DOCSLIB.ORG

Monogenic Disorders of Glucose-Stimulated Insulin Secretion: Massively Parallel Sequencing Approaches

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Monogenic Disorders of Glucose-Stimulated Insulin Secretion: Massively Parallel Sequencing Approaches Monogenic Disorders of Glucose-Stimulated Insulin Secretion: Massively Parallel Sequencing Approaches Stephanie R. Johnson FRACP, MBBS, BSc A thesis submitted for the degree of Doctor of Philosophy at The University of Queensland in 2017 Faculty of Medicine i Abstract Maturity-onset diabetes of the young (MODY) and congenital hyperinsulinism (CHI) are monogenic disorders of glucose-stimulated insulin secretion. MODY is the commonest form of monogenic diabetes and results from a heterozygous mutation in one of thirteen identified genes. It is estimated that up to 90% of cases of MODY are not diagnosed, possibly due to the insufficient awareness of or access to genetic screening. CHI is the commonest cause of persistent hypoglycemia in infancy and childhood, for which there are ten identified genes (mutation-positive disease found in 45% of all CHI, and 88% of diazoxide-unresponsive cases). Genetic confirmation of MODY and CHI traditionally involved Sanger sequencing of each exon of each gene individually; however, the development of massively parallel sequencing (MPS) allows simultaneous sequencing of multiple target regions efficiently and cost-effectively. This thesis assessed two forms of MPS - whole exome sequencing and targeted sequencing, in population and small family cohorts. An entire paediatric diabetes clinic underwent targeted MPS for MODY genes, which showed the prevalence of pathogenic/likely pathogenic MODY variants to be 2.1%. Based on these results, a cost-effectiveness analysis was performed, demonstrating that routine MPS screening for MODY in all paediatric cases with presumed type 1 diabetes was cost-saving. The utility of WES was evaluated in MODY and CHI in a proof-of-concept study. Subjects with known mutations in MODY and CHI genes were re-sequenced using two “off-the-shelf” WES capture technologies. All mutations were confirmed; however, the study showed the importance of careful selection of appropriate capture technology to ensure targeting of relevant genes and the use of bioinformatics tools appropriate to mutation type. Seven probands diagnosed with MODY but without a previously identified mutation were also assessed using WES. Three novel variants in known MODY genes were identified by WES, which variants segregated with diabetes in the families. A potential novel MODY gene was also identified through sequencing of one extended family. ii Six probands with CHI but without a known mutation were also sequenced. Two probands and their affected family members had the same variant in a potential novel CHI gene. MPS is an extremely useful addition to the diagnostic workup for paediatric cases of diabetes overall, in addition to its utility in screening clinically diagnosed MODY/CHI cases. Additionally, MPS can lead to novel gene identification for these conditions. iii Declaration by author This thesis is composed of my original work, and contains no material previously published or written by another person except where due reference has been made in the text. I have clearly stated the contribution by others to jointly-authored works that I have included in my thesis. I have clearly stated the contribution of others to my thesis as a whole, including statistical assistance, survey design, data analysis, significant technical procedures, professional editorial advice, and any other original research work used or reported in my thesis. The content of my thesis is the result of work I have carried out since the commencement of my research higher degree candidature and does not include a substantial part of work that has been submitted to qualify for the award of any other degree or diploma in any university or other tertiary institution. I have clearly stated which parts of my thesis, if any, have been submitted to qualify for another award. I acknowledge that an electronic copy of my thesis must be lodged with the University Library and, subject to the policy and procedures of The University of Queensland, the thesis be made available for research and study in accordance with the Copyright Act 1968 unless a period of embargo has been approved by the Dean of the Graduate School. I acknowledge that copyright of all material contained in my thesis resides with the copyright holder(s) of that material. Where appropriate I have obtained copyright permission from the copyright holder to reproduce material in this thesis. iv Publications during candidature Conference presentations Stephanie R. Johnson, Elizabeth Davis, Louise S. Conwell, Mark Harris, Lisa Anderson, Paul Leo, Matthew Brown, Emma L. Duncan (2016) “Massively parallel sequencing to determine the prevalence of maturity onset diabetes of the young in an entire paediatric diabetes clinic, including antibody positive type 1 diabetes” . Australasian Paediatric Endocrine Group annual scientific meeting, Alice Springs, (oral presentation in Young Investigators) S.R. Johnson , E.A. Davis, L.S. Conwell, M. Harris, E.L. Duncan. Prevalence of maturity onset diabetes of the young in a Western Australian paediatric diabetes clinic using targeted massively parallel sequencing. [Abstract O2] Pediatric Diabetes 2015;16(Suppl 21):24-49. [Awarded APEG Young Investigator Award] Stephanie R. Johnson , Louise S Conwell , Mark Harris , Emma L. Duncan. Next generation Sequencing for Maturity Onset Diabetes of the Young. [Abstract P105] Pediatric Diabetes 2014;15(Suppl.19)1-137 Stephanie R Johnson , Ivan N. McGown, Louise Conwell, Mark Harris, Emma L. Duncan. Next Generation Sequencing (NGS) to identify a novel INS gene mutation in a family with Maturity Onset Diabetes of the Young (MODY). [Abstract P2-d1-857] Hormone Research 2013;80(Suppl 1):267 Stephanie R Johnson , Ivan McGown, Mark Williams, Joyce Wu, Mark Harris, Emma Duncan, David Cowley. (2012) “Genetic Testing in Patients with a Clinical Diagnosis of Maturity-Onset Diabetes of the Young (MODY) – The Australian Experience” Monogenic Disorders of Insulin Secretion: Congenital Hyperinsulinism and Neonatal Diabetes, Philadelphia v Publications included in this thesis No publications included. vi Contributions by others to the thesis Chapter 3: Professor Emma Duncan supervised the study design, along with Assoc. Professor Louise Conwell, Dr Mark Harris, Professor Matthew Brown and Assoc. Professor Elizabeth Davis. Ms Janelle MacFarlane, Assoc. Professor Elizabeth Davis and Ms Nirubasini Paramalingam assisted with ethics submission. Dr Uma Ganti, Dr Jacqueline Curran, Professor Timothy Jones and Assoc. Professor Elizabeth Davis assisted with sample acquisition. Ms Lisa Anderson, Ms Xiaoxia Song and Ms Jessica Harris performed library preparation for the massively parallel sequencing and Ms Jessica Harris performed Sanger sequencing. Dr Paul Leo and Dr Jonathan Ellis performed the bioinformatics analysis. Dr McInerney-Leo and Professor Emma Duncan assisted in analysis of the massively parallel sequencing data. Chapter 4 : Dr Hannah Carter, Professor Nicholas Graves, Dr Samantha Hollingworth, Professor Matthew Brown and Professor Emma Duncan had a role in study design. Dr Hannah Carter and Dr Samantha Hollignworth provided data from the literature for the model. Dr Hannah Carter created the Markov Model and Dr Hannah Carter and Dr Samantha Hollingworth ran the model supervised by Professor Nicholas Graves. Professor Timothy Jones and Assoc. Professor Elizabeth Davis provided additional data from the WACDD database. Chapter 5 : Mr Ivan McGown performed the massively parallel sequencing and analysis. Professor Udo Oppermann provided the in-silico modelling. Chapter 6 : Professor Emma Duncan assisted with study design. Ms Janelle MacFarlane, Dr Aideen McInerney-Leo and Professor Emma Duncan assisted with ethics submission. Mr Ivan McGown assisted with sample acquisition. Ms Xiaixia Song and Ms Lisa Anderson performed the whole exome sequencing. Dr Paul Leo, Ms Mhairi performed the bioinformatics analysis. Dr Felicity Newell analysed the data to detect deletions. Ms Mhairi Marshall also provided the R script to calculate target and capture regions vii Chapter 7 : Professor Emma Duncan assisted with study design. Ms Janelle MacFarlane and Professor Emma Duncan assisted with ethics submission. Mr Ivan McGown assisted with sample acquisition. Ms Xiaixia Song and Ms Lisa Anderson performed the whole exome sequencing. Dr Paul Leo, Ms Mhairi performed the bioinformatics analysis. Dr Felicity Newell analysed the data to detect deletions. Ms Jessica Harris assisted with Sanger sequencing Chapter 8 : Professor Emma Duncan assisted with study design. Ms Janelle MacFarlane and Professor Emma Duncan assisted with ethics submission. Mr Ivan McGown assisted with sample acquisition. Ms Xiaoxia Song and Ms Lisa Anderson performed the whole exome sequencing. Dr Paul Leo, Ms Mhairi performed the bioinformatics analysis. Ms Jessica Harris assisted with Sanger sequencing Professor Emma Duncan provided the majority of the assistance with conceptualising the studies and with study design, data analysis and performed the majority of the paper editing. Professor Matthew Brown assisted with study design as provided all the laboratory equipment and laboratory space as well as staff to assist me and teach me. viii Statement of parts of the thesis submitted to qualify for the award of another degree None ix Acknowledgements I would firstly like to thank my wonderful supervisors, Emma, Mark and Louise, who had the belief in me and in this project from the start. You allowed me enough free rein to re-design the thesis as new opportunities arose,
Load more

Recommended publications
  • Liver Glucose Metabolism in Humans
    Biosci. Rep. (2016) / 36 / art:e00416 / doi 10.1042/BSR20160385 Liver glucose metabolism in humans Mar´ıa M. Adeva-Andany*1, Noemi Perez-Felpete*,´ Carlos Fernandez-Fern´ andez*,´ Cristobal´ Donapetry-Garc´ıa* and Cristina Pazos-Garc´ıa* *Nephrology Division, Hospital General Juan Cardona, c/ Pardo Bazan´ s/n, 15406 Ferrol, Spain Synopsis Information about normal hepatic glucose metabolism may help to understand pathogenic mechanisms underlying obesity and diabetes mellitus. In addition, liver glucose metabolism is involved in glycosylation reactions and con- nected with fatty acid metabolism. The liver receives dietary carbohydrates directly from the intestine via the portal vein. Glucokinase phosphorylates glucose to glucose 6-phosphate inside the hepatocyte, ensuring that an adequate flow of glucose enters the cell to be metabolized. Glucose 6-phosphate may proceed to several metabolic path- ways. During the post-prandial period, most glucose 6-phosphate is used to synthesize glycogen via the formation of glucose 1-phosphate and UDP–glucose. Minor amounts of UDP–glucose are used to form UDP–glucuronate and UDP– galactose, which are donors of monosaccharide units used in glycosylation. A second pathway of glucose 6-phosphate metabolism is the formation of fructose 6-phosphate, which may either start the hexosamine pathway to produce UDP-N-acetylglucosamine or follow the glycolytic pathway to generate pyruvate and then acetyl-CoA. Acetyl-CoA may enter the tricarboxylic acid (TCA) cycle to be oxidized or may be exported to the cytosol to synthesize fatty acids, when excess glucose is present within the hepatocyte. Finally, glucose 6-phosphate may produce NADPH and ribose 5-phosphate through the pentose phosphate pathway.
    [Show full text]
  • Hyperinsulinemic Hypoglycemia in Infancy: Current Concepts in Diagnosis and Management
    RRR EEE VVV III EEE WWW AAA RRR TTT III CCC LLL EEE Hyperinsulinemic Hypoglycemia in Infancy: Current Concepts in Diagnosis and Management SHRENIK VORA, SURESH CHANDRAN, VICTOR SAMUEL RAJADURAI AND #KHALID HUSSAIN From Department of Neonatology, KK Women’s and Children’s Hospital, Singapore; and #Genetics and Epigenetics in Health and Disease Genetics and Genomic Medicine Programme, UCL Institute of Child Health, Great Ormond Street Hospital for Children, 30 Guilford Street, London, UK. Correspondence to: Dr Shrenik Vora, Senior Staff Registrar, Department of Neonatology, KK Women’s and Children’s Hospital, 100, Bukit Timah Road, Singapore 229899. [email protected] Purpose: Molecular basis of various forms of hyperinsulinemic hypoglycemia, involving defects in key genes regulating insulin secretion, are being increasingly reported. However, the management of medically unresponsive hyperinsulinism still remains a challenge as current facilities for genetic diagnosis and appropriate imaging are limited only to very few centers in the world. We aim to provide an overview of spectrum of clinical presentation, diagnosis and management of hyperinsulinism. Methods: We searched the Cochrane library, MEDLINE and EMBASE databases, and reference lists of identified studies. Conclusions: Analysis of blood samples, collected at the time of hypoglycemic episodes, for intermediary metabolites and hormones is critical for diagnosis and treatment. Increased awareness among clinicians about infants “at-risk” of hypoglycemia, and recent advances in genetic diagnosis have made remarkable contribution to the diagnosis and management of hyperinsulinism. Newer drugs like lanreotide (long acting somatostatin analogue) and sirolimus (mammalian target of rapamycin (mTOR) inhibitor) appears promising as patients with diffuse disease can be treated successfully without subtotal pancreatectomy, minimizing the long-term sequelae of diabetes and pancreatic insufficiency.
    [Show full text]
  • MODY 2 Presenting As Neonatal Hyperglycaemia: a Need to Reshape the Definition of ªneonatal Diabetesº?
    Letters 1331 MODY 2 presenting as neonatal sidered a stringent criterium for selecting patients. We ob- served that the moderate low birth weight of patients No. 1 hyperglycaemia: a need to reshape and No. 3 was probablyinfluenced bya mutation of paternal the definition of ªneonatal diabetesº? origin and subsequent low fetal insulin secretion. Mutations in the GK represent the more frequent cause of Dear Sir, MODY in some series [3] and it has been calculated that up Neonatal diabetes mellitus is defined as a rare condition to 6% of familial Type II non-insulin-dependent) diabetes 1:400.000 live births) of unknown origin, characterised byhy- mellitus could bear a MODY 2 allele [3]. We and others have perglycaemia occurring in the first month of life that requires ex- frequentlyobserved that the mutation-carryingparent of chil- ogenous insulin therapy[1, 2]. The derangement of glucose me- dren with MODY 2 is unaware of his or her hyperglycaemia. tabolism in newborns affected bythis condition could be perma- As a consequence, some previouslydescribed cases of perma- nent permanent neonatal diabetes mellitus, PNDM) or could nent neonatal diabetes of unknown origin might be linked to orcouldnot)recurafterremissionofhyperglycaemiatransient GK mutations in both alleles, even in the absence of consan- neonataldiabetesmellitus,TNDM)[1,2]. Inapedigreewith ma- guineityloop i.e. compound heterozygous). Neonatal, insu- turityonset diabetes of the youngMODY) harbouring the glu- lin-requiring, permanent diabetes mellitus is the main feature cokinase GK, MODY 2) mutation Q38P, we followed a new of GK knock-out mice models [5]. pregnancyofthenon-mutantmotheroftheindexcase.Wefound We believe that the definition of neonatal diabetes currently that 15 days after delivery her child presented hyperglycaemia inuse[1,2]needstoberevisedinorderto:firstly,permanently in- 11.7 mmol) with dehydration and mild ketosis Table 1, patient corporate the concept that TNDM in some cases linked to pa- No.
    [Show full text]
  • Mechanisms of Disease: Advances in Diagnosis and Treatment of Hyperinsulinism in Neonates Diva D De León and Charles a Stanley*
    REVIEW www.nature.com/clinicalpractice/endmet Mechanisms of Disease: advances in diagnosis and treatment of hyperinsulinism in neonates Diva D De León and Charles A Stanley* SUMMARY INTRODUCTION Etiology of neonatal hypoglycemia Hyperinsulinism is the single most common mechanism of hypoglycemia Hypoglycemia is a frequent problem in newborn in neonates. Dysregulated insulin secretion is responsible for the infants that must be diagnosed and treated effi- transient and prolonged forms of neonatal hypoglycemia, and congenital ciently to avoid seizures and permanent brain genetic disorders of insulin regulation represent the most common damage. Management is complicated by the fact of the permanent disorders of hypoglycemia. Mutations in at least five genes have been associated with congenital hyperinsulinism: they that neonates present a remarkably wide range of encode glucokinase, glutamate dehydrogenase, the mitochondrial possible causes for hypoglycemia. These include enzyme short-chain 3-hydroxyacyl-CoA dehydrogenase, and the two transient forms of hypoglycemia that can occur components (sulfonylurea receptor 1 and potassium inward rectifying in normal infants, more prolonged forms of channel, subfamily J, member 11) of the ATP-sensitive potassium hypoglycemia that are associated with compli- channels (K channels). K hyperinsulinism is the most common and cations of gestation and delivery, and neonatal ATP ATP presentation of permanent hypo glycemia severe form of congenital hyperinsulinism. Infants suffering from KATP hyperinsulinism present shortly after birth with severe and persistent dis orders due to endocrine or metabolic diseases hypoglycemia, and the majority are unresponsive to medical therapy, thus (Box 1). The risks of these various forms change requiring pancreatectomy. In up to 40–60% of the children with KATP hyperinsulinism, the defect is limited to a focal lesion in the pancreas.
    [Show full text]
  • Hypoglycemia Diagnosis and Management
    HYPOGLYCEMIA DIAGNOSIS AND MANAGEMENT Beckwith-Wiedemann syndrome (BWS) is a rare disorder involving changes on a region of chromosome 11p15 that influence pre- and postnatal growth. These changes disrupt the normal balance of growth gene expression and lead to the overgrowth seen in patients with BWS. Depending on which parts of the body are affected, children with BWS can have different features. These include overgrowth of the pancreas leading to hypoglycemia or hyperinsulinism. What is hypoglycemia? Hypoglycemia is low blood sugar. It is important for the body to have normal blood sugar levels because low sugar levels can cause problems including seizures and brain damage. Approximately half of infants with BWS will have hypoglycemia. In most of these children, hypoglycemia will only last for a few days and can easily be treated with frequent feedings and/or medical doses of sugar (dextrose infusions). However, in 5-10% of children with BWS, hypoglycemia can persist, requiring additional monitoring and medical care. What causes hypoglycemia in children with BWS? Although hypoglycemia can happen for many reasons, in children with BWS, it is usually caused when the pancreas makes too much insulin (hyperinsulinism). Insulin lowers blood sugar (also called glucose) levels. We suspect that low blood sugar levels in BWS have to do with the genes on chromosome 11p15 that control growth. Additionally, there are some other genes on chromosome 11 that affect how some of the cells in the pancreas work. How do we test for hypoglycemia? A normal blood sugar level is 70 – 120 mg/dL. Low glucose levels are common in the first 24 hours of life; after the second day of life, glucose levels should normalize.
    [Show full text]
  • Maternal Or Paternal Diabetes and Its Crucial Role in Offspring Birth
    H OH metabolites OH Editorial Maternal or Paternal Diabetes and Its Crucial Role in Offspring Birth Weight and MODY Diagnosis Valeria Calcaterra 1,2,* , Angela Zanfardino 3 , Gian Vincenzo Zuccotti 2,4 and Dario Iafusco 3 1 Pediatric and Adolescence Unit, Department of Internal Medicine and Therapeutics, University of Pavia, 27100 Pavia, Italy 2 Pediatric Unit, “V. Buzzi” Children’s Hospital, 20154 Milano, Italy; [email protected] 3 Department of Pediatrics, Regional Center of Pediatric Diabetology “G. Stoppoloni”, University of Campania “Luigi Vanvitelli”, 80138 Napoli, Italy; [email protected] (A.Z.); [email protected] (D.I.) 4 Department of Biomedical and Clinical Sciences “L. Sacco”, University of Milan, 20157 Milano, Italy * Correspondence: [email protected] Received: 17 September 2020; Accepted: 26 September 2020; Published: 28 September 2020 Abstract: Maturity-onset diabetes of the young (MODY) represents a heterogenous group of monogenic autosomal dominant diseases, which accounts for 1–2% of all diabetes cases. Pregnancy represents a crucial time to diagnose MODY forms due to the 50% risk of inheritance in offspring of affected subjects and the potential implications on adequate fetal weight. Not only a history of maternal diabetes may affect the birth weight of offspring, paternal diabetes should also be taken into consideration for a correct pathogenetic diagnosis. The crucial role of maternal and paternal diabetes inheritance patterns and the impact of this inherited mutation on birthweight and the MODY diagnosis was discussed. Keywords: mother; father; diabetes; birthweight; MODY Maturity-onset diabetes of the young (MODY) represents a heterogenous group of monogenic autosomal dominant diseases, which accounts for 1–2% of all diabetes cases [1].
    [Show full text]
  • Congenital Hyperinsulinism
    Winner of the Case of the Year SWISS SOCIETY OF NEONATOLOGY Award 2012 Congenital hyperinsulinism FEBRUARY 2012 2 Morgillo D, Berger TM, Caduff JH, Barthlen W, Mohnike K, Mohnike W, Neonatal and Pediatric Intensive Care Unit (MD, BTM), Department of Pediatric Radiology (CJH), Children‘s Hospital of Lucerne, Lucerne, Switzerland, Department of Pediatric Surgery, University Medicine Greifswald (BW), Greifswald, Germany, Department of Pediatrics, University Hospital of Magdeburg (MK), Magdeburg, Germany, Diagnostic Therapeutic Centre Frankfurter Tor (MW), Berlin, Germany © Swiss Society of Neonatology, Thomas M Berger, Webmaster 3 Congenital hyperinsulinism (CHI) is characterized by INTRODUCTION inappropriate secretion of insulin by the ß cells of the islets of Langerhans and is an extremely heterogene- ous condition in terms of clinical presentation, histo- logical subgroups and underlying molecular biology. Histologically, CHI has been classified into two major subgroups: diffuse (affecting the whole pancreas) and focal (being localized to a single region of the pancreas) disease. Advances in molecular genetics, radiological imaging techniques (such as fluorine-18 L-3,4-dihydroxyphenylalanine-PET-CT (18FDOPA-PET-CT) scanning) and surgical techniques have completely changed the clinical approach to infants with severe congenital forms of hyperinsulinemic hypoglycemia. This male infant was born to a healthy 35-year-old G3/P3 CASE REPORT by spontaneous vaginal delivery at 38 4/7 weeks. His birth weight was 3530 g (P 50-75), his head circum- ference was 35 cm (P 25) and his length was 50 cm (P 25-50). Postnatal adaptation was normal with an arterial cord pH of 7.28 and Apgar scores of 8, 9, and 9 at 1, 5, and 10 minutes, respectively.
    [Show full text]
  • Monogenic Diabetes: a New Pathogenic Variant of HNF1A Gene Raquel Vilela Oliveira, Teresa Bernardo, Sandrina Martins, Ana Sequeira
    Case report BMJ Case Rep: first published as 10.1136/bcr-2019-231837 on 20 January 2021. Downloaded from Monogenic diabetes: a new pathogenic variant of HNF1A gene Raquel Vilela Oliveira, Teresa Bernardo, Sandrina Martins, Ana Sequeira Pediatrics, Unidade Local de SUMMARY hepatic nuclear factor 1 alpha (HNF1A), respec- Saúde do Alto Minho EPE, Viana Maturity onset diabetes of the young defines a tively.2–4 6 7 10 These subtypes account for up to 90% do Castelo, Portugal diabetes mellitus subtype, with no insulin resistance of all MODY cases.10 or autoimmune pancreatic β-cells dysfunction, that One of the challenges is to differentiate MODY Correspondence to from other forms of diabetes. The diagnosis of Dr Raquel Vilela Oliveira; occurs by mutation in a single gene. A 13- year- old Raquelvoliveira07@ gmail. com girl hospitalised due to hyperglycemia plus glycosuria MODY should be suspected in patients with family without ketosis, and with normal glycated haemoglobin history of diabetes of any type, diagnosed at a young Accepted 11 November 2019 of 6.8%. She started a sugar- free fast- absorption diet age (<25 years), lack of type 1 DM characteris- and no insulin therapy was required. Fasting glucose tics (the absence of islet autoantibodies, preserved was normal, but 2 hours after lunch she presented β-cell function with low or no insulin requirements hyperglycemia as after 2 hours of an oral glucose and detectable C- peptide over an extended partial tolerance test, with 217 mg/dL. Family history was remission phase), the absence of typical type 2 DM positive for type 2 diabetes mellitus with an autosomal characteristics (severe obesity, acanthosis nigricans dominant pattern.
    [Show full text]
  • (12) Patent Application Publication (10) Pub. No.: US 2016/0281166 A1 BHATTACHARJEE Et Al
    US 20160281 166A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2016/0281166 A1 BHATTACHARJEE et al. (43) Pub. Date: Sep. 29, 2016 (54) METHODS AND SYSTEMIS FOR SCREENING Publication Classification DISEASES IN SUBJECTS (51) Int. Cl. (71) Applicant: PARABASE GENOMICS, INC., CI2O I/68 (2006.01) Boston, MA (US) C40B 30/02 (2006.01) (72) Inventors: Arindam BHATTACHARJEE, G06F 9/22 (2006.01) Andover, MA (US); Tanya (52) U.S. Cl. SOKOLSKY, Cambridge, MA (US); CPC ............. CI2O 1/6883 (2013.01); G06F 19/22 Edwin NAYLOR, Mt. Pleasant, SC (2013.01); C40B 30/02 (2013.01); C12O (US); Richard B. PARAD, Newton, 2600/156 (2013.01); C12O 2600/158 MA (US); Evan MAUCELI, (2013.01) Roslindale, MA (US) (21) Appl. No.: 15/078,579 (57) ABSTRACT (22) Filed: Mar. 23, 2016 Related U.S. Application Data The present disclosure provides systems, devices, and meth (60) Provisional application No. 62/136,836, filed on Mar. ods for a fast-turnaround, minimally invasive, and/or cost 23, 2015, provisional application No. 62/137,745, effective assay for Screening diseases, such as genetic dis filed on Mar. 24, 2015. orders and/or pathogens, in Subjects. Patent Application Publication Sep. 29, 2016 Sheet 1 of 23 US 2016/0281166 A1 SSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSSS S{}}\\93? sau36 Patent Application Publication Sep. 29, 2016 Sheet 2 of 23 US 2016/0281166 A1 &**** ? ???zzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzzz??º & %&&zzzzzzzzzzzzzzzzzzzzzzz &Sssssssssssssssssssssssssssssssssssssssssssssssssssssssss & s s sS ------------------------------ Patent Application Publication Sep. 29, 2016 Sheet 3 of 23 US 2016/0281166 A1 23 25 20 FG, 2. Patent Application Publication Sep. 29, 2016 Sheet 4 of 23 US 2016/0281166 A1 : S Patent Application Publication Sep.
    [Show full text]
  • Download an Example File with Diabetes Mellitus (PDF)
    Legal Disclaimer All diagnostic and therapeutic procedures in the field of science and medicine are continuously evolving: Therefore the herein presented medical evidence is state of the technical and scientific art at the time of the editorial deadline for the respective edition of the book. All details provided herein related to a particular therapeutic mode of administration and dosing of drugs are screened employing utmost measures of accuracy and precision. Unless explicitly specified otherwise, all drug administration and dosing regimens presented herein are for healthy adults with normal renal and hepatic function. Liability cannot be assumed for any type of dosing and administration regimen presented herein. Each reader is advised to carefully consult the respective market authorization holders directions for use of the recommended drugs, medical devices and other means of therapy and diagnosis. This applies in particular also to market authorization holders summaries of product characteristics for pharmaceutical products. Despite all care, there may be translation errors. The reader is advised to carefully check information related to the indication for use, contraindications, dosing recommendations, side effects and interactions with other medications! All modes of medication use and administration are at the consumers risk. The author and his team cannot be held responsible for any damage caused by wrong therapy or diagnosis. Please refer to, i.e: www.leitlinien.de or www.guideline.gov The trade name of a trade name registered product does not provide the legal privilege to employ the trade name as a free trade mark even if it is not specifically marked as such. Drugs which are sold as generics are referred to throughout the book by their generic name and not necessarily by a particular brand name Remark: ICD 10 code version 1.3 has been employed for the index.
    [Show full text]
  • Functional Imaging of Insulitis in Type 1 Diabetes
    FUNCTIONAL IMAGING OF INSULITIS IN TYPE 1 DIABETES Teemu Kalliokoski TURUN YLIOPISTON JULKAISUJA – ANNALES UNIVERSITATIS TURKUENSIS Sarja - ser. D osa - tom. 1135 | Medica - Odontologica | Turku 2014 University of Turku Faculty of Medicine Department of Clinical Medicine Pediatrics and Turku PET Centre Doctoral Programme of Clinical Investigation Supervised by Professor emeritus Olli Simell (MD, PhD) Adjunct professor Merja Haaparanta-Solin (MSc, PhD) University of Turku, Faculty of Medicine University of Turku, Faculty of Medicine Department of Pediatrics Turku PET Centre Reviewed by Professor Timo Otonkoski (MD, PhD) Docent Olof Eriksson (MSc, PhD) University of Helsinki Uppsala University Children’s Hospital Department of Medicinal Chemistry Opponent Professor Alberto Signore (MD, PhD) Sapienza University Nuclear Medicine Unit Rome, Italy The originality of this thesis has been checked in accordance with the University of Turku quality assurance system using the Turnitin OriginalityCheck service. ISBN 978-951-29-5859-7 (PRINT) ISBN 978-951-29-5860-3 (PDF) ISSN 0355-9483 Painosalama Oy - Turku, Finland 2014 Adjunct professor Merja Haaparanta-Solin (MSc, PhD) University of Turku, Faculty of Medicine Turku PET Centre To Mervi, and our children Elias and Iiris 4 Abstract ABSTRACT Teemu Kalliokoski FUNCTIONAL IMAGING OF INSULITIS IN TYPE 1 DIABETES Department of Pediatrics and Turku PET Centre, Faculty of Medicine, University of Turku Annales Universitatis Turkuensis, 2014, Turku, Finland Type 1 diabetes (T1D) is an immune-mediated disease characterized by autoimmune inflammation (insulitis), leading to destruction of insulin-producing pancreatic β-cells and consequent dependence on exogenous insulin. Current evidence suggests that T1D arises in genetically susceptible children who are exposed to poorly characterized environmental triggers.
    [Show full text]
  • Two MODY 2 Families Identified in Brazilian Subjects
    case report Incidental mild hyperglycemia in children: two MODY 2 families identified in Brazilian subjects Hiperglicemia incidental em crianças: duas famílias com MODY 2 identificadas em brasileiros Lílian A. Caetano1,2, Alexander A. L. Jorge1, Alexsandra C. Malaquias1, Ericka B. Trarbach1, Márcia S. Queiroz2, Márcia Nery2, Milena G. Teles1,2 SUMMARY Maturity-onset diabetes of the young (MODY) is characterized by an autosomal dominant mode 1 Unidade de Endocrinologia of inheritance, early onset of hyperglycemia, and defects of insulin secretion. MODY subtypes Genética e Laboratório de described present genetic, metabolic, and clinical differences. MODY 2 is characterized by mild Endocrinologia Molecular e Celular/LIM25, Disciplina de asymptomatic fasting hyperglycemia, and rarely requires pharmacological treatment. Hence, Endocrinologia, Faculdade precise diagnosis of MODY is important for determining management and prognosis. We report de Medicina da Universidade two heterozygous GCK mutations identified during the investigation of short stature. Case 1: a de São Paulo (FMUSP), São Paulo, SP, Brazil prepubertal 14-year-old boy was evaluated for constitutional delay of growth and puberty. During 2 Unidade de Diabetes, follow-up, he showed abnormal fasting glucose (113 mg/dL), increased level of HbA1c (6.6%), and Hospital das Clínicas, FMUSP, negative β-cell antibodies. His father and two siblings also had slightly elevated blood glucose le- São Paulo, SP, Brazil vels. The mother had normal glycemia. A GCK heterozygous missense mutation, p.Arg191Trp, was identified in the proband. Eighteen family members were screened for this mutation, and 11 had the mutation in heterozygous state. Case 2: a 4-year-old boy investigated for short stature revealed no other laboratorial alterations than elevated glycemia (118 mg/dL); β-cell antibodies were nega- tive.
    [Show full text]