Persistently low natural killer cell activity in normal adults: immunological, hormonal and mood correlates

This current study is concerned with the identification of the incidence of persistently low natural killer (NK) cell activity in an essentially healthy population of young adults, and the characterization of the psychological profile associated with this pattern of NK activity. Eighty-eight individuals, between the ages of 18 and 44 years, were accrued to this project. Subjects were tested serially, at baseline induction into the study, and at 2 and 4 weeks follow-up. Baseline assessment included a complete physical examination and laboratory work-up in order to exclude anyone ill at the time of enrollment, and to have complete physical status information on all subjects at the time of accrual. NK activity, urinary catecholamine levels and psychological status were also assessed at baseline, and at the specified follow-up intervals. Individuals with low NK activity were operationally defined as those having NK function either below the group mean, or below the group lowest quartile, at baseline and at two follow-up assessments. Results showed that in both univariate analyses, as well as in logistic regression models, age and the perception of environmental stressors or 'hassles' predicted persistently low NK activity. Younger subjects, who perceived environmental events to which they were exposed as more serious in nature, were more likely to exhibit a persistently low NK profile over time than older individuals who perceived daily events as less important to them.     

Inorganic phosphate speeds loaded shortening in rat skinned cardiac myocytes

Force generation in striated muscle is coupled with inorganic phosphate (P_i) release from myosin, because force falls with increasing P_i concentration ([P_i]). However, it is unclear which steps in the cross-bridge cycle limit loaded shortening and power output. We examined the role of P_i in determining force, unloaded and loaded shortening, power output, and rate of force development in rat skinned cardiac myocytes to discern which step in the cross-bridge cycle limits loaded shortening. Myocytes (n = 6) were attached between a force transducer and position motor, and contractile properties were measured over a range of loads during maximal Ca²⁺ activation. Addition of 5 mM P_i had no effect on maximal unloaded shortening velocity (V_o) (control 1.83 ± 0.75, 5 mM added P_i 1.75 ± 0.58 muscle lengths/s; n = 6). Conversely, addition of 2.5, 5, and 10 mM P_i progressively decreased force but resulted in faster loaded shortening and greater power output (when normalized for the decrease in force) at all loads greater than 10% isometric force. Peak normalized power output increased 16% with 2.5 mM added P_i and further increased to a plateau of 35% with 5 and 10 mM added P_i. Interestingly, the rate constant of force redevelopment (k_tr) progressively increased from 0 to 10 mM added P_i, with k_tr 360% greater at 10 mM than at 0 mM added P_i. Overall, these results suggest that the P_i release step in the cross-bridge cycle is rate limiting for determining shortening velocity and power output at intermediate and high relative loads in cardiac myocytes. muscle mechanics; force-velocity relationship; cross-bridge cycle     

Construction, expression and identification of a recombinant streptococcal protein G

化学合成链球菌蛋白G的C3D基因片段,通过分子生物学的方法对蛋白G(proteinG)的C3[1]片段进行PCR扩增,拼接形成含有两个和三个重复C3片段的重组链球菌蛋白G,C3片段间以链接区D连接,即形成C3DC3和C3DC3DC3的形式,进而克隆到质粒pET21中,在大肠杆菌BL21(DE3)中表达.重组表达的蛋白经过DEAE - Sepharose和IgG- Sepharose纯化,得到纯化的重组蛋白.采用非竞争性酶免疫法对重组蛋白与不同来源IgG的结合常数进行测定,实验结果显示两种重组链球菌蛋白G均可有效地与小鼠、兔及山羊等多种不同来源抗体特异性结合.这些实验结果为下一步研究奠定了基础.     

High-pressure freezing, cellular tomography, and structural cell biology

Structural cell biology, which we define as electron microscopic analysis of intact cells, suffered a loss of interest and activity following the advances in light microscopy beginning in the 1990s. Interestingly, it is the wealth of detailed observation in the light microscope that is one of the driving forces for the current renewed interest in electron microscopy (EM). A great many cellular details are simply beyond the resolving power of the light microscope. In this article, we describe how electron microscopists are responding to the demands for better preservation of cells and for ways to view cell ultrastructure in three dimensions at high resolution. We discuss how low temperature methods, especially high-pressure freezing and freeze substitution, reduce the artifacts of conventional EM specimen preparation. We also give a brief introduction to cellular electron tomography, a powerful analytical method that can give near-atomic resolution of cell ultrastructure in three-dimensional (3-D) models.     

Pulsatile urea excretion in the gulf toadfish: mechanisms and controls

Opsanus beta expresses a full complement of ornithine–urea cycle (OUC) enzymes and is facultatively ureotelic, reducing ammonia-N excretion and maintaining urea-N excretion under conditions of crowding/confinement. The switch to ureotelism is keyed by a modest rise in cortisol associated with a substantial increase in cytosolic glutamine synthetase for trapping of ammonia-N and an upregulation of the capacity of the mitochondrial OUC to use glutamine-N. The entire day's urea-N production is excreted in 1 or 2 short-lasting pulses, which occur exclusively through the gills. The pulse event is not triggered by an internal urea-N threshold, is not due to pulsatile urea-N production, but reflects pulsatile activation of a specific branchial excretion mechanism that rapidly clears urea-N from the body fluids. A bidirectional facilitated diffusion transporter, with pharmacological similarity to the UT-A type transporters of the mammalian kidney, is activated in the gills, associated with an increased trafficking of dense-cored vesicles in the pavement cells. An 1814 kB cDNA (‘tUT’) coding for a 475–amino acid protein with approximately 62% homology to mammalian UT-A's has been cloned and facilitates phloretin-sensitive urea transport when expressed in Xenopus oocytes. tUT occurs only in gill tissue, but tUT mRNA levels do not change over the pulse cycle, suggesting that tUT regulation occurs at a level beyond mRNA. Circulating cortisol levels consistently decline prior to a pulse event and rise thereafter. When cortisol is experimentally clamped at high levels, natural pulse events are suppressed in size but not in frequency, an effect mediated through glucocorticoid receptors. The cortisol decline appears to be permissive, rather than the actual trigger of the pulse event. Fluctuations in circulating AVT levels do not correlate with pulses; and injections of AVT (at supraphysiological levels) elicit only minute urea-N pulses. However, circulating 5-hydroxytryptamine (5-HT) levels fluctuate considerably and physiological doses of 5-HT cause large urea-N pulse events. When the efferent cranial nerves to the gills are sectioned, natural urea pulse events persist, suggesting that direct motor output from the CNS to the gill is not the proximate control.     

First record of Renibacterium salmoninarum in the sea lamprey (Petromyzon marinus)

Bacterial kidney disease (BKD), caused by Renibacterium salmoninarum, is a widespread problem with major implications for salmonid fish species. The mechanisms by which the bacterium has reached high levels of infection previously unrecorded in the Laurentian Great Lakes are presently unknown. Research involving reservoirs and mechanisms of R. salmoninarum transmission in fish is lacking because of the ecologic complexity of heterogeneous habitats and the lack of adequate funding. Herein, we report on the isolation of R. salmoninarum from the kidneys of the sea lamprey (Petromyzon marinus). The bacterium was cultured from kidneys of 16% and 4% of lampreys collected from two locations within the Lake Ontario watershed in 2003 and 2004, respectively. The identity of bacterial colonies was verified with the nested polymerase chain reaction and quantitative enzyme-linked immunosorbent assay.     

Phylogenetic and population genetic analyses of Phaeosphaeria nodorum and its close relatives indicate cryptic species and an origin in the Fertile Crescent

The origin of the fungal wheat pathogen Phaeosphaeria nodorum remains unclear despite earlier intensive global population genetic and phylogeographical studies. We sequenced 1683 bp distributed across three loci in 355 globally distributed Phaeosphaeria isolates, including 74 collected in Iran near the center of origin of wheat. We identified nine phylogenetically distinct clades, including two previously unknown species tentatively named P1 and P2 collected in Iran. Coalescent analysis indicates that P1 and P2 are sister species of P. nodorum and the other Phaeosphaeria species identified in our analysis. Two species, P. nodorum and P. avenaria f. sp. tritici 1 (Pat1), comprised ～85% of the sampled isolates, making them the dominant wheat-infecting pathogens within the species complex. We designed a PCR-RFLP assay to distinguish P. nodorum from Pat1. Approximately 4% of P. nodorum and Pat1 isolates showed evidence of hybridization. Measures of private allelic richness at SSR and sequence loci suggest that the center of origin of P. nodorum coincides with its host in the Fertile Crescent. We hypothesize that the origin of this species complex is also in the Fertile Crescent, with four species out of nine found exclusively in the Iranian collections.     

Activating and inhibiting connections in biological network dynamics

Background  

Many studies of biochemical networks have analyzed network topology. Such work has suggested that specific types of network wiring may increase network robustness and therefore confer a selective advantage. However, knowledge of network topology does not allow one to predict network dynamical behavior – for example, whether deleting a protein from a signaling network would maintain the network's dynamical behavior, or induce oscillations or chaos.     

农牧交错带土地沙化的本质及其形成研究

通过对陕北农牧交错带不同类型沙化土地土体构型、土壤质地、元素组成和理化性质的分析,研究了土地沙化的本质。结果表明：受风沙作用影响,土壤中细粒物质逐渐减少,颗粒组成变粗;表层消失,最终被流沙所取代,原土壤剖面被覆盖在沙层之下;土壤有机质及养分含量减少,保水保肥性能降低,生产力不断下降;现代土壤形成过程以侵蚀和风沙沉积为主,物质淋溶和化学风化微弱。研究区土地沙化可划分为：风沙侵蚀为主、风沙蚀积平衡、风沙沉积为主和土壤形成发育4个阶段;沙化土地的类型有：肥力衰退质地粗化、表层剥蚀、片沙覆盖、流动沙丘与固定沙丘等5种。     

两种落叶分解过程中养分和碳素的行为

利用分解袋法对日本亚高山针叶林的针叶(Abies veitchii Lindl,and A.mariesi Mast.)和阔叶(Betula ermanii Cham.and B.corylifolia Regal.et Maxim．)凋落物进行了分解实验研究。结果表明尽管分解初期的两种凋落物的养分以及分解后期凋落物剩余重量差异很大,但两种凋落物养分浓度在分解后期(30个月以后)趋于一致。这种趋同现象在不同养分中有不同的趋同机制。氮元素浓度升高到分解后期浓度差变小,这种现象是由于分解菌的固持作用及受木质素的束缚所致;钾和镁在分解初期浓度急剧下降,进而浓度差变小,是由于淋溶作用所致。在分解过程中这些元素非常容易被淋溶掉,直到和土壤中的浓度达到一致为止。钙是结构元素,它的行踪和有机物组分有密切关系。由于分解后期有机组分木质化和腐殖质化进而浓度趋同,所以钙的浓度也相应趋同。