Cell signaling by the type IV pili of pathogenic Neisseria

Neisseria gonorrhoeae and Neisseria meningitidis are Gram-negative bacterial pathogens that infect human mucosal epithelia. Type IV pilus-mediated adherence of these bacteria is a crucial early event for establishment of infection. In this work, we show that the type IV pili transduce a signal into the eucaryotic host cell. Purified adherent pili, but not pili from a low binding mutant, trigger an increase in the cytosolic free calcium ([Ca2+]i) in target epithelial cells, a signal known to control many cellular responses. The [Ca2+]i increase was blocked by antibodies against CD46, a putative pilus receptor, suggesting a role for this protein in signal transduction. Pilus-mediated attachment was inhibited by depletion of host cell intracellular Ca2+ stores but not by removal of extracellular Ca2+. Further, kinase inhibition studies showed that pilus-mediated adherence is dependent on casein kinase II. In summary, these data reveal a novel function of the type IV pili, namely induction of signal transduction pathways in host cells.     

13C solid-state NMR of gramicidin A in a lipid membrane

The natural-abundance 13C NMR spectrum of gramicidin A in a lipid membrane was acquired under magic-angle spinning conditions. With fast sample spinning (15 kHz) at approximately 65 degrees C the peaks from several of the aliphatic, beta-, alpha-, aromatic, and carbonyl carbons in the peptide could be resolved. The resolution in the 13C spectrum was superior that observed with 1H NMR under similar conditions. The 13C linewidths were in the range 30-100 Hz, except for the alpha- and beta-carbons, the widths of which were approximately 350 Hz. The beta-sheet-like local structure of gramicidin A was observed as an upfield shift of the gramicidin alpha and carbonyl resonances. Under slow sample spinning (500 Hz), the intensity of the spinning sidebands from 13C in the backbone carbonyls was used to determine the residual chemical shift tensor. As expected, the elements of the residual chemical shift tensor were consistent with the single-stranded, right-handed beta6.3 helix structure proposed for gramicidin A in lipid membranes.     

Conformational stability of factor VIIa: biophysical studies of thermal and guanidine hydrochloride-induced denaturation

The binding of the multidomain protein factor VIIa (fVIIa) to tissue factor provides the interprotein communication necessary to make fVIIa an efficient catalyst of the initial event in the extrinsic pathway of blood coagulation. We have investigated the stability of individual domains in fVIIa and the influence of Ca2+ and an irreversible active-site inhibitor (FFR-chloromethyl ketone). Equilibrium guanidine hydrochloride (GuHCl)-induced unfolding monitored by tryptophan fluorescence and far-UV circular dichroism (CD) demonstrated that the gamma-carboxyglutamic acid (Gla) domain unfolds at 0.3 M GuHCl and the serine protease (SP) domain at 3 M GuHCl and that Ca2+ is a prerequisite for the formation of an ordered, compact structure in the Gla domain. The loss of amidolytic activity coincides with the first transition, which is stabilized by the active-site inhibitor, and a change in the environment of the active site is demonstrated using a fluorescent inhibitor (DEGR-chloromethyl ketone). Thermal unfolding monitored by differential scanning calorimetry (DSC) reveals that Ca2+ stabilizes the SP domain slightly, increasing the unfolding temperature by 2.7 degrees C. In addition, Ca2+ is required for a large enthalpy change concomitant with unfolding of the Gla domain, and this unfolding enthalpy is only detectable in the presence of the SP domain, indicating some kind of interaction between these domains. Thermal unfolding measured by CD indicates secondary structural changes at the same temperature as the heat absorption in the DSC but only when both the Gla domain and the SP domain are present together with Ca2+ ions. Taken together, these results indicate a Ca2+-dependent interaction between the Gla domain and the SP domain, implying a high degree of flexibility of the domains in free fVIIa. It is also shown that the epidermal growth factor-like domains are stable at elevated temperatures and high GuHCl concentrations. Moreover, already at physiological temperature, subtle structural changes take place which influence the overall shape of fVIIa and are detrimental to its enzymatic activity.     

Enhancing encoding capacity of combinational test stimulus decompressors

While scan-based compression is widely utilized in order to alleviate the test time and data volume problems,the overall compression level is dictated not only by the chain to channel ratio but also the ratio of encodable patterns.Aggressively increasing the number of scan chains in an effort to raise the compression levels may reduce the ratio of encodable patterns,degrading the overall compression level.In this paper,we present various methods to improve the ratio of encodable patterns.These methods are b...     

哈锅670 t/h锅炉炉内冷态流场数值模拟

对 HG-670 /1 40 -9煤粉炉燃烧器改造后的炉内冷态流场进行数值模拟 ,分析了炉内燃烧区烟气的速度分布及压力分布。结果表明该型号锅炉改造后在额定工况下运行时炉内可形成良好的切圆 ,无一次风贴壁现象 ,不会发生壁面结渣 ,同时避免了壁面处高温腐蚀现象的发生     

大鼠放射性皮肤溃疡组织中c-Fos、Rb蛋白表达研究

为研究大鼠放射性溃疡组织中ｃ－Ｆｏｓ、Ｒｂ表达。采用１４０只Ｗｉｓｔａｒ大鼠进行局部照射（γ射线）制备放射性皮肤溃疡动物模型,观察病变１年,然后采用ｃ－Ｆｏｓ、Ｒｂ多克隆抗体及免疫组化方法检测溃疡组织中ｃ－Ｆｏｓ、Ｒｂ的表达情况。结果表明：ｃ－Ｆｏｓ、Ｒｂ的表达阳性率分别为４５．８％（３３／７２）、６３．９％（４６／７２）,两者阳性部位主要见于增生肥大的鳞状上皮细胞胞核、增生的间质成纤维细胞胞质及