Mohammed Aejaz Habeeb

Human Fetal Liver-Derived Stem Cell Transplantation as Supportive Modality in the Management of End-Stage Decompensated Liver Cirrhosis:

Liver transplantation is the only existing modality for treating decompensated liver cirrhosis. Several factors, such as nonavailability of donors, combined with operative risks, complications associated with rejection, usage of immunosuppressive agents, and cost intensiveness, make this strategy available to only a few people. With a tremendous upsurge in the mortality rate of patients with liver disorders worldwide, there is a need to search for an alternative therapeutic tool that can combat the above limitations and serve as a supportive therapy in the management of liver diseases. Cell therapy using human fetal liver-derived stem cells can provide great potential to conservatively manage end-stage liver diseases. Therefore, the present investigation aimed to study and prove the safety and efficacy of human fetal liver-derived stem cell transplantation in patients with end-stage liver cirrhosis. Twenty-five patients with liver cirrhosis of different etiologies were infused with human fetal liver-derived stem cells (EpCAM+ve) labeled with Tc-HMPAO through hepatic artery. Our high throughput analysis using flow cytometry, RT-PCR, and cellular characterization exemplifies fetal liver cells with their high proliferation rate could be the best source for rejuvenating the diseased liver. Further, no episodes related to hepatic encephalopathy recurred in any of the subjects following hepatic stem cell transplantation. There was marked clinical improvement observed in terms of all clinical and biochemical parameters. Further, there was decrease in mean MELD score (p < 0.01) observed in 6 months follow-up in all patients. Therapy using human fetal liver stem/progenitor cells offers a potentially supportive modality to organ transplantation in the management of liver diseases.

In vitro quantitative and relative gene expression analysis of pancreatic transcription factors Pdx-1, Ngn-3, Isl-1, Pax-4, Pax-6 and Nkx-6.1 in trans-differentiated human hepatic progenitors.

Diabetes is a major health concern throughout the world because of its increasing prevalence in epidemic proportions. β‐Cell deterioration in the pancreas is a crucial factor for the progression of diabetes mellitus. Therefore, the restoration of β‐cell mass and its function is of vital importance for the development of effective therapeutic strategies and most accessible cell sources for the treatment of diabetes mellitus.

Simplified and versatile method for bisulfite‐based DNA methylation analysis of small amounts of DNA

Epigenetic alterations of gene function play a central role in the pathogenesis of many tumors and in the process of aging. Abnormal methylation at transcriptional sites of genes results in epigenetic silencing of the genes that protect against tumor formation or that repair DNA. To date, several studies have analyzed methylation status by oligonucleotide arrays, restriction analysis (COBRA), methylation‐specific amplification, and sequence analysis. Thisrequires high concentration of bisulfite‐treated DNA, which mandates use of commercially available expensive kits, and is an often laborious and time‐consuming task. In this article, we report a simplified high‐throughput method, which can serve as a surrogate for screening methylation profiles of various genes and has high sensitivity compared with the other methods described previously. J. Clin. Lab. Anal. 23:172–174, 2009.

Association of Helicobacter pylori restriction endonuclease-replacing gene, hrgA with overt gastrointestinal diseases

BACKGROUND AND AIM Helicobacter pylori has been proven to be responsible for causing various gastrointestinal disorders including gastric adenocarcinoma. Several genes of pathogen (the genes of the cag-PAI, vacA, iceA, and babA) either in combination or independently have been reported to significantly increase the risk of ulceration/gastric carcinoma, with the cagA gene having the strongest predictive value. Pursuit to identify new genes which could serve as a marker of overt disease progression, lead to the discovery of hrgA gene. METHODS Fifty-six indigenous strains of H. pylori from subjects with various gastric disorder were screened to assess the status of hrgA gene along with the cagA gene using simple polymerase chain reaction using specific oligonucleotide primers. Post-amplification, amplicons were subjected for sequencing to identify any strain specific variations in sequences from the H. pylori isolated from different disease manifestations. Histopathological analysis was done to ascertain any significant change in the histological scores of subjects infected with cagA+/hrgA+ and cagA-/hrg+ strains. RESULTS All the 56 (100%) subjects amplified with the oligonucleotide primers specific to hrgA gene, whereas 81.71% subjects showed the presence of cagA gene. Sequencing of the amplimers showed 99% homology. Histology of the cagA+/hrgA+ and cagA-/hrg+ subjects did not show any significant difference. CONCLUSION hrgA gene of Helicobacter pylori is not a ideal surrogate marker for identifying individuals with higher risk of developing overt gastro-duodenal diseases such as neoplasia of the stomach.

Prevalence, risk factors and genotype distribution of HCV and HBV infection in the tribal population: a community based study in south India

Prevalence, risk factors and genotype distribution of HCV and HBV infection in the tribal population: a community based study in south India

Effect of ultraviolet B (302 nm) irradiation on viability, metabolic and detoxification functions of goat hepatocytes--in vitro study.

The object of the present study was to investigate the effect(s) of UV-B irradiation on the functional integrity, metabolic and detoxifying capacity of the isolated goat hepatocytes. Isolated goat hepatocytes were subjected to UV-B irradiation invitro for 0, 250, 500, 1250, 2500 and 7500 Joules/m2 which correspond to the irradiation time of 0, 1, 2, 5, 10 and 30 min. Cells were then analysed for Viability (Trypan blue exclusion test [TBE], 3-[4,5-dimethylthiozol-2yl]-2,5-diphenyltetrazolium bromide [MTT] assay, Membrane integrity (Lactate dehydrogenase [LDH] leakage, Lipid peroxidation) Detoxification (Ureagenesis, Cytochrome P450 activity [CYP450, Diazepam metabolism] and Glutathione-S-Transferase [GST] activity. The results show that there was no difference in functional, metabolic as well as detoxifying parameters of the hepatocytes when irradiated from 0–1250 Joules/m2, whereas a significant alteration was appreciable in the parameters such as LDH leakage, lipid peroxidation, and CYP450 activity when irradiated beyond 1250 Joules/m2. Our present findings suggest that the biologically compatible and feasible dose of UV-B irradiation for xenotransplantation appears to be 1250 Joules/m2.

Repopulation of Cirrhotic Liver by Hepatic Stem/Progenitor Cells: A Promising Strategy Alternative to Liver Transplantation

Liver cirrhosis is one of the major causes of morbidity and mortality worldwide. Liver transplantation is the only successful and curative option for the management of this disease. However, cost effectiveness, timely availability, operative risks, need of life-long immunosuppressant, and shortage of donor organs are major challenges to fulfill the demand. Stem cells transplantation has emerged as a bridge to liver transplantation for the repopulation of cirrhotic liver due to its potential for long-term proliferation. Human fetal liver–derived stem/progenitor cells (fLSPCs) are emerging as safe and effective therapeutic possibility in the management of liver cirrhosis due to their low immunogenicity and high proliferative ability. Combination of mesenchymal stem cells with fLSPCs could be the best option to ameliorate immunomodulation, fibrotic reconstruction, and repopulation of lost hepatocytes to replenish the deficient liver functions. Merging of nanotechnology and whole-liver bioengineering approaches could provide several unanswered questions of regenerative mechanisms and developing extracorporeal liver systems.

Risk of developing liver disease in chronic HBV infected patients — a 7 year followup study

Risk of developing liver disease in chronic HBV infected patients — a 7 year followup study

Determination of Helicobacter pylori vaca genotypes and caga gene in relationship to gastroduodenal disease in South India

Determination of Helicobacter pylori vaca genotypes and caga gene in relationship to gastroduodenal disease in South India

Patterns of seroconversion from hbe antigen to antibody in patients with chronic liver disease

Background: Azathioprine (AZA) is used for the induction and maintenance of remission in Crohn’s disease (CD) and ulcerative colitis. AZA metabolism is complex and the presumed active metabolites are 6-thioguanine nucleotides (6TGN), with inactive metabolites 6-methylmercaptopurine (6MMP) (via thiopurine methyltransferase) (TPMT) and 6-thiouracil (6TU) (via xanthine oxidase) (XO). Allopurinol is a potent inhibitor of XO and shifts metabolism towards elevated 6TGN levels and associated myelosuppression and therefore allopurinol use has been considered a contraindication to AZA therapy in patients. Mesalamine has an inhibitory effect on TPMT activity and shifts AZA metabolism away from 6MMP. We present the case of a man with previously poorly controlled CD and concomitant use of allopurinol for gout treated successfully with AZA using 6TGN monitoring as a guide to therapy. Case: A 45 year-old male with CD and gout (treated with allopurinol 300 mg/day) was referred after failing mesalamine, prednisone, budesonide, cyclosporine, and infliximab. Colonoscopy and small bowel follow through showed moderately active Crohn’s ileocolitis. The patient declined ileocolectomy, and methotrexate was not offered because of elevated transaminases. AZA therapy with 6TGN monitoring was offered. Mesalamine 4.8 grams/ day was co-administered to inhibit 6MMP formation by blocking TPMT, prednisone was tapered, and infliximab 5 mg/kg at 0, 2, and 6 weeks and then every 8 weeks was initiated. TPMT activity was normal. AZA was initiated at a reduced dose of 25mg on alternate days for 2 weeks and then daily for 2 weeks. The week 4 6TGN level was 22 pmol/8 10 RBC. AZA was increased to 25 mg/50 mg on alternating days for 2 weeks then 50 mg daily for one month. The week 10 6TGN was 176 pmol/8 10 RBC. AZA was increased to 50 mg/100 mg on alternating days (average 1.0 mg/kg). The week 14 6TGN was “therapeutic” at 265 pmol/8 10 RBC and the week 14 6TGN was 278 pmol/8 10 RBC. 6MMP level were undetectable at all time points. The patient discontinued steroids and is in symptomatic remission. Conclusion: Azathioprine and allopurinol can be safely co-administered with 6TGN monitoring.