Indigenous Production of Bovine/Bubaline Reproductive Hormones

Abstract: The research work of our laboratory on buffalo pituitary hormones is summarized here in the context of MOET programme of our country. All the anterior pituitary protein hormones of this species (i.e. Follicle stimulating hormone (FSH), Growth Hormone (GH) and others) have been purified from freshly frozen pituitaries. These hormones have been extensively characterized with regard to physico chemical, immunochemical and biological features. We have also produced buffalo Prolactin (PRL), Luteinizing Hormone (LH), FSH and Thyroid Stimulating Hormone (TSH) by recombinant DNA techniques

Biology as an integrating natural science domain: a proposal for BSc (Hons) in integrated biology

In this article, I present arguments in support of teaching biology as one of the integrating natural science domains at the higher secondary school and the undergraduate levels and not as phylogenic group-based subdisciplines like botany, zoology and microbiology or functional subdisciplines like genetics and biochemistry. This is possible only if we conceptualize biology and try to understand its philosophical underpinnings

Production of antibodies specific to human chorionic gonadotropin in mice immunized against its chemical analogs

Mice immunized against DS5-hCG-β and DS6-hCG-β, chemical analogs of β-subunit of human choriogonadotropin (hCG-β) in which 5 and 6 disulphide bonds respectively were reduced and alkylated, were found to produce antibodies specific to hCG without significant crossreactivity with human lutropin (hLH) as tested in a radioimmunoassay. In contrast, mice immunized against the native hCG-β subunit produced hLH crossreacting antibodies. While the anti-DS5, DS6-hCG-β serum was capable of selectively blocking the binding of [125I]-hCG to rat testicular LH/hCG receptors, it failed to inhibit the binding of [125I]-hLH to the same receptors. The radioimmunoassay for hCG using the mouse anti-DS5, DS6-hCG-β serum was not as sensitive as that employing rabbit anti-DS5, DS6-hCG-β serum. The minimal detection limit was 5 ng/ml for the mouse antibody as compared to 1 ng/ml for the rabbit antibody

Inhibitory activity of the peptides derived from buffalo prolactin on angiogenesis

The peptide fragments obtained by cathepsin digestion of purified buffalo prolactin (buPRL) monomer have been characterized using SDS-PAGE and FPLC with regard to size and pI. Their antiangiogenic activity was tested in chick embryo chorioallantoic membrane (CAM) assay and the human endothelial cells wound healing assay. Antiangiogenic activity was observed in cathepsin-cleaved fragments from buPRL. Further, a peptide sequence 45A- 46Q-47G-48K-49G-50F-51I-52T-53M-54A-55L-56N-57S-58C, which matched with human somatostatin (hSST), a known antiangiogenic factor, was located in the second loop between the first and second a-helices in the threedimensional structure of buPRL, obtained by homology modelling. The synthetic peptide matching with SST sequence was found to exhibit antiangiogenic activity in both in vitro and ex vivo assays. It was also observed that all the peptides related to buPRL could antagonize the vascular endothelial growth factor (VEGF) and bradykinin (BK)- dependent production of endothelial nitric oxide (NO), which is a pre-requisite for endothelial tube formation. It is concluded therefore that an internal sequence in buPRL and peptide fragments derived from cathepsin-digested buPRL exhibit antiangiogenic activities

Association of polymorphisms in pulmonary surfactant protein A1 and A2 genes with high-altitude pulmonary edema

Study objectives: A potential pathogenetic cofactor for the development of high-altitude pulmonary edema (HAPE) is an increase in capillary permeability, which could occur as a result of an inflammatory reaction and/or free-radical-mediated injury to the lung. Pulmonary surfactant protein A (SP-A), the most abundant surfactant protein, has potent antioxidant properties and protects unsaturated phospholipids and growing cells from oxidative injury. Single-nucleotide polymorphisms (SNPs) in SP-A1 and SP-A2, genes encoding SP-A, have been associated with susceptibility to respiratory distress syndrome, COPD, and pulmonary infections. In view of the protective role of SP-A against inflammatory reactions and oxidative damage, the two underlying mechanisms in development of HAPE, we examined the association of constitutional susceptibility to HAPE with polymorphisms in SP-A1 and SP-A2. Design: A cross-sectional case-control study. Setting: Blood samples were collected at an altitude (≥ 3,500 m). Participants: Twelve low-altitude native (LAN) subjects with a history of HAPE, 15 healthy LAN sojourners without a history of HAPE (LAN control subjects), and 19 healthy high-altitude natives (HANs) without a history of HAPE (HAN control subjects). Measurements: The SNPs in four exons and intermediate introns of the SP-A1 and SP-A2 were screened by polymerase chain reaction and sequencing. Biochemical parameters related to oxidative stress (malondialdehyde and reduced glutathione in RBC) and membrane permeability (circulating levels of lactate dehydrogenase) were measured in plasma. Results: Allele frequencies of three loci in SP-A1 and one in SP-A2 were significantly different between LAN HAPE patients (SP-A1 C1101T: C allele, 36.4% and T allele, 63.6%; SP-A1 T3192C: T allele, 61.1% and C allele, 38.9%; SP-A1 T3234C: T allele, 61.1% and C allele, 38.9%; and SP-A2 A3265C: A allele, 21.4% and C allele, 78.6%) and LAN control subjects (SP-A1 C1101T: C allele, 8.3% and T allele, 91.7%; SP-A1 T3192C: T allele, 15% and C allele, 85%; SP-A1 T3234C: T allele, 15% and C allele, 85%; and SP-A2 A3265C: A allele, 37.5% and C allele, 62.5%) [C1101T odds ratio [OR], 6.3 with 95% confidence interval (CI), 2.8 to 14.3; T3192C OR, 8.9 with 95% CI, 4.5 to 17.6; T3234C OR, 8.9 with 95% CI, 4.5 to 17.6; and A3265C OR, 2.2 with 95% CI, 1.2 to 4.1 (p ≤ 0.01)]. Heterozygous individuals, with respect to SP-A1 C1101T and SP-A2 A3265C, showed less severity in oxidative damage in comparison with homozygous subjects (SP-A1 T1101 and SP-A2 C3265). Conclusion: The polymorphisms in SP-A1 (C1101T, T3192C, and T3234C) and SP-A2 (A3265C) might be one of the genetic factors contributing to susceptibility to HAPE

Purification and biological characterization of bacterially expressed recombinant buffalo prolactin

Recombinant prolactin (PRL) from water buffalo (Bubalus bubalis) has been cloned and expressed in a prokaryotic expression system. The hormone was also successfully refolded into a biologically active form. Total RNA was purified from buffalo pituitaries and the buPRL cDNA was synthesized using primers designed on bovine PRL sequence. This prolactin cDNA was cloned in a pET 28a vector and expressed in Escherichia coli strain BL21(DE3)pLysS. Most of the expressed protein was present as insoluble inclusion bodies. The inclusion bodies were solubilized and buPRL was purified by Ni-NTA column. The purified protein was refolded by gradually decreasing the concentration of denaturant during dialysis. Total yield of the refolded and soluble prolactin was 22 mg/L from 100 mL bacterial culture in LB medium. The recombinant prolactin was as active as native prolactin in stimulating growth of Nb2 lymphoma cells

Anti-angiogenic activity of naturally occurring lower size isoforms of Buffalo pituitary prolactin

The lower size (14 kDa-18 kDa) isoforms of buffalo pituitary prolactin (PRL), separated from the monomer and higher size forms, has anti-angiogenic activity in human endothelial cell migration assay and chick embryo chorioallantoic membrane (CAM) analysis. The isoforms also showed 35 times less immunoreactivity than the hormone monomer. There was no 16 kDa isoform of buffalo PRL. This is the first demonstration that the PRL fragments, which are physiologically cleaved in pituitary glands and biochemically purified, inhibit angiogenesis