Removal of estrogenic activity from endocrine-disrupting chemicals by purified laccase of Phlebia tremellosa

A white-rot basidiomycete, Phlebia tremellosa, produced a laccase that showed increased activity during degradation of phthalates. A laccase was purified through the ion exchange chromatography and preparative gel electrophoresis, and the estimated molecular weight was 75 kDa. The optimum pH and temperature of the purified laccase was pH 4.0 and 20 degrees C, respectively. The K(m) value of the enzyme was 55.7 microM, and the V(max) was 0.0541 OD min(-1) U(-1) for o-tolidine. Purified laccase reduced the estrogenic activity of four different endocrine-disrupting chemicals. However, this effect was reduced by a laccase inhibitor, kojic acid, which confirmed that the laccase was involved in the removal of estrogenic activity.     

Monteiro's Storm‐petrel Oceanodroma monteiroi: a new species from the Azores

The existence of two seasonally distinct breeding populations of Oceanodroma storm‐petrels in the Azores islands was first documented in 1996. The discovery of morphological differences between the populations led to the suggestion that they may represent cryptic sibling species. Recent mtDNA and microsatellite analysis from storm‐petrel populations has considerably advanced our understanding of their taxonomic relationships. Here we present new information on the timing of breeding and moult of the two Azores populations, the extent of exchange of individuals between seasons, and diet from feather isotopes. We conclude that the hot‐season Azores population should be considered a new species for which we propose the name Oceanodroma monteiroi, Monteiro's Storm‐petrel. The species is both genetically distinct and genetically isolated from the sympatric cool‐season population of Madeiran Storm‐petrel Oceanodroma castro, and from all other populations of Oceanodroma castro in the Atlantic and Pacific Oceans examined to date. Differences in the vocalizations permit species recognition, and the extent of primary feather wear and stage of moult aids separation of the two species in the Azores, which is especially valuable during August when both attend the breeding colonies in large numbers. Feather carbon and nitrogen isotopes reveal that the diet of Monteiro's Storm‐petrel differs from that of the sympatric Madeiran Storm‐petrel during both breeding and non‐breeding seasons, and unlike the Madeiran Storm‐petrel, Monteiro's Storm‐petrel appears to maintain the same foraging environment during the summer and winter months, though it shows a dietary shift to higher trophic levels during the non‐breeding season. Monteiro's Storm‐petrel is thought to be confined to the Azores archipelago, where it is currently known to nest on just two small neighbouring islets. The total population size was estimated at 250–300 pairs in 1999.     

Haematozoa infections in a Great Tit Parus major population in Central Portugal: relationships with breeding effort and health

Blood parasites may act as modulators of their hosts' ecology, life histories and fitness. We studied the prevalence of Plasmodium sp., Haemoproteus sp. and Leucocytozoon sp. and their effects on morphological, biochemical and haematological variables and on breeding effort of Great Tits Parus major . Total prevalence (percentage of individuals infected by any parasite) ranged from 7.7% to 61.1%. There was an overall positive association in prevalence between the three haematozoan parasites. No effect of sex or age on infection status was observed. Negative impacts of infection on physiological condition depended largely on year and/or season and included effects on body condition index, plasma protein and haemoglobin index. There were also indications that parasite infection increased immune response and stress levels and activated antioxidant defence mechanisms. Males with higher fledging success had a higher probability of Haemoproteus infection, and females laying heavier eggs had a higher probability of Plasmodium infection. However, clutch size was negatively associated with the probability of infection by Leucocytozoon and Haemoproteus . Surprisingly, males raising second broods had a lower prevalence of both Haemoproteus and Leucocytozoon . Only 5.7% of first-brood nestlings were infected, but those in infected nestboxes had a lower heterophil/lymphocyte ratio. This study confirms the pathogenicity of blood parasites to the host by demonstrating negative effects of infection on both physiology and breeding performance.     

Environment-friendly adhesives for surface bonding of wood-based flooring using natural tannin to reduce formaldehyde and TVOC emission

The objective of this research was to develop environment-friendly adhesives for face fancy veneer bonding of engineered flooring using the natural tannin form bark in the wood. The natural wattle tannin adhesive were used to replace UF resin in the formaldehyde-based resin system in order to reduce formaldehyde and volatile organic compound (VOC) emissions from the adhesives used between plywoods and fancy veneers. PVAc was added to the natural tannin adhesive to increase viscosity of tannin adhesive for surface bonding. For tannin/PVAc hybrid adhesives, 5%, 10%, 20% and 30% of PVAc to the natural tannin adhesives were added. tannin/PVAc hybrid adhesives showed better bonding than the commercial natural tannin adhesive with a higher level of wood penetration. The initial adhesion strength was sufficient to be maintained within the optimum initial tack range. The standard formaldehyde emission test (desiccator method), field and laboratory emission cell (FLEC) and VOC analyzer were used to determine the formaldehyde and VOC emissions from engineered flooring bonded with commercial the natural tannin adhesive and tannin/PVAc hybrid adhesives. By desiccator method and FLEC, the formaldehyde emission level of each adhesive showed the similar tendency. All adhesives satisfied the E(1) grade (below 1.5 mg/L) and E(0) grade (below 0.5 mg/L) with UV coating. VOC emission results by FLEC and VOC analyzer were different with the formaldehyde emission results. TVOC emission was slightly increased as adding PVAc.     

犬细小病毒VP2蛋白在真核细胞中的分泌表达及特性

摘要:【目的】利用真核细胞分泌表达犬细小病毒VP2蛋白和研究其特性。【方法】为构建犬细小病毒（Canine parvovirus, CPV）VP2基因的真核分泌型表达载体,首先通过酶切从含有人CD5信号肽序列的质粒中将CD5信号肽基因片段切出,将其连接到真核表达载体pcDNA3.1A的多克隆位点上,构建成pcDNA3.1-CD5sp质粒。然后再通过PCR方法从含有犬细小病毒VP2基因的质粒中扩增VP2基因,并将其插入到pcDNA3.1- CD5sp载体中CD5信号肽的下游,构建成VP2基因的真核分泌型表达载体pcDNA-CD5sp-VP2。经磷酸钙介导转染293T细胞,使其在真核细胞中进行分泌表达,并通过ELISA检测表达的VP2蛋白与犬转铁蛋白受体（TfR）结合的活性。【结果】序列分析结果表明,本实验构建的犬细小病毒VP2基因真核分泌型表达载体结构正确,将该表达载体转染的293T细胞,在培养基中通过Western-blot检测到有VP2重组蛋白的存在。经ELISA检测表明表达的重组VP2蛋白具有与犬转铁蛋白受体结合的活性。【结论】 利用人的CD5信号肽实现了犬细小病毒VP2蛋白在真核细胞中的分泌表达,表达的VP2蛋白具有与犬转铁蛋白受体结合的活性。     

Carbon accumulation at depth in Ferralsols under zero‐till subtropical agriculture

Conservation agriculture can provide a low‐cost competitive option to mitigate global warming with reduction or elimination of soil tillage and increase soil organic carbon (SOC). Most studies have evaluated the impact of zero till (ZT) only on surface soil layers (down to 30 cm), and few studies have been performed on the potential for C accumulation in deeper layers (0–100 cm) of tropical and subtropical soils. In order to determine whether the change from conventional tillage (CT) to ZT has induced a net gain in SOC, three long‐term experiments (15–26 years) on free‐draining Ferralsols in the subtropical region of South Brazil were sampled and the SOC stocks to 30 and 100 cm calculated on an equivalent soil mass basis. In rotations containing intercropped or cover‐crop legumes, there were significant accumulations of SOC in ZT soils varying from 5 to 8 Mg ha^?1 in comparison with CT management, equivalent to annual soil C accumulation rates of between 0.04 and 0.88 Mg ha^?1. However, the potential for soil C accumulation was considerably increased (varying from 0.48 to 1.53 Mg ha^?1 yr^?1) when considering the soil profile down to 100 cm depth. On average the estimate of soil C accumulation to 100 cm depth was 59% greater than that for soil C accumulated to 30 cm. These findings suggest that increasing sampling depth from 30 cm (as presently recommended by the IPCC) to 100 cm, may increase substantially the estimates of potential CO₂ mitigation induced by the change from CT to ZT on the free‐draining Ferralsols of the tropics and subtropics. It was evident that that legumes which contributed a net input of biologically fixed N played an important role in promoting soil C accumulation in these soils under ZT, perhaps due to a slow‐release of N from decaying surface residues/roots which favored maize root growth.     

Membrane topological structure of neutral system N/A amino acid transporter 4 (SNAT4) protein

Members of system N/A amino acid transporter (SNAT) family mediate transport of neutral amino acids, including l-alanine, l-glutamine, and l-histidine, across the plasma membrane and are involved in a variety of cellular functions. By using chemical labeling, glycosylation, immunofluorescence combined with molecular modeling approaches, we resolved the membrane topological structure of SNAT4, a transporter expressed predominantly in liver. To analyze the orientation using the chemical labeling and biotinylation approach, the "Cys-null" mutant of SNAT4 was first generated by mutating all five endogenous cysteine residues. Based on predicted topological structures, a single cysteine residue was introduced individually into all possible nontransmembrane domains of the Cys-null mutant. The cells expressing these mutants were labeled with N-biotinylaminoethyl methanethiosulfonate, a membrane-impermeable cysteine-directed reagent. We mapped the orientations of N- and C-terminal domains. There are three extracellular loop domains, and among them, the second loop domain is the largest that spans from amino acid residue ～242 to ～335. The orientation of this domain was further confirmed by the identification of two N-glycosylated residues, Asn-260 and Asn-264. Together, we showed that SNAT4 contains 10 transmembrane domains with extracellular N and C termini and a large N-glycosylated, extracellular loop domain. This is the first report concerning membrane topological structure of mammalian SNAT transporters, which will provide important implications for our understanding of structure-function of the members in this amino acid transporter family.     

Tyr-301 Phosphorylation Inhibits Pyruvate Dehydrogenase by Blocking Substrate Binding and Promotes the Warburg Effect

The mitochondrial pyruvate dehydrogenase complex (PDC) plays a crucial role in regulation of glucose homoeostasis in mammalian cells. PDC flux depends on catalytic activity of the most important enzyme component pyruvate dehydrogenase (PDH). PDH kinase inactivates PDC by phosphorylating PDH at specific serine residues, including Ser-293, whereas dephosphorylation of PDH by PDH phosphatase restores PDC activity. The current understanding suggests that Ser-293 phosphorylation of PDH impedes active site accessibility to its substrate pyruvate. Here, we report that phosphorylation of a tyrosine residue Tyr-301 also inhibits PDH α 1 (PDHA1) by blocking pyruvate binding through a novel mechanism in addition to Ser-293 phosphorylation. In addition, we found that multiple oncogenic tyrosine kinases directly phosphorylate PDHA1 at Tyr-301, and Tyr-301 phosphorylation of PDHA1 is common in EGF-stimulated cells as well as diverse human cancer cells and primary leukemia cells from human patients. Moreover, expression of a phosphorylation-deficient PDHA1 Y301F mutant in cancer cells resulted in increased oxidative phosphorylation, decreased cell proliferation under hypoxia, and reduced tumor growth in mice. Together, our findings suggest that phosphorylation at distinct serine and tyrosine residues inhibits PDHA1 through distinct mechanisms to impact active site accessibility, which act in concert to regulate PDC activity and promote the Warburg effect.     

A 14 000-year record of paleoenvironmental change in the western basin of China's third largest lake,Lake Taihu

Effect of leaf extract of Ocimum sanctum on (i) the specific and non-specific immune responses and (ii) disease resistance against Aeromonas hydrophila was investigated in Oreochromis mossambicus. Sheep red blood cells (SRBC) and Heat aggregated bovine serum albumin (HA-BSA) were used as antigens for specific and non-specific immune response studies, respectively. Antigens were administered through intraperitoneal route. Anti-SRBC antibody titres were determined by direct haemagglutination and anti-bacterial antibodies were determined by bacterial agglutination. Nitroblue tetrazolium (NBT) assay was used to determine neutrophil activity. Disease resistance was measured in terms of relative percent survival (RPS). The immunostimulatory effect of the leaf extract of O. sanctum, when administered through intraperitoneal and oral routes was obvious. Leaf extract of O. sanctum, when administered intraperitoneally, stimulated both antibody response and neutrophil activity. Dietary intake of O. sanctum also enhanced the antibody response and disease resistance against A. hydrophila. Possibility of using O. sanctum as immunostimulant in the maintenance of finfish health in intensive freshwater aquaculture is suggested.