Differential Effect of DDT,DDE, and DDD on COX‐2 Expression in the Human Trophoblast Derived HTR‐8/SVneo Cells

The purpose of this study was to investigate the effect of 1,1,1‐trichloro‐2,2‐bis‐(chlorophenyl)ethane (DDT), 1,1‐bis‐(chlorophenyl)‐2,2‐dichloroethene (DDE), and 1,1‐dichloro‐2,2‐bis(chlorophenyl)ethane (DDD) isomers on COX‐2 expression in a human trophoblast‐derived cell line. Cultured HTR‐8/SVneo trophoblast cells were exposed to DDT isomers and its metabolites for 24 h, and COX‐2 mRNA and protein expression were assessed by RT‐PCR, Western blotting, and ELISA. Prostaglandin E₂ production was also measured by ELISA. Both COX‐2 mRNA and protein were detected under control (unexposed) conditions in the HTR‐8/SVneo cell line. COX‐2 protein expression and prostaglandin E₂ production but not COX‐2 mRNA levels increased only after DDE and DDD isomers exposure. It is concluded that DDE and DDD exposure induce the expression of COX‐2 protein, leading to increased prostaglandin E2 production. Interestingly, the regulation of COX‐2 by these organochlorines pesticides appears to be at the translational level. © 2012 Wiley Periodicals, Inc. J Biochem Mol Toxicol 26:454‐460, 2012; View this article online at wileyonlinelibrary.com . DOI 10:1002/jbt.21444     

Kilovoltage beam model for flat panel imaging system with bow-tie filter for scatter prediction and correction

PurposeKilovoltage flat-panel imaging systems are used for cone-beam Computed Tomography (CBCT) and digital Tomosynthesis (DTS). Hereby, the presence of scatter and relatively large dose from imaging are challenging factors. In this study a phenomenological beam model was developed to characterize imager response to imaging beams with a bow-tie filter (Varian OBI system).Materials and methodThe kilovoltage beam model was based on dose ratio formalism and thus was using standard concepts of megavoltage dose calculation such as scatter factors, tissue maximum ratio and off-axis ratio. Primary and scatter (head and phantom scatter) were modeled with three Gaussian kernels. Parameters were based on measured transmission images for slabs of solid water of different total thickness and various jaw settings.ResultsThe beam model was used to evaluate contributions from primary, secondary and tertiary contributions for different geometrical objects such as cylinders and step-like phantoms. Theoretical predictions of radiographs using the model for known objects are consistent with the measurements.ConclusionSecondary and tertiary contributions were interpreted as scatter and can be subtracted from CBCT projections based on the analytical model. Therefore our model can provide a basis for improvement of image quality (less artifacts due to scatter, better contrast and resolution) in CBCT reconstruction.     

Usefulness of combining intermittent hypoxia and physical exercise in the treatment of obesity

Obesity is an important public health problem worldwide and is a major risk factor for a number of chronic diseases such as type II diabetes, adverse cardiovascular events and metabolic syndrome-related features. Different treatments have been applied to tackle body fat accumulation and its associated clinical manifestations. Often, relevant weight loss is achieved during the first 6 months under different dietary treatments. From this point, a plateau is reached, and a gradual recovery of the lost weight may occur. Therefore, new research approaches are being investigated to assure weight maintenance. Pioneering investigations have reported that oxygen variations in organic systems may produce changes in body composition. Possible applications of intermittent hypoxia to promote health and in various pathophysiological states have been reported. The hypoxic stimulus in addition to diet and exercise can be an interesting approach to lose weight, by inducing higher basal noradrenalin levels and other metabolic changes whose mechanisms are still unclear. Indeed, hypoxic situations increase the diameter of arterioles, produce peripheral vasodilatation and decrease arterial blood pressure. Furthermore, hypoxic training increases the activity of glycolytic enzymes, enhancing the number of mitochondria and glucose transporter GLUT-4 levels as well as improving insulin sensitivity. Moreover, hypoxia increases blood serotonin and decreases leptin levels while appetite is suppressed. These observations allow consideration of the hypothesis that intermittent hypoxia induces fat loss and may ameliorate cardiovascular health, which might be of interest for the treatment of obesity. This new strategy may be useful and practical for clinical applications in obese patients.     

Involvement of astrocytes in transmissible spongiform encephalopathies: a confocal microscopy study

Astroglial proliferation associated with pathological prion protein (PrPsc) deposition is widely described in Transmissible Spongiform Encephalopathies (TSEs). However, little is known of the actual role played by glia in their pathogenesis. The aim of the study has been to determine whether PrPsc is located exclusively in neurons or in both neurons and glial cells present in the central nervous system in a natural Scrapie model. Samples of cerebellum from 25 Scrapie sheep from various flocks were sectioned. Following epitope retrieval with formic acid, proteinase K and heat treatment, primary antibody L42 and primary antibodies against glial fibrillary acidic protein were applied as prion- and astrocytic-specific markers, respectively. For visualization, a suitable mixture of fluorochrome-conjugated secondary antibodies was used. Relevant controls were processed in the same manner. As determined by confocal microscopy, PrPsc deposits co-localized with glial cells in all samples. Our results suggest that these cells can sustain active prion propagation, in agreement with similar findings from other studies of primary cell cultures and inoculated mice. Furthermore, despite ongoing debate regarding whether varied TSE sources show differences in their tropism for different cell lineages in the brains of affected animals, no differences in co-localization results were seen.     

RNA self-cleavage activated by ultraviolet light-induced oxidation

A novel UV-C-light-induced ribozyme activity was discovered within the highly structured 5'-genomic regions of both Hepatitis C Virus (HCV) and the related Classic Swine Fever Virus (CSFV). Cleavage is mediated by exposure to UV-C light but not by exogenous oxygen radicals. It is also very selective, occurring at base positions HCV C(79) and CSFV A(45) in some molecules and at the immediately adjacent 5'-positions HCV U(78) and CSFV U(44) in others. Among other reaction products, the majority of biochemically active products detected contained 3'-phosphate and 5'-phosphate-end groups at the newly generated termini, along with a much lower amount of 3'-hydroxyl end group. While preservation of an E-loop RNA structure in the vicinity of the cleavage site was a requisite for HCV RNA self-cleavage, this was not the case for CSFV RNA. The short size of the reactive domains (~33 nt), which are compatible with primitive RNA motifs, and the lack of sequence homology, indicate that as-yet unidentified UV-activated ribozymes are likely to be found throughout structured RNAs, thereby providing clues to whether early RNA self-cleavage events were mediated by photosensitive RNA structures.     

Microscopic origin of gating current fluctuations in a potassium channel voltage sensor

Voltage-dependent ion channels open and close in response to changes in membrane electrical potential due to the motion of their voltage-sensing domains (VSDs). VSD charge displacements within the membrane electric field are observed in electrophysiology experiments as gating currents preceding ionic conduction. The elementary charge motions that give rise to the gating current cannot be observed directly, but appear as discrete current pulses that generate fluctuations in gating current measurements. Here we report direct observation of gating-charge displacements in an atomistic molecular dynamics simulation of the isolated VSD from the KvAP channel in a hydrated lipid bilayer on the timescale (10-μs) expected for elementary gating charge transitions. The results reveal that gating-charge displacements are associated with the water-catalyzed rearrangement of salt bridges between the S4 arginines and a set of conserved acidic side chains on the S1-S3 transmembrane segments in the hydrated interior of the VSD.