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MicroRNA exhibit altered expression in the inflamed colonic mucosa of ulcerative colitis patients

World Journal of Gastroenterology 2017年第29期23卷 5324-5332页

作者： Swati Valmiki Vineet Ahuja jaishree paul School of Life Sciences Jawaharlal Nehru University Department of Gastroenterology All India Institute of Medical Sciences

To investigate the miRNA expression in colonic mucosal biopsies from endoscopically inflamed and non inflamed regions of ulcerative colitis (UC) patients. METHODSColonic mucosal pinch biopsies were analyzed from the inflamed and non inflamed regions of same UC patient. Total RNA was isolated and differential miRNA profiling was done using microarray platform. Quantitative Real Time PCR was performed in colonic biopsies from inflamed (n = 8) and non-inflamed (n = 8) regions of UC and controls (n = 8) to validate the differential expression of miRNA. Potential targets of dysregulated miRNA were identified by using in silico prediction tools and probable role of these miRNA in inflammatory pathways were predicted. RESULTSThe miRNA profile of inflamed colonic mucosa differs significantly from the non-inflamed. Real time PCR analysis showed that some of the miRNA were differentially expressed in the inflamed mucosa as compared to non inflamed mucosa and controls (miR-125b, miR-223, miR-138, and miR-155), while (miR-200a) did not show any significant changes. In contrast to microarray, where miR-378d showed downregulation in the inflamed mucosa, qRT-PCR showed a significant upregulation in the inflamed mucosa as compared to the non inflamed. The in silico prediction analysis revealed that the genes targeted by these miRNAs play role in the major signaling pathways like MAPK pathway, NF-κB signaling pathway, cell adhesion molecules which are all assciated with UC. CONCLUSIONThe present study reports disease specific alteration in the expression of miR-125b, miR-155, miR-223 and miR-138 in UC patients and also predict their biological significance.

关键词： Ulcerative colitis Colon mucosa MicroRNA Microarray qRT-PCR In silico analysis

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Fluctuations in butyrate-producing bacteria in ulcerative colitis patients of North India

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World Journal of Gastroenterology 2013年第22期19卷 3404-3414页

作者： Reena Kumari Vineet Ahuja jaishree paul School of Life Sciences Jawaharlal Nehru University New Delhi 110067 India Department of Gastroenterology All India Institute of Medical Sciences New Delhi 110029 India

AIM: To study the interplay between butyrate concentration and butyrate-producing bacteria in fecal samples of ulcerative colitis (UC) patients vs control individuals. METHODS: Fecal samples were collected from 14 control individuals (hemorrhoid patients only) and 26 UC patients (severe: n = 12, moderate: n = 6, remission: n = 8), recruited by the gastroenterologist at the Department of Gastroenterology, All India Institute of Medical Sciences, New Delhi, India. Disease activity in UC patients was determined by clinical colitis activity index. We employed fluorescent in situ hybridization in combination with flow cytometry to enumerate the clostridium cluster population targeted by 16S rRNA gene probe. Major butyrate-producing species within this cluster were quantified to see if any change existed in control vs UC patients with different disease activity. This observed change was further validated by quantitative polymerase chain reaction. In addition to this,we carried out gas chromatography to evaluate the changes in concentration of major short chain fatty acids (SCFAs), namely acetate, n -butyrate, iso -butyrate, in the above samples. Student t test and Graph pad prism-6 were used to compare the data statistically. RESULTS: There was a significant decrease of Clostridium coccoides (control, 25.69% ± 1.62% vs severe, 9.8% ± 2.4%, P = 0.0001) and Clostridium leptum clusters (control, 13.74% ± 1.05% vs severe, 6.2% ± 1.8%, P = 0.0001) in fecal samples of UC patients. Furthermore, we demonstrated that some butyrateproducing members of the clostridial cluster, like Fecalibacterium prausnitzii (control, 11.66% ± 1.55% vs severe, 6.01% ± 1.6%, P = 0.0001) and Roseburia intestinalis (control, 14.48% ± 1.52% vs severe, 9% ± 1.83%, P = 0.02) were differentially present in patients with different disease activity. In addition, we also demonstrated decreased concentrations of fecal SCFAs, especially of n -butyrate (control, 24.32 ± 1.86 mmol/μL vs severe, 12.74 ± 2.75 mmol

关键词： Fecal microbiota Ulcerative colitis Short chain fatty acids Clostridial cluster Fluorescent in situ hybridization-flow cytometry Quantitative polymerase chain reaction

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Neuroimmunomodulation by gut bacteria:Focus on inflammatory bowel diseases

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World Journal of Gastrointestinal Pathophysiology 2021年第3期12卷 25-39页

作者： Surbhi Aggarwal Raju Ranjha jaishree paul Department of Biochemical Engineering and Biotechnology Indian Institute of TechnologyDelhi 110016India School of Life Sciences Jawaharlal Nehru UniversityDelhi 110067India Field Unit Raipur ICMR-National Institute of Malaria ResearchRaipur 492015ChhattisgarhIndia

Microbes colonize the gastrointestinal tract are considered as highest complex ecosystem because of having diverse bacterial species and 150 times more genes as compared to the human *** or dysbiosis in gut bacteria can cause dysregulation in gut homeostasis that subsequently activates the immune system,which leads to the development of inflammatory bowel disease(IBD).Neuromediators,including both neurotransmitters and neuropeptides,may contribute to the development of aberrant immune *** are emerging as a regulator of inflammatory processes and play a key role in various autoimmune and inflammatory *** may influence immune cell’s function via the receptors present on these *** cytokines secreted by the immune cells,in turn,regulate the neuronal functions by binding with their receptors present on sensory *** bidirectional communication of the enteric nervous system and the enteric immune system is involved in regulating the magnitude of inflammatory *** in gut bacteria influence the level of neuromediators in the colon,which may affect the gastrointestinal inflammation in a disease *** neuromediators concentration via dysbiosis in gut microbiota is one of the novel approaches to understand the pathogenesis of *** this article,we reviewed the existing knowledge on the role of neuromediators governing the pathogenesis of IBD,focusing on the reciprocal relationship among the gut microbiota,neuromediators,and host *** the neuromediators and host-microbiota interactions would give a better insight in to the disease pathophysiology and help in developing the new therapeutic approaches for the disease.

关键词： Inflammation Gut microbiota Inflammatory bowel disease Enteric nervous system Neuroimmunomodulation Neuromediator

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Antioxidant enzyme profile of two clinical isolates of Entamoeba histolytica varying in sensitivity to antiamoebic drugs

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World Journal of Clinical Infectious Diseases 2017年第2期7卷 21-31页

作者： Lakshmi Rani Iyer Neha Banyal Shailendra Naik jaishree paul School of Life Sciences Jawaharlal Nehru University

AIM To study the sensitivity and antioxidant enzyme response in two clinical isolates of Entamoeba histolytica(E. histolytica) during treatment with antiamoebic drugs,auranofin and metronidazole. METHODS E. histolytica were isolated from stool samples and maintained in Robinson's biphasic culture medium. Clinial isolates were maintained in xenic culture medium,and harvested for determination of minimum inhibitory concentrations to the two antiamoebic drugs,Metronidazole and Auranofin using microtiter plate tests. The percent survival of the two isolates were determined using the trypan blue cell count. Isolate 980 was treated with 70 μmol/L and 2 μmol/L while isolate 989 was treated with 20 μmol/L and 0.5 μmol/L of metronidazole and auranofin respectively for 24 h. Fifty thousand cells of each isolate were harvested after 24 h of treatment for analysis of the mRNA expressions of the antioxidant enzymes,thioredoxin reductase,peroxiredoxin and FeSOD using the specific primers. Cell lysate was used for determination of enzyme activity of thioredoxin reductase by measuring DTNB reduction spectrophotometrically at412 *** Minimum inhibitory concentration of the clinical isolates 980 and 989 for auranofin was 3 μmol/L and 1 μmol/L respectively while that for metronidazole was 80 μmol/L and 30 μmol/L respectively. Thioredoxin reductase,peroxiredoxin and FeSOD expression levels were significantly reduced in the isolate 980 when treated with Auranofin. Metronidazole treatment showed a down regulation of thioredoxin reductase. Though not significant both at the mRNA and the enzyme activity levels. Peroxiredoxin and FeSOD however remained unchanged. Auranofin treatment of isolate 989,showed an upregulation in expression of thioredoxin reductase while Peroxiredoxin and FeSOD did not show any change in expression. Upon treatment with metronidazole,isolate 989 showed an increase in thioredoxin reductase expression. Peroxiredoxin and FeSOD expressions however remain unchange

关键词： Metronidazole Entamoeba histolytica Amoebiasis Thioredoxin reductase Minimum inhibitory concentrations Auranofin

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Disseminated cryptosporidiosis:Case report and literature review

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World Journal of Clinical Infectious Diseases 2017年第2期7卷 32-37页

作者： Shehla Khalil Bijay R Mirdha jaishree paul Ashutosh Panda Yogita Singh Department of Microbiology All India Institute of Medical Sciences School of Life Sciences Jawaharlal Nehru University

Cryptosporidiosis,better known as an intestinal disease may disseminate to infect other sites including the respiratory tract. Little information however is available on respiratory cryptosporidiosis that may largely be due to lower frequency of respiratory cryptosporidiosis. Respiratory cryptosporidiosis has been majorly reported in immunocompromised individuals and children. Here we report a case of respiratory and intestinal cryptosporidiosis in a fifteen months old child with CD8+ deficiency. The patient in spite of treatment with Nitazoxanide and Azithromycin followed by Intravenous immunoglobulin and Bovine colostrum had a fatal outcome. The Cryptosporidium spp. isolate was subjected to molecular characterization. The Cryptosporidium spp. was identified both in stool specimen and Endotracheal aspirate(ETA). The blood sample was negative for Cryptosporidium spp. The Cryptosporidium spp. isolate from stool as well as ETA was identified as Cryptosporidium hominis(C. hominis) using Multiplex Allele Specific Polymerase Chain Reaction assay and was subtyped as Ia A23G1R1 subtype using gp60 gene polymerase chain reaction assay followed by sequencing.

关键词： Cryptosporidiosis Disseminated disease CD8+deficiency Cryptosporidium hominis Subtyping

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