Md. Aejaz Habeeb

Pluripotent stem cells - a review of the current status in neural regeneration.

Pharmacological or neurosurgical therapies currently in practice to treat the damage in various neurodegenerative disorders are not efficient in preventing progression or cure of these progressive neurodegenerative processes. Recently, a new approach, cell therapy using stem cell, is being evaluated. However, the use of this therapy in the treatment of these neurological diseases is highly restricted, mainly owing to several technical difficulties and limitations. The strategy of isolation and characterization of neural stem cells from various sources will probably provide a major impetus and open up an interesting, novel therapeutic modality for several neurodegenerative disorders. The high regenerative potential of damaged neural tissues suggests that various embryonic/adult sources serve as a proxy for neural stem cells for cell-based therapy.

Phylogenetic analysis, based on EPIYA repeats in the cagA gene of Indian Helicobacter pylori, and the implications of sequence variation in tyrosine phosphorylation motifs on determining the clinical outcome.

The population of India harbors one of the world’s most highly diverse gene pools, owing to the influx of successive waves of immigrants over regular periods in time. Several phylogenetic studies involving mitochondrial DNA and Y chromosomal variation have demonstrated Europeans to have been the first settlers in India. Nevertheless, certain controversy exists, due to the support given to the thesis that colonization was by the Austro-Asiatic group, prior to the Europeans. Thus, the aim was to investigate pre-historic colonization of India by anatomically modern humans, using conserved stretches of five amino acid (EPIYA) sequences in the cagA gene of Helicobacter pylori. Simultaneously, the existence of a pathogenic relationship of tyrosine phosphorylation motifs (TPMs), in 32 H. pylori strains isolated from subjects with several forms of gastric diseases, was also explored. High resolution sequence analysis of the above described genes was performed. The nucleotide sequences obtained were translated into amino acids using MEGA (version 4.0) software for EPIYA. An MJ-Network was constructed for obtaining TPM haplotypes by using NETWORK (version 4.5) software. The findings of the study suggest that Indian H. pylori strains share a common ancestry with Europeans. No specific association of haplotypes with the outcome of disease was revealed through additional network analysis of TPMs.

Isolation and Characterization of Stem Cells Sub Population within the Human Fetal Liver

Isolation and Characterization of Stem Cells Sub Populationwithin the Human Fetal Liver Human fetal liver is the potential source of both hematopoietic and non-hematopoietic stem cells which can be identified using phenotypic markers. Isolation of homogenous populations of hepatic progenitor cells and their sub-populations is an essential prerequisite for investigating specific markers and appropriate cell types for their possible clinical applications. Several studies have demonstrated the presence of a variety of stem cell populations within the fetal liver. The present study was undertaken to identify specialized cell populations, their valuable growth potential and bi-potential differentiation capability derived from human fetal liver using CD133.

Real-time cellular and molecular dynamics of bi-metallic self-therapeutic nanoparticle in cancer cells

Since last decades various kinds of nanoparticles have been functionalized to improve their biomedical applications. However, the biological effect of un-modified/non-functionalized bi-metallic magnetic nanoparticles remains under investigated. Herein we demonstrate a multifaceted non-functionalized bi-metallic inorganic Gd-SPIO nanoparticle which passes dual high MRI contrast and can kill the cancer cells through several mechanisms. The results of the present study demonstrate that Gd-SPIO nanoparticles have potential to induce cancer cell death by production of reactive oxygen species and apoptotic events. Furthermore, Gd-SPIO nanoparticles also enhance the expression levels of miRNA-199a and miRNA-181a-7p which results in decreased levels of cancer markers such as C-met, TGF-β and hURP. One very interesting finding of this study reveals side scatter-based real-time analysis of nanoparticle uptake in cancer cells using flow cytometry analysis. In conclusion, this study paves a way for future investigation of un-modified inorganic nanoparticles to purport enhanced therapeutic effect in combination with potential anti-tumor drugs/molecules in cancer cells.

Haplotype Analyses of DNA Repair Gene Polymorphisms and Their Role in Ulcerative Colitis

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.

Molecular dynamics of pancreatic transcription factors in bioengineered humanized insulin producing neoorgan

BACKGROUND The present study has been aimed to identify molecular dynamics of pancreatic transcription factors (pTFs) during events of directed trans-differentiation of human hepatic progenitor cells (hHPCs) into insulin producing cells (InPCs) within bioengineered humanized neoorgan. The study demonstrates applicability of acellularized whole splenic scaffold (ASOS) to generate insulin producing humanized transplantable neoorgan through activation of pancreatic transcription factors. METHODS An efficient acellularization process was developed for xenogeneic rat spleen using change in different gradients of reagents perfusion through splenic artery for varying time points. The acellularized xenogeneic spleen scaffold was characterized thoroughly for preservation of extra-cellular matrix and retention of organ specific vasculature and mechanical properties. Further scaffolds were sterilized and repopulated with hHPCs which were triggered using a stage wise induction with growth factors and hyperglycemic challenge for trans-differentiation into InPCs. Dynamics of pTFs alone or simultaneously during induction process was identified using gene expression analysis and immunological staining. RESULTS The cells within the engineered neoorgan respond to growth factors and extrinsic hyperglycemic challenge and generate large number of InPCs under controlled dynamic regulation of pTFs. Highly controlled regulation of pTFs generates higher percentage of Nkx-6.1+/C-peptide+ cells within the engineered splenic scaffolds. Generation of high percentage of insulin and C-peptide positive cells in three-dimensional organ architecture responded better to hyperglycemic stimuli and produced higher quantity of insulin than 2D-culture system. CONCLUSION The present study provides a novel platform for designing effective regenerative strategies using whole organ scaffolds to control hyperglycemia under tight regulation of pTFs using humanized neoorgan system.

Bimetallic redox nanoprobe enhances the therapeutic efficacy of hyperthermia in drug-resistant cancer cells

Cancer nanotheranostics has emerged as one of the most promising fields of medicine wherein nano-sized molecules/agents are used for combined diagnosis and treatment of cancer. Despite promises of novel cancer therapeutic approaches, several crucial challenges have remained to be overcome for successful clinical translation of such agents. Hence, the present study has been aimed to investigate the therapeutic efficacy of bimetallic gadolinium super-paramagnetic iron oxide nanoformulation of ascorbic acid in synergism with hyperthermia on ascorbic acid-resistant breast cancer cells. This particular strategy provides real-time MRI-based non-invasive imaging of drug loading in resistant cancer cells along with highly enhanced therapeutic efficacy. This unique redox nanoprobe is capable of reversing drug-resistance mechanism in cancer cells and offers better therapeutic possibilities in targeted and effective destruction of drug-resistant cancer cells.

1114 TRANSPLANTATION OF EpCAM+ve HUMAN HEPATIC STEM CELLS IN LIVER CIRRHOSIS AND CELLULAR IMMUNE RESPONSE

Introduction: Liver transplantation is the only effective treatment for decompensated liver-cirrhosis. Several factors, such as nonavailability of donors, operative-risks, complications associated with rejection, usage of immunosuppressive agents, and high cost of treatment, make this strategy available to only a few people. Human foetal liver derived hepatic progenitor cell transplantation (HSCT) have shown to be a good alternative to manage end-stage liver diseases. In this retrospective study, we investigated safety and efficacy of HSCT by monitoring the T-cell, NK-cell and cytokines which play major role in cellular immune response and rejection of chronic decompensated liver cirrhosis patients. Materials and Methods: A total of 5 patients with decompensated liver cirrhosis were enrolled in the study. After giving human foetal liver-derived EpCAM positive cell transplantation, T-cell (CD3, CD4 and CD8), NK-cells (CD16) by flowcytometry and cytokine-levels (IL2, TNFb, IFNa, IFNb and IFNg) by ELISA were monitored four times within a month. Result: Present study demonstrated that after HSCT patient showed marked clinical recovery and decline in the MELD score and there was no significant variation found in cell mediated response and cytokine levels between pre and post transplantation. Conclusion: Hence this preliminary study demonstrated human foetal liver-derived EpCAM positive stem cell transplantation is safe for end stage liver cirrhosis.