Disruption of a single copy of the SERCA2 gene results in altered Ca2+ homeostasis and cardiomyocyte function

A mouse model carrying a null mutation in one copy of the sarcoplasmic reticulum (SR) Ca(2+)-ATPase isoform 2 (SERCA2) gene, in which SERCA2 protein levels are reduced by approximately 35%, was used to investigate the effects of decreased SERCA2 level on intracellular Ca(2+) homeostasis and contractile properties in isolated cardiomyocytes. When compared with wild-type controls, SR Ca(2+) stores and Ca(2+) release in myocytes of SERCA2 heterozygous mice were decreased by approximately 40-60% and approximately 30-40%, respectively, and the rate of myocyte shortening and relengthening were each decreased by approximately 40%. However, the rate of Ca(2+) transient decline (tau) was not altered significantly, suggesting that compensation was occurring in the removal of Ca(2+) from the cytosol. Phospholamban, which inhibits SERCA2, was decreased by approximately 40% in heterozygous hearts, and basal phosphorylation of Ser-16 and Thr-17, which relieves the inhibition, was increased approximately 2- and 2.1-fold. These results indicate that reduced expression and increased phosphorylation of phospholamban provides compensation for decreased SERCA2 protein levels in heterozygous heart. Furthermore, both expression and current density of the sarcolemmal Na(+)-Ca(2+) exchanger were up-regulated. These results demonstrate that a decrease in SERCA2 levels can directly modify intracellular Ca(2+) homeostasis and myocyte contractility. However, the resulting deficit is partially compensated by alterations in phospholamban/SERCA2 interactions and by up-regulation of the Na(+)-Ca(2+) exchanger.     

Species differences between rat and mouse CCKA receptors determine the divergent acinar cell response to the cholecystokinin analog JMV-180

The cholecystokinin (CCK) analog JMV-180 acts as a partial agonist in rats and a full agonist in mice. Whether this functional variability is due to species differences in CCK receptor structure or to alterations in the cellular environment is unknown. To address this question, an adenoviral construct encoding the rat CCK(A) receptor (AdCCK(A)R) was used to express the rat receptor in acini from CCK(A) receptor-deficient mice (CCK(A)R -/-). Infection of CCK(A)R -/- acini in vitro with pAdCCK(A)R led to a time-dependent increase in (125)I-CCK(8) binding. The affinity for JMV-180 of the adenovirally transferred rat and the endogenous mouse CCK(A) receptors was not different. In native mouse acini, JMV-180 acted as a full agonist (both stimulation and inhibition of amylase release). In contrast, in mouse acini expressing pAdCCK(A)R JMV-180 acted as a partial agonist (only stimulation of amylase release). In addition, the pattern of protein synthesis induced by JMV-180 in CCK(A)R -/- mouse acini infected with AdCCK(A)R resembled the pattern observed in wild-type rats (lack of inhibition) rather than the respective pattern in wild-type mice (inhibition). These data suggest that species differences in the CCK(A) receptor of rats and mice account for the observed divergence in the acinar cell response to JMV-180.     

Interactions between HIV-1 gp41 core and detergents and their implications for membrane fusion

The gp41 envelope protein mediates entry of human immunodeficiency virus type 1 (HIV-1) into the cell by promoting membrane fusion. The crystal structure of a gp41 ectodomain core in its fusion-active state is a six-helix bundle in which a N-terminal trimeric coiled coil is surrounded by three C-terminal outer helices in an antiparallel orientation. Here we demonstrate that the N34(L6)C28 model of the gp41 core is stabilized by interaction with the ionic detergent sodium dodecyl sulfate (SDS) or the nonionic detergent n-octyl-beta-D-glucopyranoside (betaOG). The high resolution x-ray structures of N34(L6)C28 crystallized from two different detergent micellar media reveal a six-helix bundle conformation very similar to that of the molecule in water. Moreover, N34(L6)C28 adopts a highly alpha-helical conformation in lipid vesicles. Taken together, these results suggest that the six-helix bundle of the gp41 core displays substantial affinity for lipid bilayers rather than unfolding in the membrane environment. This characteristic may be important for formation of the fusion-active gp41 core structure and close apposition of the viral and cellular membranes for fusion.     

Infection of human dendritic cells by a sindbis virus replicon vector is determined by a single amino acid substitution in the E2 glycoprotein

The ability to target antigen-presenting cells with vectors encoding desired antigens holds the promise of potent prophylactic and therapeutic vaccines for infectious diseases and cancer. Toward this goal, we derived variants of the prototype alphavirus, Sindbis virus (SIN), with differential abilities to infect human dendritic cells. Cloning and sequencing of the SIN variant genomes revealed that the genetic determinant for human dendritic cell (DC) tropism mapped to a single amino acid substitution at residue 160 of the envelope glycoprotein E2. Packaging of SIN replicon vectors with the E2 glycoprotein from a DC-tropic variant conferred a similar ability to efficiently infect immature human DC, whereupon those DC were observed to undergo rapid activation and maturation. The SIN replicon particles infected skin-resident mouse DC in vivo, which subsequently migrated to the draining lymph nodes and upregulated cell surface expression of major histocompatibility complex and costimulatory molecules. Furthermore, SIN replicon particles encoding human immunodeficiency virus type 1 p55(Gag) elicited robust Gag-specific T-cell responses in vitro and in vivo, demonstrating that infected DC maintained their ability to process and present replicon-encoded antigen. Interestingly, human and mouse DC were differentially infected by selected SIN variants, suggesting differences in receptor expression between human and murine DC. Taken together, these data illustrate the tremendous potential of using a directed approach in generating alphavirus vaccine vectors that target and activate antigen-presenting cells, resulting in robust antigen-specific immune responses.     

The blocking effect of BmP02, one novel short-chain scorpion peptide on transient outward K(+) channel of adult rat ventricular myocyte

Two components F-2-7-4 and F-2-7-5, each composed of 28 amino acid residues, were purified from the venom of Buthus martensi Karsch by an opportune procedure with cation-exchange column chromatography and repeated HPLC. Both components were totally accounted to about 0.88% dry weight of the crude venom.The molecular weights of both components were determined to be 2950 and 2935 by mass spectrometry, which were fully coincidence with that of the known novel short-chain peptides BmP02 and BmP03, respectively [Romi-Lebrun R, Martin-Eauclaire M-F, Escoubas P, Wu FQ, Lebrun B, Hisada M, Nakajima T. Characterization of four toxins from Buthus martensi scorpion venom, which act on apamin-sensitive Ca²⁺-activated K⁺ channels. Eur J Biochem 1997;145:457–464]. In addition, the sequence of component F-2-7-4 was analyzed to be the same as that of BmP02. The components F-2-7-4 and F-2-7-5 purified in this study were, thus, finally distinguished to be BmP02 and BmP03 from the same venom. Using whole cell patch-clamp recording, it was found that BmP02 diminished the current of transient outward K⁺ channel in adult rat ventricular myocyte in a concentration-dependent manner. The inhibitory effect was reversible. Dynamic studies showed that the activation, inactivation and recovery processes of the transient outward K⁺ channel were not changed significantly after applying of BmP02. In addition, when BmP02 was applied to guinea pig ventricular myocyte, both delayed and inward rectified K⁺ currents showed no change compared with the control. The results suggest strongly that BmP02 or -like peptides from scorpion venom may provide a useful probe for the studying of transient outward K⁺ channel in rat ventricular myocyte.     

四部体小麦“简阳矮兰麦”与黑麦可杂交性及其在六倍体水平上的遗传特性

中国六倍体普通小麦地方品种有一个比较独特的特征-存在丰富的高亲和性材料。因此,研究中国四倍体小麦的亲和性问题具有特殊的意义。“简阳矮兰麦”是来源于四川省的一个四倍体小麦地方品种,它与黑麦有高的可杂交性,其杂交结实率达60%。遗传分析表明,“简阳矮兰麦”与黑麦的高可杂交性是受2-3对隐性基因所控制,但3对基因的可能性更大。而且,这些隐性基因的作用在合成六倍体小麦后仍能比较完全的表达。通过与六倍体普通小麦相比较,结果表明四倍体小麦的可杂交性系统与六倍体小麦的可杂交性系统的作用方式是类似的。 Abstract:It is a special characteristic that many Chinese common wheat landraces showed a high crossability with rye.Thus,it is important that elucidate the genetic control of the crossability of Chinese tetraploid wheat with rye.Triticum turgidum cv.Jianyangailanmai native to Sichuan,China has high crossability with rye,up to 60%.In this study,it is indicated that the crossability of Jianyangailanmai with rve is controlled by two or more probably three recessive genes,which was almost totally expressed in the hexaploid wheat level.The operation of these recessive genes influencing crossability with rye was similar to that of hexaploid common wheat.     

中国荷斯坦牛IL8基因遗传多态性与泌乳性状以及体细胞评分的关联

以上海某奶牛场30个公牛家系的610头中国荷斯坦牛为试验材料,采用聚合酶链式反应-单链构象多态性(PCR-SSCP)技术对Interleukin-8(IL8)基因的遗传多态性进行了分析,采用混合动物模型分析了IL8基因突变位点与测定日产奶量、测定日乳脂率、测定日乳蛋白率、305d校正产奶量、305d乳脂量、305d乳蛋白量及测定日体细胞评分7个性状的相关性,寻找可用于生产实际的分子标记。共检测到KK、KA和AA3种基因型,频率分别为0.187、0.451和0.362,等位基因K和A的频率分别为0.412和0.588。该位点突变对测定日产奶量、305d乳蛋白量、305d校正产奶量和305d乳脂量以及体细胞评分影响达到极显著水平(P0.01),对测定日乳蛋白率的影响达到显著水平(P0.05),对测定日乳脂率影响不显著(P0.05)。多重比较表明:KK基因型对测定日产奶量、305d校正产奶量、305d乳蛋白量和305d乳脂量极显著高于AA和KA基因型(P0.01)。KK基因型的体细胞评分(SCS)最小二乘均值极显著低于KA、AA基因型(P0.01)。对于测定日的乳蛋白率AA基因型显著低于KA、KK型(P0.05)。IL8基因遗传突变对中国荷斯坦牛泌乳性状和乳房炎抗性有较大的遗传效应,可用于中国荷斯坦牛的分子标记辅助选择。     

Multilocus sequence typing and virulence factors analysis of Escherichia coli O157 strains in China

Escherichia coli O157:H7, an important food-borne pathogen, has become a major public health concern worldwide. The aim of this study was to investigate the molecular epidemiologic feature of E. coli O157:H7 strains in China. 105 E. coli O157:H7 isolates were collected from various hosts and places over 9 years. A multilocus sequence typing scheme (MLST) was applied for bacteria genotyping and polymerase chain reaction (PCR) was used for virulence factor identification. Seven new MLST sequence types (STs), namely ST836, ST837, ST838, ST839, ST840, ST841, and ST842 were identified, which grouped into two lineages. Phylogenetic analysis suggested that the most two frequent STs in China, ST837 and ST836, may be the derivatives of E. coli O157:H7 Sakai or E. coli O157:H7 EDL933. Geographical diversity and host variety of E. coli O157:H7 were observed in China. In addition, the different distribution of tccp was detected. The data presented herein provide new insights into the molecular epidemiologic feature of E. coli O157:H7, and aid in the investigation of the transmission regularity and evolutionary mechanism of E. coli O157:H7.     

JH biosynthesis by reproductive tissues and corpora allata in adult longhorned beetles, Apriona germari

We report on juvenile hormone (JH) biosynthesis from long‐chain intermediates by specific reproductive tissues and the corpora allata (CA) prepared from adult longhorned beetles, Apriona germari. The testes, male accessory glands (MAGs), ovaries, and CA contained the long‐chain intermediates in the JH biosynthetic pathway, farnesoic acid (FA), methyl farnesoate (MF), and JH III. The testes and ovaries, but not CA, produced radioactive JH III after the addition of ³H‐methionine and, separately, unlabeled methionine, to the incubation medium. We inferred that endogenous FA is methylated to MF in the testes and ovaries. Addition of farnesol led to increased amounts of FA in the testes, MAGs, ovaries, and CA, indicating oxidation of farnesol to FA. Addition of FA to incubation medium yielded increased JH III, again indicating methylation of FA to MF in the testes, MAGs, ovaries, but not CA. Addition of MF to incubation medium also led to JH III, from which we inferred the epoxidation of MF to JH III. JH biosynthesis from farnesol in the testes, MAGs, and ovaries of A. germari proceeds via oxidation to FA, methylation to MF, and epoxidation to JH III. This is a well‐known pathway to JH III, described here for the first time in reproductive tissues of longhorned beetles. © 2010 Wiley Periodicals, Inc.     

Development of seven novel EST-SSR markers from Cycas panzhihuaensis (Cycadaceae)

? Premise of the study: Cycas panzhihuaensis L. Zhou & S. Y. Yang is a vulnerable gymnosperm endemic to China, where its range represents the northernmost occupation of Cycas L. We developed EST-derived SSR markers to investigate its genetic diversity and population structure. ? Methods and Results: Based on the expressed sequence tag (EST) database for Cycas rumphii Miq., seven simple sequence repeat (SSR) markers were identified and screened in 55 individuals from seven wild populations of C. panzhihuaensis. Alleles numbered 2 to 5, and their observed heterozygosity and expected heterozygosity ranged from 0.0000 to 0.6545 and from 0.0535 to 0.6966, respectively. ? Conclusions: These new EST-SSR markers will enhance further studies of the population genetics of C. panzhihuaensis, allowing researchers to design reasonable conservation and management protocols.