Identification of actin as an estradiol 17-beta-stimulated protein in the human placenta

Addition of estradiol 17-beta to first trimester human placental minces resulted in an increased synthesis of a protein of apparent molecular weight 45 kDa. The specific involvement of estrogen in the stimulation of this protein was established by demonstrating a reduction in the level of this protein by the addition of CCS 16949 A, an inhibitor of aromatase, a key enzyme in the biosynthesis of estradiol 17-beta and ICI 182,780, an estrogen receptor antagonist. The protein was purified to homogeneity and N-terminal sequencing of two of the internal peptides obtained by enzymatic digestion of the protein, as well as the absence of a free N-terminal indicated that it could be actin. This was confirmed by Western blotting using commercially available actin antiserum. The role of estradiol 17-beta in the stimulation of actin synthesis in human placenta was also established by monitoring the quantitative inhibition of DNase I by actin.

Expression of the extracellular domain of the human heat-stable enterotoxin receptor in Escherichia coli and generation of neutralizing antibodies

The entire extracellular domain of the human heat-stable enterotoxin (ST) receptor as well as a truncated N-terminal domain were cloned as glutathione S-transferase fusion proteins and expressed in Escherichia coli. The recombinant fusion proteins were purified from both the cytosol and the inclusion body fractions by selective detergent extraction followed by glutathione-agarose affinity chromatography. The purified protein, corresponding to the entire extracellular domain, bound the stable toxin peptide with an affinity comparable to that of the native receptor characterized from the human colonic T84 cell line. No binding was observed with the N-terminal truncated fragment of the receptor under similar conditions, Polyclonal antibodies were raised to the entire extracellular domain fusion protein as well as the truncated extracellular domain fusion protein, and the antibodies were purified by affinity chromatography. Addition of the purified antibodies to T84 cells inhibited ST binding and abolished ST-mediated cGMP production, indicating that critical epitopes involved in ligand interaction are present in the N-terminal fragment of the receptor, Purified antibodies recognized a single protein of M(r) 160,000 Da on Western blotting with T84 membranes, corresponding to a size of the native glycosylated receptor in T84 cells. These studies are the first report of the expression, purification, and characterization of any member of the guanylyl cyclase family of receptors in E. coli and show that binding of the toxin to the extracellular domain of the receptor is possible in the absence of any posttranslational modifications such as glycosylation. The recombinant fusion proteins as well as the antibodies that we have generated could serve as useful tools in the identification of critical residues of the extracellular domain involved in ligand interaction.

Regional variation in the composition and structure of mixed-species bird flocks in the Western Ghats and Sri Lanka

Mixed-species bird flocks are attractive models for the investigation of geographical variation in animal communities, as they represent a subset of the avifauna in most forested regions of the world. Yet studies of the regional variation in flock size and the composition of flocks are few, due to the predominance of studies carried out at single study site. Here, we review nine studies of mixed-species flocks conducted at 16 sites along the Western Ghats in India and in Sri Lanka. We find that flock size varies as much within this region as it does globally, with observation time being a confounding variable. Flock composition, however, is predictably related to elevation. Flocks at high elevations (>1200 m) in the Western Ghats strongly resemble flocks at high elevations in the mountain ranges of Sri Lanka in their composition, especially at the family level. We compare these flocks to flocks of other regions and make recommendations on study methodology that can facilitate comparisons across studies.

Promotion of fibronectin independent invasion by C5a peptidase into epithelial cells in group A Streptococcus

Background & objectives: Group A Streptococcus, causative agent of several clinical manifestations codes for multiple protein invasins which help the bacterium to enter non-phagocytic cells. C5a peptidase (SCPA) is a surface protein conserved among different serotypes of M1 strain. The present study was taken up to study SCPA promoted fibronectin independent entry of GAS into epithelial cells. Methods: An isogenic 90226 emm1DeltaAB (M1(-)) mutant was constructed, with thermosensitive pGhost vector. This isogenic M1(-) mutant expressed SCPA on the surface as determined by Western blotting and immunofluorescence. Results: On preincubation with anti-SCPA serum, the isogenic M1(-) strain exhibited 54 per cent decreased invasion as compared to the bacteria incubated with control serum. Also, purified recombinant SCPA proteins blocked internalization of M1(-) streptococci into HEp-2 cells. The M1(-) strain invaded at the same efficiency in the presence or absence of fibronectin. Interpretation & conclusion: These results suggested that SCPA acted as a potential invasin of group A streptococcus and promoted invasion independent of fibronectin.

Road kills of amphibians in different land use areas from Sharavathi river basin, central Western Ghats, India

A survey of amphibian mortality on roads was carried out in the Sharavathi river basin in the central Western Ghats. Road kills in three different land use areas: agricultural fields, water bodies and forests were recorded for four days along three 100m stretches in each type of area. One-hundred-and-forty-four individuals belonging to two orders, eight families, 11 genera and 13 species were recorded in the survey. Kills/km observed were: in forest 55, agricultural fields 38 and water bodies 27, for an overall average of 40 kills/km. Kill species compositions varied significantly between land use areas, but not overall kill rates.

Rainfall changes over tropical montane cloud forests of southern Western Ghats, India

The southern Western Ghats tropical montane cloud forest sites (Gavi, Periyar, High wavys and Venniyar), which are characterized by frequent or seasonal cloud cover at the vegetation level, are considered one of the most threatened ecosystems in India and the world. Three out of four montane cloud forest sites studied in the southern Western Ghats had experienced diminishing trends of seasonal average and total rainfall, especially during summer monsoon season. The highest level of reduction for summer monsoon season was observed at Gavi rainforest station (>20 mm/14 years) in Kerala followed by Venniyar (>20 mm/20 years) site in Tamil Nadu. Average annual and total precipitation increased during the study period irrespective of the seasons over Periyar area, and the greatest values were recorded for season 2 (>25 mm/28 years). Positive trends for winter monsoon rainfall has been observed for three stations (Periyar, High wavys and Venniyar) except Gavi, and the trend was positive and significant (90%) for Periyar and High wavys. Increase in summer monsoon rainfall was observed for Periyar site and the trend was found to be significant (95%).

Practising Western Science Outside the West: Personal Observations on the Indian Scene

Modern science, which was an indigenous product of Western culture, is now being practised in many non-Western countries. This paper discusses the peculiar social, cultural and intellectual problems which scientists of these non-Western countries face in adopting Western science in their situations, with special reference to India. It is pointed out that, in addition to money and communication, it is necessary to have a proper psychological gestalt to practise science satisfactorily. The author analyzes his experience as a physics student in India and in the United States to clarify the nature of this psychological gestalt, and to explain what makes it difficult for non-Western scientists to acquire it.

Heat-shock protein 70-2 (HSP70-2) expression in bladder urothelial carcinoma is associated with tumour progression and promotes migration and invasion

Purpose: Testis specific heat-shock protein 70-2 (HSP70-2), a member of HSP70 chaperone family, is essential for the growth of spermatocytes and cancer cells. We investigated the association of HSP70-2 expression with clinical behaviour and progression of urothelial carcinoma of bladder. Experimental design: We assessed the HSP70-2 expression by RT-PCR and HSP70-2 protein expression by immunofluorescence, flow cytometry, immunohistochemistry and Western blotting in urothelial carcinoma patient specimens and HTB-1, UMUC-3, HTB-9, HTB-2 and normal human urothelial cell lines. Further, to investigate the role of HSP70-2 in bladder tumour development, HSP70-2 was silenced in the high-grade invasive HTB-1 and UMUC-3 cells. The malignant properties of urothelial carcinoma cells were examined using colony formation, migration assay, invasion assay in vitro and tumour growth in vivo. Results: Our RT-PCR analysis and immunohistochemistry analysis revealed that HSP70-2 was expressed in both moderate to well-differentiated and high-grade invasive urothelial carcinoma cell lines studied and not in normal human urothelial cells. In consistence with these results, HSP70-2 expression was also observed in superficially invasive (70%) and muscle-invasive (90%) patient's tumours. Furthermore, HSP70-2 knockdown significantly suppressed cellular motility and invasion ability. An in vivo xenograft study showed that inhibition of HSP70-2 significantly suppressed tumour growth. Conclusions: In conclusion, our data suggest that the HSP70-2 expression is associated with early spread and progression of urothelial carcinoma of bladder cancer and that HSP70-2 can be the potential therapeutic target for bladder urothelial carcinoma. (C) 2009 Elsevier Ltd. All rights reserved.

Road kills of amphibians in different land use areas from Sharavathi river basin, central Western Ghats, India

A survey of amphibian mortality on roads was carried out in the Sharavathi river basin in the central Western Ghats. Road kills in three different land use areas: agricultural fields, water bodies and forests were recorded for four days along three 100m stretches in each type of area. One-hundred-and-forty-four individuals belonging to two orders, eight families, 11 genera and 13 species were recorded in the survey. Kills/km observed were: in forest 55, agricultural fields 38 and water bodies 27, for an overall average of 40 kills/km. Kill species compositions varied significantly between land use areas, but not overall kill rates.

Hills, dams and forests. Some field observations from the Western Ghats

Man’s attempts to intensify the use of natural resources can often result in the exhaustion of the resource or deterioration of other interacting resources. The single-minded pursuit of the development of the water resources of the rivers of the Western Ghats shows many examples of this view, particularly in the unnecessary destruction of the dwindling forest resources. This destruction may be caused by (i) problems o f rehabilitation, e.g. the Ramanagar settlement of the Kalinadi project (ii) the impact of labourers, e.g. the destruction of evergreen sholas on the Upper Nilgiri plateau (iii) the access to encroachers and poachers, e.g. Panshet and Kalinadi (iv) faulty planning, e.g. Linganamakki and Kalinadi. This destruction of forest cover has had a number of deleterious consequences in (i) worsening the shortages of forest resources, (ii) hastening the siltation of the reservoirs, (iii) ecological imbalances as in the rapid spread ofEupatorium in the Kalinadi project area and (iv) the decimation of biological diversity, as in the great reduction of evergreen forests in the Western Ghats, threatening the survival of lion-tailed macaque and the extinction of grass species,Hubbardia heptaneuron. It is stressed that the only sustainable and therefore true development is environmentally sound development. The interests of the weaker sections of the society often provide a good index of the soundness of the development from an environmental point of view. The planning of the development process with this perspective is a great scientific and technological challenge that must be taken up.

Ecology of the swampy relic forests of Kathalekan from central Western Ghats, India

Introduction of agriculture three millennia ago in Peninsular India’s Western Ghats altered substantially ancient tropical forests. Early agricultural communities, nevertheless, strived to attain symbiotic harmony with nature as evident from prevalence of numerous sacred groves, patches of primeval forests sheltering biodiversity and hydrology. Groves enhanced heterogeneity of landscapes involving elements of successional forests and savannas favouring rich wildlife. A 2.25 km2 area of relic forest was studied at Kathalekan in Central Western Ghats. Interspersed with streams studded with Myristica swamps and blended sparingly with shifting cultivation fallows, Kathalekan is a prominent northernmost relic of southern Western Ghat vegetation. Trees like Syzygium travancoricum (Critically Endangered), Myristica magnifica (Endangered) and Gymnacranthera canarica (Vulnerable) and recently reported Semecarpus kathalekanensis, are exclusive to stream/swamp forest (SSF). SSF and non-stream/swamp forest (NSSF) were studied using 18 transects covering 3.6 ha. Dipterocarpaceae, its members seldom transgressing tropical rain forests, dominate SSF (21% of trees) and NSSF (27%). The ancient Myristicaceae ranks high in tree population (19% in SSF and 8% in NSSF). Shannon-Weiner diversity for trees is higher (>3) in six NSSF transects compared to SSF (<3). Higher tree endemism (45%), total endemic tree population (71%) and significantly higher above ground biomass (349 t/ha) cum carbon sequestration potential (131 t/ha) characterizes SSF. Faunal richness is evident from amphibians (35 species - 26 endemics, 11 in IUCN Red List). This study emphasizes the need for bringing to light more of relic forests for their biodiversity, carbon sequestration and hydrology. The lives of marginal farmers and forest tribes can be uplifted through partnership in carbon credits, by involving them in mitigating global climatic change through conservation and restoration of high biomass watershed forests.

Novel reproductive mode in a torrent frog Micrixalus saxicola (Jerdon) from the Western Ghats, India

Reproductive modes in anurans are highly diverse despite external fertilization being a constraint. There are 39 reproductive modes documented so far (Wells, 2007). An apparently new reproductive mode is reported in a torrent frog, Micrixalus saxicola, an endemic and ancient anuran frog of the Western Ghats, considering the type of cavity made inside the lotic water body, involvement of the female in digging the cavity and concealing the eggs.

A possible role for a low molecular weight peptide in regulation of testosterone production by rat Leydig cells

Sodium dodecyl sulphate-polyacrylamide gel electrophoresis of Percoll purified Leydig cell proteins from 20- and 120-day-old rats revealed a significant decrease in a low molecular weight peptide in the adult rats. Administration of human chorionic gonadotropin to immature rats resulted in a decrease in the low molecular weight peptide along with increase in testosterone production. Modulation of the peptide by human chorionic gonadotropin could be confirmed by Western blotting. The presence of a similar peptide could be detected by Western blotting in testes of immature mouse, hamster, guinea pig but not in adrenal, placenta and corpus luteum. Administration of testosterone propionate which is known to inhibit the pituitary luteinizing hormone levels in adult rats resulted in an increase in the low molecular weight peptide, as checked by Western blotting. It is suggested that this peptide may have a role in regulation of acquisition of responsiveness to luteinizing hormone by immature rat Leydig cells.

Development of a probe-based blotting technique for the detection of Tobacco streak virus

Digoxigenin (DIG)-labeled DNA probe was developed for a sensitive and rapid detection of the Tobacco streak virus (TSV) isolates in India by dot-blot and tissue print hybridization techniques. DIG-labeled DNA probe complementary to the coat protein (CP) region of TSV sunflower isolate was designed and used to detect the TSV presence at field levels. Dot-blot hybridization was used to check a large number of TSV isolates with a single probe. In addition, a sensitivity of the technique was examined with the different sample extraction methods. Another technique, the tissue blot hybridization offered a simple, reliable procedure and did not require a sample processing. Thus, both non-radioactively labeled probe techniques could facilitate the sample screening during TSV outbreaks and offer an advantage in quarantine services.