Evidence for a dinuclear active site in the metallo-beta-lactamase BcII with substoichiometric Co(II). A new model for metal uptake

Metallo-beta-lactamases are zinc-dependent enzymes that constitute one of the main resistance mechanisms to beta-lactam antibiotics. Metallo-beta-lactamases have been characterized both in mono- and dimetallic forms. Despite many studies, the role of each metal binding site in substrate binding and catalysis is still unclear. This is mostly due to the difficulties in assessing the metal content and site occupancy in solution. For this reason, Co(II) has been utilized as a useful probe of the active site structure. We have employed UV-visible, EPR, and NMR spectroscopy to study Co(II) binding to the metallo-beta-lactamase BcII from Bacillus cereus. The spectroscopic features were attributed to the two canonical metal binding sites, the 3H (His(116), His(118), and His(196)) and DCH (Asp(120), Cys(221), and His(263)) sites. These data clearly reveal the coexistence of mononuclear and dinuclear Co(II)-loaded forms at Co(II)/enzyme ratios as low as 0.6. This picture is consistent with the macroscopic dissociation constants here determined from competition binding experiments. A spectral feature previously assigned to the DCH site in the dinuclear species corresponds to a third, weakly bound Co(II) site. The present work emphasizes the importance of using different spectroscopic techniques to follow the metal content and localization during metallo-beta-lactamase turnover.     

Trehalose and trehalose-hydrolyzing enzyme in the haemolymph of Locusta migratoria infected with Metarhizium anisopliae strain CQMa102

Topical application of the Metarhizium anisopliae var. acridum specialist strain CQMa 102 to the locust Locusta migratoria manilensis results in changes of the concentrations of trehalose and glucose in the haemolymph. Micrographs of the locust haemolymph shows Metarhizium anisopliae can effectivly penetrate the external skeleton of locust and after 2 days infection, the hyphae body will appear in the haemolymph of infected insects. The time in decrease of trehalose concentration coincided with that in increase of trehalose-hydrolysing enzyme activity in the haemolymph of the fungus-infected insects. Overlay gel analysis indicated there was considerably more trehalose-hydrolysing activity in the haemolymph of locusts infected by fungus than in controls. A comparable isoform was identified in in vitro culture of the fungus, suggesting a fungal origin for the in vivo enzyme. Haemolymph trehalose decreased significantly during mycosis of locusts by M. anisopliae. All these results suggested that this fungus may take advantage of competing nutrient utilization against the insect by its trehalose-hydrolyzing enzyme secretion. It may provide fundamental knowledge for fungal pathogenesis.     

Temporal genetic differentiation in cultured and natural beds of the brown mussel Perna perna (Mytilidae)

Perna perna is the most important cultivated mussel of Santa Catarina, Brazil, sustaining an important economic input for many local families. Natural stocks of P. perna are depleted by the extraction of adults and seeds for consumption and culture. The aim of the present study was to use the microsatellite locus pms-2 to study the variation of the genetic composition and diversity between natural and cultured stocks in samples of 2001 and 2005 from Penha, Santa Catarina. DNA was extracted from adductor muscle by Chelex/proteinase-K and phenol/chloroform protocols. Amplification by polymerase chain reaction was performed using specific primers for analyzing the pms-2 locus. Polymerase chain reaction products were submitted to vertical denatured 6% polyacrylamide gel electrophoresis and horizontal 2% agarose gel electrophoresis, and visualized by silver staining and ethidium bromide, respectively. Allele diversity and heterozygote deficiency were higher for samples of 2005 than for those of 2001. No significant genetic differentiation was found between natural and cultured stocks of 2001 by the chi(2) test, but G(2) (likelihood ratio) detected slight differences (I = 0.949; chi(2), P = 0.147; G(2), P = 0.046), while cultured and natural stocks of 2005 were very different (I = 0.798, P = 0.006). Between the years of 2001 and 2005, a large change in genetic composition was observed (I = 0.582; P < 0.001). Although nothing is known about natural changes in the genetic composition of this species with time, the results suggest a strong impact of human activities on natural stocks of P. perna, which is expected to be related to heavy extraction and farming.     

长江源区同域分布兔狲、藏狐和赤狐的时空重叠度

兔狲(Otocolobus manul)、藏狐(Vulpes ferrilata)和赤狐(V. vulpes)是青藏高原三江源区域分布的重要小型食肉兽。本研究于2014年6月至2019年9月在青海省长江源区沱沱河和通天河沿岸选取208个位点布设红外相机, 通过所获取的时空分布数据比较了上述3种同域分布小型食肉兽的时空利用情况。通过空间重叠度系数的比较分析, 兔狲和藏狐、兔狲和赤狐以及藏狐和赤狐之间的空间重叠度系数分别为0.25、0.48和0.17, 这表明兔狲、藏狐和赤狐三者在空间利用上存在一定的差异。通过核密度估计方法分析, 兔狲和藏狐属典型的昼行性动物, 而赤狐以夜行性活动为主。兔狲、藏狐和赤狐每个物种在冷暖两季的日活动节律重叠指数分别为0.83、0.78和0.88。两两比较分析表明, 兔狲和藏狐二者的日活动节律重叠指数最高(0.84), 兔狲和赤狐在夜间活动时段存在一定重叠(0.63), 而藏狐和赤狐的时间生态位分化最明显, 重叠指数最低(0.48)。此外, 在暖季, 两两物种之间的日活动节律重叠指数均小于其冷季的重叠指数。综上所述, 长江源区兔狲、藏狐和赤狐3种小型食肉兽可通过空间和时间资源的利用差异来降低物种间的干扰和竞争, 从而达到同域物种共存的目的。     

稻飞虱天敌螯蜂研究进展

稻飞虱（褐飞虱Nilaparvata lugens、白背飞虱Sogatella furcifera、灰飞虱Laodelphax striatellus）是世界性重要的水稻Oryza sativa L.害虫之一,给水稻生产造成了重大经济损失。化学防治一直是控制稻飞虱的主要途径,但长期使用化学药剂使稻飞虱产生抗药性,并引起害虫增殖等诸多弊端,迫切需要有效的生物防控手段进行控害。螯蜂是稻飞虱若虫和成虫期重要天敌,兼具捕食与寄生的双重习性,在控制稻飞虱种群数量方面发挥着重要的作用,然而,国内外有关螯蜂的研究报道仍然偏少。本文综述了我国稻飞虱天敌螯蜂常见种类、生物学特性、控害效果及其影响因子,分析了存在的问题,旨在为进一步开发利用螯蜂资源提供参考。     

Leber congenital amaurosis and retinitis pigmentosa with Coats-like exudative vasculopathy are associated with mutations in the crumbs homologue 1 (CRB1) gene

Mutations in the crumbs homologue 1 (CRB1) gene cause a specific form of retinitis pigmentosa (RP) that is designated "RP12" and is characterized by a preserved para-arteriolar retinal pigment epithelium (PPRPE) and by severe loss of vision at age <20 years. Because of the early onset of disease in patients who have RP with PPRPE, we considered CRB1 to be a good candidate gene for Leber congenital amaurosis (LCA). Mutations were detected in 7 (13%) of 52 patients with LCA from the Netherlands, Germany, and the United States. In addition, CRB1 mutations were detected in five of nine patients who had RP with Coats-like exudative vasculopathy, a relatively rare complication of RP that may progress to partial or total retinal detachment. Given that four of five patients had developed the complication in one eye and that not all siblings with RP have the complication, CRB1 mutations should be considered an important risk factor for the Coats-like reaction, although its development may require additional genetic or environmental factors. Although no clear-cut genotype-phenotype correlation could be established, patients with LCA, which is the most severe retinal dystrophy, carry null alleles more frequently than do patients with RP. Our findings suggest that CRB1 mutations are a frequent cause of LCA and are strongly associated with the development of Coats-like exudative vasculopathy in patients with RP.     

Electric and magnetic fields of the closed depolarization front in the anisotropic myocardium

On the basis of the bidomain model of anisotropic myocardium, mathematical relationships for calculating the characteristics of the extracellular electric and magnetic fields outside a closed or open depolarization front were derived on the assumption that the generator and fields are axially symmetric with respect to the longitudinal direction of the fibres making up the excitable tissue. The spatial configuration of the fields was determined and quantitative estimates were obtained for the electric potential and magnetic induction caused solely by the macroscopic anisotropy of the myocardium tissue as a volume conductor.