Journal of Microbiology

Journal Article

Comprehensive Analysis of Gut Microbiota Alteration in the Patients and Animal Models with Polycystic Ovary Syndrome: Jing Zhou , Xuemei Qiu , Xuejing Chen , Sihan Ma , Zhaoyang Chen , Ruzhe Wang , Ying Tian , Yufan Jiang , Li Fan , Jingjie Wang; J. Microbiol. 2023;61(9):821-836. Published online October 12, 2023; DOI: https://doi.org/10.1007/s12275-023-00079-9

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Polycystic ovary syndrome (PCOS) is a common disease of endocrine–metabolic disorder, and its etiology remains largely unknown. The gut microbiota is possibly involved in PCOS, while the association remains unclear. The comprehensive analysis combining gut microbiota with PCOS typical symptoms was performed to analyze the role of gut microbiota in PCOS in this study. The clinical patients and letrozole-induced animal models were determined on PCOS indexes and gut microbiota, and fecal microbiota transplantation (FMT) was conducted. Results indicated that the animal models displayed typical PCOS symptoms, including disordered estrous cycles, elevated testosterone levels, and ovarian morphological change; meanwhile, the symptoms were improved after FMT. Furthermore, the microbial diversity exhibited disordered, and the abundance of the genus Ruminococcus and Lactobacillus showed a consistent trend in PCOS rats and patients. The microbiota diversity and several key genera were restored subjected to FMT, and correlation analysis also supported relevant conclusions. Moreover, LEfSe analysis showed that Gemmiger, Flexispira, and Eubacterium were overrepresented in PCOS groups. Overall, the results indicate the involvement of gut microbiota in PCOS and its possible alleviation of endocrinal and reproductive dysfunctions through several special bacteria taxa, which can function as the biomarker or potential target for diagnosis and treatment. These results can provide the new insights for treatment and prevention strategies of PCOS.

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Gut Microbes in Polycystic Ovary Syndrome and Associated Comorbidities; Type 2 Diabetes, Non-Alcoholic Fatty Liver Disease (NAFLD), Cardiovascular Disease (CVD), and the Potential of Microbial Therapeutics
Vineet Singh, Kanika Mahra, DaRyung Jung, Jae-Ho Shin
Probiotics and Antimicrobial Proteins.2024; 16(5): 1744. CrossRef
Potential therapeutic application and mechanism of gut microbiota-derived extracellular vesicles in polycystic ovary syndrome
Liangliang Yang, Tingxiu Liu, Yan Liao, Yuehan Ren, Zheng Zheng, Mingyue Zhang, Yue Yu, Chang Liu, Chaoying Wang, Tong Chen, Lili Zhang, Dongxue Zheng, Haidan Zhao, Zhexin Ni, Xinmin Liu
Biomedicine & Pharmacotherapy.2024; 180: 117504. CrossRef
Research Advance on the Prevention and Treatment of Polycystic Ovary Syndrome Based on Dysbiosis of Gut Microbiota
钰炜王
Advances in Clinical Medicine.2024; 14(08): 895. CrossRef

Observational Study

Early gut microbiota in very low and extremely low birth weight preterm infants with feeding intolerance: a prospective case-control study: Ling Liu , Dang Ao , Xiangsheng Cai , Peiyi Huang , Nali Cai , Shaozhu Lin , Benqing Wu; J. Microbiol. 2022;60(10):1021-1031. Published online August 19, 2022; DOI: https://doi.org/10.1007/s12275-022-2180-2

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Abstract

The potential role of the gut microbiota in the pathogenesis of feeding intolerance (FI) remains unclear. Understanding the role of the gut microbiota could provide a new avenue for microbiota-targeted therapeutics. This study aimed to explore the associations between aberrant gut microbiota and FI in very low or extremely low birth weight (VLBW/ELBW) preterm infants. In this observational case-control study, VLBW/ ELBW infants were divided into two groups: FI group and feeding tolerance (FT) group. 16S rRNA gene sequencing was performed to analyze the gut microbial diversity and composition of the infants. The differences in the gut microbiota of the two groups were compared. In total, 165 stool samples were obtained from 44 infants, among which, 31 developed FI and 13 served as controls. Alpha diversity was the highest in the meconium samples of the two groups. LEfSe analysis revealed that the abundances of Peptostreptococcaceae, Clostridiales and Clostridia in the FT group were significantly higher than in the FI group. At the phylum level, the FI group was dominated by Proteobacteria, and the FT group was dominated by Firmicutes. The meconium samples of the FI group had higher proportions of γ-proteobacteria and Escherichia-Shigella and a lower proportion of Bacteroides compared with the FT group. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis demonstrated that aberrant gut bacteria in the FI group were strongly associated with dysregulation of C5-Brancheddibasic- acid-metabolism, protein kinases, and sporulation. These findings reveal candidate microbial markers to prevent FI. Increased relative abundances of γ-proteobacteria and Escherichia-Shigella and decreased abundance of Bacteroides in meconium were associated with an increased risk of FI, while Peptostreptococcaceae, Clostridiales and Clostridia reduced the risk of FI in VLBW/ELBW infants.

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Reduced Gut Bacterial Diversity in Early Life Predicts Feeding Intolerance in Preterm Neonates
Maria Di Chiara, Alessandro Lazzaro, Daniela Scribano, Maria Trancassini, Valeria Pietropaolo, Michele Sonnessa, Chiara De Luca, Rita Prota, Elisa Onestà, Gianluigi Laccetta, Gianluca Terrin
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Calorie restriction during gestation impacts maternal and offspring fecal microbiome in mice
Stephanie P. Gilley, Meghan L. Ruebel, Sree V. Chintapalli, Clyde J. Wright, Paul J. Rozance, Kartik Shankar
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Dynamics alteration of the gut microbiota and faecal metabolomes in very low or extremely low birth weight infants: a Chinese single-center, prospective cohort study
Ling Liu, Chaohong Chen, YeShan Li, Dang Ao, Jiayuan Wu, Nali Cai, Wen Li, Min Xiang
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Metabolic and fecal microbial changes in adult fetal growth restricted mice
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Investigating prenatal and perinatal factors on meconium microbiota: a systematic review and cohort study
Jenni Turunen, Mysore V. Tejesvi, Niko Paalanne, Tytti Pokka, Sajeen Bahadur Amatya, Surbhi Mishra, Anna Kaisanlahti, Justus Reunanen, Terhi Tapiainen
Pediatric Research.2024; 95(1): 135. CrossRef
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Brigitta I.R.V. Corebima, Rinawati Rohsiswatmo, Dewi Santosaningsih, Wisnu Barlianto, Kusworini Handono
Archives of Medical Science.2023; 20(2): 444. CrossRef
Dynamics and Crosstalk between Gut Microbiota, Metabolome, and Fecal Calprotectin in Very Preterm Infants: Insights into Feeding Intolerance
Luyang Hong, Yihuang Huang, Junyan Han, Shujuan Li, Lan Zhang, Siyuan Jiang, Qi Zhou, Xincheng Cao, Weiyin Yu, Yi Yang, Shangyu Hong, Yufeng Zhou, Weili Yan, Yun Cao
Nutrients.2023; 15(22): 4849. CrossRef
Characteristics of Gut Microbiota in Small for Gestational Age Infants with Very Low Birth Weight
Hung-Yang Chang, Jen-Shiu Chiang Chiau, Jui-Hsing Chang, Chyong-Hsin Hsu, Chia-Ying Lin, Mary Hsin-Ju Ko, Hung-Chang Lee
Nutrients.2022; 14(23): 5158. CrossRef
Compositional Differences of Meconium Microbiomes of Preterm and Term Infants, and Infants That Developed Necrotizing Enterocolitis or Feeding Intolerance
Hyun Mi Kang, Sol Kim, Seok Hwang-Bo, In Hyuk Yoo, Yu-Mi Seo, Moon Yeon Oh, Soo-Ah Im, Young-Ah Youn
Pathogens.2022; 12(1): 55. CrossRef

Journal Article

Description of Nocardioides piscis sp. nov., Sphingomonas piscis sp. nov. and Sphingomonas sinipercae sp. nov., isolated from the intestine of fish species Odontobutis interrupta (Korean spotted sleeper) and Siniperca scherzeri (leopard mandarin fish): Dong-Wook Hyun , Yun-Seok Jeong , Jae-Yun Lee , Hojun Sung , So-Yeon Lee , Jee-Won Choi , Hyun Sik Kim , Pil Soo Kim , Jin-Woo Bae; J. Microbiol. 2021;59(6):552-562. Published online April 20, 2021; DOI: https://doi.org/10.1007/s12275-021-1036-5

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Abstract

A polyphasic taxonomic approach was used to characterize three novel bacterial strains, designated as HDW12AT, HDW- 15BT, and HDW15CT, isolated from the intestine of fish species Odontobutis interrupta or Siniperca scherzeri. All isolates were obligate aerobic, non-motile bacteria, and grew optimally at 30°C. Phylogenetic analysis based on 16S rRNA sequences revealed that strain HDW12AT was a member of the genus Nocardioides, and closely related to Nocardioides allogilvus CFH 30205T (98.9% sequence identities). Furthermore, strains HDW15BT and HDW15CT were members of the genus Sphingomonas, and closely related to Sphingomonas lutea JS5T and Sphingomonas sediminicola Dae 20T (97.1% and 97.9% sequence identities), respectively. Strain HDW12AT contained MK-8 (H4), and strains HDW15BT and HDW15CT contained Q-10 as the respiratory quinone. Major polar lipid components of strain HDW12AT were diphosphatidylglycerol, phosphatidylglycerol, and phosphatidylinositol, and those of strains HDW15BT and HDW15CT were sphingoglycolipid, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, and phosphatidylcholine. The G + C content of strains HDW12AT, HDW15BT, and HDW15CT were 69.7, 63.3, and 65.5%, respectively. The results of phylogenetic, phenotypic, chemotaxonomic, and genotypic analyses suggest that strain HDW12AT represents a novel species within the genus Nocardioides, and strains HDW15BT and HDW15CT represent two novel species within the genus Sphingomonas. We propose the names Nocardioides piscis for strain HDW12AT (= KACC 21336T = KCTC 49321T = JCM 33670T), Sphingomonas piscis for strain HDW15BT (= KACC 21341T = KCTC 72588T = JCM 33738T), and Sphingomonas sinipercae for strain HDW15CT (= KACC 21342T = KCTC 72589T = JCM 33739T).

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Sphingomonas flavescens sp. nov., isolated from soil
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Review

A comprehensive review of SARS-CoV-2 genetic mutations and lessons from animal coronavirus recombination in one health perspective: Woonsung Na , Hyoungjoon Moon , Daesub Song; J. Microbiol. 2021;59(3):332-340. Published online February 23, 2021; DOI: https://doi.org/10.1007/s12275-021-0660-4

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Abstract

SARS-CoV-2 was originated from zoonotic coronaviruses and confirmed as a novel beta-coronavirus, which causes serious respiratory illness such as pneumonia and lung failure, COVID-19. In this review, we describe the genetic characteristics of SARS-CoV-2, including types of mutation, and molecular epidemiology, highlighting its key difference from animal coronaviruses. We further summarized the current knowledge on clinical, genetic, and pathological features of several animal coronaviruses and compared them with SARSCoV- 2, as well as recent evidences of interspecies transmission and recombination of animal coronaviruses to provide a better understanding of SARS-CoV-2 infection in One Health perspectives. We also discuss the potential wildlife hosts and zoonotic origin of this emerging virus in detail, that may help mitigate the spread and damages caused by the disease.

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Journal Articles

N-acetylcysteine prevents the development of gastritis induced by Helicobacter pylori infection: Sungil Jang , Eun-Jung Bak , Jeong-Heon Cha; J. Microbiol. 2017;55(5):396-402. Published online April 29, 2017; DOI: https://doi.org/10.1007/s12275-017-7089-9

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Abstract

Helicobacter pylori (H. pylori) is a human gastric pathogen, causing various gastric diseases ranging from gastritis to gas-tric adenocarcinoma. It has been reported that combining N-acetylcysteine (NAC) with conventional antibiotic therapy increases the success rate of H. pylori eradication. We evalu-ated the effect of NAC itself on the growth and coloniza-tion of H. pylori, and development of gastritis, using in vitro liquid culture system and in vivo animal models. H. pylori growth was evaluated in broth culture containing NAC. The H. pylori load and histopathological scores of stomachs were measured in Mongolian gerbils infected with H. pylori strain 7.13, and fed with NAC-containing diet. In liquid culture, NAC inhibited H. pylori growth in a concentration-depen-dent manner. In the animal model, 3-day administration of NAC after 1 week from infection reduced the H. pylori load; 6-week administration of NAC after 1 week from infection prevented the development of gastritis and reduced H. pylori colonization. However, no reduction in the bacterial load or degree of gastritis was observed with a 6-week administ-ration of NAC following 6-week infection period. Our results indicate that NAC may exert a beneficial effect on reduction of bacterial colonization, and prevents the development of severe inflammation, in people with initial asymptomatic or mild H. pylori infection.

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Cot kinase plays a critical role in Helicobacter pylori-induced IL-8 expression: Sungil Jang , Jinmoon Kim , Jeong-Heon Cha; J. Microbiol. 2017;55(4):311-317. Published online March 31, 2017; DOI: https://doi.org/10.1007/s12275-017-7052-9

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Abstract

Helicobacter pylori is a major pathogen causing various gastric diseases including gastric cancer. Infection of H. pylori induces pro-inflammatory cytokine IL-8 expression in gastric epithelial cells in the initial inflammatory process. It has been known that H. pylori can modulate Ras-Raf-Mek-Erk signal pathway for IL-8 induction. Recently, it has been shown that another signal molecule, cancer Osaka thyroid oncogene/tumor progression locus 2 (Cot/Tpl2) kinase, activates Mek and Erk and plays a role in the Erk pathway, similar to MAP3K signal molecule Raf kinase. Therefore, the objective of this study was to determine whether Cot kinase might be involved in IL-8 induction caused by H. pylori infection. AGS gastric epithelial cells were infected by H. pylori strain G27 or its isogenic mutants lacking cagA or type IV secretion system followed by treatment with Cot kinase inhibitor (KI) or siRNA specific for Cot kinase. Activation of Erk was assessed by Western blot analysis and expression of IL-8 was measured by ELISA. Treatment with Cot KI reduced both transient and sustained Erk activation. It also reduced early and late IL-8 secretion in the gastric epithelial cell line. Furthermore, siRNA knockdown of Cot inhibited early and late IL-8 secretion induced by H. pylori infection. Taken together, these results suggest that Cot kinase might play a critical role in H. pylori type IV secretion apparatus-dependent early IL-8 secretion and CagA-dependent late IL-8 secretion as an alternative signaling molecule in the Erk pathway.

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Helicobacter pylori outer membrane protein, HomC, shows geographic dependent polymorphism that is influenced by the Bab family: Aeryun Kim , Stephanie L. Servetas , Jieun Kang , Jinmoon Kim , Sungil Jang , Yun Hui Choi , Hanfu Su , Yeong-Eui Jeon , Youngmin A. Hong , Yun-Jung Yoo , D. Scott Merrell , Jeong-Heon Cha; J. Microbiol. 2016;54(12):846-852. Published online November 26, 2016; DOI: https://doi.org/10.1007/s12275-016-6434-8

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Abstract

The array of outer membrane proteins (OMPs) found in Helicobacter pylori provides a crucial component for persistent colonization within the gastric niche. Not only does H. pylori harbor a wide number of OMPs, but these OMPs often vary across strains; this likely contributes to immune evasion, adaptation during long term colonization, and potentially differential disease progression. Previous work from our group described OMP differences among the Bab family (babA, babB, and babC) and Hom family (homA and homB) from 80 American H. pylori clinical isolates (AH) and 80 South Korean H. pylori clinical isolates (KH). In the current study, we expanded our investigation to include the less well characterized Hom family member, HomC. Overall, we identified and genotyped three homC variants: homCS, homCL, and homCM, in both populations. Similar to other polymorphic genes, the KH group showed less overall diversity, with 97.5% of strains harboring homCL. In contrast, a more heterogeneous profile was observed in strains derived from an American population; we found nearly equal distribution of homCS and homCL. Further analysis of the AH group identified associations between homC polymorphism and bab genotype; in AH strains, there was a significant association between homCL and carriage of babA at locus A. Since babA is an important virulence factor for the development of severe gastric disease, these data may suggest that homC polymorphism plays a role in H. pylori pathogenesis.

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Research Support, Non-U.S. Gov't

Novel nuclear targeting coiled-coil protein of Helicobacter pylori showing Ca2+-independent, Mg2+-dependent DNase I activity: Young Chul Kwon , Sinil Kim , Yong Seok Lee , Je Chul Lee , Myung-Je Cho , Woo-Kon Lee , Hyung-Lyun Kang , Jae-Young Song , Seung Chul Baik , Hyeon Su Ro; J. Microbiol. 2016;54(5):387-395. Published online April 20, 2016; DOI: https://doi.org/10.1007/s12275-016-5631-9

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Abstract

HP0059, an uncharacterized gene of Helicobacter pylori, encodes a 284-aa-long protein containing a nuclear localization sequence (NLS) and multiple leucine-rich heptad repeats. Effects of HP0059 proteins in human stomach cells were assessed by incubation of recombinant HP0059 proteins with the AGS human gastric carcinoma cell line. Wild-type HP0059 proteins showed cytotoxicity in AGS cells in a concentrationdependent manner, whereas NLS mutant protein showed no effect, suggesting that the cytotoxicity is attributed to host nuclear localization. AGS cells transfected with pEGFP-HP0059 plasmid showed strong GFP signal merged to the chromosomal DNA region. The chromosome was fragmented into multiple distinct dots merged with the GFP signal after 12 h of incubation. The chromosome fragmentation was further explored by incubation of AGS chromosomal DNA with recombinant HP0059 proteins, which leaded to complete degradation of the chromosomal DNA. HP0059 protein also degraded circular plasmid DNA without consensus, being an indication of DNase I activity. The DNase was activated by MgCl2, but not by CaCl2. The activity was completely blocked by EDTA. The optimal pH and temperature for DNase activity were 7.0–8.0 and 55°C, respectively. These results indicate that HP0059 possesses a novel DNase I activity along with a role in the genomic instability of human gastric cells, which may result in the transformation of gastric cells.

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Journal Article

Use of Selected Lactic Acid Bacteria in the Eradication of Helicobacter pylori Infection: Jin-Eung Kim , Min-Soo Kim , Yeo-Sang Yoon , Myung-Jun Chung , Do-Young Yum; J. Microbiol. 2014;52(11):955-962. Published online October 3, 2014; DOI: https://doi.org/10.1007/s12275-014-4355-y

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Abstract

Helicobacter pylori is among the major pathogenic bacteria that cause chronic gastritis and peptic ulcer disease and is related to the development of gastric cancer. Several chemicals, including antibiotics, have been used to eradicate H. pylori; however, they do not always curb the infection. Ten representative type strains of lactic acid bacteria (LAB) were screened for antagonism toward H. pylori via inhibition of urease activity. Strains inhibiting the binding of H. pylori to human gastric cell line cells and suppressing H. pylori-induced interleukin-8 (IL-8) production were also screened. Of these, Pediococcus pentosaseus (SL4), which inhibited the adhesion of H. pylori to MKN-45 gastric cancer cells, Bifidobacterium longum (BG7), with urease inhibiting activity, and Lactococcus lactis (SL3), and Enterococcus faecalis (SL5), which suppressed H. pylori-induced IL-8 production within MKN-45 and AGS cells, were selected. In mouse model, these LAB stains in combination significantly suppressed IL-8 levels in serum. Gastric pH also recovered to normal values after the administration of these LAB. These stains effectively suppressed H. pylori viability, although not to the extent of antibiotic treatment. When used as probiotics, LAB may help decrease the occurrence of gastritis and reduce the risk of H. pylori infection without, inducing side effects.

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Helicobacter pylori outer membrane protein, HomC, shows geographic dependent polymorphism that is influenced by the Bab family
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Research Support, Non-U.S. Gov't

An Easy Way for the Rapid Purification of Recombinant Proteins from Helicobacter pylori Using a Newly Designed Expression Vector: Hyung-Lyun Kang , Jin-Sung Jo , Soon-Uck Kwon , Jae-Young Song , Ji-Hyun Seo , Myung-Je Cho , Seung-Chul Baik , Hee-Shang Youn , Kwang-Ho Rhee , Woo-Kon Lee; J. Microbiol. 2014;52(7):604-608. Published online June 28, 2014; DOI: https://doi.org/10.1007/s12275-014-3679-y

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Abstract

We constructed a H. pylori expression vector which consisted of both a His-tag and a GST tag as purification tools for recombinant protein and a chloramphenicol resistant cat gene as a reporter. The backbone of the vector pBK contained an ColEI origin of replication and a kanamycin resistant gene. A set of oligos for the His-tag and the PCR product of gst (glutathione S-transferase) gene were inserted sequentially in frame in the multi-cloning site of pBK. The orf of cat was inserted downstream of the gst to generate pBKHGC. The 3' part of H. pylori clpB and flaA were cloned into the vector which was introduced into H. pylori. Recombinant proteins were purified by GSH affinity column, digested with thrombin and were analyzed by western blotting. The final recombinant proteins were successfully purified.

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Gene polymorphisms of pathogenic Helicobacter pylori in patients with different types of gastrointestinal diseases
Yu-Li Chen, Xiao-Qiang Mo, Gan-Rong Huang, Yan-Qiang Huang, Juan Xiao, Li-Juan Zhao, Hong-Yu Wei, Qian Liang
World Journal of Gastroenterology.2016; 22(44): 9718. CrossRef

Review

MINIREVIEW] Nontraditional Therapies to Treat Helicobacter pylori Infection: Morris O. Makobongo , Jeremy J. Gilbreath , D. Scott Merrell; J. Microbiol. 2014;52(4):259-272. Published online March 29, 2014; DOI: https://doi.org/10.1007/s12275-014-3603-5

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Abstract

The Gram-negative pathogen Helicobacter pylori is increasingly more resistant to the three major antibiotics (metronidazole, clarithromycin and amoxicillin) that are most commonly used to treat infection. As a result, there is an increased rate of treatment failure; this translates into an overall higher cost of treatment due to the need for increased length of treatment and/or the requirement for combination or sequential therapy. Given the rise in antibiotic resistance, the complicated treatment regime, and issues related to patient compliance that stem from the duration and complexity of treatment, there is clearly a pressing need for the development of novel therapeutic strategies to combat H. pylori infection. As such, researchers are actively investigating the utility of antimicrobial peptides, small molecule inhibitors and naturopathic therapies. Herein we review and discuss each of these novel approaches as a means to target this important gastric pathogen.

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Amin Talebi Bezmin Abadi
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Menghua Xiong, Yan Bao, Xin Xu, Hua Wang, Zhiyuan Han, Zhiyu Wang, Yeqing Liu, Songyin Huang, Ziyuan Song, Jinjing Chen, Richard M. Peek, Lichen Yin, Lin-Feng Chen, Jianjun Cheng
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Helicobacter pylori outer membrane protein, HomC, shows geographic dependent polymorphism that is influenced by the Bab family
Aeryun Kim, Stephanie L. Servetas, Jieun Kang, Jinmoon Kim, Sungil Jang, Yun Hui Choi, Hanfu Su, Yeong-Eui Jeon, Youngmin A. Hong, Yun-Jung Yoo, D. Scott Merrell, Jeong-Heon Cha
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Research Support, Non-U.S. Gov'ts

Phenotypic and Genotypic Analysis of Clarithromycin-Resistant Helicobacter pylori from Bogotá D.C., Colombia: Alba A. Trespalacios , William Otero , Jorge E. Caminos , Marcela M. Mercado , Jenny Ávila , Liliana E. Rosero , Azucena Arévalo , Raúl A. Poutou-Piñales , David Y. Graham; J. Microbiol. 2013;51(4):448-452. Published online August 30, 2013; DOI: https://doi.org/10.1007/s12275-013-2465-6

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Abstract: Resistance of Helicobacter pylori to clarithromycin is the most common cause of treatment failure in patients with H. pylori infections. This study describes the MICs and the presence of 23S rRNA mutations of H. pylori isolates from Bogotá, D.C., Colombia. H. pylori were isolated from gastric biopsies from patients with functional dyspepsia. Clarithromycin susceptibility was investigated by agar dilution and strains were considered resistant if the MIC was ≥1 μg/ml. DNA sequences of the 23S rRNA gene of strains resistant and sensitive to clarithromycin were determined to identify specific point mutations. Clarithromycin resistance was present in 13.6% of patients by agar dilution. The A2143G, A2142G and A2142C mutations were found in 90.5, 7.1, and 2.4% of H. pylori strains with resistance genotype.The resistant phenotype was associated with 23S rRNA resistance genotype in 85.7% of isolates. The point mutations in 23S rRNA were well correlated with MICs values for clarithromycin.

Genetic Organization and Conjugal Plasmid DNA Transfer of pHP69, a Plasmid from a Korean Isolate of Helicobacter pylori: Jung-Soo Joo , Jae-Young Song , Seung-Chul Baik , Woo-Kon Lee , Myung-Je Cho , Kon-Ho Lee , Hee-Shang Youn , Ji-Hyun Seo , Kwang-Ho Rhee , Hyung-Lyun Kang; J. Microbiol. 2012;50(6):955-961. Published online December 30, 2012; DOI: https://doi.org/10.1007/s12275-012-2580-9

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Abstract: We isolated pHP69, a 9,153 bp plasmid from Helicobacter pylori with a 33.98% (G+C) content. We identified 11 open reading frames (ORFs), including replication initiation protein A (repA), fic (cAMP-induced filamentation protein), mccC, mccB, mobA, mobD, mobB, and mobC, as well as four 22 bp tandem repeat sequences. The nucleic acid and predicted amino acid sequences of these ORFs exhibited significant homology to those of other H. pylori plasmids. pHP69 repA encodes a replication initiation protein and its amino acid sequence is similar to those of replicase proteins from theta-type plasmids. pHP69 contains two types of repeat sequences (R1 and R2), a MOBHEN family mobilization region comprising mobC, mobA, mobB, and mobD, and genes encoding microcin B and C. Among the 36 H. pylori strains containing plasmids, mobA or mccBC are present in 12 or 6, respectively and 3 contain both genes. To examine intrinsic capability of H. pylori for conjugative plasmid transfer, a shuttle vector pBHP69KH containing pHP69 and replication origin of pBR322 was constructed. It was shown that this vector could stably replicate and be mobilized among clinical H. pylori strains and demonstrated to gene transfer by natural plasmid.

Research Support, U.S. Gov't, Non-P.H.S.

NOTE] The Helicobacter pylori Ferric Uptake Regulator (Fur) Is Essential for Growth Under Sodium Chloride Stress: Hanan Gancz , D. Scott Merrell; J. Microbiol. 2011;49(2):294-298. Published online May 3, 2011; DOI: https://doi.org/10.1007/s12275-011-0396-7

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Abstract: Epidemiological data and animal models indicate that Helicobacter pylori and dietary NaCl have a synergistic ill effect on gastric maladies. Here we show that the Ferric Uptake Regulator (Fur), which is a crucial regulatory factor required for H. pylori colonization, is essential for growth in the presence of high NaCl concentrations. Moreover, we demonstrate that the transcriptional response induced by sodium chloride stress exhibits similarities to that seen under iron depletion.

Research Support, Non-U.S. Gov't

Identification of S-Nitrosylation of Proteins of Helicobacter pylori in Response to Nitric Oxide Stress: Wei Qu , Yabin Zhou , Yundong Sun , Ming Fang , Han Yu , Wenjuan Li , Zhifang Liu , Jiping Zeng , Chunyan Chen , Chengjiang Gao , Jihui Jia; J. Microbiol. 2011;49(2):251-256. Published online May 3, 2011; DOI: https://doi.org/10.1007/s12275-011-0262-7

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Abstract: Innate and adaptive immune responses are activated in humans when Helicobacter pylori invades the gastric mucosa. Nitric oxide (NO) and reactive nitrogen species are important immune effectors, which can exert their functions through oxidation and S-nitrosylation of proteins. S-nitrosoglutathione and sodium nitroprusside were used as NO donors and H. pylori cells were incubated with these compounds to analyze the inhibitory effect of NO. The suppressing effect of NO on H. pylori has been shown in vitro. Furthermore, the proteins modified by S-nitrosylation in H. pylori were identified through the biotin switch method in association with matrix-assisted laser desorption ionization/time-of-flight tandem mass spectrometry (MALDITOF- MS/MS). Five S-nitrosylated proteins identified were a chaperone and heat-shock protein (GroEL), alkyl hydroperoxide reductase (TsaA), urease alpha subunit (UreA), HP0721, and HP0129. Importantly, S-nitrosylation of TsaA and UreA were confirmed using purified recombinant proteins. Considering the importance of these enzymes in antioxidant defenses, adherence, and colonization, NO may exert its antibacterial actions by targeting enzymes through S-nitrosylation. Identification of protein S-nitrosylation may contribute to an understanding of the antibacterial actions of NO. Our findings provide an insight into potential targets for the development of novel therapeutic agents against H. pylori infection.

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