Noninvasive genetic monitoring of tiger (Panthera tigris tigris) population of Orang National Park in the Brahmaputra floodplain, Assam, India

The Brahmaputra Valley of Assam, India, is one of the prime habitats for the endangered Royal Bengal tiger Panthera tigris tigris. With dwindling global population, estimation of the minimum number of tigers has always been a curiosity to wildlife researchers as well as to protected area managers. In the present study, DNA-based techniques were used for identifying individual tigers present in Orang National Park of Assam, from 57 faecal samples collected during February 2009. Orang National Park stands as an island of a single forest patch along the north bank of river Brahmaputra. The present study confirms the presence of 17 individual tigers in Orang National Park, with five male and 12 female. DNA-based capture–recapture analysis yielded minimum range estimate of 18 and 19 individuals, with possible overestimates of population size following two models of capture probability in CAPWIRE. The results of our genetic counting of tigers are compared with the estimates of 19 tigers based on pugmark analysis by the state Forest Department in 2000 and an independent capture–recapture estimate of 14 (±3.6) individuals based on photographic identity study in 2009. Looking at high mortality of tigers in the area, with 19 reported deaths during 2000 to 2009, our results indicate high individual turnover in the area. This study shows that Orang National Park harbours a healthy breeding population of tigers. However, the possibility of a source-sink dynamics operating in the landscape could not be ruled out, with possible immigration from nearby Kaziranga National Park on the south bank of Brahmaputra, which has the highest reported density of the species in the world.     

Records of chironomids of the tribe Pentaneurini (Diptera: Chironomidae) in the Eastern Himalayas of India

Larva, pupa and female imago of Paramerina ampliseta n. sp. and Rheopelopia tuberculata (Chaudhuri & Debnath 1987), pupa and male imago of Paramerina clara n. sp. and pupa of Ablabesmyia (Ablabesmyia) alba Chaudhuri, Debnath & Nandi 1983 and Paramerina inficia Chaudhuri & Debnath 1985 are described from the Eastern Himalayas of India.     

On some biochemical physiognomies of two common Darjeeling tea cultivars in relation to blister blight disease

Abstract

Tea, Camellia sinensis (L.) O. Kuntze, an agro-based industry, is considered as one of the prime sector of exporting resources and thus considered as “cash-crop”. Earlier report shows that tea is native to eastern and northern India, which was cultivated and consumed there since long back. Presently, more scientific reports confirmed the health-benefit traits of tea and awareness increased to a greater extent and in this regard, tea has gained its best worldwide popularity. Darjeeling Tea attained its highest acceptance globally for its pre-eminence in flavour, colour and taste and thus crop improvement is the prime interest to the Indian Scientific Community. Blister blight disease, a common disorder of tea bushes (Exobasidium vexans, a Basidiomycetes fungus) causes drastic damage of tea plantation. Depending on quality production, two common cultivars were released by TRA, Jorhat, Assam viz. Bannockburn – 157 (B-157) and Ambei Valai - 2 (AV-2), of which B-157 is susceptible to the Blister Blight and AV-2 is fairly resistant cultivar. Some biochemical considerations between the two cultivars have been made here for understanding the biochemical reasons behind the resistant characteristics. Plant secondary metabolites, like total phenol, proanthocynadin, total soluble protein provide defending feature against disease onset. AV-2 cultivar shows advantage over B-157 in these regard. Depending on band intensity analysis of native gels, acid phosphatase, catechol oxidase, peroxidase and superoxide dismutase occur in superior amount in AV-2 cultivar than that of B-157. The specific role of these enzymes in blister blight disease compatibility of two cultivars studied has been discussed.     

Comparative molecular dynamics simulation studies for determining factors contributing to the thermostability of chemotaxis protein “CheY”

Comparative molecular dynamics simulations of chemotaxis protein “CheY” from thermophilic origin Thermotoga maritima and its mesophilic counterpart Salmonella enterica have been performed for 10?ns each at 300 and 350?K, and 20?ns each at 400 and 450?K. The trajectories were analyzed in terms of different factors like root-mean-square deviation, root-mean-square fluctuation, radius of gyration, solvent accessible surface area, H-bonds, salt bridge content, and protein–solvent interactions which indicate distinct differences between the two of them. The two proteins also follow dissimilar unfolding pathways. The overall flexibility calculated by the trace of the diagonalized covariance matrix displays similar flexibility of both the proteins near their optimum growth temperatures. However, at higher temperatures mesophilic protein shows increased overall flexibility than its thermophilic counterpart. Principal component analysis also indicates that the essential subspaces explored by the simulations of two proteins at different temperatures are nonoverlapping and they show significantly different directions of motion. However, there are significant overlaps within the trajectories and similar direction of motions are observed for both proteins at 300?K. Overall, the mesophilic protein leads to increased conformational sampling of the phase space than its thermophilic counterpart. This is the first ever study of thermostability of CheY protein homologs by using protein dynamism as a main impact. Our study might be used as a model for studying the molecular basis of thermostability of two homologous proteins from two organisms living at different temperatures with less visible differences.     

Pirfenidone Nanoparticles Improve Corneal Wound Healing and Prevent Scarring Following Alkali Burn

Purpose

To evaluate the effects of pirfenidone nanoparticles on corneal re-epithelialization and scarring, major clinical challenges after alkali burn.

Methods

Effect of pirfenidone on collagen I and α-smooth muscle actin (α-SMA) synthesis by TGFβ induced primary corneal fibroblast cells was evaluated by immunoblotting and immunocytochemistry. Pirfenidone loaded poly (lactide-co-glycolide) (PLGA) nanoparticles were prepared, characterized and their cellular entry was examined in primary corneal fibroblast cells by fluorescence microscopy. Alkali burn was induced in one eye of Sprague Dawley rats followed by daily topical treatment with free pirfenidone, pirfenidone nanoparticles or vehicle. Corneal re-epithelialization was assessed daily by flourescein dye test; absence of stained area indicated complete re-epithelialization and the time for complete re-epithelialization was determined. Corneal haze was assessed daily for 7 days under slit lamp microscope and graded using a standard method. After 7 days, collagen I deposition in the superficial layer of cornea was examined by immunohistochemistry.

Results

Pirfenidone prevented (P<0.05) increase in TGF β induced collagen I and α-SMA synthesis by corneal fibroblasts in a dose dependent manner. Pirfenidone could be loaded successfully within PLGA nanoparticles, which entered the corneal fibroblasts within 5 minutes. Pirfenidone nanoparticles but not free pirfenidone significantly (P<0.05) reduced collagen I level, corneal haze and the time for corneal re-epithelialization following alkali burn.

Conclusion

Pirfenidone decreases collagen synthesis and prevents myofibroblast formation. Pirfenidone nanoparticles improve corneal wound healing and prevent fibrosis. Pirfenidone nanoparticles are of potential value in treating corneal chemical burns and other corneal fibrotic diseases.     

Mis-sizing of stent promotes intimal hyperplasia: impact of endothelial shear and intramural stress

Stent can cause flow disturbances on the endothelium and compliance mismatch and increased stress on the vessel wall. These effects can cause low wall shear stress (WSS), high wall shear stress gradient (WSSG), oscillatory shear index (OSI), and circumferential wall stress (CWS), which may promote neointimal hyperplasia (IH). The hypothesis is that stent-induced abnormal fluid and solid mechanics contribute to IH. To vary the range of WSS, WSSG, OSI, and CWS, we intentionally mismatched the size of stents to that of the vessel lumen. Stents were implanted in coronary arteries of 10 swine. Intravascular ultrasound (IVUS) was used to size the coronary arteries and stents. After 4 wk of stent implantation, IVUS was performed again to determine the extent of IH. In conjunction, computational models of actual stents, the artery, and non-Newtonian blood were created in a computer simulation to yield the distribution of WSS, WSSG, OSI, and CWS in the stented vessel wall. An inverse relation (R(2) = 0.59, P < 0.005) between WSS and IH was found based on a linear regression analysis. Linear relations between WSSG, OSI, and IH were observed (R(2) = 0.48 and 0.50, respectively, P < 0.005). A linear relation (R(2) = 0.58, P < 0.005) between CWS and IH was also found. More statistically significant linear relations between the ratio of CWS to WSS (CWS/WSS), the products CWS × WSSG and CWS × OSI, and IH were observed (R(2) = 0.67, 0.54, and 0.56, respectively, P < 0.005), suggesting that both fluid and solid mechanics influence the extent of IH. Stents create endothelial flow disturbances and intramural wall stress concentrations, which correlate with the extent of IH formation, and these effects were exaggerated with mismatch of stent/vessel size. These findings reveal the importance of reliable vessel and stent sizing to improve the mechanics on the vessel wall and minimize IH.     

Controllable self-assembly from fibrinogen-gold (fibrinogen-Au) and thrombin-silver (thrombin-Ag) nanoparticle interaction

Fibrinogen conjugated gold nanoparticles (fibrinogen-Au) and thrombin conjugated silver nanoparticles (thrombin-Ag) were synthesized by heating (90 degrees C) the proteins (50 microg protein/ml) with 1mM AgNO(3) or AuCl(3). The resultant particles were harvested and examined by flow cytometry, scanning electron microscopy (SEM), transmission emission microscopy (TEM), optical microscopy and dynamic light scattering. SEM and TEM images revealed that the fibrinogen-Au and thrombin-Ag particles interacted. The emergent bio-nanoconjugate population could be controlled by addition of thrombin-Ag. The method may be exploited in parametrizing coagulation factors and other clinically important protein-protein interactions.     

Synthesis and antiproliferative activity of some novel derivatives of diospyrin, a plant-derived naphthoquinonoid

Derivatisation of diospyrin, a bisnaphthoquinonoid isolated from Diospyros montana Roxb., led to the modification of its inhibitory activity, in vitro, towards a murine tumour model, Ehrlich ascites carcinoma (EAC), and two human cancer cell lines, viz., malignant skin melanoma (A375) and epidermoid laryngeal carcinoma (Hep2). Among the novel derivatives, an epoxide exhibited the maximum antiproliferative activity (IC(50) values in the range of 0.03-0.21 microM) and a comparatively lower toxicity (IC(50) approximately 98 microM) in normal human peripheral blood mononuclear cells (PBMC). This compound might provide a novel 'lead' for the development of clinically effective antiproliferative agents against cancer.