Molecular cloning,characterization and functional assessment of the myosin light polypeptide chain 2 (mylz2) promoter of farmed carp,Labeo rohita

We cloned the 5′-flanking region (1.2 kb) of a muscle-specific gene, encoding myosin light chain 2 polypeptide (mylz2) of a farmed carp, Labeo rohita (rohu). Sequence analysis using TRANSFAC-database search identified the consensus cis acting regulatory elements of TATA-box and E (CANNTG)-box, including the monocyte enhancer factor 2 motif, implying that it is likely to be a functional promoter. The proximal promoter (~620 bp) was highly homologous with that of Danio rerio (zebrafish) as compared to Channa striatus (snakehead murrel) counterparts and showed less identity with Sparus auratus (gilthead sea bream), Xenopus laevis (African clawed frog) and Rattus norvegicus (Norway rat). Direct muscular (skeletal) injection of the construct containing the mylz2 promoter (0.6 kb) fused to a green fluorescent protein (GFP) reporter gene showed efficient expression in L. rohita, validating its functional activity. Further, the functional activity was confirmed by the observation that this promoter drove GFP expression in the skeletal muscle of transgenic rohu. The promoter may have potential applications for value-addition in ornamental fishes and studying gene regulatory functions.     

Climate‐driven uncertainties in modeling terrestrial gross primary production: a site level to global‐scale analysis

We used a land surface model to quantify the causes and extents of biases in terrestrial gross primary production (GPP) due to the use of meteorological reanalysis datasets. We first calibrated the model using meteorology and eddy covariance data from 25 flux tower sites ranging from the tropics to the northern high latitudes and subsequently repeated the site simulations using two reanalysis datasets: NCEP/NCAR and CRUNCEP. The results show that at most sites, the reanalysis‐driven GPP bias was significantly positive with respect to the observed meteorology‐driven simulations. Notably, the absolute GPP bias was highest at the tropical evergreen tree sites, averaging up to ca. 0.45 kg C m^?2 yr^?1 across sites (ca. 15% of site level GPP). At the northern mid‐/high‐latitude broadleaf deciduous and the needleleaf evergreen tree sites, the corresponding annual GPP biases were up to 20%. For the nontree sites, average annual biases of up to ca. 20–30% were simulated within savanna, grassland, and shrubland vegetation types. At the tree sites, the biases in short‐wave radiation and humidity strongly influenced the GPP biases, while the nontree sites were more affected by biases in factors controlling water stress (precipitation, humidity, and air temperature). In this study, we also discuss the influence of seasonal patterns of meteorological biases on GPP. Finally, using model simulations for the global land surface, we discuss the potential impacts of site‐level reanalysis‐driven biases on the global estimates of GPP. In a broader context, our results can have important consequences on other terrestrial ecosystem fluxes (e.g., net primary production, net ecosystem production, energy/water fluxes) and reservoirs (e.g., soil carbon stocks). In a complementary study (Barman et al., 2013 ), we extend the present analysis for latent and sensible heat fluxes, thus consistently integrating the analysis of climate‐driven uncertainties in carbon, energy, and water fluxes using a single modeling framework.     

Cardiac ion channel current modulation by the CFTR inhibitor GlyH-101

The role in the heart of the cardiac isoform of the cystic fibrosis transmembrane conductance regulator (CFTR), which underlies a protein kinase A-dependent Cl⁻ current (I_Cl.PKA) in cardiomyocytes, remains unclear. The identification of a CFTR-selective inhibitor would provide an important tool for the investigation of the contribution of CFTR to cardiac electrophysiology. GlyH-101 is a glycine hydrazide that has recently been shown to block CFTR channels but its effects on cardiomyocytes are unknown. Here the action of GlyH-101 on cardiac I_Cl.PKA and on other ion currents has been established. Whole-cell patch-clamp recordings were made from rabbit isolated ventricular myocytes. GlyH-101 blocked I_Cl.PKA in a concentration- and voltage-dependent fashion (IC₅₀ at +100 mV = 0.3 ± 1.5 μM and at −100 mV = 5.1 ± 1.3 μM). Woodhull analysis suggested that GlyH-101 blocks the open pore of cardiac CFTR channels at an electrical distance of 0.15 ± 0.03 from the external membrane surface. A concentration of GlyH-101 maximally effective against I_Cl.PKA (30 μM) was tested on other cardiac ion currents. Inward current at −120 mV, comprised predominantly of the inward-rectifier background K⁺ current, I_K1, was reduced by ∼43% (n = 5). Under selective recording conditions, the Na⁺ current (I_Na) was markedly inhibited by GlyH-101 over the entire voltage range (with a fractional block at −40 mV of ∼82%; n = 8). GlyH-101 also produced a voltage-dependent inhibition of L-type Ca²⁺ channel current (I_Ca,L); fractional block at +10 mV of ∼49% and of ∼28% at −10 mV; n = 11, with a ∼−3 mV shift in the voltage-dependence of I_Ca,L activation. Thus, this study demonstrates for the first time that GlyH-101 blocks cardiac I_Cl.PKA channels in a similar fashion to that reported for recombinant CFTR. However, inhibition of other cardiac conductances may limit its use as a CFTR-selective blocker in the heart.     

Malonato complexes of oxidovanadium(IV): Synthesis, structural characterization and exploration of their insulin mimetic properties

Several bis-malonatooxidovanadium(IV) complexes of the general type [M₂(H₂O)_n][VO(mal)₂(H₂O)] (where M = Li(1), Na(2), K(3), Cs(4) and NH₄(5); n = 3.5, 1, 3, 1 and 1, respectively) were isolated in good yield and high purity. These complexes were fully characterized by various physicochemical techniques (elemental analysis, UV-Vis, IR, EPR, CV, etc.) complexes 1, 2 and 3 were structurally characterized by single crystal X-ray diffraction technique. In vivo antidiabetic properties of bis-malonato complexes 1, 2, 3 and 5 have been studied using Streptozotocin induced diabetic rats. Significant lowering of blood sugar level has been noticed. At the same time these complexes were found to regulate secondary pathophysiological complications like liver damage and lowering of the total antioxidant status (TAS) in diabetic rats. Results of these study are expected to a expand the possibility of designing new oxidovanadium(IV) complexes of O,O chelating ligands with significant antidiabetic properties.     

Sulphate is a competitive inhibitor of the binding of nucleotide to myosin. A comparison with phosphate

By the use of rapid reaction methods (rapid flow quench and stopped flow) it has been shown that sulphate is a competitive inhibitor of the binding of epsilon-ATP and ATP to myosin. At low ionic strengths, the Ki was in the micromolar range. Under several conditions used sulphate was more effective than phosphate. Neither anion was very effective in inhibiting the binding of epsilon-ATP to actomyosin.     

Impacts of flood control schemes on inland fisheries in Bangladesh: guidelines for mitigation

Flood control, drainage and irrigation (FCDI) schemes are widespread in Bangladesh. They are built to control water levels to improve agricultural production based on high yielding varieties (HYV) of rice that cannot tolerate rapid inundation or that require irrigation, and to provide protection from extreme flood events. The benefits to the agricultural sector can be significant. At some sites in Bangladesh, farmers report up to 80% more agricultural production inside the schemes than outside. However, fish production and species richness is typically lowered by these structures. Fish yields inside a typical flood control compartment can be 50% lower than outside, with up to 25 species of fish absent or less abundant. Lower rates of recruitment of migratory whitefish species, whose lateral migrations are obstructed by the embankments, were found to be largely responsible for these differences. With a risk of more extreme flooding during the monsoon season but hotter and more arid dry season conditions predicted as a consequence of climate change, more FCDI schemes may need to be constructed to provide flood protection and to meet increasing irrigation needs. Based on fisheries monitoring and mark-recapture studies undertaken at 3 sluice gates, nine recommendations for operating sluice gates to mitigate the impacts of FCDI schemes on fish production and biodiversity are described. These recommendations aim to improve the access of migratory whitefish to modified floodplains and to improve or sustain the production of resident (non-migratory) blackfish whilst minimising agricultural sector losses. Guest editors: R. L. Welcomme & G. Marmulla Hydropower, Flood Control and Water Abstraction: Implications for Fish and Fisheries     

The exo- or endonucleolytic preference of bovine pancreatic ribonuclease A depends on its subsites structure and on the substrate size

The cleavage pattern of oligocytidylic acid substrates by bovine pancreatic ribonuclease A (RNase A) was studied by means of reversed-phase HPLC. Oligocytidylic acids, ranging from dinucleotides to heptanucleotides, were obtained by RNase A digestion of poly(C). They were identified by MALDI-TOF mass spectrometry; it was confirmed that all of them corresponded to the general structure (Cp)(n)C>p, in which C>p indicates a 2',3'-cyclic phosphate. This is a confirmation of the proposed mechanism for RNase A, wherein the so-called hydrolytic (or second) step is in fact a special case of the reverse of transphosphorylation (first step). The patterns of cleavage for the oligonucleotide substrates show that the native enzyme has no special preference for endonucleolytic or exonucleolytic cleavage, whereas a mutant of the enzyme (K7Q/R10Q-RNase A) lacking p(2) (a phosphate binding subsite adjacent, on the 3' side, to the main phosphate binding site p(1)) shows a clear exonucleolytic pattern; a mutant (K66Q-RNase A) lacking p(0) (a phosphate binding subsite adjacent, on the 5' side, to the main phosphate binding site p(1)) shows a more endonucleolytic pattern. This indicates the important role played by the subsites on the preference for the bond cleaved. Molecular modeling shows that, in the case of the p(2) mutant, the amide group of glutamine can form a hydrogen bond with the 2',3'-cyclic terminal phosphate, whereas the distance to a 3',5'-phosphodiester bond is too long to form such a hydrogen bond. This could explain the preference for exonucleolytic cleavage shown by the p(2) mutant.     

Chromosomal and plasmid encoded drug resistances of a <Emphasis Type="Italic">Klebsiella pneumoniae</Emphasis> UTI 2 strain isolated from urine of a post-operative patient

The multi drug resistance Klebsiella pneumoniae in urinary tract infection is a common clinical problem in developing country like India. Use of random antibiotics, resulting multi drug resistance development, creates difficulties for treatment. In our present study, we investigated a strain of Klebsiella pneumoniae UTI 2 with multiple drug resistance, which was isolated from urine of a post operative woman patient (50 years) suffering from urinary tract infection with high fever. This strain is resistant to 36 antibiotics and sensitive to cefotaxime (Ce) and imipenem (I). After curing of plasmids, we observed that, 55% of drug resistant loci of K. pneumoniae UTI 2 are chromosomal and 40% are plasmid encoded. The organism is sensitive to 5% of drugs tested, i.e. Ce and I. This study contributes to understand the drug resistance of Klebsiella pneumoniae, which will enable better clinical management of catheter-associated urinary tract infections, a major health problem.     

Effect of protein kinase C inhibition on hypoxic pulmonary vasoconstriction

The current study was done to test the hypothesis that protein kinase C (PKC) inhibitors prevent the increase in pulmonary vascular resistance and compliance that occurs in isolated, blood-perfused dog lungs during hypoxia. Pulmonary vascular resistances and compliances were measured with vascular occlusion techniques. Hypoxia significantly increased pulmonary arterial resistance, pulmonary venous resistance, and pulmonary capillary pressure and decreased total vascular compliance by decreasing both microvascular and large-vessel compliances. The nonspecific PKC inhibitor staurosporine (10(-7) M), the specific PKC blocker calphostin C (10(-7) M), and the specific PKC isozyme blocker G?-6976 (10(-7) M) inhibited the effect of hypoxia on pulmonary vascular resistance and compliance. In addition, the PKC activator thymeleatoxin (THX; 10(-7) M) increased pulmonary vascular resistance and compliance in a manner similar to that in hypoxia, and the L-type voltage-dependent Ca(2+) channel blocker nifedipine (10(-6) M) inhibited the response to both THX and hypoxia. These results suggest that PKC inhibition blocks the hypoxic pressor response and that the pharmacological activation of PKC by THX mimics the hypoxic pulmonary vasoconstrictor response. In addition, L-type voltage-dependent Ca(2+) channel blockade may prevent the onset of the hypoxia- and PKC-induced vasoconstrictor response in the canine pulmonary vasculature.