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Published OnlineFirst October 29, 2018; DOI: 10.1158/1055-9965.EPI-18-0542 Research Article Cancer Epidemiology, Biomarkers The Microbiomes of Pancreatic and Duodenum & Prevention Tissue Overlap and Are Highly Subject Specific but Differ between Pancreatic Cancer and Noncancer Subjects Erika del Castillo1,4, Richard Meier2, Mei Chung1, Devin C. Koestler2,3, Tsute Chen4, Bruce J. Paster4,5, Kevin P. Charpentier6, Karl T. Kelsey7, Jacques Izard8,9, and Dominique S. Michaud1 Abstract Background: In mice, bacteria from the mouth can trans- cavity. Bacterial DNA across different sites in the pancreas and locate to the pancreas and impact pancreatic cancer progres- duodenum were highly subject specific in both cancer and sion. In humans, oral bacteria associated with periodontal noncancer subjects. Presence of genus Lactobacillus was signif- disease have been linked to pancreatic cancer risk. It is not icantly higher in noncancer subjects compared with cancer known if DNA bacterial profiles in the pancreas and duode- subjects and the relative abundance of Fusobacterium spp., num are similar within individuals. previously associated with colorectal cancer, was higher in Methods: Tissue samples were obtained from 50 subjects cancer subjects compared with noncancer subjects. with pancreatic cancer or other conditions requiring foregut Conclusions: Bacterial DNA profiles in the pancreas were surgery at the Rhode Island Hospital (RIH), and from 34 similar to those in the duodenum tissue of the same subjects, organs obtained from the National Disease Research Inter- regardless of disease state, suggesting that bacteria may be change. 16S rRNA gene sequencing was performed on 189 migrating from the gut into the pancreas. Whether bacteria tissue samples (pancreatic duct, duodenum, pancreas), 57 play a causal role in human pancreatic cancer needs to be swabs (bile duct, jejunum, stomach), and 12 stool samples. further examined. Results: Pancreatic tissue samples from both sources (RIH Impact: Identifying bacterial taxa that differ in cancer and National Disease Research Interchange) had diverse bac- patients can provide new leads on etiologically relevant terial DNA, including taxa typically identified in the oral bacteria. Introduction high fatality rate, and the silent progression of early disease, identifying risk factors for the prevention and early detection of In 2018, an estimated 55,440 individuals will be diagnosed pancreatic cancer is critical to reducing its mortality. To date, with pancreatic cancer in the United States, and only 8% of these known risk factors for pancreatic cancer, including smoking, individuals are expected to survive the next 5 years (1). Given this obesity, diabetes, heavy alcohol consumption, family history, and markers of genetic susceptibility, cannot, even collectively, be used for early detection and risk stratification of pancreatic 1Department of Public Health & Community Medicine, Tufts University School of 2 cancer in the general population (2). Medicine, Tufts University, Boston, Massachusetts. Department of Biostatistics, Studies have suggested a link between bacteria and pancreatic The University of Kansas Medical Center, Kansas City, Kansas. 3University of Kansas Cancer Center, The University of Kansas Medical Center, Kansas City, cancer risk (3), highlighting the need to more critically explore the Kansas. 4The Forsyth Institute, Cambridge, Massachusetts. 5Harvard School of underlying factors that affect the microbiome of the oral cavity Dental Medicine, Boston, Massachusetts. 6Department of Surgery, Rhode Island and upper digestive tract in both cancer patients and cancer-free Hospital, Providence, Rhode Island. 7Department of Epidemiology and Pathol- individuals. The current research on oral bacteria and pancreatic ogy and Laboratory Medicine, Brown University, Providence, Rhode Island. 8 cancer risk stems from a number of observational studies that Food Science and Technology Department, University of Nebraska, Lincoln, reported a higher risk of pancreatic cancer among individuals with Nebraska. 9School of Biological Sciences, University of Nebraska, Lincoln, Nebraska. periodontitis, when compared with those without periodontitis (3, 4). Periodontitis, an inflammatory disease of the gums, is Note: Supplementary data for this article are available at Cancer Epidemiology, largely driven by keystone pathogens and pathobionts (5). Two Biomarkers & Prevention Online (http://cebp.aacrjournals.org/). large prospective cohort studies have reported positive associa- fi E. del Castillo and R. Meier share rst authorship. tions between periodontal disease pathogens and subsequent Corresponding Author: Dominique S. Michaud, Tufts University, 136 Harrison pancreatic cancer risk (6, 7); in these two studies, detection of Avenue, Boston, MA 02111. Phone: 617-636-0482; Fax: 617-636-6807; E-mail: elevated antibodies to Porphyromonas gingivalis, measured in [email protected] blood collected prior to cancer diagnosis, was associated with a doi: 10.1158/1055-9965.EPI-18-0542 two-fold higher risk of pancreatic cancer (6), and presence (vs. Ó2018 American Association for Cancer Research. absence) of P. gingivalis in saliva collected prior to cancer diagnosis 370 Cancer Epidemiol Biomarkers Prev; 28(2) February 2019 Downloaded from cebp.aacrjournals.org on October 1, 2021. © 2019 American Association for Cancer Research. Published OnlineFirst October 29, 2018; DOI: 10.1158/1055-9965.EPI-18-0542 Microbiomes in Pancreatic Tissue and Duodenum was associated with a 60% increase in risk of pancreatic cancer of cancer, use of other over-the-counter medications were also (7). Aggregatibacter actinomycetemcomitans, another periodontal included on the questionnaire. Stool collection kits with ethanol pathogen, was also associated with pancreatic cancer risk in the as a fixative [95% (w/w) ethanol] were provided prior to surgery prospective study using saliva (7). (23); participants were asked to return the samples using a pre- Few investigations to date have attempted to detect bacteria in paid box. pancreatic tissue. Earlier studies reported the presence of bacteria A protocol was established for processing tissue samples in pancreatic ducts of subjects with chronic pancreatitis or bile collected during surgery to reduce contamination. A technician duct obstruction (8–10). Other studies have investigated the from the Pathology Department was informed in advance of presence of specific bacterial DNA in the pancreatic tissue of the surgery date and time, and was paged as soon as the pancreatic cancer subjects, namely species of Helicobacter (11) specimens had been obtained. Surgical tissue samples were and Fusobacterium (12). The most comprehensive molecular frozen within 1 hour of the surgery time, as well as tissues swab microbiome studies to date reported the presence of a diverse samples from the stomach, jejunum, and bile duct that were bacterial populations in fluids collected from the bile duct, collected using DNA-free forensic sterile swabs whenever pos- pancreas, and jejunum of subjects undergoing pancreaticoduo- sible. During surgery, the surgeon also recorded (on a surgery denectomy (13), and in pancreatic cyst fluid removed endoscop- form for the study) if the patient had received prior pre-OP ically from pancreatic cysts (14). In mice, bacteria have been endoscopic ultrasound (EUS), had previously had their gall- shown to translocate from the mouth to the pancreas, and germ- bladderremoved,orhadreceivedpriorplacementofastent free mice have reduced progression of pancreatic ductal adeno- (for treatment of symptoms); all subjects received a single dose carcinoma (15). of perioperative antibiotics immediately prior skin incision at Metagenomics studies on DNA isolated from tissue samples thetimeoftheoperation.Tissuesamples(pancreatictumor from cancer subjects have been conducted for lung (16), colo- tissue, pancreatic cysts, normal pancreatic tissue, pancreatic rectal (17), esophageal (18), stomach (19), and breast cancer ducts, and duodenums) were prepared by a Rhode Island (20). These studies demonstrate that 16S rRNA gene sequencing Hospital pathologist; cancerous and noncancerous tissues were can be effectively conducted on fresh tissue samples where the identified, separated and labeled. All samples were de-identi- ratio of bacterial to human DNA is much lower than at other fiedandstoredatÀ80C until processing. human sites (e.g., stool or oral cavity; ref. 21). Moreover, these Upon review of pathology records, ICD10 codes were studies have shown that bacterial profiles at different organ sites assigned to each subject; 39 subjects had pancreatic cancer are often unique (16, 20) and that changes may be associated with (ICD10 codes C25.0–C25.9; the majority of cases were adeno- cancer (18, 19). In two recent studies, bacterial DNA was mea- carcinomas, only two subjects had neuroendocrine tumors of sured in tumor tissue samples obtained from patients with the pancreas), 12 subjects had periampullary cancer (ICD10 pancreatic ductal adenocarcinoma (PDAC) using 16S rRNA gene codes C24.0–C24.1), 18 subjects had other pancreatic condi- sequencing (15, 22); however, comparison of microbiota in tions (ICD10 codes K86.0–K86.3), and the remaining eight pancreas and different gastrointestinal tissue was not conducted had other gastrointestinal conditions. The study was approved in these patients. by Lifespan's Research Protection Office for recruitment at RIH, To date, no study to our knowledge has characterized the as
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  • Hickman Catheter-Related Bacteremia with Kluyvera

    Hickman Catheter-Related Bacteremia with Kluyvera

    Jpn. J. Infect. Dis., 61, 229-230, 2008 Short Communication Hickman Catheter-Related Bacteremia with Kluyvera cryocrescens: a Case Report Demet Toprak, Ahmet Soysal, Ozden Turel, Tuba Dal1, Özlem Özkan1, Guner Soyletir1 and Mustafa Bakir* Department of Pediatrics, Section of Pediatric Infectious Diseases and 1Department of Microbiology, Marmara University School of Medicine, Istanbul, Turkey (Received September 10, 2007. Accepted March 19, 2008) SUMMARY: This report describes a 2-year-old child with neuroectodermal tumor presenting with febrile neu- tropenia. Blood cultures drawn from the peripheral vein and Hickman catheter revealed Kluyvera cryocrescens growth. The Hickman catheter was removed and the patient was successfully treated with cefepime and amikacin. Isolation of Kluyvera spp. from clinical specimens is rare. This saprophyte microorganism may cause serious central venous catheter infections, especially in immunosuppressed patients. Clinicians should be aware of its virulence and resistance to many antibiotics. Central venous catheters (CVCs) are frequently used in confirmed by VITEK AMS (VITEK Systems, Hazelwood, Mo., patients with hematologic and oncologic disorders. Along with USA) and by API (Analytab Inc., Plainview, N.Y., USA). Anti- their increased use, short- and long-term complications of microbial susceptibility was assessed by the disc diffusion CVCs are more often being reported. The incidence of CVC method. K. cryocrescens was sensitive for cefotaxime, cefepime, infections correlates with duration of catheter usage, immuno- carbapenems, gentamycine, amikacin and ciprofloxacin. logic status of the patient, type of catheter utilized and mainte- Intravenous cefepime and amikacin were continued and the nance techniques employed. A definition of CVC infection CVC was removed. His echocardiogram was normal and has been difficult to establish because of problems differen- a repeat peripheral blood culture was sterile 48 h after the tiating contaminant from pathogen microorganisms.