SPG also award three annual Lorne Travelling PhD/honours students: Scholarships to help the best and brightest PhD students 1) The annual Thompson Prize is in recognition of the attend the Lorne conferences. eminent local protein scientist, EOP (Ted) Thompson. The The Lorne Travel awards went to Neil Robertson (USyd) Thompson Prize was inaugurated in 1992 and is awarded winning the ATA Scientific Lorne Travel Award and for the best oral presentation by a young local scientist in Claudia Kielkopf (UOW) winning the SPG Lorne Travel the field of protein structure and function. Award. The Greg Ralston Memorial Award (Honours The Thompson Prize moved further afield in 2016: it was student continuing on to do a PhD) went to Gabrielle held as a half-day meeting at the Illawarra Health and McClymont (USyd). Medical Research Institute, University of Wollongong 3) Lastly, look out for the East Coast Protein Meeting (UOW), hosted by Heath Ecroyd and Justin Yerbury. Tony (ECPM) this year in Coffs Harbour (14–16 July), organised Ngo (Victor Chang Cardiac Research Institute) kicked off by the SPG. The ECPM is a joint initiative with the the talks by describing his work on CoINPocket and how Queensland Protein Group that is held every two years. he has used this bioinformatics tool to predict ligands The focus of this meeting is on Early Career Protein for orphan GPCRs. Emma Dawson (UTS) described her Researchers who present their work alongside a few work on newly characterised dynamin-like dlp proteins keynote speakers. and how they contributes to membrane integrity of Jason Low and Liza Cubeddu Helicobacter pylori, suggesting a possible future drug website: www.mmb.usyd.edu.au/spg/

Vol 48 No 1 April 2017 AUSTRALIAN BIOCHEMIST Page 37 Metabolism and Molecular Medicine: an ASBMB Special Interest Group

The Metabolism and Molecular Medicine Special vagal afferents from the stomach control satiety and how Interest Group (MMM-SIG) is one of the largest in sensitivity of these neurons changes in a diurnal fashion the ASBMB, with more than 120 members. Since and in response to nutrient excess. Dr Frances Byrne from 2014, the group has been under the direction of Nigel the University of NSW then gave an excellent overview Turner (University of NSW) and Sean McGee (Deakin of the role that a specific glucose transporter, GLUT6, University). The goal of the MMM-SIG is to support plays in the development of endometrial cancer. The final activities that showcase the best research in this field speaker of the symposium (short talk selected from the from Australia and abroad. This has included active abstracts) was Stephen Fairweather from the Australian contributions to the ComBio meetings, as well as National University, who discussed the physiology and financial support for other conferences that are relevant biochemistry of amino acid transporters. to members of the MMM-SIG. In 2015, the MMM-SIG was a sponsor for the 1st Australian Cancer and Metabolism Meeting (ACMM 2015). This meeting was initiated to provide an opportunity for scientists with expertise in nutrient metabolism to engage with cancer biologists and discuss the latest developments and advances in this rapidly expanding field. The meeting was attended by approximately 180 delegates and the broad topics covered across the meeting were the involvement of lipid metabolism, nutrient uptake, obesity and metabolic signalling in the regulation of cancer cell growth, as well as a session on therapeutic targeting of metabolic pathways to treat cancer. The keynote speaker for the Enjoying the welcome mixer at ComBio 2016. From left: meeting was Professor Brendan Manning from the international speaker Matthew Hirschey (Duke University), School of Public Health at Harvard University. He gave Arthe Raajendiran, Matthew Watt and Sean McGee. an excellent presentation on the control of nutrient metabolism and cancer cell growth by the mechanistic In 2017, the MMM-SIG will be supporting the 2nd target of rapamycin (mTOR). Throughout the meeting, Australian Cancer and Metabolism Meeting scheduled for there were a number of other stimulating talks from 15–17 May at the Victorian Comprehensive Cancer Centre, invited and selected speakers, a dedicated session for Melbourne. This is sure to be an outstanding highlight on student presentations and a lively poster session in the conference calendar for 2017, with a plenary lecture which there was very active engagement and discussion to be delivered by one of the world leaders in this field, across the 50 poster presentations. Overall, this inaugural Professor Ralph Deberardinis from the University of meeting was a great success and showcased the breadth Texas Southwestern Medical Center. The meeting will and depth of outstanding cancer metabolism research cover areas including immuno-metabolism and cancer, being performed by Australian researchers. metabolic disease, lifestyle interventions and cancer, novel In 2016, the MMM-SIG organised a symposium on metabolic pathways in cancer, and cancer metabolism and ‘Nutrient Sensing Mechanisms and Pathways’ at the therapeutics. The MMM-SIG will also continue to support ComBio meeting in Brisbane. We were lucky to attract the attraction of world-class speakers at the upcoming Dr Matthew Hirschey from Duke University as an ComBio2017 meeting in Adelaide. international speaker for this symposium. Dr Hirschey Given the increasing recognition of the role of aberrant opened the symposium with a fantastic presentation that metabolism in many major diseases, the MMM-SIG is described how metabolites generated from the metabolism committed to supporting relevant meetings in this field. of fatty acids can directly alter histone acetylation. This While this support may not always be financial, we can represents a novel epigenetic mechanism linking dietary assist you to enhance the quality of your meeting by fuels to the reprogramming of gene expression. Associate connecting you with our network of enthusiastic scientists Professor Zane Andrews from Monash University then from across the country. Please feel free to contact us if presented new findings on the metabolic role of the you have proposals for future ComBio symposia or if we enzyme carnitine acetyltransferase and how its deletion can support you in any other way. from specific hypothalamic neurons can influence whole- body glucose and lipid metabolism. The third speaker in Nigel Turner and Sean McGee the symposium was Professor Amanda Page from the email: [email protected] University of Adelaide, who presented research on how

Page 38 AUSTRALIAN BIOCHEMIST Vol 48 No 1 April 2017 Science Teachers’ Association of Victoria Science Talent Search The Victorian Branch of the ASBMB continues to be a well represented in the finals with two of our young proud sponsor of the Science Talent Search, which was budding scientists placing in the top three of the Inventions held in October last year at La Trobe University. This (Engineering) category. popular annual event founded by the Science Teachers’ We are extremely fortunate to be in the position to be Association of Victoria in 1952, was established with three able to contribute to the development of young aspiring broad aims: scientists and we look forward to the opportunity to 1. To stimulate ongoing interest in serious study of the support this initiative for many years to come. sciences by encouraging independent, self-motivated Erinna Lee, ASBMB Victorian State Representative project work amongst students of science, giving students www.sciencevictoria.com.au/sts the opportunity to communicate their achievements to a wider audience and recognising effort and achievement Left: A St Kevin’s College in their scientific enterprise. senior school student receives 2. To promote the direct involvement of the students in the the first place award in the process of science and its communication. Engineering Inventions 3. To give the public an opportunity to see the quality of category of the BHP Billiton work being achieved in science, by both primary and Science and Engineering secondary school students. Awards. His project was As part of the event, students enrolled in Prep through titled “Synth-etic: The to Year 12 are invited to submit projects in ten categories: world’s first ‘Hood-Wind’ Computer Programs, Games, Science Photography, instrument”. He is pictured Posters and Scientific Wall Charts, Working Models, with the Science Teachers’ Inventions, Experimental Research, Creative Writing, Association of Victoria’s Video Productions and Class Experimental Research President, Soula Bennett. Project. The theme for 2016 was ‘Drones, Droids and Robots’. There were a total of 2154 entries from 3089 student participants from 153 Victorian schools. The Victorian Branch of the ASBMB supported the 2016 Science Talent Search with a $1,000 donation, which was awarded as major and minor bursaries to 25 students from both Victorian Primary and High Schools, including: Box Hill High School, Brighton Primary School, Eltham College, Fintona Girls School, Firbank Grammar School, Lab Rats Science Club, Plenty Valley Christian College, Shelford Girls’ Grammar, St Leonard’s College, St Margaret’s School and Wesley College. The major Experimental Research and Inventions Right: Images of a banana’s winners were then provided the fantastic opportunity to chromosome clumps at attend the National BHP Billiton Science and Engineering resolution of 0.5 microns. Awards at the end of 2016. Here, Victorian students were

Opening ceremony. Vol 48 No 1 April 2017 AUSTRALIAN BIOCHEMIST Page 39 So I Filed a Provisional Patent Application – What Next?

A series of regular articles Content of the Complete Application on intellectual property. It is important to understand that your involvement with In this issue, Sarah Hennebry the patent application does not end after the filing of the Patent Attorney, FPA Patent document. Attorneys, outlines the steps The preparation of the complete application is the last following the filing of a opportunity for a patent application to include data and patent application. a description of how to perform the invention. After filing the provisional application, you should continue to Introduction develop the invention and perform experiments which My previous article in the December 2016 Australian are relevant for inclusion in the complete application. Biochemist discussed the type of information you might It is increasingly difficult to obtain a claim that is broader wish to provide to your patent attorney when you are first than exactly what is shown in the examples of a patent considering filing a patent application, or when you have specification. If the claims of a patent are restricted to the engaged your attorney to draft a patent application. specific examples provided in the specification, then third This article looks at what you may need to do after your parties may be able to design around a patent claim, and application has been filed, and what happens to your this may reduce the commercial value of the patent. application should you decide to continue pursuing the Some types of data can assist in broadening the scope of application after 12 months. a patent claim. Ideally, these data should be included in the provisional application, but can still be of significant First Steps benefit if obtained during the 12 months following the Generally, the first application that is filed in the patenting filing of the provisional, and included in the complete process is a ‘provisional’ (sometimes called ‘basic’) application. Some experiments and data to consider are application. This type of application is not examined by shown in Table 1. the patent office, and lapses after 12 months. In some instances, it is possible to skip the step of filing Complete Applications and International the provisional application and proceed straight to filing a Applications complete application. This, however, is not typical. The complete application can be one or more individual national applications or alternatively, an ‘international 12 Months Later application’ (also called a ‘PCT application’). It is important If, after 12 months from filing the provisional to understand that there is no such thing as an ‘international application, you still wish to pursue patent protection, a patent’; an international patent application is simply a ‘complete’application must be filed. bundle of national applications and enables the applicant to • the complete application must be filed on or before file a single application and then designate a large number the 12 month anniversary of filing the provisional of countries at a later point in time (see Fig. 1). application; One benefit of filing an international application is • the complete application will be given a ‘priority that more time is provided to obtain relevant funds for date’ which corresponds to the date on which the pursuing patent protection (including time to find a provisional application was filed. commercial partner, if needed). Furthermore, filing an If you don’t want to pursue patent protection, or are international application buys you more time to decide on concerned that more time is needed to develop the the countries in which patent protection will be sought. invention, no action is required and the provisional patent The international application will be published application will lapse (and the contents of the application approximately 18 months after you filed your provisional will not be published, other than the title and applicant application (or 18 months after filing your international/ details). complete application if you skipped filing a provisional

Fig. 1. Timeline of patent filing and prosecution.

IP Formalities, examination, grant Development

0 12 months 18 months 30 months up to 7-10 years

Page 40 AUSTRALIAN BIOCHEMIST Vol 48 No 1 April 2017

Document Title Position and Style So I Filed a Provisional Patent Application – What Next? application). This is another significant deadline to The tests for patentability that are applied in each consider, particularly if you have not yet published details jurisdiction vary and it is important to recognise that of your invention elsewhere. It is possible to withdraw the just because one Patent Office will grant a patent, not all international or complete application prior to publication, Patent Offices will do the same. if you want to avoid disclosing your invention to the It is common for patent attorneys to require the input public. Once this deadline has passed, your patent of the inventors during the examination process. application will be in the public domain and it will no For example, the inventors’ technical expertise and longer be possible to keep your invention secret. understanding of the prior art may be required for the Approximately 18 months after filing a PCT application purposes of rebutting any assertions made by the patent (or 30 months after the provisional application was filed), examiners. Further, declarations made by the inventors it is necessary to designate the specific countries in which can also be of use to clarify the state of the art prior to a patent is desired. It is not possible to obtain patent the invention, and for highlighting difficulties in arriving protection in all countries through a PCT application and at the invention (supporting, for example, inventive step you should discuss this with your patent attorney so that arguments). appropriate action is taken at an appropriate time. What Happens After Examination? Examination After examination, and once the patent examiner In most jurisdictions, examination of the patent considers that the claims are allowable, the patent application is performed before a decision is made application will proceed to ‘grant’. to ‘grant’ a patent. This examination is performed by In most countries it is necessary to pay periodic fees to examiners employed by the national patent offices in each maintain the patent application and any patent granted country. The technical background of patent examiners on that application up until expiry of the patent. The cost varies from country to country. of these fees varies considerably between countries and There is significant variation in the time taken for are payable up until expiry of the patent. If the fees are not examination in each jurisdiction, as well as the start date of paid, it is possible that the patent application will lapse (or examination. While in some cases it is possible to expedite in the case of a patent, it will cease). the examination process, in extreme cases it can take up to ten years after filing an application for examination to How Long Does a Patent Last? be complete. During this time, your patent attorney may In most jurisdictions, the term of a patent is 20 years. require your input at different stages, to assist them in This is calculated from the filing date of the complete prosecuting your application through the examination application. There are variations to this term, for example, process. where the patent is directed to certain pharmaceutical The examination process includes: products that require registration through a regulatory • an assessment of the claims made in the patent body (such as the Therapeutics Goods Administration in application to determine if they are eligible for patent Australia). protection under the patent laws of that country; – are the claims novel (new) and inventive (not What Rights Do I Have Once My Patent Is obvious)? – this assessment will be made by comparison with Granted? any disclosures that are in the ‘prior art base’ (see Once a patent has been granted, a patentee may sue for Glossary below). infringement of the patent, if they believe that there is • (in most jurisdictions) an assessment of the quality of an existing (or sometimes threatened) infringement of a the patent specification, including patent. Because patents are granted on a national basis, – whether the specification provides sufficient infringement of a patent must also be established on a instructions for the person skilled in the art (PSA; national basis, and with reference to the specific claims see Glossary below) to perform (would ‘create’ or that are granted in each relevant jurisdiction. ‘recreate’ be a better word?) the invention; – whether the skilled person was ‘in possession of Summary the invention’ at the time of filing the application. It is clear from the above that obtaining patent protection • if a patent examiner considers that there is a lawful is a lengthy process and the involvement of the inventors reason to object to one or more claims made in the does not end after the filing of the initial provisional patent application, a report will be issued application. Obtaining patent protection requires – the patent applicant may respond to the report, planning and forward thinking to ensure that applicants either by amending the claims, submitting aren’t caught out or limited to claiming a subset of their arguments or a combination of both approaches. true invention. Discussing the overall process with your • if the examiner’s objections cannot be overcome within patent attorney in the early stages of patent drafting will the allocated time, the patent application will lapse. help to make the process as smooth as possible.

Vol 48 No 1 April 2017 AUSTRALIAN BIOCHEMIST Page 41 So I Filed a Provisional Patent Application – What Next?

Table 1. Data to include in a patent application. Generate a class Consider the case where a single amino acid substitution in an enzyme results in increased Try to create a group or enzymatic activity. If only one example is provided in the application (for example, ‘genus’ from a specific substitution to a tyrosine) then the patent claims may be limited to the specific substitution. example. Data showing that mutation to any aromatic residue also gives rise to the altered activity, may provide a generalisation that enables the applicant to obtain a broader claim. 80% solution Inventors tend to try to solve a problem completely and perfectly. When briefing their This is a question of attorney, inventors often focus on the ‘100% solution’ (as discussed in the December 2016 how someone might Australian Biochemist). Consider a small molecule inhibitor of an enzyme. Consider the core follow an inventive idea, structure of the molecule and the options for substituents. Do the data show that only one without replicating the variation in the molecule’s structure is useful of inhibiting enzyme function or is it possible invention exactly and to make certain alterations to the substituent, without significantly affecting the inhibitory avoiding infringement. function of the molecule? Considering the answers to these questions, and providing data showing that modifications to the structure do not affect function, may enable the patentee to claim a broader class of structures. Test drive other models Consider the scenario where initial experimental data show efficacy of a molecule in Extrapolate from one inhibiting tumour growth and reduction in tumour size, but only in one animal model of a context or disease type specific cancer. Test the molecule on related cancer animal models so that the results can be to another. extrapolated to the use of the molecule in treating more than one type of cancer. Negative results Although negative results typically don’t make it into journal articles, these data can be Even negative results helpful in obtaining patent protection. For example, if a number of experiments were can be informative and conducted but didn’t work, this may support subsequent inventive step arguments (i.e., helpful in obtaining that the solution to the problem being solved was not necessarily obvious or the skilled a patent, even if they person did not have ‘an expectation of success’). Negative results can also be useful if trying don’t make it into the to claim the synergistic combination of known compounds. For example, if synergism final publication in a only occurs within the specific range of concentrations of the actives, then negative data scientific journal. can show that simply combining the known compounds would not necessarily lead to the result (again assisting in an inventive step argument).

Table 2. Patent glossary. Term Definition Inventive A subjective test is applied to the patent claims and asks the question of whether the claims are obvious. step This question is answered by considering the knowledge of the ‘person skilled in the art’ as at the priority date of the patent application. There may be variation in the way this test is applied in different jurisdictions. A determination of whether the patent claim relates to a ‘new’ invention, by reference to publically Novelty available information, including patent literature and non-patent literature. In some jurisdictions, ‘actions’ and trade displays may also be considered when assessing novelty.

An international application that allows a patent applicant to make use of an administrative process to PCT file the same application in multiple countries at the same time. application A hypothetical person adopted for the purposes of determining whether a patent claim involves an Person inventive step. This person is said to have the skills and background knowledge of someone working in skilled a particular technical field, but does not necessarily have inventive capacity. When considering whether in the art a claim is inventive, examiners (or Courts) will put themselves in the shoes of the PSA. (PSA) The patent and non-patent literature that is considered against the claims of a patent application. The Prior art rules for constructing the prior art base may differ between jurisdictions. For example, in some countries base the disclosures of a few documents can be combined, particularly for inventive step determination. Usually this is the date that the provisional application is filed and must be no more than 12 months Priority earlier than the filing date of the complete application. In some cases, where no provisional application date is filed, the priority date will be the same date as the filing date of the complete application.

Page 42 AUSTRALIAN BIOCHEMIST Vol 48 No 1 April 2017 2015 LabGear Australia Discovery Science Award On the Road with Intracellular Trafficking I was fortunate now clear from many studies incorporating genetics, to be awarded the cell biology and animal models, that dysfunction in LabGear Australia membrane trafficking events associated with endosomes Discovery Science and the Golgi underpin the development of a number Award in 2015. The of neurological diseases. The work by Cheryl Chia, Wei award is a travelling Hong Toh and Anson Tan in my lab have helped to map lectureship to the intracellular trafficking pathways of both BACE1 enable the awardee and APP and define where the two membrane proteins to present his/her converge to promote APP processing and amyloid beta work at centres production. Both Cheryl and Wei Hong have previously within Australia been awarded ASBMB Fellowships for their achievements. and New Zealand The PhD student presentations at the APG were of a very Paul Gleeson receives the 2015 and to also present high standard. The three students gave excellent talks: Heidi ASBMB LabGear Australia Discovery a symposium talk at Neubauer (Centre for Cancer Biology), on the oncogenic Science Award from Derek Brown, ComBio. role of sphingosine kinase 2; Shee-Chee Ong (Flinders LabGear Australia. Over my career, University) on novel insulin analogues with therapeutic I have been lucky to be able to explore research topics benefits; and Stephanie Begg (University of Adelaide) in a number of different areas across cell biology and on the metalloproteome of Streptococcus pneumoniae. The immunology. One constant in my wandering research session was held at the attractive Adelaide University life has been a fascination with the form and function of Graduates Clubhouse, and hosted in a very engaging the Golgi apparatus, which has followed me throughout manner by Mark Corbett, Neurogenetics Research my career. The LabGear Award was for the contributions Program, University to this topic. Our historical understanding of the of Adelaide. functions of the Golgi has traditionally been restricted to My second visit the regulation of glycosylation and membrane transport. sponsored by the However, I believe there are many dark secrets to the LabGear Award was Golgi apparatus and indeed some are now beginning to the Institute for to be revealed. For example, advances over the past Molecular Bioscience few years have provided clear evidence that the Golgi (IMB), University contributes to the regulation of higher order functions of Queensland, in such as cell polarisation, directed migration, directed July/August 2016 secretion, metabolism and autophagy. The symposium where I spent a Super-resolution image of the Golgi I presented at ComBio2015 in Melbourne discussed our very enjoyable and ribbon structure stained with cis- work on these higher functions and on the emerging role productive two and trans-Golgi markers, of membrane tethers in regulating Golgi morphology. Our day visit hosted by by Prajakta Gosavi. findings were also beginning to reveal some unexpected Rohan Teasdale functions associated with the ribbon structure of the and Jenny Stow. Here I gave a seminar titled ‘The Golgi Golgi apparatus, which is the typical structure found in apparatus: a regulator of higher order cell functions?’ vertebrate cells. An exciting possibility is that the Golgi where I presented work from Prajakta Gosavi in my lab, acts as a major sensor of the cell. demonstrating that the Golgi ribbon structure controls The timing of signalling pathways and autophagy. Over the two days, the Award was I had very lively and stimulating discussions with Jenny particularly apt as and Rohan and a number of other cell biology colleagues I was nearing the including Alpha Yap, Rob Parton, Brett Collins, Dominic end of my ten-year Ng (School of Biomedical Sciences) and Fred Meunier stint as Head of (Queensland Brain Institute). It was very exciting to Department (mid- hear about the latest developments at UQ and to discuss 2016) and would be with Jenny the lattice light sheet images of macrophage Guest lecture by Paul Gleeson able to focus more on ruffling she had captured on her visit to Janelia Research to the Adelaide Protein Group. research activities. Campus, Virginia, USA. Photo: Houng Taing. My first visit funded I would like to thank LabGear for their generosity by the LabGear Award was to Adelaide in June 2015 to in providing funding for this award. It allowed many give a guest lecture to the Adelaide Protein Group (APG) productive interactions across states with PhD students, and to judge the 2015 APG PhD student award. My talk early career researchers and senior colleagues. focused on our work on the trafficking of the b-secretase Professor Paul Gleeson, Department of Biochemistry BACE1, and amyloid precursor protein (APP) and the and Molecular Biology, Bio21 Molecular Science and production of amyloid beta in Alzheimer’s disease. It is Biotechnology Institute, University of Melbourne.

Vol 48 No 1 April 2017 AUSTRALIAN BIOCHEMIST Page 43 FAOBMB annual conference held in the Philippines

Paul Gleeson, FAOBMB Representative from Australia, reports on the FAOBMB Conference and Council Meeting held in Manila, the Philippines, during December 2016.

The 25th FAOBMB included a presentation on the history of the discovery International Conference of conotoxins in the Philippines by Dr Lourdes Cruz; and 43rd Annual Meeting an exhilarating presentation by Dr Danilo Tagle (NIH) of the Philippine Society of on the advances in microfabricated devices to engineer Biochemistry and Molecular ‘organs on chips’ for drug testing, a field which has now Biology (PSBMB) was held progressed to include multiple organs on a chip to analyse in Manila at the Philippine physiological systems; and an elegant presentation by International Conference Professor Ricky Johnson (Peter MacCallum Institute, Centre from 5–7 December, Melbourne) on epigenetic regulation and the potential of 2016. The convenor was HDAC inhibitors for treatment of leukemia. Professor Gracia Fe Yu from the University of the Philippines, and the meeting’s theme was focused on Biochemistry and Molecular Biology in Professor Gracia Fe Yu in Health and Wellness, with formal Filipino attire at the an emphasis on translation opening of the Conference. of natural products. The local organising committee, comprising Professor Gracia Fe Yu, Dr Apolinario Yambot (President of PSBMB) and Dr John Donnie Ramos (Immediate Past President of PSBMB) delivered a lively and highly interactive Chinese colleagues at the Conference. From left: Professor meeting. There were more than 575 participants, Cong-Zhao Zhou (delegate of CSBMB), Professor Rongwu including 60 foreign delegates, of which there were nine Yang (winner of FAOBMB Education Award), Professor Feng plenary speakers. Particularly notable were the vibrant Shao (winner of FAOBMB Award for Research Excellence) and interactions between students and international delegates, Professor Zengyi Chang (incoming FAOBMB President). and the organisers should be congratulated for facilitating sessions to maximise student involvement. The winner of the 2016 FAOBMB Award for Research The meeting commenced with welcome addresses Excellence was Professor Feng Shao (National Institute from the FAOBMB President Professor Kiyoshi Fukui of Biological Sciences, Beijing, China) who gave (Japan), the FAOBMB President-Elect, Professor Zengyi an outstanding plenary lecture on host–pathogen Chang (China) and the IUBMB President-Elect Professor interactions, in which he described his contribution to Andrew Wang (Taipei, China). All three highlighted unravelling the mechanisms underlying the necrotic the importance of the regional alliances promoted by cell death pathway, pyroptosis, and the major role of the FAOBMB and enthusiastically promoted further this pathway in innate inflammatory responses against expansion of the organisation. bacteria. He described his very elegant approaches to Highlights from the scientific program included the define the intracellular sensors and effector machinery nine plenary lectures and Award presentations. These associated with pyroptosis. His studies have also revealed the unexpected finding that pyroptosis may well be the underlying mechanism for the non-classical secretion of IL-1b. The winner of the 2016 FAOBMB Education Award was Professor Rongwu Yang from Nanjing University, China. In his lecture, Professor Yang presented his personal experience on teaching biochemistry with his very enthusiastic style. His vibrant presentation stressed the importance of a passionate approach to teaching and he highlighted multiple mechanisms for engagement with students during lectures. He also presented examples of a range of creative prompts he uses to promote visual understanding of the concepts of biochemistry. Professor Feng Shao receiving the FAOBMB Award for Professor Yang has kindly consented to his talk being Research Excellence (including certificate and trophy) made available at the following link: http://faobmb. from Professor Kiyoshi Fukui (FAOBMB President), with com/2017/01/09/iubmb-education-symposium-at-25th- Professor Zengyi Chang (FAOBMB President-Elect) at left. faobmb-conference-held-in-manila-in-december-2016/ Page 44 AUSTRALIAN BIOCHEMIST Vol 48 No 1 April 2017 FAOBMB annual conference held in the Philippines

conference program (Sunday 4 December, 2016), with Paul Gleeson as the Australian delegate (ASBMB representative). The meeting was attended by delegates from 15 of the 21 constituent member countries and was chaired by the FAOBMB President, Professor Kiyoshi Fukui (Japan) (2014–2016) and the Secretary-General, Professor Phillip Nagley (Australia). The President highlighted in his report the initiatives by the FAOBMB, particularly in education and in strengthening the relationship with IUBMB. Three FAOBMB members are now serving on the Executive Committee of IUBMB, namely Professor Andrew Wang (Taipei, China) as President-Elect of IUBMB, Professor Janet Macaulay (Australia) with the portfolio of education Paul Gleeson with a group of young Filipino scientists. and training and Dr Avadhesha Surolia (India) as member There were other excellent talks in the Education for publications. Professor Zengyi Chang, President-Elect Symposium, including those given by Associate Professor for FAOBMB at the time of the Council meeting, took over Susan Rowland (Australia) and Professor Hoon Eng Khoo as President from January 2017. Paul Gleeson indicated (Singapore); their presentations can also be found on the the considerable enthusiasm of the ASBMB to bid for the link above. IUBMB congress in Melbourne in 2024 and the FAOBMB Dr Lahiru Gangoda (Department of Biochemistry and Council was supportive of this potential bid. Genetics, La Trobe University) was one of eight Young The 26th FAOBMB Scientists who were successful in obtaining FAOBMB Conference will be held in Travel Fellowships to attend the conference and to deliver Kobe, Japan, 6–9 December oral presentations of their work. A unique feature of the 2017, as a joint meeting program was the special young scientist forum, in which with a large consortium of outstanding Filipino undergraduate research students biological science societies presented their laboratory-based research projects. Five (ConBio2017). The next finalists were selected from over 30 applications for triennial IUBMB Congress oral presentations in a very well attended session and will be held as a joint meeting all talks were of a very high standard. The international with the FAOBMB Congress FAOBMB judges, including Wayne Patrick (New in Seoul, Korea, 4–9 June Zealand) and Paul Gleeson (Australia), awarded the best 2018. In 2019, the 27th presentation to Arman Ali Ghodsinia (National Institute FAOBMB Conference will be of Molecular Biology and Biotechnology, University of held in Malaysia. Traditional Filipino the Philippines) for his research on PI3 kinase mutations Thanks to Phillip Nagley dancers at the cultural and oncogenesis. for his input and some of the evening of the Conference. The FAOBMB Council meeting was held prior to the photographs.

Members of FAOBMB Executive Committee, Delegates to FAOBMB Council and observers at the Council meeting held in Manila, Philippines.

Vol 48 No 1 April 2017 AUSTRALIAN BIOCHEMIST Page 45 OUR SUSTAINING MEMBERS

ASBMB welcomes the following new Disclaimer Sustaining Members: The Australian Biochemist is Cleaver Scientific Ltd, United Kingdom published by the Australian Society GeneWorks Pty Ltd, SA for Biochemistry and Molecular Isolab Laborgeräte GmbH, Germany Biology Inc. The opinions expressed in this magazine do not necessarily represent the views of the Australian Society for Biochemistry and Molecular Biology Inc. 95% QE Back Illuminated Scientific CMOS Camera Add Some Spark® to Your Research The new Prime 95B Back Illuminated Increase productivity for advanced Scientific CMOS (sCMOS) camera research and drug discovery from Photometrics with 95% Quantum applications with Tecan’s Spark® Efficiency (QE) is an ideal choice multimode microplate reader. This for Super-Resolution Microscopy, freely configurable system is designed Confocal Microscopy, Single Molecule to enhance and streamline biochemical Fluorescence and Light Sheet AAVPrime™ Adeno-associated and cell-based workflows, both today Microscopy. Viral Particles and in the future. The Photometrics Prime 95B sCMOS GeneCopoeia’s AAVPrime™ Adeno- At the heart of the instrument camera is the first sCMOS camera to associated virus (AAV) products are the are Spark’s unique Fusion Optics, offer 95% Quantum Efficiency (QE) ideal tools for inserting genes into a broad which combine the flexibility of and Back Illumination (BI) in the range of cell types with high efficiency monochromators with the sensitivity same camera and it now outperforms and enhanced safety. of filters. This set-up allows on-the-fly EMCCD cameras. GeneCopoeia’s optimised helper- switching between optics, increasing The Prime 95B’s sensor converts productivity without compromising free human AAV system allows up to 95% of incident photons into a viral packaging without potentially on data quality. This is complemented measurable signal. by patent-pending High Speed pathogenic helper adenovirus. Many pre- Monochromators offering full The back illuminated sensor brings made particle options are available in 3 absorbance scans – from 200 to 1,000 nm light into the pixel photodiode from different serotypes, including fluorescent – in under five seconds. And to ensure behind, avoiding structures that reflect reporters or you can request custom complete confidence in your data, the or absorb light. Combined with large AAV particles with genes up to 3 kb in optional Te-Cool™ module uniquely 11μm pixels, the Prime 95B camera length. Particle preparations are available allows the measurement temperature to can deliver over 300% more signal in either standard or purified (suitable for be set below the ambient temperature, than other sCMOS cameras at 100X in vivo use) formats. ensuring more accurate and reliable magnification. Advantages results. The extreme sensitivity not only • High titers. Titer of purified 14 Spark® has also been developed with allows fainter signals to be detected, particles can be up to 10 GC/ml cell-based assays in mind, and includes it provides the flexibility to increase (genome copies/ml) a host of software and hardware features frame rates, or turn down the excitation • Versatile. Usable in a broad range of – including precise temperature and gas intensity to reduce cellular photo- host cell types regulation, built-in cell counting and damage. • Low toxicity. Does not integrate into viability assessment, and automated the host genome. Key features include 95% QE, 16 imaging and confluence measurements • Low immunogenicity. Minimal host bit dynamic range, large 11μm x – to simplify cell biology protocols and immune response. 11μm Pixel Area, 1.3e- Read Noise enable long-term, walkaway experiments. • Safe. Not associated with any (rms), 41fps @ 16-bit/82fps @ 12-bit, These options, combined with the ability human disease. regulated air cooling to -10 deg C, to read up to 1,536-well microplates, single cable connection rather than a For further information please contact ensure there is a Spark® configuration to dual camera link, large field of view, C United Bioresearch, GeneCopoeia’s increase the productivity of your lab. Mount interface, multiple expose out Australian partner. www.tecan.com/spark triggering and SMART streaming. United Bioresearch Products Scitech Pty Ltd Kirrily Smith (03) 9480 4999 Phone (02) 4575 0309 [email protected] [email protected] www.scitech.com.au www.genecopoeia.com

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Chirascan™-plus ACD is the world’s medicine researchers and clinicians are first and only truly automated CD now looking for GMP quality products, spectrometer. Unattended operation which provide a smoother regulatory increases the productivity of CD approval process. measurements markedly, with up to The Cytori Celase® Enzyme is 200 samples per day now routinely currently used in US FDA approved achievable. The improved productivity clinical trials. transforms the number and type of Worthington is excited to offer a experiment that can be undertaken and foundational and versatile enzyme Chirascan Circular Dichroism addresses key needs in formulation and for advancing adipose-based research Spectrometers bio-comparability applications. programs from pre-clinical to clinical levels, while eliminating the need to Chirascan sets new standards for steady- ‘qCD’ (quantitative CD) reflects a perform costly and time consuming state circular dichroism spectroscopy. It number novel capabilities that improve bridging studies. incorporates innovative optical design performance and enable absolute features to maximise light throughput, CD measurements (a first in CD ScimaR particularly in the far-UV wavelength spectrometry). Email [email protected] region, and a sophisticated digital data For further information contact:- Freecall 1800 639 364 acquisition system that facilitates the Scientex Pty Ltd Phone 03 9842 3386 rapid collection of more accurate and Tel 03 9899 6100 Fax 03 9842 3407 precise CD spectra. Chirascan’s digital Fax 03 9899 6122 CD spectra acquisition approach ensures Email [email protected] that unmodified CD spectra are collected www.scientex.com.au and any post-acquisition smoothing of the CD spectra will be non-distorting and completely reversible. This approach jetMESSENGER™ from also simplifies the operation – Chirascan PolyPlus is as straightforward to use as a single- beam spectrophotometer. A new mRNA transfection reagent for hard to transfect cells. Other key features include: • Able to collect thermal denaturation mRNA transfection is as easy as DNA CD spectra in a single experiment; transfection, with the advantage that enabling identification of the Celase® GMP Enzyme Launch mRNA does not need to reach the cell secondary structural changes Celase® GMP is a proprietary nucleus for expression nor require cell associated with each phase transition enzyme containing a unique blend of division for efficient gene expression. • 5 detection channels: CD, collagenase, neutral protease and buffer Hence, cells that are slow-dividing or that Absorbance/Transmission, salts that are produced using avian and have developed specific mechanisms to HT, Temperature and Voltage. mammalian tissue-free raw materials, protect their genome can finally be used Simultaneous multi-channel data aseptically processed, sterile filtered and for gene expression. jetMESSENGER™ acquisition ensures that all key highly purified under GMP guidelines. gives high transfection efficiency in all information is recorded with every Manufactured by Cytori Therapeutics, usually difficult to transfect cells, e.g. measurement you make this product line is ideal for cell isolation primary cells, neurons, suspension cells • Very low nitrogen usage. Rapid and studies for laboratories aiming to and various cancer cell lines. efficient nitrogen purging combined facilitate a smooth transition from bench With jetMESSENGER™ researchers with a sealed monochromator and animal research to downstream can now achieve unmatched housing ensures that just 5 l/min is clinical applications. transfection efficiency, and outperform required for far-UV work A single, sterile, ready-to-use enzyme DNA transfection by switching to • Moveable detector. The detector containing both collagenase and neutral mRNA. This is extremely gentle on cells, position is easily adjustable and can protease is ideal for a wide range of with no risk of genome integration, and be set close to the cell to optimised adipose stem cell, biomedical and it is perfectly suited for CRISPR/Cas9 performance with highly scattering bioprocessing applications. gene editing, iPS generation, stem cell samples e.g. membrane proteins Not all research applications require differentiation and immunotherapy A large range of accessories are the use of a GMP grade enzyme in assays. available, ensuring the researcher can be early phase studies. However, the VWR International Pty Ltd confident of a highly effective and future- recent FDA guidance issued for tissue Tel: 1300 727 696 proof spectrometer that can be adapted and cell products specifically cites that Fax: 1300 135 123 as research interests evolve. These are GMP grade reagents should be utilized Email: [email protected] best explored with the comprehensive in drug-type validated processes. Web: au.vwr.com brochure available on request. Subsequently, both regenerative

Vol 48 No 1 April 2017 AUSTRALIAN BIOCHEMIST Page 47 OUR SUSTAINING MEMBERS compatible with widely used analytical DAINTREE software (e.g., UltraScan, SEDFIT/ SEDPHAT, SEDANAL, DCDT+), and scientific the same cells and rotors used with the AUSTRALIA ProteomeLab XL-A/XL-I, the Optima New Products to the DAINTREE AUC surpasses many features of its Range from Coyote Bioscience forerunner. Coyote’s products are affordable solutions for any laboratory. For field work or in laboratories where Advances in Unique Protein power sockets are sparse, the G10 and and Macromolecule Analysis G20 Cordless Tissue Grinders from Technology Coyote can work cordlessly for up to 10 LI-COR Biosciences is a leading hours. The G50 corded Tissue Grinder is AUC is a unique technique that innovator in systems for plant research, adjustable from 3K to 8K rpm. enables molecules to float freely and gas analysis, drug discovery, protein unbound so they can be characterized research, and small animal imaging. 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From studies ranging For information please contact detection systems. Moina Macaskill from global climate change to cancer Daintree Scientific Australia All system optics are contained research, scientists rely on LI-COR Phone (03) 6376 3335 outside the rotor chamber, making the products and services to address Email [email protected] Optima AUC easier to clean. A 15-inch important biological and environmental www.daintreescientific.com.au (38 cm) touchscreen display indicates challenges. More information at the progression of experiments, while www.licor.com. remote monitoring lets researchers set up, monitor and extract experiment data from virtually any location. Though

Page 48 AUSTRALIAN BIOCHEMIST Vol 48 No 1 April 2017 OUR SUSTAINING MEMBERS GeneWorks will be exhibiting at ComBio 2017 in Adelaide. GeneWorks [email protected] www.geneworks.com.au LATEX Bead Conjugation Kits FreeCall 1800 882 555 Innova Biosciences has introduced Phone 08 8159 6250 a range of LATEX Bead Conjugation Kits. These simple to use, one-step kits for covalently conjugating antibodies, proteins and peptides (or any other GeneWorks is a proudly independent biomolecule with an amine group) to supplier of molecular and cell biology specially treated latex beads without the products and associated services based need for extensive optimisation. in Adelaide. Our mission is to facilitate access to innovative technology The latex conjugation reaction has been solutions and services to enable developed using Innova Biosicences successful research or commercial expertise in simple and quick one- outcomes for our clients. step antibody conjugations such as our InnovaCoat® GOLD kits (covalent New introductions to GeneWorks’ Selecting the Right Gel conjugation to gold nanoparticles) product range include Alvéole’s Documentation System and Lightning-Link® kits (covalent PRIMO, the world’s first multi- conjugation of antibodies, proteins and protein patterning system. PRIMO Ease of use and reliability have always peptides to enzymes and fluorophores) allows researchers to study the been the biggest considerations in to produce a kit unlike any latex bead influence of the microenvironment choosing a gel documentation system. conjugation product available. on intracellular and intercellular Recently, gel documentation systems mechanisms. We now also supply with alternatives to UV have become Quick and easy to use: products from BRANDEL, including increasingly popular and required as • 30 seconds to set up the one-step a novel Gradient Fractionator system researchers shift their preferences to conjugation reaction used for the separation of organelles, the use of safer fluorescent dyes instead • 3 minutes hands-on time and RNA, DNA and the study of protein of toxic ethidium bromide. The use of 35 minutes total time until the interactions. BRANDEL also make alternative light sources for excitation, conjugates are ready to use systems for cell harvesting and tissue such as blue light, also serves to reduce Specially treated latex is resistant to perfusion. the risk of UV exposure. aggregation: Organisations looking for workflow Axygen® Gel Documentation • High yields of functional conjugates automation should consider unique systems are easy to use and reliable for can be made without the need for instruments from Art Robbins the fast gel imaging operations required harsh resuspension methods like Instruments (ARI) such as the by researchers today. It deploys a sonication and vortexing Cobra non-contact nanoliter-capable high resolution 5.4 MP camera which Only two buffers are used to test for dispensers, and the Scorpion Screen captures publication quality images and optimal activity: Builder high speed single channel features an Auto Exposure tool which • No extensive pH optimisations as pipettor. enables the calculation of the optimal exposure time with just a single click. is typically required for traditional Excitement continues to build around passive conjugation methods Clearbridge Biomedics’ ClearCell Axygen® Gel Documentation Choice of bead colour and kit sizes: system for viable CTC isolation, and systems also feature a Darkroom • Available 400nm latex beads in red, Bioo Scientific’s NextPrep-Mag cfDNA control which allows the selection blue or black Isolation and NEXTflex NGS Library from UV 302, UV 365, Epi White, or • Two kit sizes available: Mini kits preparation kits and barcodes. Epi Blue light sources, as well as an are ideal for antibody screening or optional Trans White light illumination GeneWorks’ expanding range of tray. With many added features and ‘proof of principle’ experiments, services include custom Genotyping and Midi kits which are 10 times the options available in the market today, (including assay design), Gene selecting the right gel documentation size of the Mini kits. Bulk material is Expression Analysis and Sample also available for further scale up system is important. Axygen® Gel Preparation which we provide to Documentation systems assist the user BioNovus Life Sciences researchers and companies wishing to in capturing hassle-free images in the Ph: (02) 9484-0931 to outsource their wet lab work. least amount of time. Please contact us Email: [email protected] GeneWorks also provides expert on [email protected] Web: www.bionovuslifesciences.com.au Instrumentation Services to support all the equipment we supply.

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platform of the known brands available your data is analysed automatically at today. Seamless transition to using the the end. CleanNA PCR clean up and isolation HUNK fully automates the process kits on your existing automated liquid of chemical denaturation and analyses handling platforms mean less time the data giving you DGs for two-state for optimisation of the protocol and and three-state proteins. Just add your reduced cost of reagents. protein, formulations and denaturants, and the HUNK performs the liquid SSI Vertex Pipette Tips For your cost effective, automation friendly, high quality sequencing results, handling and intrinsic fluorescence LabGear Australia are pleased to efficient purification of PCR products detection to create and measure your announce its appointment as the new and Size Exclusion requirements for chemical denaturation curve. You exclusive distributor for the SSI Vertex Next Generation Sequencing Library can measure up to 96 different DGs, range of pipette tips. The range comes Preps, please contact LabGear Australia unattended. in filtered and non-filtered versions on 1800 LabGear (1800 522 432) or email UNcle and HUNK have your stability and are offered with the NoStick® [email protected] and aggregation studies covered. resin formulation as standard which For more information, visit www.axt. enables viscous liquids to be dispensed com.au or email [email protected] completely. All tips are certified free from RNase, DNase, RNA, DNA, and PCR inhibitors and are manufactured from US FDA approved medical grade virgin polypropylene. LabGear also offers the full range of SSI Ultraflux PCR products including PCR tubes, strip tubes, caps, PCR plates, and sealing film. Completing the offering are CryoFreeze Isolab Laborgeräte GmbH is a global tubes, racks and screw cap tubes. UNcle and HUNK – the Perfect laboratory supplier company that Pair for Studying Protein specialises in manufacturing laboratory GC Biotech Aggregation and Stability consumables. LabGear Australia are the exclusive Do you need to rank your constructs Isolab sterile production plants are well Australian distributors for GC Biotech. or optimise your formulations? AXT, equipped (Class 10,000 sterile) clean GC Biotech manufactures a range of partnering with Unchained Labs, rooms (validated by international DIN magnetic bead based products for can help you do this through either and ISO standards) with HEPA filters genomics, Sanger and Next Generation temperature-induced stress, disrupting and positive pressure areas which are sequencing applications. The products the 3D protein structure, or through supplied with air locked doors. Isolab are: CleanPCR, CleanNGS, CleanDTR chemical denaturation, determining certified plastics are DNase, RNase and clean up kits, Clean Circulating DNA the Gibbs free energy (DG) required for pyrogen-free, and BioClean certificated. from plasma or serum, CleanNA DNA protein unfolding. and RNA isolation kits from a wide The UNcle stability platform uses Some of our life science products are: range of sample and tissue types like, fluorescence, static light scattering sterile tubes (centrifuge, micro, cryo, blood and tissue, plant, pathogen, FFPE (SLS) and dynamic light scattering PCR, K3 EDTA), caps, flasks, plates, and plasmids. (DLS) to characterise protein stability. racks, swabs, boxes, pestles, slides, petri dishes, pipettes and workstations. CleanNGS delivers efficient PCR and With temperature control from 15–95oC Next Generation Library Prep clean up and sealed samples, you can even do Our ‘one stop shopping for whilst CleanDTR offers efficient PCR long term studies to fully understand laboratories’ catalogue includes: and Sanger sequencing reaction clean protein stability. Measurements include glassware, consumables, equipment, up based on the unique paramagnetic thermal melting, thermal aggregation, instruments and chemicals. isothermal stability, thermal recovery, bead technology. The Clean Circulating Catalogue: http://catalog.isolab.de/ sizing, polydispersity and viscosity, as DNA Kit has significant uses in research, 2016/index.html Cancer Diagnostics, prenatal screening well as determination of the diffusion (NIPT), Organ transplantation and interaction parameter and second virial Please see us: www.isolab.de Post Trauma Monitoring to produce coefficient to help predict colloidal scalable, isolation of high quality stability and likelihood of aggregation and high recovery of targeted DNA and protein-protein interactions. fragments. This innovative magnetic UNcle can also measure DG – prep bead technology can be used in a your samples offline with denaturants manual setting in 96 well plates or on and incubate at ambient temperature your current automated liquid handling and then read them on the UNcle, where

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FORTHCOMING MEETINGS

ComBio2017 • Plant Biology 2–5 October 2017 • Biotechnology and Sustainable Futures Combined ASBMB, ASPS and • Developmental, Stem Cell and ANZSCDB Annual Meetings Regenerative Biology Adelaide Convention Centre, Adelaide • Proteins and Proteomics • Genomes, Epigenetics and Early Registration and Abstract Bioinformatics Deadline: Friday, 30 June 2017 • Cell Biology • Cell Signalling and Metabolism ComBio2017 will be held at the new The keynote speaker for this conference state of the art Adelaide Convention is Emmanuelle Charpentier of the Max Centre located in the heart of the city Planck Institute for Infection Biology, Further information: on the Torrens River, and overlooking Berlin. Professor Charpentier is the co- www.combio.org.au/combio2017 the magnificent Adelaide Oval Precinct. inventor of CRISP-Cas9. The program The Convention Centre and hotels are will feature a number of other overseas Conference Chair: located a stone’s throw from the many plenary presentations from some of the Michael Michael restaurants and cultural activities that best international scientists together with [email protected] make Adelaide such an engaging and a number of society speciality lectures. enchanting destination. Several poster sessions are also planned. Registration/Exhibition The scientific program of the conference Sally Jay will include the themes: [email protected]

New Horizons in 26th FAOBMB 24th IUBMB–15th FAOBMB Biochemistry & Molecular Conference Congress Biology Education 6–9 December 2017 4–9 June 2018 5–8 September 2017 Kobe, Japan Seoul, Korea The 26th FAOBMB Conference will be This is a triennial Congress of IUBMB, Rehovot, Israel held as a combined meeting with the combined with the FAOBMB Congress This education conference is supported Japanese Consortium of Biological when held in our region. A Young Scientist by IUBMB and FEBS. The conference Sciences (ConBio 2017). FAOBMB Travel Program will be held in conjunction with proceeds the 42nd FEBS Congress, 10– Fellowships will be available for young the Congress, 2–4 June 2018. 14 September in Jerusalem, Israel. scientists to attend this conference. Further information: Further information: Further information: Email: [email protected] Email: [email protected] Email: [email protected] Website: www.iubmb2018.org Website: www.weizmann.ac.il/ Website: www.aeplan.co.jp/conbio2017 conferences/NHBMB2017

Vol 48 No 1 April 2017 AUSTRALIAN BIOCHEMIST Page 51 DIRECTORY COUNCIL FOR 2017 STATE SPECIAL INTEREST PRESIDENT REPRESENTATIVES GROUPS Professor Leann Tilley Department of Biochemistry and AUSTRALIAN CAPITAL TERRITORY ADELAIDE PROTEIN GROUP Molecular Biology Dr Peter Mabbitt Contact: Dr Christopher McDevitt University of Melbourne ANU College of Physical and Mathematical Research Centre for Infectious Diseases PARKVILLE VIC 3010 Sciences University of Adelaide Ph (03) 8344 2227 Australian National University ADELAIDE SA 5005 Email: [email protected] CANBERRA ACT 0200 Ph (08) 8313 0413 Email: [email protected] Email: [email protected] PAST PRESIDENT Professor Michael Ryan NEW SOUTH WALES AUSTRALIAN YEAST GROUP Department of Biochemistry and Dr Katharine Michie Chair: Dr Alan Munn Molecular Biology School of Physics Griffith University Gold Coast Monash University University of New South Wales PMB 50, Gold Coast Mail Centre CLAYTON VIC 3800 SYDNEY NSW 2052 SOUTHPORT QLD 9726 Ph (03) 9902 4909 Ph (02) 9385 4587 Ph (07) 07 5552 9307 Email: [email protected] Email: [email protected] Email: [email protected] TREASURER QUEENSLAND BIOCHEMICAL EDUCATION Associate Professor Terrence Piva Dr Dominic Chi Hiung Ng Chair: Associate Professor Susan Rowland School of Medical Sciences School of Biomedical Sciences School of Chemistry and Molecular Biosciences RMIT University, PO Box 71 University of Queensland University of Queensland BUNDOORA VIC 3083 ST LUCIA QUEENSLAND 4072 ST LUCIA QLD 4072 Ph (03) 9925 6503 Ph (07) 3365 3077 Ph: (07) 3365 4615 Email: [email protected] Email: [email protected] Email: [email protected] SECRETARY SOUTH AUSTRALIA MELBOURNE PROTEIN GROUP Associate Professor Briony Forbes Dr Steven Polyak President: Dr Douglas Fairlie Medicinal Biochemistry School of Molecular and Biomedical Science Olivia Newton John Cancer Research Institute Flinders University University of Adelaide Austin Hospital BEDFORD PARK SA 5042 ADELAIDE SA 5005 HEIDELBERG VIC 3084 Ph (08) 8204 4221 Ph (08) 8313 6042 Email: [email protected] Email: [email protected] Email: [email protected] METABOLISM AND MOLECULAR EDITOR TASMANIA MEDICINE GROUP Dr Suresh Mathivanan Dr Kate Brettingham-Moore Chair: Dr Nigel Turner La Trobe Institute for Molecular Science School of Medicine School of Medical Science Department of Biochemistry University of Tasmania University of New South Wales La Trobe University HOBART TAS 7008 KENSINGTON NSW 2052 BUNDOORA VIC 3086 Ph (03) 6226 4609 Ph (02) 9385 2548 Ph (03) 9479 2565 Email: [email protected] Email: [email protected] Email: [email protected] VICTORIA QUEENSLAND PROTEIN GROUP COMMUNICATIONS DIRECTOR Dr Erinna Lee Chair: Dr Brett Collins Dr Suresh Mathivanan Olivia Newton-John Cancer Research Institute Institute for Molecular Bioscience La Trobe Institute for Molecular Science 145 Studley Rd University of QLD, ST LUCIA QLD 4072 Department of Biochemistry HEIDELBERG VIC 3084 Ph (07) 3346 2043 La Trobe University Ph (03) 9496 5726 Email: [email protected] BUNDOORA VIC 3086 Email: [email protected] Ph (03) 9479 2565 RNA NETWORK AUSTRALASIA Email: [email protected] WESTERN AUSTRALIA Chair: Dr Archa Fox Associate Professor Nicolas Taylor Harry Perkins Institute of Medical Research EDUCATION REPRESENTATIVE ARC Centre of Excellence in Plant Energy 6 Verdun Street Associate Professor Susan Rowland Biology NEDLANDS WA 6009 Institute for Teaching and Learning University of Western Australia Ph (08) 6151 0762 Innovation (ITaLI) CRAWLEY WA 6009 Email: [email protected] University of Queensland Ph (08) 6488 7005 ST LUCIA QLD 4072 Email: [email protected] SYDNEY PROTEIN GROUP Ph (07) 3365 3089 Chair: Dr Liza Cubeddu Email: [email protected] School of Science and Health, University of Western Sydney, PENRITH NSW 2751 FAOBMB REPRESENTATIVE Ph (02) 4620 3343 Professor Paul Gleeson Email: [email protected] Department of Biochemistry and Molecular Biology University of Melbourne PARKVILLE VIC 3010 Ph (03) 8344 2354 Email: [email protected] SECRETARY FOR SUSTAINING MEMBERS ASBMB NATIONAL OFFICE Sally Jay PO Box 2331 COPY DEADLINE FOR c/- ASBMB National Office KENT TOWN SA 5071 NEXT ISSUE: PO Box 2331 Ph (08) 8362 0009 Monday, 12 June 2017 KENT TOWN SA 5071 Fax (08) 8362 0009 Ph (08) 8362 0009 Email: [email protected] Fax (08) 8362 0009 http://www.asbmb.org.au Email: [email protected]

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