BMPR2 expression is suppressed by signaling through the estrogen receptor

Background

Low endogenous testosterone levels have been shown to be a risk factor for the development of cardiovascular disease and cardiovascular benefits associated with testosterone replacement therapy are being advocated; however, the effects of endogenous testosterone levels on acute coronary vasomotor responses to androgen administration are not clear. The objective of this study was to compare the effects of acute androgen administration on in vivo coronary conductance and in vitro coronary microvascular diameter in intact and castrated male swine.

Methods

Pigs received intracoronary infusions of physiologic levels (1?C100 nM) of testosterone, the metabolite 5??-dihydrotestosterone, and the epimer epitestosterone while left anterior descending coronary blood flow and mean arterial pressure were continuously monitored. Following sacrifice, coronary arterioles were isolated, cannulated, and exposed to physiologic concentrations (1?C100 nM) of testosterone, 5??-dihydrotestosterone, and epitestosterone. To evaluate effects of the androgen receptor on acute androgen dilation responses, real-time PCR and immunohistochemistry for androgen receptor were performed on conduit and resistance coronary vessels.

Results

In vivo, testosterone and 5??-dihydrotestosterone produced greater increases in coronary conductance in the intact compared to the castrated males. In vitro, percent maximal dilation of microvessels was similar between intact and castrated males for testosterone and 5??-dihydrotestosterone. In both studies epitestosterone produced significant increases in conductance and microvessel diameter from baseline in the intact males. Androgen receptor mRNA expression and immunohistochemical staining were similar in intact and castrated males.

Conclusions

Acute coronary vascular responses to exogenous androgen administration are increased by endogenous testosterone, an effect unrelated to changes in androgen receptor expression.     

Spectrophotometric estimation of cobalt with ninhydrin

A violet coloured complex was developed when cobalt metal reacts with ninhydrin at pH 8.2, using sodium acetate buffer solution. Absorbance of the complex was measured at 395 nm. Various factors, such as volume of the ligand used, solution pH, stability of the complex with time and interference of other metals, which effect the complex formation have been studied in detail. Present developed method can be used for the spectrophotometric estimation of cobalt with ninhydrin complex. The method is simple, selective and cheap for the determination of cobalt in very less time.     

Effects of recombinant gonadotropin hormones on the expression of vitellogenin, gonadotropin subunits and gonadotropin receptors in cinnamon clownfish, Amphiprion melanopus

Gonadotropins (GTHs) are the key regulators of reproduction in vertebrates. The present study investigated autoregulatory effects of gonadotropins, using recombinant FSH (rFSH) and LH (rLH) in cinnamon clownfish (Amphiprion melanopus). Experiments were carried out to investigate the actions of cinnamon clownfish rFSH and rLH on expression of GTH subunits, GTH receptors, and vitellogenin (Vtg) mRNA in vivo and in vitro. Plasma estradiol-17β (E(2)) level was also measured in immature fish following treatments with rFSH and rLH. The results demonstrate increasing levels of GTH subunits, GTH-receptors, Vtg mRNA levels, as well as plasma E(2) levels following injection with rFSH and rLH. The findings support the hypothesis that LH and FSH stimulate reproduction, in part, by autoregulatory mechanisms leading to upregulation of GTH receptors and GTH hormone production in cinnamon clownfish. The results provide a framework for better understanding of the mechanisms of GTH-mediated control of reproduction in cinnamon clownfish and other vertebrates.     

In silico mutation on a mutant lipase from Acinetobacter haemolyticus towards enhancing alkaline stability

Abstract

Alkaline-stable lipases are highly valuable biocatalysts that catalyze reactions under highly basic conditions. Herein, computational predictions of lipase from Acinetobacter haemolyticus and its mutant, Mut-LipKV1 was performed to identify functionally relevant mutations that enhance pH performance under increasing basicity. Mut-LipKV1 was constructed by in silico site directed mutagenesis of several outer loop acidic residues, aspartic acid (Asp) into basic ones, lysine (Lys) at positions 51, 122 and 247, followed by simulation under extreme pH conditions (pH 8.0–pH 12.0). The energy minimized Mut-LipKV1 model exhibited good quality as shown by PROCHECK, ERRAT and Verify3D data that corresponded to 79.2, 88.82 and 89.42% in comparison to 75.2, 86.15, and 95.19% in the wild-type. Electrostatic surface potentials and charge distributions of the Mut-LipKV1 model was more stable and better adapted to conditions of elevated pHs (pH 8.0???10.0). Mut-LipKV1 exhibited a mixture of neutral and positive surface charge distribution compared to the predominantly negative charge in the wild-type lipase at pH 8.0. Data of molecular dynamics simulations also supported the increased alkaline-stability of Mut-LipKV1, wherein the lipase was more stable at a higher pH 9.0 (RMSD = ～0.3?nm, RMSF = ～0.05–0.2?nm), over the optimal pH 8.0 of the wild-type lipase (RMSD = 0.3?nm, RMSF = 0.05–0.20?nm). Thus, the adaptive strategy of replacing surface aspartic acid to lysine in lipase was successful in yielding a more alkaline-stable Mut-LipKV1 under elevated basic conditions.

Communicated by Ramaswamy H. Sarma     

Non-isotopic RNA probes

Summary Several studies have shown the use of non-radioactive labelled DNA probes for in situ hybridisation, mainly to identify cellular DNA. In this study mRNA in situ hybridisation was performed on rat pituitary with biotinylated complementary (c) RNA probes for rat prolactin and growth hormone (GH), and compared with radioactive ³⁵S-radiolabelled probes. Biotinylated cRNA probes were labelled with either biotin-11-UTP or with allylamine-UTP, the latter method being able to produce a higher yield of labelled RNA. Different detection systems were tested, and hybridisation signal was seen in cells of anterior pituitary with both types of biotinylated probes. The signals were detected using either avidin-biotin-complex with peroxidase (ABC), peroxidase-anti-peroxidase (PAP) or gold-silver methods. ABC peroxidase detected using glucose oxidase-diaminobenzidine (DAB)-nickel solution appeared to be the best method for detecting labelled RNA probes, with very strong signal and low background. The biotinylated probes were comparable in sensitivity to the radiolabelled probes in detecting prolactin and GH mRNAs in the anterior lobe of the rat pituitary. These results indicate an alternative methods of labelling and detection of biotinylated probes which could have a potential role in research and diagnostic techniques.     

Inappropriate Shock Delivered By Implantable Cardioverter Defibrillator - Cardiac Resynchronization Therapy (ICD-CRT) Due To Myopotential Oversensing

The clinical efficacy of ICD-CRT therapy depends on accurate sensing of intracardiac signals and sensing algorithms. We report the occurrence of sensing abnormality in a patient with ICD-CRT. In this patient, oversensing of myopotentials during strenuous muscular activity resulted in an inappropriate ICD-CRT discharge. Although modern ICDs are highly effective in detecting and terminating malignant tachyarrhythmias, their detection specificity must be improved. It is possible to find the mechanism of arrhythmia by EGM. Simple device reprogramming make it possible to avoid the oversensing of myopotentials.     

Effects of raw and diluted municipal sewage effluent with micronutrient foliar sprays on the growth and nutrient concentration of foxtail millet in southeast Iran

In this study, the effect of irrigation with raw or diluted municipal sewage effluent accompanied by foliar micronutrient fertilizer sprays was examined on the growth, dry matter accumulation, grain yield, and mineral nutrients in foxtail millet plants. The experimental design was a split plot with three irrigation sources: raw sewage, 50% diluted sewage, and well water comprising the main treatments, and four combinations of Mn and Zn foliar sprays as sub-treatments that were applied with four replications. The experiment was conducted in 2009 at the Zabol University research farm in Zabol, south Iran. The applied municipal sewage effluent contained higher levels of micronutrients and macronutrients and exhibited greater degrees of electrical conductivity compared to well water. Because of the small scale of industrial activities in Zabol, the amount of heavy metals in the sewage was negligible (below the limits set for irrigation water in agricultural lands); these contaminants would not be severely detrimental to crop growth. The experimental results indicated that irrigation of plants with raw or diluted sewage stimulates the measured growth and productivity parameters of foxtail millet plants. The concentrations of micronutrients and macronutrients were also positively affected. These stimulations were attributed to the presence of high levels of such essential nutrients as N, P, and organic matter in wastewater. Supplied in sewage water alone, Mn and Zn were not able to raise the productivity of millet to the level obtained using fertilizers at the recommended values; this by itself indicated that additional nutrients from fertilizers are required to obtain higher levels of millet productivity with sewage farming. Despite the differences in nutrient concentrations among the different irrigation water sources, the micronutrient foliar sprays did not affect the concentrations of micronutrients and macronutrients in foxtail millet plants. These results suggested that municipal sewage effluent could be utilized efficiently as an important source of water, and that the nutrients used in growing foxtail millet with sewage water irrigation did not have any significant harmful effect on crop productivity. In contrast, the nutrients proved beneficial to soil fertility and millet productivity and quality.     

Some studies of the quality of irrigation water and the germination and growth of wheat at different concentrations of boron

Summary The irrigation water in use at the Punjab University New Campus is of good quality except the water of one tubewell which is relatively poor (high-salinity and low-sodium water) and has 0.45 ppm boron. Water with a boron content of upto 3.00 ppm did not affect the germination of the seeds of wheat (Triticum vulgare L. var. Mexi-Pak) and there was some stimulation of the growth of seedlings by an increase in the concentration of boron beyond 0.45 ppm. However, these seedlings, when transplanted to the soils supplied with waters containing different amounts of boron, showed a decrease in their growth at higher concentrations of boron with their ageing, and when these plants were about 100 days old and almost mature, their growth decreased significantly with an increase in the concentration of boron to 2.00 or 3.00 ppm in the water supplied to them. The growth of the plants of Mexi-Pak wheat is inversely related to the boron content of their tissues when it is more than 0.60 ppm. Mexi-Pak wheat is semi-tolerant to boron. re]19750605