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Helicobacter pylori and other Helicobacter species DNA in human bile samples from patients with various hepato-biliary diseases

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World Journal of Gastroenterology 2006年第14期12卷 2181-2186页

作者： Santosh K Tiwari Aleem A Khan Mohd Ibrahim Mohd Aejaz Habeeb C M Habibullah center for liver research and diagnostics deccan college of medical sciences KanchanbaghHyderabad 500 058Andhra PradeshIndia center for liver research and diagnostics deccan college of medical sciences KanchanbaghHyderabad 500 058Andhra PradeshIndia Department of GastroenterologyDeccan College of Medical Sciences Hyderabad 500 058Andhra PradeshIndia

AIM： To investigate the presence of Helicobacter species by nested PCR of 16S rRNA genes followed by the presence of Flelicobacter pylori （H pylori） 16S rRNA, ureA, cagA genes in bile obtained at endoscopic retrograde cholangio-pancreatography （ERCP） from 60 Indian subjects. METHODS： Sixty bile samples were obtained from patients diagnosed with various hepato-biliary diseases and control subjects at ERCP. PCR analysis was carried out using primers for Helicobacter genus 16S rRNA gene and H pylori （16S rRNA, urea and cagA） genes. Gastric H pylori status was also assessed from biopsies obtained at endoscopy from patients with various hepatobiliary diseases and controls. The control group mainly consisted of subjects with gastric disorders. Sequencing analysis was performed to confirm that PCR products with 16S rRNA and cagA primers were derived from H pylori. RESULTS No Helicobacters were grown in culture from the bile samples. Helicobacter DNA was detected in bile of 96.7% and 6.6% of groups Ⅰ and Ⅱ respectively. Ten from group Ⅰ were positive for 16S rRNA and ureA and 9 were positive for cagA gene. In contrast of the 2 from the control, 1 amplified with 16S rRNA, ureA and cagA primers used. The sequences of the 16S rRNA genes and cagA were 99% similar to Helicobacter pylori. CONCLUSION： Helicobacters are associated with the pathogenesis of various hepato-biliary disorders.

关键词： Helicobacter pylori Bile Hepato-biliary diseases PCR, Sequence analysis

Antimicrobial activity of Sapindus mukorossi and Rheum emodi extracts against H pylori: In vitro and in vivo studies

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World Journal of Gastroenterology 2006年第44期12卷 7136-7142页

作者： Mohammed Ibrahim Aleem A Khan Santosh K Tiwari Mohammed Aejaz Habeeb MN Khaja CM Habibullah Centre for liver research and diagnostics Deccan college of Medical Sciences and Allied Hospitals Kanchanbagh Hyderabad India

AIM： To evaluate the antibacterial activity of Sapindus mukorossi （S. mukorossl） and Rheum emodi （R. emodl）. METHODS： Powders of S. mukorossi and R. emodi were extracted successively with petroleum ether, benzene, chloroform and ethanol and were concentrated in vacuum. The disk diffusion method was used for in vitro studies and in vivo studies were performed on male Wister rats. Thirty resistant clinical isolates of H pylon, as determined by their antibiotic sensitivity patterns by E-test, along with two Gram ＋ve （S. aureus, B. subtilis） and two Gram -ve （E. coli, P. vugaris） organisms were screened for their susceptibility patterns against these extracts. RESULTS： In our screening, all 30 resistant isolates and the other four organisms （two Gram ＋ve S. aureus, B. subtilis and two Gram -ve, E. coli, P. vugaris） were sensitive to the test compounds. It was found that ethanol and chloroform extracts of S. mukorossi and ethanol and benzene extracts of R. emodi inhibited Hpylori at very low concentrations, In the in vitro study, the isolates showed a considerable zone of inhibition at very low concentrations （10 μg/mL） for both the extracts. In the in vivo study, the H pylori infection was cleared with minimal doses of extracts of S. mukorossi （2.5 mg/mL） and R. emodi （3.0 mg/mL） given orally for seven days. CONCLUSION： We can conclude from this study that the extracts of S. mukorossi and R. emodi inhibited the growth of pylori in vitro and, in in vivo studies, the H pylori infection cleared within seven days at very low concentrations. We also found that H pylori did not acquire resistance against these herbal extracts even after 10 consecutive passages.

关键词： H pylori Sapindus mukorossi Rheum emodi

Characterization of hepatic progenitors from human fetal liver using CD34 as a hepatic progenitor marker

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World Journal of Gastroenterology 2007年第16期13卷 2319-2323页

作者： Parveen Nyamath Ayesha Alvi Aejaz Habeeb Sanjeev Khosla Aleem A Khan CM Habibullah center for liver research and diagnostics Deccan College of Medical Sciences and Allied Hospitals Kanchanbagh Hyderabad-58 Andhra Pradesh India Centre for DNA Fingerprinting and diagnostics Nacharam Hyderabad 500072 India

AIM: To enrich putative hepatic progenitors from the developing human fetal liver using CD34 as a marker. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

关键词： Progenitor cells Fetal liver Magnetic cell sorting Flow cytometry Immunohistochemistry

Bioengineered humanized livers as better three-dimensional drug testing model system

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World Journal of Hepatology 2018年第1期10卷 22-33页

作者： Sandeep Kumar Vishwakarma Avinash Bardia Chandrakala Lakkireddy Raju Nagarapu Md Aejaz Habeeb Aleem Ahmed Khan Central Laboratory for Stem Cell research and Translational Medicine Centre for Liver Research and Diagnostics Deccan College of Medical Sciences

AIM To develop appropriate humanized three-dimensional ex-vivo model system for drug testing. METHODS Bioengineered humanized livers were developed in this study using human hepatic stem cells repopulation within the acellularized liver scaffolds which mimics with the natural organ anatomy and physiology. Six cytochrome P-450 probes were used to enable efficient identification of drug metabolism in bioengineered humanized livers. The drug metabolism study in bioengineered livers was evaluated to identify the absorption, distribution, metabolism, excretion and toxicity *** The bioengineered humanized livers showed cellular and molecular characteristics of human livers. The bioengineered liver showed three-dimensional natural architecture with intact vasculature and extra-cellular matrix. Human hepatic cells were engrafted similar to the human liver. Drug metabolism studies provided a suitable platform alternative to available ex-vivo and in vivo models for identifying cellular and molecular dynamics of pharmacological *** The present study paves a way towards the development of suitable humanized preclinical model systems for pharmacological testing. This approach may reduce the cost and time duration of preclinical drug testing and further overcomes on the anatomical and physiological variations in xenogeneic systems.

关键词： Acellularization Repopulation Drug testing Humanized liver Bioengineering

Hepatoprotective activity of Sapindus mukorossi and Rheum emodi extracts: In vitro and in vivo studies

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World Journal of Gastroenterology 2008年第16期14卷 2566-2571页

作者： Mohammed Ibrahim Mohammed Nane Khaja Anjum Aara Aleem Ahmed Khan Mohammed Aejaz Habeeb Yalavarthy Prameela Devi Mangamoori Lakshmi Narasu Chitoor Mohammed Habibullah Centre for liver research and diagnostics Deccan college of Medical Sciences and Allied Hospitals Kanchanbagh Nizam Institute of Pharmacy Deshmukhi pochampally (Mandal) Near Ramoji Film City Centre for Biotechnology Jawaharlal Nehru Technological University Kukatpally

AIM: To study the hepatoprotective capacity of Sapindus mukorossi (S. mukorossi) and Rheum emodi (R. emodi) extracts in CCl4 treated male rats. METHODS: The dried powder of S. mukorossi and R. emodi was extracted successively with petroleum ether, benzene, chloroform, and ethanol and concentrated in vacuum. Primary rat hepatocyte monolayer cultures were used for in vitro studies. In vivo, the hepatoprotective capacity of the extract of the fruit pericarp of S. mukorossi and the rhizomes of R. emodi was analyzed in liver injured CCl4-treated male rats. RESULTS: In vitro: primary hepatocytes monolayer cultures were treated with CCl4 and extracts of S. mukorossi & R. emodi. A protective activity could be demonstrated in the CCl4 damaged primary monolayer culture. In vivo : extracts of the fruit pericarp of S. mukorossi (2.5 mg/mL) and rhizomes of R. emodi (3.0 mg/mL) were found to have protective properties in rats with CCl4 induced liver damage as judged from serum marker enzyme activities. CONCLUSION: The extracts of S. mukorossi and R. emodi do have a protective capacity both in vitro on primary hepatocytes cultures and in in vivo in a rat model of CCl4 mediated liver injury.

关键词： Hepatoprotective activity Sapindus mukorossr Rheum emodi

Hepatic stem cells: A viable approach for the treatment of liver cirrhosis

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World Journal of Stem Cells 2015年第5期7卷 860-865页

作者： Md Aejaz Habeeb Sandeep Kumar Vishwakarma Avinash Bardia Aleem Ahmed Khan center for liver research and diagnostics Deccan College of Medical Sciences

liver cirrhosis is characterized by distortion of liver architecture, necrosis of hepatocytes and regenerative nodules formation leading to cirrhosis. Various types of cell sources have been used for the management and treatment of decompensated liver *** of stem cells has offered a new dimension for regenerative therapy and has been considered as one of the potential adjuvant treatment modality in patients with end stage liver diseases(ESLD). Human fetal hepatic progenitor cells are less immunogenic than adult ones. They are highly propagative and challenging to cryopreservation. In our earlier studies we have demonstrated that fetuses at 10-18 wk of gestation age contain a large number of actively dividing hepatic stem and progenitor cells which possess bipotent nature having potential to differentiate into bile duct cells and mature hepatocytes. Hepatic stem cell therapy for the treatment of ESLD is in their early stage of the translation. The emerging technology of decellularization and recellularization might offer a significant platform for developing bioengineered personalized livers to come over the scarcity of desired number of donor organs for the treatment of ESLD. Despite these significant advancements long-term tracking of stem cells in human is the most important subject nowadays in order to answer several unsettles issues regarding the route of delivery, the choice of stem cell type(s), the cell number and the timepoint of cell delivery for the treatment in a chronic setting. Answering to these questions will further contribute to the development of safer, noninvasive, and repeatable imaging modalities that could discover better cell therapeutic approaches from bench to bedside. Combinatorial approach of decellularization and nanotechnology could pave a way towards the better understanding in determination of cell fate posttransplantation.

关键词： Hepatic stem cells Bioengineering livercirrhosis Labeling and tracking

Current progress and emerging technologies for generating extrapancreatic functional insulin-producing cells

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World Journal of Translational Medicine 2022年第1期10卷 1-13页

作者： Md Aejaz Habeeb Sandeep Kumar Vishwakarma Safwaan Habeeb Aleem Ahmed Khan Centre for liver research and diagnostics Deccan College of Medical SciencesHyderabad 500058TelanganaIndia

Diabetes has been one of the major concerns in recent years,due to the increasing rate of morbidity and mortality *** available treatment strategies for uncontrolled diabetes mellitus(DM)are pancreas or islet ***,these strategies are limited due to unavailability of quality pancreas/islet donors,life-long need of immunosuppression,and associated *** therapy has emerged as a promising alternative options to achieve the clinical benefits in the management of uncontrolled *** the last few years,various sources of cells have been used to convert into insulin-producingβ-like *** extrapancreatic sources of cells may play a significant role inβ-cell turnover and insulin secretion in response to environmental ***/progenitor cells from liver have been proposed as an alternative choice that respond well to glucose stimuli under strong transcriptional *** liver is one of the largest organs in the human body and has a common endodermal origin with pancreatic ***,liver has been proposed as a source of a large number of insulinproducing *** merging of nanotechnology and 3D tissue bioengineering has opened a new direction for producing islet-like cells suitable for in vivo transplantation in a cordial *** review summarizes extrapancreatic sources for insulin-secreting cells with reference to emerging technologies to fulfill the future clinical need.

关键词： Diabetes mellitus Cell-based therapy Insulin producing cells Extrapancreatic sources Biomaterials Tissue engineering

Long-term and non-invasive in vivo tracking of DiD dye-labeled human hepatic progenitors in chronic liver disease models

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World Journal of Hepatology 2022年第10期14卷 1884-1898页

作者： Chaturvedula Tripura Srinivas Gunda Sandeep Kumar Vishwakarma Avinash Raj Thatipalli Jedy Jose Mahesh Kumar Jerald Aleem Ahmed Khan Gopal Pande Cell and Stem Cell Biology CSIR-Centre for Cellular and Molecular BiologyHyderabad 500007TelanganaIndia Central Laboratory for Stem Cell research and Translational Medicine Centre for Liver Research and DiagnosticsDeccan College of Medical SciencesHyderabad 500058TelanganaIndia

BACKGROUND Chronic liver diseases(CLD)are the major public health burden due to the continuous increasing rate of global morbidity and *** inherent limitations of organ transplantation have led to the development of stem cell-based therapy as a supportive and promising therapeutic ***,identifying the fate of transplanted cells in vivo represents a crucial *** To evaluate the potential applicability of DiD dye as a cell labeling agent for longterm,and non-invasive in vivo tracking of transplanted cells in the *** Magnetically sorted,epithelial cell adhesion molecule positive(1×106 cells/mL)fetal hepatic progenitor cells were labeled with DiD dye and transplanted into the livers of CLD-severe combined immunodeficiency(SCID)***-infrared(NIR)imaging was performed for in vivo tracking of the DiD-labeled transplanted cells along with colocalization of hepatic markers for up to 80 *** existence of human cells within mouse livers was identified using Alu polymerase chain reaction and *** NIR fluorescence imaging of CLD-SCID mice showed a positive fluorescence signal of DiD at days 7,15,30,45,60,and 80 ***,positive staining of cytokeratin,c-Met,and albumin colocalizing with DiD fluorescence clearly demonstrated that the fluorescent signal of hepatic markers emerged from the DiD-labeled transplanted *** of liver function was also observed with serum levels of glutamic-oxaloacetic transaminase,glutamate-pyruvate transaminase,and *** detection of human-specific Alu sequence from the transplanted mouse livers provided evidence for the survival of transplanted cells at day *** DiD-labeling is promising for long-term and non-invasive in vivo cell tracking,and understanding the regenerative mechanisms incurred by the transplanted cells.

关键词： Chronic liver diseases Cell transplantation Cell tracking and imaging DiD Hepatic progenitors

Bioengineered functional humanized livers： An emerging supportive modality to bridge the gap of organ transplantation for management of end-stage liver diseases

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World Journal of Hepatology 2018年第11期10卷 822-836页

作者： Sandeep Kumar Vishwakarma Chandrakala Lakkireddy Avinash Bardia Syed Ameer Basha Paspala Chaturvedula Tripura Md Aejaz Habeeb Aleem Ahmed Khan Central Laboratory for Stem Cell research and Translational Medicine Centre for Liver Research and Diagnostics Deccan College of Medical Sciences Kanchanbagh Hyderabad 500058 Telangana India Dr Habeebullah Life sciences Attapur Hyderabad 500058 Telangana India CSIR-Centre for Cellular and Molecular Biology Habsiguda Hyderabad 500007 Telangana India

End stage liver diseases （ESLD） represent a major, neglected global public health crisis which requires an urgent action towards fnding a proper cure. Orthotro-pic liver transplantation has been the only definitive treatment modality for ESLD. However, shortage of donor organs, timely unavailability, post-surgery related complications and financial burden on the patients li-mits the number of patients receiving the transplants. Since last two decades cell-based therapies have revolu-tionized the feld of organ/tissue regeneration. However providing an alternative organ source to address the donor liver shortage still poses potential challenges. The developments made in this direction provide useful futuristic approaches, which could be translated into preclinical and clinical settings targeting appropriate applications in specific disease conditions. Earlier studies have demonstrated the applicability of this particular approach to generate functional organ in rodent system by connecting them with portal and hepatic circulatory networks. However, such strategy requires very high level of surgical expertise and also poses the technical and financial questions towards its future applicability. Hence, alternative sites for generating secondary organs are being tested in several types of disease conditions. Among different sites, omentum has been proved to be more appropriate site for implanting several kinds of functional tissue constructs without eliciting much immunological response. Hence, omentum may be con-sidered as better site for transplanting humanized bio-engineered ex vivo generated livers, thereby creating a secondary organ at intra-omental site. However, the expertise for generating such bioengineered organs are limited and only very few centres are involved for inve-stigating the potential use of such implants in clinical practice due to gap between the clinical transplant surgeons and basic scientists working on the concept evolution. Herein we discuss t

关键词： Bioengineered liver Omentum Secondary organ Transplantation End stage liver diseases