从害虫马尾松毛虫中提取甲壳素的初步研究

采用酸碱法对马尾松毛虫蛹中甲壳素的提取方法进行了初步研究。重点分析了N aOH不同处理条件对脱有机物质和甲壳素产率的影响,确定了从马尾松毛虫蛹中提取甲壳素的基本工艺:(1)脱矿物质:盐酸浓度为2.5%,浸泡时间为20 h,浸泡温度为30℃;(2)脱有机物质:N aOH浓度为6%,浸泡温度为75℃,浸泡时间为10 h;(3)脱色:双氧水浓度为9%,浸泡时间为2.5 h,浸泡温度为80℃。在此条件下,获得的甲壳素产品为白色粉状固体,其N含量为6.87%,灰分为1.19%,水分为8.37%。产率为29.97%。产品的N含量达到食品级甲壳素标准,灰分含量达到工业级甲壳素标准。实验有助于后续深入研究马尾松毛虫蛹中甲壳素的提取,也为今后进一步制备虫蛹壳聚糖打下了基础。     

Secreted kinase phosphorylates extracellular proteins that regulate biomineralization

Protein phosphorylation is a fundamental mechanism regulating nearly every aspect of cellular life. Several secreted proteins are phosphorylated, but the kinases responsible are unknown. We identified a family of atypical protein kinases that localize within the Golgi apparatus and are secreted. Fam20C appears to be the Golgi casein kinase that phosphorylates secretory pathway proteins within S-x-E motifs. Fam20C phosphorylates the caseins and several secreted proteins implicated in biomineralization, including the small integrin-binding ligand, N-linked glycoproteins (SIBLINGs). Consequently, mutations in Fam20C cause an osteosclerotic bone dysplasia in humans known as Raine syndrome. Fam20C is thus a protein kinase dedicated to the phosphorylation of extracellular proteins.     

Increase of resistance of <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> plants to phytopathogenic fungi expressing hevein-like peptides from weed plant <Emphasis Type="Italic">Stellaria media</Emphasis>

It is shown that constitutive hyperexpression of new hevein-like peptides from the weed plant chickweed (Stellaria media)in mouse-ear cress (Arabidopsis thaliana) plants leads to a substantial increase of their resistance to phytopathogenic fungi Botrytis cinerea and Bipolaris sorokiniana. Thus, common chickweed peptides can play a definite role in protecting this weed plant and be useful as a new genetic tool for producing plants resistant to fungal diseases.     

Creation of a protein vector construct including an SSBTne DNA-binding domain and VirD2 nuclear localization signal

A genetic engineering construct is created which encodes a chimeric protein for nonviral transgenesis that includes the SSB DNA-binding domain and VirD2 nuclear localization signal. A method is developed for purifying the target protein.     

Effect of a Single Dose of Cadmium on Pregnant Wistar Rats and their Offspring

Cadmium (Cd) is a well‐known toxicant targeting many organs, among them placenta. This heavy metal also has embryonary and foetal toxicity. This study was undertaken to analyse the effect of a single Cd dose administered at 4, 7, 10 or 15 days of gestation on the offspring of pregnant rats sacrificed at 20 days of gestation. Cadmium chloride was administered subcutaneously at 10 mg/kg body weight to Wistar pregnant dams; control animals received a proportionate volume of sterile normal saline by the same route. Maternal uteri, livers, kidneys and lungs, and foetuses were examined at necropsy. Samples of maternal organs and whole foetuses were collected for histopathologic examination, determination of Cd levels and staining by the Alizarin red S technique. Results revealed a clear embryotoxic and a teratogenic effect of this heavy metal, the former as a significant increase in the number of resorptions, and the latter as significant decrease of the gestational sac weight, and the size and weight of foetuses of Cd‐treated dams as well as induced malformations in skull bones, vertebrae and thoracic, and pelvian limbs. The deleterious effects found were similar to those previously reported for other animal models suggesting a high conservation of the pathogenic mechanisms of Cd. Additionally, many of the addressed aspects showed a slight dependence on the time of administration of the toxic that might be due to the accumulation of the metal in different organs, as we were able to demonstrate by the analysis of its concentration.     

Polypeptide Modulators of TRPV1 Produce Analgesia without Hyperthermia

Transient receptor potential vanilloid 1 receptors (TRPV1) play a significant physiological role. The study of novel TRPV1 agonists and antagonists is essential. Here, we report on the characterization of polypeptide antagonists of TRPV1 based on in vitro and in vivo experiments. We evaluated the ability of APHC1 and APHC3 to inhibit TRPV1 using the whole-cell patch clamp approach and single cell Ca²⁺ imaging. In vivo tests were performed to assess the biological effects of APHC1 and APHC3 on temperature sensation, inflammation and core body temperature. In the electrophysiological study, both polypeptides partially blocked the capsaicin-induced response of TRPV1, but only APHC3 inhibited acid-induced (pH 5.5) activation of the receptor. APHC1 and APHC3 showed significant antinociceptive and analgesic activity in vivo at reasonable doses (0.01–0.1 mg/kg) and did not cause hyperthermia. Intravenous administration of these polypeptides prolonged hot-plate latency, blocked capsaicin- and formalin-induced behavior, reversed CFA-induced hyperalgesia and produced hypothermia. Notably, APHC3’s ability to inhibit the low pH-induced activation of TRPV1 resulted in a reduced behavioural response in the acetic acid-induced writhing test, whereas APHC1 was much less effective. The polypeptides APHC1 and APHC3 could be referred to as a new class of TRPV1 modulators that produce a significant analgesic effect without hyperthermia.     

Autosomal recessive hypercholesterolemia caused by mutations in a putative LDL receptor adaptor protein

Atherogenic low density lipoproteins are cleared from the circulation by hepatic low density lipoprotein receptors (LDLR). Two inherited forms of hypercholesterolemia result from loss of LDLR activity: autosomal dominant familial hypercholesterolemia (FH), caused by mutations in the LDLR gene, and autosomal recessive hypercholesterolemia (ARH), of unknown etiology. Here we map the ARH locus to an approximately 1-centimorgan interval on chromosome 1p35 and identify six mutations in a gene encoding a putative adaptor protein (ARH). ARH contains a phosphotyrosine binding (PTB) domain, which in other proteins binds NPXY motifs in the cytoplasmic tails of cell-surface receptors, including the LDLR. ARH appears to have a tissue-specific role in LDLR function, as it is required in liver but not in fibroblasts.