[Show abstract][Hide abstract] ABSTRACT: Ulcerative colitis (UC) is a major clinical form of inflammatory bowel disease. UC is
characterized by mucosal inflammation limited to the colon, always involving the
rectum and a variable extent of the more proximal colon in a continuous manner.
Genetic variations in DNA repair genes may influence the extent of repair functions,
DNA damage, and thus the manifestations of UC. This study thus evaluated the role of
polymorphisms of the genes involved in DNA repair mechanisms.
A total of 171 patients and 213 controls were included. Genotyping was carried out by
ARMS PCR and PCR-RFLP analyses for RAD51, XRCC3 and hMSH2 gene
polymorphisms. Allelic and genotypic frequencies were computed in both control &
patient groups and data was analyzed using appropriate statistical tests. The
frequency of 'A' allele of hMSH2 in the UC group caused statistically significant
increased risk for UC compared to controls (OR 1.64, 95% CI 1.16-2.31, p =0.004).
Similarly, the CT genotype of XRCC3 gene was predominant in the UC group and
increased the risk for UC by 1.75 fold compared to controls (OR 1.75, 95% CI 1.15-
2.67, p=0.03), further confirming the risk of 'T' allele in UC. The GC genotype
frequency of RAD51 gene was significantly increased (p= 0.02) in the UC group
(50.3%) compared to controls (38%). The GC genotype significantly increased the risk
for UC compared to GG genotype by 1.73 fold (OR 1.73, 95% CI 1.14- 2.62, p =0.02)
confirming the strong association of 'C' allele with UC. Among the controls, the SNP
loci combination of hMSH2:XRCC3 were in perfect linkage. The GTC and ACC
haplotypes were found to be predominant in UC than controls with a 2.28 and 2.93 fold
significant increase risk of UC.
PLoS ONE 09/2014; 9(9). DOI:10.1371/journal.pone.0108562 · 3.23 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: In Indian population, hepatitis C virus (HCV) genotypes 1 and 3 are prevalent and predominant with the highest frequency. However, other genotypes are seldom reported, and among them the HCV genotype 5a is exceptionally rare. The presented case had no history for either blood transfusion or using any type of IV drugs and never traveled to any other country. He was serologically positive with HCV antibodies and HCV RNA. 5'UTR-specific amplification and sequencing of infected viral genome confirmed that he had been infected with HCV genotype 5a which is not closely related to other common prevalent genotypes like 1a, 1b, 3a, and 3b in India. Patient's wife and children tested negative for anti-HCV and HCV-RNA. This unique case report could be attributed to circulation of HCV genotype 5a from other geographic area at very low frequency in India as determined by phylogenetic analysis and nucleic acid-sequencing methods.
[Show abstract][Hide abstract] ABSTRACT: Pegylated-interferon-alfa (PEG-IFN-α) with ribavirin is an established treatment in chronic hepatitis due to hepatitis C virus (HCV) (CH-C). Such treatment is expensive and in resource-poor countries such as India, alternative less expensive therapy is needed.
Multicenter randomized controlled trial comparing two treatment regimens (interferon-alfa-2b [IFN-α-2b] 3 million unit/day [MU/day] and ribavirin 1000 mg/day [I+R] vs IFN-α-2b 3 MU/day and glycyrrhizin 250 mg [I+G]) in CH-C. Viral, host characteristics and therapeutic responses were assessed (ICMR-6 months trial for chronic hepatitis-CTRI/2008/091/000105).
One hundred and thirty-one patients meeting the inclusion criteria were randomized to I + G (n=64) or I+R (n=67) during the period February 2002 to May 2005. About 85% (I+G=53, I+R=58) completed 6 months of treatment and 89% of them (I+G=46, I+R=53) completed 6 months of follow-up after completion of treatment. Hepatitis C virus genotype 3 was the major type detected (71% patients). The mean log10 viral load (copies/mL), histological activity index, and fibrosis stage for all patients were 5.1 ± 0.98, 5 ± 2, and 2± 1.5, respectively. Sustained viral response (SVR) was significantly higher in I + R group than in I + G group (65.7% vs 46.9%, OR=2.2, P = 0.03). Treatment with I + G was associated with significantly lower frequencies of leukopenia (2% vs 17%, P <0.01) and anemia (8% vs 40%, P <0.001) as compared to treatment with I + R.
Genotype 3 HCV infection with low viral load is prevalent in India. Daily IFN with ribavirin showed significantly better responses. Leukopenia and anemia were significantly more in ribavirin group. Responses observed with IFN + ribavirin were similar to the reported response rates with PEG-IFN suggesting that this modality may be considered as a cheaper alternative of treatment for chronic hepatitis C.
[Show abstract][Hide abstract] ABSTRACT: Ulcerative colitis is a multifactorial disease in which genetic factors play a major role. Functional mutations in the genes related to innate immune response exacerbate mucosal damage coupled with persistent inflammation. The cytokine macrophage migration inhibitory factor (MIF), CD14, and Toll-like receptor 4 (TLR4) are the central players with clearly defined roles in inflammation. The aim of this study was to investigate the association between MIF-173G > C, CD14-159C > T, and TLR4-299A > G polymorphisms and mononuclear cell expression in patients with ulcerative colitis (UC). Genotyping of MIF-173G > C, CD14-159C > T, and TLR4-299A > G polymorphisms was performed by amplification refractory mutation system-polymerase chain reaction and allele-specific amplification in 139 and 176 patients with UC and controls, respectively. Simultaneously, the expression levels of intracellular MIF, mCD14, and mTLR4 were determined in mononuclear cells using a flow cytometer. Polymorphisms in CD14-159C > T and TLR4-299A > G significantly affected mCD14 and mTLR4 expression levels and also increased susceptibility to UC. Although intracellular MIF expression levels differed among patient and control groups, the polymorphism in MIF 173G > C was not observed to be associated with a risk of UC.
Human immunology 12/2011; 73(2):201-5. DOI:10.1016/j.humimm.2011.12.006 · 2.14 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Purpose: Orthotopic liver transplantation (OLT) is the ultimate treatment
for end-stage liver failure and for numerous liver based inborn errors of
metabolism. However, several limitations such as shortage of organ donors,
high cost and several complications/risks of OLT. Liver cell transplantation
(LCT) is emerging as successful treatment. Stem cells from different sources
are eligible for liver cell therapy according to their hepatic potential, for instance
mesenchymal stem cells (MSCs), hematopoietic stem cells (HSCs)
and adult liver stem/progenitor cells. However, the current research on the
hepatic fate of the stem cells is still facing difficulties to demonstrate the acquisition
of a functional hepatocyte, both in vitro and in vivo. In experimental
studies heaptic progenitors from human fetal is shown promising results in
the engraftment and proliferation. Recently Hisaya azuma et al. demonstrated
the transplantation of serial human hepatic progenitors completely
repopulated the whole liver. In this study we are reporting transplantation
of hepatic progenitors in 5 patients with cirrhosis of liver (Child Pugh B, C)
with 6 months follow-up.
Methods: This clinical study was approved by Institutional Ethics Committee.
Informed consent was taken from patient for carrying out the study.
Briefly, liver cells were isolated by two step collagenase digestion method,
from the aborted fetus of 14–20 weeks gestation. Total liver cells were subjected
to magnetic activated cell sorting (MACS) with CD326 antibody
directly labeled to microbeads (MACS, Miltenyi Biotec) and collected hepatic
progenitor cells. Cell viability and cell activity were assessed by trypan
blue dye exclusion method and MTT assay respectively and reported 90%
of viability with higher cellular activity. Cells were infused by trans femoral
catheterization technique, Hepatic artery was selectively cannulated with 8
F guage catheter. 0.1 × 108 fetal liver progenitor cells in 10–12 ml of DNS
solution were infused at the rate of 1 ml/per minute to achieve reconstitution
of the liver with transplanted cells.
Results: All the procedures were performed safely without any post infusion
complications. Follow up results after 6 months showed oveall improvement
in geneal condition. All the patients showed improvement in serum albumin,
bilirubin, ALT after first month of cell infusion. Ultrasonography findings
showed decrease in ascitis.
Conclusion: This study demonstrates the efficacy of hepatic progenitor cell
in management of cirrhosis liver. As the procedure is very simple, infusion
of hepatic progenitors can be repeated to improve the liver functions.
The American Journal of Gastroenterology 09/2008; 103:S136-S137. DOI:10.1111/j.1572-0241.2008.02139_6.x · 10.76 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: This study was performed to determine the safety and tolerability of injecting autologous bone marrow stem cells (BMC) (CD34+) into four patients with liver insufficiency. The study was based on the hypothesis that the CD34+ cell population in granulocyte colony stimulating factor (G-CSF) mobilized blood and autologous bone marrow contains a subpopulation of cells with the potential for regenerating damaged tissue. We separated the CD34+ stem cell population from the bone marrow. The potential of the BMC to differentiate into hepatocytes and other cell lineages has already been reported. Several reports have also demonstrated the plasticity of hematopoietic stem cells to differentiate into hepatocytes. Recently Sakaida demonstrated reduction in fibrosis in chemically induced liver cirrhosis following BMC transplantation. From a therapeutic point of view, chronic liver cirrhosis is one of the targets for BMC transplantation. In this condition, there is excessive deposition of extracellular matrix and hepatocyte necrosis. Encouraged by this evidence that the CD34+ cell population contains cells with the potential to form hepatocyte-like elements, four patients with liver insufficiency were given G-CSF to mobilize stem cells. CD34+ cells (0.1 x 10(8)) were injected into the hepatic artery. No complications or specific side effects related to the procedure were observed; four patients showed improvements in serum albumin, bilirubin and ALT after one month from the cell infusion.
[Show abstract][Hide abstract] ABSTRACT: Cholangiodestruction of bile ducts leads to biliary atresia, a rare disease characterized by intrahepatic and extrahepatic biliary inflammation. If the intrahepatic biliary tree is unaffected, surgical reconstruction by the Kasai procedure of hepatoportoenterostomy of the extra hepatic biliary tract is possible. Untreated, this condition leads to cirrhosis and death within the first year of the life. If the atresia is complete, liver transplantation is the only option. As a result of the shortage of donor livers, hepatocytes have been infused over the past two decades, providing proof of the concept that cell therapy can be effective for the treatment of liver diseases. In the present study, we report a confirmed case of a girl of 1 year of age with increased bilirubin of 28.5 mg/dL and pediatric end-stage liver disease score 20. Biochemical liver function tests showed cholestasis (elevated cholesterol and gamma-GTs) and increased ALT, total bilirubin, conjugated bilirubin, and ALP. The patient was treated with hepatic progenitor cell infusion through the hepatic artery. The total bilirubin and conjugated bilirubin started decreasing during the first month after cell infusion. The level of total bilirubin maintained a threefold decrease after months of cell infusion. The conjugated bilirubin was 16.35 mg/dL before cell infusion, decreasing to eightfold after cell infusion. After 2 months of cell infusion, hepatobiliary scintigraphy showed increased liver cell function. This case demonstrated the efficacy and functionality of hepatic progenitor cells for the management of biliary atresia. Further, as there was a decrease in serum bilirubin, it showed that there was some percentage of the engraftment of the infused cells. As the procedure is simple and the patient has tolerated the infusion therapy, it might be repeated to manage biliary atresia.
[Show abstract][Hide abstract] ABSTRACT: Crigler-Najjar Syndrome (CNS) is characterized by mild, chronic unconjugated hyperbilirubinemia resulting from an autosomal-recessive inherited deficiency of hepatic uridine/diphosphoglucuronate-glucuronosyl transferase 1Al since birth. Herein we have reported a confirmed case of CNS type 1 in a 2-year-old girl with an unconjugated hyperbilirubinemia (>30 mg/dL) treated by hepatic progenitor cell infusion through the hepatic artery. No procedure-related complications were encountered. No kernicterus was observed. The total bilirubin started falling at 10 days after cell infusion. Two months after cell infusion the bilirubin fell from 29.0 to 16 mg/dL, with the conjugated bilirubin increasing approximately fivefold, the unconjugated bilirubin decreasing nearly twofold, and the SGPT also decreasing from 210 U/L to 64 U/L. This study demonstrated the efficacy of hepatic progenitor cells to manage hyperbilirubinemia in these patients. As the procedure is simple and the patient has tolerated the cell therapy, infusion can be repeated as required to manage hyperbilirubinemia, which often causes lethal kernicterus. This study was developed to assess the safety, feasibility, and efficacy of hepatic progenitor cell transplantation in a child with CNS type 1.
[Show abstract][Hide abstract] ABSTRACT: To evaluate and develop a multiplex polymerase chain reaction (PCR) assay for diagnosing and specific identification of virulent Helicobacter pylori strains and their main virulence genes cagA, cagE, cagT, vacA and hrgA.
Genomic DNA from 82 gastric tissues was screened. A master pool of all the ingredients of multiplex reaction was prepared for amplification. Amplicons were sequenced to confirm the amplification of each target genes. Multiplex PCR assay was able to detect all the five target genes in 81.7% and deletions in one or more loci among 18.3%. Genotype cagT +ve/hrgA +ve/cagA +ve/cagE +ve/vacAs1 +ve was more predominant in this study population (67.07%). hrgA, cagT, cagE and cagA genes were present in 100%, 92.7%, 85.4% and 81.7% of the subjects, respectively. The vacAs1 subtype had higher prevalence frequency in patients with overt gastrointestinal disease (78.57%) than with GERD (gastro-esophageal reflux disease) and NUD (non-ulcer dispepsia) (50%).
The multiplex PCR assay developed herein was able to genotype H. pylori isolates based on the main virulence genes.
The ability to identify H. pylori and the majority of their virulence gene markers by multiplex PCR assay represents a considerable advancement over other PCR-based methods for genotyping H. pylori from large population, and can be explored to gain insights at the genotypic variability exhibited by this pathogen.
[Show abstract][Hide abstract] ABSTRACT: In developing countries, the Helicobacter pylori (H. pylori) infection rate is high, especially in lower socioeconomic groups. The populace in developing countries lives in conditions that are highly conducive to the acquisition of microorganisms. Poor hygiene, crowded household conditions and deficient sanitation mark their day-to-day life. We aimed to find out the roles of household hygiene and water source in the prevalence and transmission of H. pylori infection among the South Indian population using polymerase chain reaction (PCR) assay.
The selected population consisted of 500 adults of varying ages ranging from 30 to 79 years, with upper gastrointestinal tract symptoms. Each participant in the study was given a questionnaire to complete. Samples to assess H. pylori infection included three gastric biopsies (two from the antrum and one from the corpus region). Infection was detected by PCR amplification of the 16S rRNA gene of H. pylori. The data was then examined statistically by univariate and multivariate analyses.
The overall prevalence of H. pylori was detected to be 80 percent. Prevalence increased with an increase in age and it was found to be 90 percent in the 70-79 year age group (p-value is less than 0.01). The prevalence of infection among people who drank water from wells was 92 percent compared with 74.8 percent of those who drank tap water (p-value is less than 0.001). H. pylori infection prevalence was found to be higher in people with low clean water index (CWI) (88.2 percent) than in those with higher CWI (33.3 percent) (p-value is less than 0.001). While the prevalence of H. pylori in the subjects with lower socioeconomic status was 86.1 percent, in higher groups, it was 70 percent (p-value is less than 0.001). The prevalence of H. pylori was also found to be higher in subjects who lived in overcrowded houses. It was 83.7 percent with high crowding index, 76.6 percent with medium crowding index, and 71.3 percent with low crowding index (p-value is less than 0.05).
The results of the present study suggest that the risk of acquisition and transmission of H. pylori can be prevented to a large extent by following improved household hygienic practices, proper waste disposal measures as well as the regular use of boiling water for drinking purposes.
Singapore medical journal 07/2007; 48(6):543-9. · 0.60 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To enrich putative hepatic progenitors from the developing human fetal liver using CD34 as a marker.
Aborted fetuses of 13-20 wk were used for the isolation of liver cells. The cells were labeled with anti CD34; a marker used for isolating progenitor population and the cells were sorted using magnetic cell sorting. The positive fractions of cells were assessed for specific hepatic markers. Further, these cells were cultured in vitro for long term investigation.
Flow cytometric and immunocytochemical analysis for alphafetoprotein (AFP) showed that the majority of the enriched CD34 positive cells were positive for AFP. Furthermore, these enriched cells proliferated in the long term and maintained hepatic characteristics in in vitro culture.
The study shows that aborted human fetal liver is a potential source for isolation of hepatic progenitors for clinical applications. The study also demonstrates that CD34 can be a good marker for the enrichment of progenitor populations.
World Journal of Gastroenterology 05/2007; 13(16):2319-23. · 2.37 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The genetic composition of the intricate cytotoxin associated gene pathogenicity island (cag PAI) of Helicobacter pylori is known to significantly influence the outcome of the disease. Hence, analysis of complete cag PAI of H. pylori isolated from saliva would be of immense importance in standardizing saliva as a reliable non-invasive diagnostic specimen and also to evaluate the type of H. pylori infection. The aim of the present study was to analyze the genes of cag PAI of H. pylori for their presence and correlating them with the disease status of the patients.
One hundred and twenty patients (55 duodenal ulcer [DU], 25 gastric ulcer and 40 non-ulcer dyspepsia [NUD]) were investigated for the present study. Eight pairs of oligonucleotide primers (cagA1, cagA2, cagAP1, cagAP2, cagE, cagT, LEC1 and LEC2) of five different loci; cagA, cagA promoter region, cagE which represents cagI region, cagT and LEC representing cagII were used to detect the presence of the cag PAI genes by polymerase chain reaction.
The comprehensive analysis of the genes constituting cag PAI showed almost equivalent prevalence of all the genes between both the study groups (ulcer and NUD) included. Little significant difference was found in the percentage distribution in both the clinical groups. cagE and cagT were found in a larger proportion of the ulcer group (92.5% and 96.2%) compared with the NUD group (77.5% and 85%), respectively.
Saliva could be efficiently used as a non-invasive source for H. pylori and cagT might be an important locus of the cag PAI, thus greatly influencing the disease condition of the subjects.
Journal of Gastroenterology and Hepatology 11/2005; 20(10):1560-6. DOI:10.1111/j.1440-1746.2005.03955.x · 3.50 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Current guidelines that recommend Helicobacter pylori eradication treatment without endoscopy in selected patients underscore the importance of non-invasive testing. The accuracy of saliva as a non-invasive specimen was compared with that of invasive tests in pretreatment diagnosis of H. pylori infection.
One hundred patients undergoing gastroscopy were grouped into 80 symptomatic and 20 asymptomatic subjects and were investigated for the presence of H. pylori in saliva and stomach. Samples tested comprised saliva and gastric biopsies collected from each patient. Exclusion criteria were history of peptic ulcer, bleeding ulcer, cancer or recent use of antibiotics, proton pump inhibitors and non-steroidal anti-inflammatory drugs. Two sets of primers homologous to 534 bp fragment of H. pylori DNA, which have been shown previously to be highly specific and sensitive, were used for the polymerase chain reaction (PCR) amplification.
72 (90 percent) of the symptomatic group and 10 asymptomatic subjects were infected with H. pylori in the stomach as determined by histology and direct PCR amplification of biopsy DNA obtained from each subject. H. pylori DNA was identified in the saliva of 70 (87.5 percent) symptomatic subjects and 12 (60 percent) asymptomatic control subjects.
High rates of detection using saliva as a specimen indicate that saliva of the infected person could serve as a reliable non-invasive alternative to detect the presence of H. pylori infection in comparison to the currently available standard diagnostic tests.
Singapore medical journal 06/2005; 46(5):224-8. · 0.60 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The genomic diversity of Helicobacter pylori from the vast Indian subcontinent is largely unknown. We compared the genomes of 10 H. pylori strains from Ladakh, North India. Molecular analysis was carried out to identify rearrangements within and outside the cag pathogenicity island (cag PAI) and DNA sequence divergence in candidate genes. Analyses of virulence genes (such as the cag PAI as a whole, cagA, vacA, iceA, oipA, babB, and the plasticity cluster) revealed that H. pylori strains from Ladakh are genetically distinct and possibly less virulent than the isolates from East Asian countries, such
as China and Japan. Phylogenetic analyses based on the cagA-glr motifs, enterobacterial repetitive intergenic consensus patterns, repetitive extragenic palindromic signatures, the glmM gene mutations, and several genomic markers representing fluorescent amplified fragment length polymorphisms revealed that
Ladakhi strains share features of the Indo-European, as well as the East Asian, gene pools. However, the contribution of genetic
features from the Indo-European gene pool was more prominent.
[Show abstract][Hide abstract] ABSTRACT: We report the case of a 26-year-old second gravida in the third trimester of pregnancy who presented with a history of nausea, repeated vomiting and jaundice. The patient was diagnosed as acute fatty liver of pregnancy. After delivery, the condition of the patient progressed to grade IV encephalopathy and did not improve despite all intensive clinical management measures. After 3 days in grade IV encephalopathy, the patient was infused 3 x 10(8) human foetal hepatocytes. The patient's level of consciousness started improving after 24 hours of foetal hepatocyte transfusion and she recovered completely within 7 days.
Tropical gastroenterology: official journal of the Digestive Diseases Foundation 01/2004; 25(3):141-3.
[Show abstract][Hide abstract] ABSTRACT: Eradication of H. pylori infection cures peptic ulcer disease and conversely, relapse is associated with reappearance of H. pylori infection. However, it is not clear whether the recurrence of ulcers following H. pylori eradication is due to recrudescence (identical strain) of the previous infection or as a result of exogenous reinfection (different strain) by another strain. The aim of the present study was to analyze the FAFLP patterns of pre and post treatment H. pylori samples to check if the recurrence was due to recrudescence or reinfection.
24 of 30 duodenal ulcer (DU) subjects screened for H. pylori infection were positive for H. pylori infection. The treatment regime included pantoprazole, ciprofloxacin and amoxicillin. The patients were called for a repeat endoscopy after one month and screened for H. pylori infection. FAFLP analysis and PCR for the cagA and vacA gene was performed for the pre and post treatment samples.
Of the 24 positive H. pylori patients, only 6 were negative after treatment and the remaining 18 were positive for H. pylori infection. The analysis of the pre and post treatment samples of the 18 patients showed that the FAFLP profiles of the initial and follow-up pools were similar to one another.
It can be concluded that in the present series of patients, reinfection was due to recrudescence of infection due to incomplete eradication. The study also suggests that DNA fingerprinting by FAFLP provides discriminatory and complementary data for identifying strains of H. pylori while monitoring therapy.
Indian Journal of Medical Microbiology 07/2003; 21(3):166-71. · 0.88 Impact Factor