Cardiovascular,Metabolic and Inflammatory Changes after Ovariectomy and Estradiol Substitution in Hereditary Hypertriglyceridemic Rats

Background: If menopause is really independent risk factor for cardiovascular disease is still under debate. We studied if ovariectomy in the model of insulin resistance causes cardiovascular changes, to what extent are these changes reversible by estradiol substitution and if they are accompanied by changes in other organs and tissues. Methods: Hereditary hypertriglyceridemic female rats were divided into three groups: ovariectomized at 8th week (n = 6), ovariectomized with 17-β estradiol substitution (n = 6), and the sham group (n = 5). The strain of abdominal aorta measured by ultrasound, expression of vascular genes, weight and content of myocardium and also non-cardiac parameters were analyzed. Results: After ovariectomy, the strain of abdominal aorta, expression of nitric oxide synthase in abdominal aorta, relative weight of myocardium and of the left ventricle and circulating interleukin-6 decreased; these changes were reversed by estradiol substitution. Interestingly, the content of triglycerides in myocardium did not change after ovariectomy, but significantly increased after estradiol substitution while adiposity index did not change after ovariectomy, but significantly decreased after estradiol substitution. Conclusion: Vascular and cardiac parameters under study differed in their response to ovariectomy and estradiol substitution. This indicates different effects of ovariectomy and estradiol on different cardiovascular but also extracardiac structures.     

Anticonvulsive Effects and Pharmacokinetic Profile of Cannabidiol (CBD) in the Pentylenetetrazol (PTZ) or N-Methyl-D-Aspartate (NMDA) Models of Seizures in Infantile Rats

In spite of use of cannabidiol (CBD), a non-psychoactive cannabinoid, in pediatric patients with epilepsy, preclinical studies on its effects in immature animals are very limited. In the present study we investigated anti-seizure activity of CBD (10 and 60 mg/kg administered intraperitoneally) in two models of chemically induced seizures in infantile (12-days old) rats. Seizures were induced either with pentylenetetrazol (PTZ) or N-methyl-D-aspartate (NMDA). In parallel, brain and plasma levels of CBD and possible motor adverse effects were assessed in the righting reflex and the bar holding tests. CBD was ineffective against NMDA-induced seizures, but in a dose 60 mg/kg abolished the tonic phase of PTZ-induced generalized seizures. Plasma and brain levels of CBD were determined up to 24 h after administration. Peak CBD levels in the brain (996 ± 128 and 5689 ± 150 ng/g after the 10- and 60-mg/kg doses, respectively) were reached 1–2 h after administration and were still detectable 24 h later (120 ± 12 and 904 ± 63 ng/g, respectively). None of the doses negatively affected motor performance within 1 h after administration, but CBD in both doses blocked improvement in the bar holding test with repeated exposure to this task. Taken together, anti-seizure activity of CBD in infantile animals is dose and model dependent, and at therapeutic doses CBD does not cause motor impairment. The potential risk of CBD for motor learning seen in repeated motor tests has to be further examined.     

Antioxidant and antiproliferative activities of mango (Mangifera indica L.) flesh and peel

The antioxidant and antiproliferative properties of flesh and peel of mango (Mangifera indica L.) were investigated. The cytoprotective effect of mango flesh and peel extracts on oxidative damage induced by H₂O₂ in a human hepatoma cell line, HepG2, were determined, and the underlying mechanism was examined by a single-cell electrophoresis assay (comet assay). Treatment of HepG2 cell with mango peel extract prior to oxidative stress was found to inhibit DNA damage. The free radical scavenging activities of mango flesh and peel extracts were evaluated by electron spin resonance (ESR). The mango peel extract exhibited stronger free radical scavenging ability on 1,1-diphenyl-2-picrylhydrazyl (DPPH) and alkyl radicals than mango flesh extract, regardless of ripeness. Similarly, peel extract exhibited significant antiproliferative effect against all tested cancer cell lines, compared to that of flesh extract, in a dose-dependent manner. The result also showed that the antiproliferative activity of mango flesh and peel extracts correlated with their phenolic and flavonoid contents. Thus, mango peel, a major by-product obtained during the processing of mango product, exhibited good antioxidant activity and may serve as a potential source of phenolics with anticancer activity.     

Structural Study of the Partially Disordered Full‐Length δ Subunit of RNA Polymerase from Bacillus subtilis

The partially disordered δ subunit of RNA polymerase was studied by various NMR techniques. The structure of the well‐folded N‐terminal domain was determined based on inter‐proton distances in NOESY spectra. The obtained structural model was compared to the previously determined structure of a truncated construct (lacking the C‐terminal domain). Only marginal differences were identified, thus indicating that the first structural model was not significantly compromised by the absence of the C‐terminal domain. Various ¹⁵N relaxation experiments were employed to describe the flexibility of both domains. The relaxation data revealed that the C‐terminal domain is more flexible, but its flexibility is not uniform. By using paramagnetic labels, transient contacts of the C‐terminal tail with the N‐terminal domain and with itself were identified. A propensity of the C‐terminal domain to form β‐type structures was obtained by chemical shift analysis. Comparison with the paramagnetic relaxation enhancement indicated a well‐balanced interplay of repulsive and attractive electrostatic interactions governing the conformational behavior of the C‐terminal domain. The results showed that the δ subunit consists of a well‐ordered N‐terminal domain and a flexible C‐terminal domain that exhibits a complex hierarchy of partial ordering.     

New applications of pHluorin—measuring intracellular pH of prototrophic yeasts and determining changes in the buffering capacity of strains with affected potassium homeostasis

pHluorin is a pH‐sensitive variant of green fluorescent protein for measuring intracellular pH (pH_in) in living cells. We constructed a new pHluorin plasmid with the dominant selection marker KanMX. This plasmid allows pH measurements in cells without auxotrophic mutations and/or grown in chemically indefinite media. We observed differing values of pH_in for three prototrophic wild‐types. The new construct was also used to determine the pH_in in strains differing in the activity of the plasma membrane Pma1 H⁺‐ATPase and the influence of glucose on pH_in. We describe in detail pHluorin measurements performed in a microplate reader, which require much less hands‐on time and much lower cell culture volumes compared to standard cuvettes measurements. We also utilized pHluorin in a new method of measuring the buffering capacity of yeast cell cytosol in vivo, shown to be ca. 52 mM /pH for wild‐type yeast and moderately decreased in mutants with affected potassium transport. Copyright © 2010 John Wiley & Sons, Ltd.     

Analysis of Causes of Falsework Failures in Concrete Structures

A study of 85 major falsework collapses of bridges and buildings in the past 23 yrs has documented the types of collapsed falseworks and failed permanent structures, the construction stages at the time of collapse, and the causes of failures. Three causes of failure were identified: triggering; enabling; and procedural causes. Most failures occurred because of the interaction of the triggering and enabling events that were, in many cases, produced by inadequacies in the procedural methods. Impact forces resulting from concreting operations have repeatedly triggered falsework failures that were enabled by deficiencies in the falsework bracings, components, connections, foundations, and design. Inadequate review of falsework design and monitoring procedures were frequent problems that facilitated the occurrernce of these events. The findings emphasize the importance of proper delineation of responsibility of each party in the building process in order to reduce falsework failures in the future.     

Plasma processes at atmospheric and low pressures

In the last few decades there has been an intense development in non-equilibrium (“cold”) plasma surface processing systems at atmospheric pressure. This new trend is stimulated mainly to decrease equipment costs by avoiding expensive pumping systems of conventional low-pressure plasma devices. This work summarizes physical and practical limitations where atmospheric plasmas cannot compete with low-pressure plasma and vice-versa. As the processing conditions for atmospheric plasma are rather different from reduced pressure systems in many cases these conditions may increase final equipment costs substantially. In this work we briefly review the main principles, advantages and drawbacks of atmospheric plasma for a better understanding of the capabilities and limitations of the atmospheric plasma processing technology compared with conventional low-pressure plasma processing.