Evolution of the CD163 family and its relationship to the bovine gamma delta T cell co-receptor WC1

Background  

The scavenger receptor cysteine rich (SRCR) domain is an ancient and conserved protein domain. CD163 and WC1 molecules are classed together as group B SRCR superfamily members, along with Spα, CD5 and CD6, all of which are expressed by immune system cells. There are three known types of CD163 molecules in mammals, CD163A (M130, coded for by CD163), CD163b (M160, coded for by CD163L1) and CD163c-α (CD163L1 or SCART), while their nearest relative, WC1, is encoded by a multigene family so far identified in the artiodactyl species of cattle, sheep, and pigs.     

Comparison of defined culture systems for feeder cell free propagation of human embryonic stem cells

There are many reports of defined culture systems for the propagation of human embryonic stem cells in the absence of feeder cell support, but no previous study has undertaken a multi-laboratory comparison of these diverse methodologies. In this study, five separate laboratories, each with experience in human embryonic stem cell culture, used a panel of ten embryonic stem cell lines (including WA09 as an index cell line common to all laboratories) to assess eight cell culture methods, with propagation in the presence of Knockout Serum Replacer, FGF-2, and mouse embryonic fibroblast feeder cell layers serving as a positive control. The cultures were assessed for up to ten passages for attachment, death, and differentiated morphology by phase contrast microscopy, for growth by serial cell counts, and for maintenance of stem cell surface marker expression by flow cytometry. Of the eight culture systems, only the control and those based on two commercial media, mTeSR1 and STEMPRO, supported maintenance of most cell lines for ten passages. Cultures grown in the remaining media failed before this point due to lack of attachment, cell death, or overt cell differentiation. Possible explanations for relative success of the commercial formulations in this study, and the lack of success with other formulations from academic groups compared to previously published results, include: the complex combination of growth factors present in the commercial preparations; improved development, manufacture, and quality control in the commercial products; differences in epigenetic adaptation to culture in vitro between different ES cell lines grown in different laboratories.     

Energy transformation coupled to formate oxidation during anaerobic fermentation

A correlation between the rate of ATP synthesis by F0F1 ATP-synthase and formate oxidation by formate hydrogen lyase (FHL) has been established in inverted membrane vesicles of Escherichia coli JW 136 mutant with double deletions (delta hya/ delta hyb) of hydrogenase 1 and 2 grown anaerobically on glucose in the absence of external electron acceptors (pH 6.5). ATP synthesis was suppressed by H+ -ATPase inhibitors N,N'-dicyclohexylcarbodiimide (DCCD) and sodium azide as well as by the protonophore carbonyl cyanide-m-chlorophenyhydrazone (CCCP). Copper ions inhibited formate-dependent hydrogenase and ATP-synthase activities but did not affect the ATPase activity of vesicles. The maximal rate of ATP synthesis (0.83 microM/min x mg protein) stimulated by K+ ions was determined when sodium formate, ADP and inorganic phosphate were applied simultaneously. The results confirm the assumption about the dual role of hydrogenase 3, formate hydrogen lyase subunit, which is able to couple the reduction of protons to H2 and their translocation through a membrane with chemiosmotic synthesis of ATP.     

Communication between accessory factors and the Cre recombinase at hybrid psi-loxP sites

By placing loxP adjacent to the accessory sequences from the Xer/psi multimer resolution system, we have imposed topological selectivity and specificity on Cre/loxP recombination. In this hybrid recombination system, the Xer accessory protein PepA binds to psi accessory sequences, interwraps them, and brings the loxP sites together such that the product of recombination is a four-node catenane. Here, we investigate communication between PepA and Cre by varying the distance between loxP and the accessory sequences, and by altering the orientation of loxP. The yield of four-node catenane and the efficiency of recombination in the presence of PepA varied with the helical phase of loxP with respect to the accessory sequences. When the orientation of loxP was reversed, or when half a helical turn was added between the accessory sequences and loxP, PepA reversed the preferred order of strand exchange by Cre at loxP. The results imply that PepA and the accessory sequences define precisely the geometry of the synapse formed by the loxP sites, and that this overcomes the innate preference of Cre to initiate recombination on the bottom strand of loxP. Further analysis of our results demonstrates that PepA can stimulate strand exchange by Cre in two distinct synaptic complexes, with the C-terminal domains of Cre facing either towards or away from PepA. Thus, no specific PepA-recombinase interaction is required, and correct juxtaposition of the loxP sites is sufficient to activate Cre in this system.     

Effect of acetylcholine on coordination of spontaneous activities registered from different areas of uterus in rats

Spontaneous electrical activities of uterine corpus, periuterine part of horn and uterine cervix in response to intravenous injections of different concentrations of acetylcholine were observed. The changes of frequency and amplitude characteristics of rhythmogenesis in these conditions, both separately and with their co-active state were analysed. Acetylcholine in concentration of 10(-3) M in the blood of the animal creates the optimal conditions for synchronization and coordination of electrical activities in the above areas of the uterus.     

Allelic polymorphism of MTHFR, MTR and MTRR genes in patients with cleft lip and/or palate and their mothers

The frequency of common MTHFR, MTR and MTRR genes polymorphisms was evaluated among patients with non-syndromic cleft lip and/or palate (CL/P), their mothers and healthy persons from West-Ukrainian region. MTHFR 677TT genotype was shown to increase more than three-fold risk of CL/P and for mothers the risk of having CL/P children may increase two-fold compared with homozygous carriers of MTHFR 677CC genotype (OR = 3.3, OR = 1.92, respectively). The heterozygous MTR 2756AG genotype was associated with 1.5-fold increased risk of CL/P compared with the AA genotype (OR = 1.48). The heterozygous genotype MTRR 66AG was associated with the 5.56-fold increased CL/P risk (OR = 5.56) and for mothers with 2.6-fold increased risk of delivering a CL/P offspring (OR = 2.6). The results showed that MTRR 66G allele is more prevalent than MTRR 66A (wild type) and the MTRR 66GG genotype frequency was significantly lower among CL/P patients and their mothers than in control group among Western Ukrainian inhabitants.     

Application of Spiroplasma melliferum proteogenomic profiling for the discovery of virulence factors and pathogenicity mechanisms in host-associated spiroplasmas

To date, no genome of any of the species from the genus Spiroplasma has been completely sequenced. Long repetitive sequences similar to mobile units present a major obstacle for current genome sequencing technologies. Here, we report the assembly of the Spiroplasma melliferum KC3 genome into 4 contigs, followed by proteogenomic annotation and metabolic reconstruction based on the discovery of 521 expressed proteins and comprehensive metabolomic profiling. A systems approach allowed us to elucidate putative pathogenicity mechanisms and to discover major virulence factors, such as Chitinase utilization enzymes and toxins never before reported for insect pathogenic spiroplasmas.     

祁连山大野口流域青海云杉种群数量动态

种群数量动态揭示了种群的结构特征及其潜在的驱动机制，有助于预测种群未来的动态，进而为森林生态系统的保护与恢复提供理论依据。本研究基于10.2 hm²青海云杉动态监测样地数据，以种群径级结构代替年龄结构，编制静态生命表，绘制径级结构图、存活曲线、死亡率曲线、消失率曲线和4个生存分析函数曲线，分析青海云杉种群数量特征，并利用种群数量动态变化指数和时间序列模型对种群数量动态进行预测。结果表明：（1）青海云杉种群的年龄结构近似于倒"J"型，幼苗和小树储量丰富；（2）种群存活曲线趋近于Deevey-Ⅱ型，为稳定型种群，死亡率曲线和消失率曲线变化趋势基本一致，均在第2、8龄级出现高峰期；（3）生存率曲线呈下降趋势，累计死亡率曲线呈上升趋势，死亡密度曲线缓慢下降，而危险率曲线逐渐上升，该种群具有：前期减少、中期稳定、后期衰退的生长特点；（4）种群数量变化动态指数V_pi>0，表明该种群属于增长型种群，V_pi''>0且趋近于0，则表明该种群趋近于稳定型；（5）时间序列预测分析表明，在未来2、4、6、8个龄级时间后，种群呈稳定增长趋势。研究显示，祁连山大野口流域青海云杉种群为稳定增长型种群，只要未来不遭受强烈干扰，种群数量会保持逐渐增长。针对该种群幼龄个体在前期的更新过程死亡率较高情况，建议在今后的经营管理中应重点加强对第1、2龄级植株生存环境的保护和改善，提高幼苗和小树的存活率。     

Protein degradation by the proteasome and dissection of its in vivo importance with synthetic inhibitors

Molecular Biology Reports -