Interaction of the human polyomavirus, JCV, with human B-lymphocytes.

The human polyomavirus, JCV, is the causative agent of the central nervous system demyelinating disease progressive multifocal leukoencephalopathy (PML). The principal target of JCV infection in the central nervous system (CNS) is the myelinating oligodendrocyte. However, the site of JCV multiplication outside of the CNS and the mechanism by which virus gains access to the brain are not known. Recently, JCV infected B-lymphocytes have been demonstrated in PML patients in several lymphoid organs, in circulating peripheral lymphocytes, and in brain, suggesting a possible role of B-lymphocytes in the dissemination of virus to the brain. The experiments reported here were undertaken to understand more about the interactions of JCV with human B-lymphocytes. The data show that JCV is able to multiply in either Epstein-Barr virus transformed (EBV) or EBV negative human B cell lines resulting in production of infectious, progeny virions. In addition, nuclear proteins extracted from these B cells bind to similar nucleotides within the JCV regulatory region that are bound by nuclear proteins extracted from human fetal glial cells, the most susceptible host and principal target cell for JCV infection in vitro. It is not known, however, whether these DNA binding proteins from susceptible B cells and glial cells are similar.     

Dementia,ageing, and the structure of human memory

The memory performance of patients suffering from senile dementia of the Alzheimer type (SDAT) (N = 29), normal subjects of equivalent age and education (N = 58), and young normal controls (N = 42) was tested using free recall and verbal and nonverbal span. Three measures were derived from the free recall task: primacy based on the first item, secondary memory based on the middle serial positions, and primary memory based on recency and the Waugh-Norman correction factor. The SDAT patients differed from the normal elderly on all free recall and span measures except for primary memory. The elderly were clearly inferior to the young on secondary memory, and were marginally poorer on primary memory and the two span measures. Three possible explanations of this pattern of results are considered, based on the dichotomous modal model of memory, levels of processing, and working memory. It is suggested that the assumption that SDAT patients suffer from a deficit in the central executive component of working memory offers the best of these interpretations.     

人类肝脏cDNA文库的构建及hHGF基因的筛选、克隆与表达

目的构建人类肝脏的cDNA文库，从中筛选hHGF基因并进行克隆与表达。方法从人类胎儿肝脏中快速分离提取出mRNA；将已提取出的mRNA构建成cDNA文库；从中筛选出hHGF（hepatocyte growth factor）基因后，进行克隆并测序确定；构建表达质粒pBV221-hHGF后转化大肠杆菌BL21（DE3α），以异丙基硫代-B-D-半乳糖苷（IPTG）诱导，收集菌体后提取蛋白样品并对其进行SDS-PAGE电泳及蛋白免疫印迹检测。结果成功地构建了人类肝脏的cDNA文库；经筛选得到约2200bp的hHGF基因；对其进行测序确认；成功地构建该基因的表达型质粒，并对表达蛋白进行蛋白免疫印迹检测，检测出其表达产生的蛋白质相对分子量在100000左右。结论人类肝脏cDNA文库成功地构建后可用于筛选获取完整的cDNA基因，从中筛选、克隆hHGF基因，并测序确认，进行表达蛋白质检测，结果正确，以上工作为进一步深入地进行与人类肝脏cDNA文库及hHGF相关的实验研究奠定了一定的基础。     

Study on dioxan separation in the radioimmunoassay of human growth hormone, human chorionic gonadotrophin, adrenocorticotrophic hormone, and insulin

The dioxan precipitation for separating free from antibody-bound hormone in a radioimmunoassay has been applied to human chorionic gonadotrophin (HCG), human growth hormone (HGH), adrenocorticotrophic hormone (ACTH), and insulin and compared with the charcoal-dextran system of separation. The dioxan separation depends on protein concentration. The behaviour of damaged labelled hormone is difficult to assess. Completeness of separation has to be tested in antigen excess. The optimum conditions must be carefully studied for each hormone.     

Adsorption studies of human serum albumin, human gamma-globulins, and human fibrinogen on the surface of p(S/PGL) microspheres

Adsorption of human serum albumin (HSA), human gamma-globulins (gammaG), and human fibrinogen (Fb) onto the surface of poly(styrene/alpha-t-butoxy-omega-vinylbenzyl-polyglycidol) microspheres (P(S/PGL)) with controlled fraction of polyglycidol in the interfacial layer was investigated. The microspheres were synthesized by the emulsifier-free radical copolymerization of styrene and alpha-t-butoxy-omega-vinylbenzyl-polyglycidol macromonomer (PGL). Macromonomers with number average molecular weights Mn = 950 and 2,700 were used in the syntheses. Fraction of polyglycidol in the microsphere surface layer was varied from 0.22 to 0.44, depending on the composition of the monomer feed. It was found that the maximal surface concentration of adsorbed proteins and the equilibrium constant of protein adsorption decreased with increased fraction of polyglycidol in the microsphere surface layer. For microspheres with the highest fraction of polyglycidol at the surface the maximal surface protein concentration was c. ten times lower and the adsorption equilibrium constant was c. one hundred times lower than for the reference polystyrene microspheres. The dependence of maximal surface concentration of adsorbed proteins on the fraction of polyglycidol in the particle interfacial layer indicated random distribution of polyglycidol chains without formation of polyglycidol and polystyrene patches at the microspheres surface.     

Comparative recombination rates in the rat, mouse, and human genomes

Levels of recombination vary among species, among chromosomes within species, and among regions within chromosomes in mammals. This heterogeneity may affect levels of diversity, efficiency of selection, and genome composition, as well as have practical consequences for the genetic mapping of traits. We compared the genetic maps to the genome sequence assemblies of rat, mouse, and human to estimate local recombination rates across these genomes. Humans have greater overall levels of recombination, as well as greater variance. In rat and mouse, the size of the chromosome and proximity to telomere have less effect on local recombination rate than in human. At the chromosome level, rat and mouse X chromosomes have the lowest recombination rates, whereas human chromosome X does not show the same pattern. In all species, local recombination rate is significantly correlated with several sequence variables, including GC%, CpG density, repetitive elements, and the neutral mutation rate, with some pronounced differences between species. Recombination rate in one species is not strongly correlated with the rate in another, when comparing homologous syntenic blocks of the genome. This comparative approach provides additional insight into the causes and consequences of genomic heterogeneity in recombination.     

The nucleus pararaphales in the human, chimpanzee, and macaque monkey

The human cerebral cortex and cerebellum are greatly expanded compared to those of other mammals, including the great apes. This expansion is reflected in differences in the size and organization of precerebellar brainstem structures, such as the inferior olive. In addition, there are cell groups unique to the human brainstem. One such group may be the nucleus pararaphales (PRa); however, there is disagreement among authors about the size and location of this nucleus in the human brainstem. The name “pararaphales” has also been used for neurons in the medulla shown to project to the flocculus in the macaque monkey. We have re-examined the existence and status of the PRa in eight humans, three chimpanzees, and four macaque monkeys using Nissl-stained sections as well as immunohistochemistry. In the human we found a cell group along the midline of the medulla in all cases; it had the form of interrupted cell columns and was variable among cases in rostrocaudal and dorsoventral extent. Cells and processes were highly immunoreactive for non-phosphorylated neurofilament protein (NPNFP); somata were immunoreactive to the synthetic enzyme for nitric oxide, nitric oxide synthase, and for calretinin. In macaque monkey, there was a much smaller oval cell group with NPNFP immunoreactivity. In the chimpanzee, we found a region of NPNFP-immunoreactive cells and fibers similar to what was observed in macaques. These results suggest that the “PRa” in the human may not be the same structure as the flocculus-projecting cell group described in the macaque. The PRa, like the arcuate nucleus, therefore may be unique to humans.     

A new subdivision,the marginal division,in the neostriatum of the human brain

<正> A new subetemtory was discovered byas at the caudal most edge of the neostriatum(St) and surrounding the rostral berder of the globus pallidus(GP) in the rat btain with neuroanato-my and immunohistochemistry methods.This area was named marginal division(MrD)based on its location.     

Histomorphometric comparison of the human,swine, and ovine collecting systems

The ovine kidney has been recently determined to be a better model than the swine kidney for the study of collecting system healing after partial nephrectomy. However, there is no histological study comparing the collecting systems of these species. To compare human, swine, and ovine collecting systems using histomorphometry. The collecting systems of 10 kidneys from each species (human, swine, and ovine) were processed for histomorphometry. The thickness of the three layers (mucosal connective tissue, submucosal muscular, and adventitial connective tissue) were measured. The densities of smooth muscle fibers, elastic system fibers, and cells were also measured. Additionally, blood vessel density in the adventitial connective tissue was measured. Analysis of the collecting systems from the three species presented several differences. The adventitial connective tissue from the swine samples was thicker, with more blood vessels and smooth muscle fibers, compared with that from the human and ovine samples. Swine also had higher density of elastic fibers on the submucosal muscular layer. Ovine and human collecting systems shared several similar features, such as blood vessel and elastic fiber density in all layers and the density of cellular and muscular fibers in the submucosal muscular and adventitial connective tissue layers. The collecting system of the ovine kidney is more similar to that of the human kidney compared with that of the swine kidney. This may explain the differences between the healing mechanisms in swine and those in humans and sheep after partial nephrectomy. Anat Rec, 299:967–972, 2016. © 2016 Wiley Periodicals, Inc.     

Cloning, characterization, and chromosome mapping of the human GlcAT-S gene

 We report on the structure, map location, and tissue expression of the human GlcAT-S gene. The gene covers approximately 85 Kb on chromosome 6 (6q13) between the D6S455 and D6S1673 markers. GlcAT-S is composed of four exons and encodes a 324-amino-acid protein, which shows 89% homology with the rat glcat-s protein and is involved in the biosynthesis of the HNK-1 carbohydrate epitope on glycoproteins. Although GlcAT-S was considered an interesting candidate gene for the RP25 locus, the absence of any pathogenic mutations in probands of RP25-linked families ruled out that candidacy. Received: May 23, 2002 / Accepted: September 6, 2002     

Endothelial cell proliferation follows the mid-cycle luteinizing hormone surge, but not human chorionic gonadotrophin rescue, in the human corpus luteum

The corpus luteum is essential for the maintenance of early pregnancy in women. Angiogenesis may be one factor involved in luteal rescue. The aim of this study was to determine the changes in endothelial cell proliferation throughout the luteal phase and in human chorionic gonadotrophin (HCG)-simulated early pregnancy. Human corpora lutea obtained throughout the luteal phase and in simulated early pregnancy were immunostained with antibodies for endothelial and proliferating cells. Number and distribution of endothelial and proliferating cells were examined. Endothelial cells were least abundant in the early luteal phase, increasing in the mid-luteal phase (P < 0.03). Endothelial numbers did not differ significantly between the late and the rescued corpora lutea. Endothelial cell proliferation was greatest in the early luteal phase and continued at a lower level during later stages. Simulated early pregnancy resulted in no change in endothelial cell proliferation. These results showed that a high degree of endothelial cell proliferation is associated with formation of the human corpus luteum. Unchanging levels of proliferation following HCG treatment (for 5-8 days from day 12 to day 16 post-ovulation, at 125 IU to 16,000 IU, following a daily doubling of dose) suggest that alternative processes are involved during luteal rescue.