检测HIV-1载量的荧光实时定量PCR技术的建立及其应用

准确测定HIV 1的前病毒载量和病毒载量的技术 ,在感染者预后和艾滋病患者药物治疗效果的评价以及艾滋病的其它研究方面 ,都具有十分重要的应用价值。以定量的HIV 1DNA和RNA为标准外参照 ,利用SYBRGreen荧光染料和GeneAmp5 70 0SequenceDetectionSystem (5 70 0系统 ) ,建立了测定HIV 1的前病毒载量和病毒载量的荧光实时定量PCR技术。以病毒感染细胞和培养上清为材料 ,测定了三种化合物 (AZT ,GL和WT)对细胞内的前病毒载量和培养上清中的病毒载量的抑制活性 ,并与合胞体形成抑制方法测定化合物抗病毒活性的结果进行了比较。根据病毒载量、前病毒载量和合胞体形成计算出的三种化合物的治疗指数均依次变小 ,提出以荧光实时定量PCR技术测定前病毒载量 ,会在评价药物在体内外根除或减少存在于CD4休止或记忆T淋巴细胞中的HIV 1前病毒方面有特别的价值。     

Relationship between worm burden and male proportion in Schistosoma mansoni experimentally infected rodents and primates. A meta-analytical approach

This paper investigates the relationship between the worm burden and the male proportion of adult Schistosoma mansoni. This relationship was tested using data from the literature on S. mansoni experimentally infected rodents and primates. The analyses were performed within each animal model (rat, mouse, hamster and monkey) and showed positive linear relationships between worm burden and male proportion. An analysis between the four animal models showed the same positive linear relationship. The role of the genetic diversity of male and female schistosomes in this relationship is discussed.     

Inference for set-based effects in genetic association studies with interval-censored outcomes

The rapid acceleration of genetic data collection in biomedical settings has recently resulted in the rise of genetic compendiums filled with rich longitudinal disease data. One common feature of these data sets is their plethora of interval-censored outcomes. However, very few tools are available for the analysis of genetic data sets with interval-censored outcomes, and in particular, there is a lack of methodology available for set-based inference. Set-based inference is used to associate a gene, biological pathway, or other genetic construct with outcomes and is one of the most popular strategies in genetics research. This work develops three such tests for interval-censored settings beginning with a variance components test for interval-censored outcomes, the interval-censored sequence kernel association test (ICSKAT). We also provide the interval-censored version of the Burden test, and then we integrate ICSKAT and Burden to construct the interval censored sequence kernel association test—optimal (ICSKATO) combination. These tests unlock set-based analysis of interval-censored data sets with analogs of three highly popular set-based tools commonly applied to continuous and binary outcomes. Simulation studies illustrate the advantages of the developed methods over ad hoc alternatives, including protection of the type I error rate at very low levels and increased power. The proposed approaches are applied to the investigation that motivated this study, an examination of the genes associated with bone mineral density deficiency and fracture risk.     

Metabolic burden in recombinant CHO cells: effect ofdhfr gene amplification andlacZ expression

Foreign protein production levels in two recombinant Chinese hamster ovary (CHO) cell lines were compared in cells transfected with different expression vectors. One vector pNL1 contained the gene for neomycin resistance (neo ^r ) and thelacZ gene which codes for intracellular -galactosidase, with both genes controlled by the constitutive simian virus (SV40) promoter. The other vector CDG contained the amplifiabledhfr gene andlacZ gene, controlled by the constitutive SV40 and cytomegalovirus (CMV) promoters, respectively. Cell growth and -galactosidase expression were compared quantitatively after cells were selected in different concentrations of the neomycin analog G418 and methotrexate, respectively. A 62% reduction in growth rate occurred in recombinant CHO cells in which thelacZ anddhfr genes were highly amplified and expressed. In contrast, the combined effects of the unamplifiedneo ^r gene andlacZ gene expression on the growth kinetics were small. Any metabolic burden caused bylacZ gene expression, which was evaluated separately from the effect ofneo ^r gene expression, must be negligible, as higher expression of -galactosidase (1.5×10^–6 units/cell) occurred in unamplified cells compared to the cells in whichlacZ was amplified by thedhfr-containing vector (3×10^–7 units/cell). Thus, the main factor causing severe growth reduction (metabolic burden) in cells containing the amplifieddhfr gene system was not overexpression of -galactosidase butdhfr andlacZ gene co-amplification anddhfr gene expression.     

Lead levels in ancient and contemporary Japanese bones

During the past few centuries, lead production, consumption and emissions, to our total environment have increased remarkably. We have determined the concentrations of lead in 41 well-preserved ancient and 11 contemporary rib bones of a mature age (40–60 y), with a view to historically evaluating lead exposure in humans. The oldest Japanese bones (1000–300b.c.) were found to contain a mean of 0.58 μg Pb/g dry wt and a mean molar ratio of lead to calcium of 0.6×10⁻⁶, compared with 4.7–5.2×10⁻⁶ in the bones of the Edo era (1600–1867a.d.) and contemporary residents in Japan. The mean molar ratios of female bones were always higher than those of male bones for each era. From this fact we may assume that facial cosmetics were one of the main routes of lead exposure among the ancient Japanese, especially those who lived during the Edo era.     

The effects of sex,age and diet of mice and gerbils on susceptibility to Microphallus pygmaeus (Digenea: Microphallidae)

Molan A. L. and James B. L. 1984. The effects of sex, age and diet of mice and gerbils on susceptibility to Microphallus pygmaeus (Digenea: Microphallidae). International Journal for Parasitology14: 521–526. Mature male mice (40–100-day-old) and gerbils (60–150-day-old) harbour significantly more Microphallus pygmaeus than their female counterparts. Adult worms from male mice and gerbils consistently contain more eggs than those from females but the differences are not significant. No sex differences in worm recovery occur in immature or ageing mice and gerbils. Seventy-day-old mice and gerbils of both sexes harbour significantly more worms with a higher egg production than immature and ageing animals. The worm burden of sucklings, however, is higher than in weanlings and mature mice and gerbils of both sexes fed on cow's milk harbour significantly more worms than those fed on a normal pellet diet.     

A Market‐Based Framework for Quantifying Displaced Production from Recycling or Reuse

The most significant environmental benefit of recycling or reusing a wide range of products and materials is typically the potential to displace primary material production; lack of displacement significantly reduces the environmental benefits of these activities. Because no consensus method to estimate displacement rate has emerged, environmental assessments have tended to assume that displacement occurs on a one‐to‐one basis. However, displaced production is a complex phenomenon governed primarily by market mechanisms, rather than physical relationships. This article advances the understanding of displacement by presenting a market‐based framework describing the displacement relationship and a methodology for quantifying displacement rate based on partial equilibrium modeling. Using this methodology, a general symbolic equation for displacement rate after an increase in recycling is derived. The model highlights the market mechanisms that govern displaced production and identifies five price response parameters that affect displacement rate. Results suggest that one‐to‐one displacement occurs only under specific parameter restrictions that are unlikely in competitive commodity markets, but zero displacement is possible if secondary materials are poor substitutes for primary materials; displacement is likely to be reduced if secondary materials have inferior technical properties. The presented methodology can be generally applied to any system in which recycled or reused materials are substitutes or complements for primary materials. Implications for improving recycling and reuse efficacy and environmental assessment methodology are discussed, and suggestions are presented for expanding the displacement methodology in future research.     

Plasmid Effects on Escherichia coli Metabolism

ABSTRACT:?

The idea that plasmids replicate within hosts at the expense of cell metabolic energy and preformed cellular blocks depicts plasmids as a kind of molecular parasites that, even when they may eventually provide plasmid-carrying strains with growth advantages over plasmid-free strains, doom hosts to bear an unavoidable metabolic burden. Due to the consistency with experimental data, this idea was rapidly adopted and used as a basis of different hypotheses to explain plasmid-host interactions. In this article we critically discuss current ideas about plasmid effects on host metabolism, and present evidence suggesting that the complex interaction between plasmids and hosts is related to the alteration of the cellular regulatory status.     

On the origin and evolution of major morphological characters

Although the mathematical principles underpinning population-level evolution are now well studied, the origin and evolution of morphological novelties has received far less attention. Here, a broad but general theory for how this sort of change takes place is outlined, relying on functional continuity, least-constrained components of morphology, redundancy and preadaptation. At least four distinct sorts of redundancy are identified: (i) redundancy arising through duplication (amplification); (ii) redundancy arising through regionalisation (parcellation); (iii) redundancy arising through functional convergence; and (iv) redundancy arising from shared function (functional degeneracy). Although organisms are here recognised to be functionally constrained ("burdened", in Riedl's terminology), these constraints can be overcome through the combination of the four principles given above. Contrary to its common treatment, functional constraint is neither an ever-increasing restriction on the scope of evolution, nor does it require drastic events to overcome or "break" it. Rather, it is an evolutionary quantity, subject to selection at some level. The rules that govern morphological evolution are the primary controls on what is allowed to happen in the evolution of the overall genotype-phenotype system, suggesting strong controls on the sorts of developmental changes that might be associated with macroevolution. This model, then, sees organism functionality as the primary control on evolvability, with exact genetic make-up being of secondary importance. It should prove possible to recast traditional notions of body-plan evolution into the formulations of complex system analysis, which in the future may prove a unifying discipline for fields as disparate as palaeontology and gene regulatory networks. In particular, understanding how morphology can evolve may provide the critical link between the ecological and morphological networks that are currently the intense focus of evolutionary investigations.