Hsp70 promotes epithelial sodium channel functional expression by increasing its association with coat complex II and its exit from endoplasmic reticulum

The epithelial sodium channel (ENaC) plays an important role in the homeostasis of blood pressure and of the airway surface liquid, and inappropriate regulation of ENaC results in refractory hypertension (in Liddle syndrome) and impaired mucociliary clearance (in cystic fibrosis). The regulation of ENaC by molecular chaperones, such as the 70-kDa heat shock protein Hsp70, is not completely understood. Building on the previous suggestion by our group that Hsp70 promotes ENaC functional and surface expression in Xenopus oocytes, we investigated the mechanism by which Hsp70 acts upon ENaC in epithelial cells. In Madin-Darby canine kidney cells stably expressing epitope-tagged αβγ-ENaC and with tetracycline-inducible overexpression of Hsp70, treatment with 1 or 2 μg/ml doxycycline increased total Hsp70 expression ~2-fold and ENaC functional expression ~1.4-fold. This increase in ENaC functional expression corresponded to an increase in ENaC expression at the apical surface of the cells and was not present when an ATPase-deficient Hsp70 was similarly overexpressed. The increase in functional expression was not due to a change in the rate at which ENaC was retrieved from the apical membrane. Instead, Hsp70 overexpression increased the association of ENaC with the Sec24D cargo recognition component of coat complex II, which carries protein cargo from the endoplasmic reticulum to the Golgi. These data support the hypothesis that Hsp70 promotes ENaC biogenesis and trafficking to the apical surface of epithelial cells.     

Radix/Rhizoma Notoginseng extract (Sanchitongtshu) for ischemic stroke: A randomized controlled study

Agents of sanchi have been widely used as a complementary medicine for stroke in China. Sanchitongshu is a new Chinese patent medicine extracted from sanchi which has stronger anti-platelet activity than other agents of sanchi. Our aim was to investigate the synergistic action of low dose of aspirin combined with sanchitongshu capsule in the treatment of patients with light and moderate ischemic stroke in acute and subacute stages.This was a multi-center, double-blinded, randomized controlled clinical trial conducted in four hospitals in China from July 2004 to 2006. 140 patients of ischemic stroke in anterior cerebral circulation within 30 days of onset were enrolled. Participants were assigned either to receive aspirin (50 mg per day) and sanchitongshu capsule (200 mg three times a day) or aspirin (50 mg per day) and placebo capsule.Low dose of aspirin combined with sanchitongshu capsule significantly ameliorated neurological deficit (increased score of ESS: t = −5.02, p < 0.0001) and activities of daily living (increased score of BI: t = −2.4, p = 0.0178) after treatment compared with aspirin alone. Adverse reaction which occurred equally in both arms, was light to moderate and disappeared without special treatment.Sanchitongshu capsule, as a complementary medicine to aspirin, was effective in improving outcomes after ischemic stroke. It was a safe drug in our trial.     

Biochemical characterisation of the D60A mutant of the elongation factor 1α from the archaeon Sulfolobus solfataricus

The D60A mutant of the elongation factor (EF) 1α from Sulfolobus solfataricus (Ss), was obtained as heterologous expressed protein and characterised. This substitution was carried out in order to analyse the involvement of this evolutionally conserved amino acid position in the interaction between the elongation factor and guanosine nucleotides and in the coordination of magnesium ions. The expression system used produced a folded protein able to catalyse, although to a slightly lower extent with respect to the wild-type enzyme, protein synthesis in vitro and NaCl-dependent intrinsic GTPase activity. The affinity for guanosine nucleotides was almost identical to that exhibited by wild-type SsEF-1α; vice versa, the GDP exchange rate was one order of magnitude faster on the mutated elongation factor, a property partially restored when the exchange reaction was analysed in the presence of the magnesium ions chelating agent EDTA. Finally, the D60A substitution only a little affected the high thermal stability of the elongation factor. From a structural point of view, the analysis of the data reported confirmed that this conserved carboxyl group belongs to a protein region differentiating the GDP binding mode among elongation factors from different organisms.     

In vitro antibacterial activities of methanol and aqueous leave extracts of selected medicinal plants against human pathogenic bacteria

People use medicinal plants as diet, and for treatment of infectious and noninfectious diseases and they use brief procedures like frying and cooking to do so. Medicinal plants; Moringa oleifera, Azadirachta indica, and Lepidium sativum which is believed to have active components that help to treat and manage various diseases were investigated for their antibacterial activities against Staphylococcus aureus, Salmonella Typhi, Streptococcus agalactiae and Shigella boydii. Solvent methanol and aqueous were used for extraction of crudes by means of maceration. Susceptibility testing was determined by using disc diffusion method and Minimum inhibitory concentration was determined by broth dilution method. Heat treated plant material activity against test pathogen was aimed to identify resistance capacity of plant material at different interval of time and temperature. All plant extracts under study was active against all tested pathogen after exposure to 45 °C for 30 min. The antibacterial activities of the non-heat-treated extracts of Azadirachta indica were relatively low; the results of this study show that extracts of Azadirachta indica have better residual antibacterial activities. Methanol extracts of all plant leaves showed highest activity as compared to the aqueous extracts. This is probably assigning the choice of extraction solvent for extracting desired active phytochemical from plants. Many of the people in the study area were illiterate and they did not have awareness about the ways use of medicinal plants. They use the medicinal plants by cooking and frying for different purposes. In the main, plant material can be affected as the temperature of treatment is increases with respect to various times of exposures.     

Heat shock induces oxidative stress in rotan Perccottus glenii tissues

The effects of temperature transition from 19 to 32 °C on oxidative stress indices and activities of the main antioxidant enzymes were investigated in the rotan, Perccottus glenii. Levels of lipid peroxides (LOOH), thiobarbituric acid-reactive substances (TBARS), low- (L-SH) and high-molecular mass (H-SH) thiols and activities of superoxide dismutase (SOD) and catalase were measured in rotan brain, liver and muscle over 1–12 h of high-temperature exposure followed by 3 or 24 h lower (19 °C) temperature recovery. Heat shock exposure during 1 h transiently increased 1.5–3.2-fold LOOH levels in rotan tissues with subsequent suppression of their content; however, 12 h exposure again increased LOOH levels in the brain. TBARS content were elevated by 2–3-fold during the entire heat shock exposure in the brain and liver. Levels of both products of lipid peroxidation were generally near control values during return to 19 °C. L-SH content was lowered during heat shock exposure in the brain, transiently increased after 6 h in the liver and almost disappeared after longer treatment in the muscle. Liver H-SH content slightly decreased under heat shock exposure, but was elevated after 6 h in the brain and muscle. In the latter case, L-SH level was below control values during recovery. SOD activities increased 2-fold in the liver after 6–12 h heat shock. Liver catalase activities decreased at the same conditions. Generally, a quick response to suppression of lipid peroxidation and possible involvement of its products in the up-regulation of antioxidant enzymes seem to be key adaptations to high temperature.     

Is Female Visual Signaling to Male Song Socially Regulated in Brown‐headed Cowbirds?

This research focused on how adult female brown‐headed cowbirds, Molothrus ater, regulate social feedback on a group level to shape the development of male song. Specifically, females produce rapid wing movements in response to male song, termed ‘wing strokes,’ which have been shown to shape male song and predict song quality. These effects have been documented in captive dyads and triads, but not in more naturalistic flocks, where song development actually occurs. Here, we studied wing stroking in small seminatural flocks of differing female‐to‐male ratios. Despite differences in the number of females and their social selectivity, the same pattern of female feedback emerged in seven of eight flocks: One female produced the majority of wing strokes to male song, making her the primary wing stroker in her flock. Previous studies on large flocks have demonstrated females to facilitate male song improvisation and development if they exhibited higher social selectivity by approaching immature males less. Here, we found that primary wing strokers were indeed more socially selective than non‐primary wing strokers. This research is the first to document social stimulation being facilitated at the group level to ensure that more highly selective females deliver the most feedback.