• Journal of Microbiology and Biotechnology
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• v.26 no.8
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• pp.1446-1451
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• 2016

Clostridium difficile toxin A causes acute gut inflammation in animals and humans. It is known to downregulate the tight junctions between colonic epithelial cells, allowing luminal contents to access body tissues and trigger acute immune responses. However, it is not yet known whether this loss of the barrier function is a critical factor in the progression of toxin A-induced pseudomembranous colitis. We previously showed that NADH:quinone oxidoreductase 1 (NQO1) KO (knockout) mice spontaneously display weak gut inflammation and a marked loss of colonic epithelial tight junctions. Moreover, NQO1 KO mice exhibited highly increased inflammatory responses compared with NQO1 WT (wild-type) control mice when subjected to DSS-induced experimental colitis. Here, we tested whether toxin A could also trigger more severe inflammatory responses in NQO1 KO mice compared with NQO1 WT mice. Indeed, our results show that C. difficile toxin A-mediated enteritis is significantly enhanced in NQO1 KO mice compared with NQO1 WT mice. The levels of fluid secretion, villus disruption, and epithelial cell apoptosis were also higher in toxin A-treated NQO1 KO mice compared with WT mice. The previous and present results collectively show that NQO1 is involved in the formation of tight junctions in the small intestine, and that defects in NQO1 enhance C. difficile toxin A-induced acute inflammatory responses, presumably via the loss of epithelial cell tight junctions.

• Korean Journal of Clinical Pharmacy
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• v.28 no.3
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• pp.174-180
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• 2018

Objective: Clostridium difficile Infection (CDI) is one of the common nosocomial infections. As elderly population increases, the proper treatment has been emphasized. We investigated the risk factors associated with CDI unimprovement in elderly patients. Furthermore, we performed drug use evaluation of old CDI patients and oldest-old CDI patients. Methods: It was a retrospective study using electronic medical record at Kangbuk Samsung Medical Center (KBSMC) from January 2016 to December 2017. Seventy three patients aged 65 years or older, diagnosed with CDI by Clostridium difficile Toxin B Gene [Xpert] were screened and they were assessed for risk factors regarding unimprovement status. We also evaluated drug use evaluation in old patients ($65{\leq}age$<80) and oldest-old patients ($80{\leq}age$) by assessing the use of initial therapy, severity, dose, route, treatment course, days of use, total days of use and treatment outcome of initial therapy. Results: Out of 73 patients aged over 65 years, four patients were excluded because they did not receive any treatment. There were 31 improved patients and 38 unimproved patients after initial therapy. We were able to find out patients with surgical comorbidity or endocrine comorbidity (especially, diabetes mellitus) had 2.885 more risk of becoming unimproved than those patients without surgical comorbidity or endocrine comorbidity. Drug use evaluation for CDI was generally fair, but vancomycin as initial therapy is more recommended than metronidazole. Conclusion: Although age, antibiotics exposure, use of antacids are all important risk factors for CDI, our result did not show statistical significance for these risk factors. However, the study is meaningful because the number of elderly population keeps increasing and recently updated guideline suggests the use of vancomycin as drug of choice for CDI.

• Journal of Microbiology and Biotechnology
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• v.26 no.4
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• pp.659-665
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• 2016

The oligosaccharides in human milk constitute a major innate immunological mechanism by which breastfed infants gain protection against infectious diarrhea. Clostridium difficile is the most important cause of nosocomial diarrhea, and the C-terminus of toxin A with its carbohydrate binding site, TcdA-f2, demonstrates specific abolishment of cytotoxicity and receptor binding activity upon diethylpyrocarbonate modification of the histidine residues in TcdA. TcdA-f2 was cloned and expressed in E. coli BL21 (DE3). A human milk oligosaccharide (HMO) mixture displayed binding with TcdA-f2 at 38.2 respond units (RU) at the concentration of 20 μg/ml, whereas the eight purified HMOs showed binding with the carbohydrate binding site of TcdA-f2 at 3.3 to 14 RU depending on their structures via a surface plasma resonance biosensor. Among them, Lacto-N-fucopentaose V (LNFPV) and Lacto-N-neohexaose (LNnH) demonstrated tight binding to TcdA-f2 with docking energy of −9.48 kcal/mol and −12.81 kcal/mol, respectively. It displayed numerous hydrogen bonding and hydrophobic interactions with amino acid residues of TcdA-f2.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.15 no.1
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• pp.29-37
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• 2012

Purpose: To examine the prevalence of $Clostridium$ $difficile$ ($C.$ $difficile$) colonization (CDC) and potential neonatal determinants of CDC in hospitalized preterm infants. Methods: Fecal samples were serially collected within 72 h after birth and at 1, 2, and 4-6 weeks of age from preterm infants in the neonatal intensive care units (NICUs) of two different university hospitals. Total bacterial DNA was extracted from each fecal sample from 49 infants, and polymerase chain reaction (PCR) was performed with primers for the 16S gene of $C.$ $difficile$ and the toxin A and toxin B genes. The correlation between the results of $C.$ $difficile$ PCR assays and the clinical characteristics of the infants was analyzed. Results: The prevalence rates of CDC were 34.7, 37.2, 41.3, and 53.1% within 72 h after birth and at 1, 2, and 4.6 weeks of age, respectively. The toxin positivity rate was significantly higher in the infants with persistent CDC than in those with transient CDC (8/12 [66.7%] vs. 6/25 [24.5%] ($p$=0.001). Among the various neonatal factors, only the feeding method during the first week after birth was significantly associated with persistent CDC. Exclusive breast-milk feeding (EBMF) significantly decreased the risk of persistent CDC compared to formula or mixed feeding (adjusted odds ratio: 0.133, 95% confidence interval: 0.02-0.898, $p$=0.038). Conclusion: The prevalence of CDC increased with the duration of hospitalization in preterm infants in the NICU. EBMF during the first week after birth in hospitalized preterm infants may protect against persistent CDC.

• Journal of Microbiology and Biotechnology
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• v.22 no.12
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• pp.1629-1635
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• 2012

Previously, we demonstrated that the erythropoietin receptor (EpoR) is present on fibroblasts, where it regulates focal contact. Here, we assessed whether this action of EpoR is involved in the reduced cell adhesion observed in colonocytes exposed to Clostridium difficile toxin A. EpoR was present and functionally active in cells of the human colonic epithelial cell line HT29 and epithelial cells of human colon tissues. Toxin A significantly decreased activating phosphorylations of EpoR and its downstream signaling molecules JAK-2 (Janus kinase 2) and STAT5 (signal transducer and activator of transcription 5). In vitro kinase assays confirmed that toxin A inhibited JAK 2 kinase activity. Pharmacological inhibition of JAK2 (with AG490) abrogated activating phosphorylations of EpoR and also decreased focal contacts in association with inactivation of paxillin, an essential focal adhesion molecule. In addition, AG490 treatment significantly decreased expression of occludin (a tight junction molecule) and tight junction levels. Taken together, these data suggest that inhibition of JAK2 by toxin A in colonocytes causes inactivation of EpoR, thereby enhancing the inhibition of focal contact formation and loss of tight junctions known to be associated with the enzymatic activity of toxin A.

• Korean Journal of Clinical Laboratory Science
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• v.46 no.4
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• pp.140-142
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• 2014

To evaluate the recovery rates to increase toxigenic C. difficile, the selective enrichment broth culture methods were compared with commonly used cytotoxin assays and toxigenic culture. First, the enrichment culture, using the selective medium broth for 2 to 5 days, was performed and then, toxigenic C. difficile was confirmed by C. difficile toxin gene-specific PCR after being cultured on C. difficile selective agar. The sensitivity of C. difficile from the enrichment culture (100%) was higher than that of C. difficile selective agar culture (93.8%), while positive predictive values (PPV) were low; 72.7% (16/22) and 88.2% (15/17). PPV of the enrichment culture are not high. Recently, combinations of C. difficile selective agar culture, C. difficile A & B assays, glutamate dehydrogenase, and nucleic acid amplification method are widely used. The enrichment culture was disadvantageous in PPV, turn-around time, and cost. So, what we performed is not considered as a common method of diagnosis of C. difficile-associated diarrhea.

• Journal of Yeungnam Medical Science
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• v.1 no.1
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• pp.171-178
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• 1984

Many reports have been made concerning underlying and associated conditions causing pseudomembranous colitis and it has been documented that occurrence of pseudomembranous colitis is related with antibiotics administration. Recent study showed that Clostridium difficile produced enterotoxin by colonization in intestinal wall and leading into pseudomembranous colitis. Diagnosis is based on positive culture of Clostridium difficile, positive test of Clostridium difficile toxin and specific histological findings after observation of whitish plaque on colonoscopic or sigmoidoscopic examination. Authors have experienced one case of pseudomembranous colitis developing after long term ampicillin administration in a case with colon cancer associated with diarrhea and diagnosis was confirmed by typical pseudomembrane on biopsy following classical whitish plaque observation on sigmoidoscopic examination. Symptoms have been ameliorated by discontinuation of antibiotics and administration of metronidazole in four days and disappearance of whitish plaque on repeated sigmoidoscopic examination and improvement of clinical symptoms after 9 days of medication.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.1 no.1
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• pp.138-143
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• 1998

Chronic diarrhea in children is a common problem with numerous causes. Although most of these causes are benign, critical illness may present as chronic diarrhea. In a patient of chronic diarrhea, gastrointestinal infections are the most common causes in children of all ages and antibiotics may cause chronic diarrhea by altering intestinal microflora, which can result in the emergence of bacterial overgrowth. Overgrowth of Clostridium difficile may cause pseudomembranous colitis. We experienced 25-month-old boy who suffered from chronic diarrhea and partially treated with antibiotics irregularly. Colonoscopic findings of this child showed multiple plaques with white to yellowish exudate which adhere to the mucosal surface of a variable length of rectum. Histollogically, each plaque comprised a pseudomembrane of mucous debris, inflammatory cells, and exudate overlying groups of partially disrupted glands. A latex agglutination test on patient's stool was positive to toxin A of Clostridium difficile. He was recovered after stopping the antibiotics he has been prescribed, and being given vancomycin for 2 weeks. We report this case with brief review of literature.

• Korean Journal of Clinical Laboratory Science
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• v.49 no.2
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• pp.141-149
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• 2017

To examine the cause of acute diarrheal disease in children and adolescents, 521 fecal samples underwent multiple-PCR for a period of two years, between March 2015 and February 20178, in the Jeju region of Korea. Based on the analysis of 179 positive samples, 102 samples were male (56.98%) and 77 were female (43.02%), and highest positive rates were shown in the age group of 3~4 years (51.96%) and 5~6 years (12.29%). When 209 cases (40.12%), including double infection were analyzed, there were 88 cases (16.89%) of norovirus-GII infection, 26 cases (4.99%) of Campylobacter spp. infection, 18 cases (3.45%) of rotavirus infection, 18 cases (3.45%) of Clostridium difficile Toxin B infection, 17 cases (3.26%) of adenovirus infection, 16 cases (3.07%) of Clostridium perfringens infection, 11 cases (2.11%) of astrovirus infection, 5 cases (0.96%) of Salmonella spp. infection, 3 cases (0.58%) of norovirus-GI, Yersinia spp. and Aeromonas spp. infections, and 1 case (0.19%) of verocytotoxin-producing E. coli infection. Based on a seasonal separation of early childhood, norovirus-GI and norovirus-GII mainly prevailed during the winter, when the temperature is low. Typical enteritis with an increased prevalence of rotavirus during the spring. Astrovirus prevailed between the months of April and October, when norovirus-GI, norovirus-GII, and rotavirus did not prevail. With increasing age, acute diarrheal disease was not only induced by a virus, but also by bacteria. Although a test for virus is an effective method when trying to identify the cause during early childhood by multiplex-PCR, it would be desirable to undergo tests for both virus and bacteria concurrently as age increases.

• Parasites, Hosts and Diseases
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• v.55 no.5
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• pp.513-521
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• 2017

Infectious diarrhea is endemic in most developing countries. We aimed to investigate the protozoan, viral, and bacterial causes of acute diarrhea in Taif, Saudi Arabia. A cross-sectional prospective 1-year study was conducted on 163 diarrheal patients of various ages. Stool samples were collected, 1 per patient, and tested for 3 protozoa, 3 viruses, and 9 bacteria with the Luminex Gastrointestinal Pathogen Panel. Overall, 53.4% (87/163) of samples were positives (20.8% protozoa, 19.6% viruses, 2.8% bacteria, and 9.8% mixed). Rotavirus (19.6%), Giardia duodenalis (16.5%), and Cryptosporidium spp. (8.5%) were the mostly detected pathogens. Adenovirus 40/41 (4.2%), Salmonella (3%), Shiga toxin-producing Escherichia coli (3%), and Entamoeba histolytica (2.4%) were also detected. Norovirus GI/II, Vibrio cholerae, Yersinia enterocolitica, and Clostridium difficile toxin A/B were not detected in any patients. All pathogens were involved in coinfections except E. histolytica. Giardia (5.5%) and rotavirus (3%) were the most commonly detected in co-infections. Enterotoxigenic E. coli (2.4%), Campylobacter spp. (2.4%), E. coli 0157 (1.8%), and Shigella spp. (1.2%) were detected in patients only as co-infections. Infections were more in children 0-4 years, less in adults <40 years, and least >40 years, with statistically significant differences in risk across age groups observed with rotavirus (P<0.001), Giardia (P=0.006), and Cryptosporidium (P=0.036) infections. Lastly, infections were not significantly more in the spring. This report demonstrates the high burden of various enteropathogens in the setting. Further studies are needed to define the impact of these findings on the clinical course of the disease.