Changes of synapsin I messenger RNA expression during rat brain development

Synapsin I is a synaptic phosphoprotein that is involved in the short-term regulation of neurotransmitter release. In this report we present the first extensive study of the developmental expression of its corresponding messenger ribonucleic acid (mRNA) by in situ hybridization and northern blot analysis in rat brain. Synapsin I mRNA showed pronounced differences in expression in different brain regions during postnatal development. The early expression of synapsin I mRNA in ontogenetically older regions such as the thalamus, the piriform cortex and the hippocampus coincides with the earlier maturation of these regions, in contrast to its later expression in ontogenetically younger areas such as the cerebellum and the neocortex. An intriguing expression pattern was found in the hippocampus. In all hippocampal subregions synapsin I mRNA expression increased from postnatal day (PND) 1 to 17. After PND 17, however, there was a marked dissociation between persisting high expression levels in CA3 and the dentate gyrus and a strong decline in synapsin I mRNA expression in CA1. The persistence of synapsin I in some adult rat brain regions indicates that it plays a part in synapse formation during plastic adaption in neuronal connectivities.     

Vertical transmission of human T-cell leukemia virus type I (HTLV-I): Detection of proviral DNA in HTLV-I carrier gravida

The seroprevalence rate of human T-cell leukemia virus type I (HTLV-I) in pregnant women in the Osaka district was determined by enzyme-linked immunosorbent assay and Western blot analysis. Twenty-one (1.0%) of 2192 samples tested were positive for both assays and the seropositive parturients were found to be integrated with HTLV-I proviral DNA in their mononuclear cells by a DNA dot blot hybridization assay using HTLV-I DNA probe or by a selective DNA amplification technique using the polymerase chain reaction (PCR). On the other hand, proviral DNA was not detected in cord blood of the neonates born to the carrier mothers, indicating that transplacental infection of HTLV-I during pregnancy could be excluded. The results support the hypothesis that postpartum infection via breast milk plays a significant role among the possible perinatal transmission routes.     

Presence and distribution of ghrelin-immunopositive cells in the chicken gastrointestinal tract

The presence and distribution patterns of ghrelin, a gastric acylated peptide, were studied in the entire gastrointestinal tract of the chicken (Gallus domesticus) using the peroxidase-antiperoxidase immunohistochemical method, western blot analysis and a specific antibody against the C-terminal region of rat ghrelin. Ghrelin-immunopositive cells were observed in the mucosal layer of all segments examined. The largest numbers of ghrelin-positive cells were located at the base of lobuli of the proventriculus gland, along villi of the intestines and in crypts of the duodenum. Lower numbers of ghrelin-immunostained cells were located in crypts of jejunum and ileum and only few ghrelin-immunostained cells were detected at the base of crypts of the large intestine. Closed and open types of cells were observed in all segments. Western blot analysis confirmed the presence of ghrelin-like protein in the entire chicken gastrointestinal tract. The anatomical distribution patterns and the morphological characteristics of chicken ghrelin-positive cells suggest that they are endocrine cells. Furthermore, it is concluded that ghrelin shows a high degree of preservation during evolution from non-mammalian vertebrates to mammals.     

抗核糖体抗体阳性判断标准的探讨

目的 :分析针对分子量为 38kD和 或 15 16 5kD多肽抗原的抗核糖体抗体 (anti ribosomalantibodies)与SLE的关系 ,探讨抗核糖体抗体的阳性判断标准及其在SLE中的检出情况。方法 :收集病人血清 2 6 2例 ,包括SLE115例、RA5 7例、硬皮病 2 0例、MCTD39例及其他免疫性疾病 31例 ,用免疫印迹法 (Western blot)检测其抗ENA抗体谱。结果 :分别以同时出现 38kD和 16 5 15kD蛋白条带以及出现 38kD蛋白条带为判断抗核糖体抗体阳性的标准 ,则该抗体对诊断SLE的敏感性和特异性结果分别为 2 8 7% (33 115 )、96 6 % (14 2 14 7)及 17 4 % (2 0 115 )、97 3% (14 3 14 7) ,两者比较敏感性有显著性差异 (P <0 0 5 ) ,特异性无显著性差异 (P >0 0 5 ) ;仅 38kD分子阳性的 14例样本中 ,SLE占大多数 ,显著多于其它疾病 (71 4 %比 2 8 6 % ,P <0 0 5 ) ;38kD分子阳性同时伴抗Sm抗体阳性者多于抗Sm抗体阴性者 ,但无显著性差异 (6 4 3%比 35 7% ,P >0 0 5 )。结论 :抗核糖体抗体 38kD分子与SLE密切相关 ,而与抗Sm抗体相关性不强。以 38kD为主要抗原多肽判断抗核糖体抗体 ,不仅可以提高该抗体的阳性检出率 ,同时也不会降低其对SLE诊断的特异性 ,该判断方法值得推广应用。     

Development of a syncytia inhibition assay for the detection of antibodies to bovine leukemia virus in naturally infected cattle; comparison with Western blot and agar gel immunodiffusion

A syncytia inhibition assay (SIA) for the detection of antibodies to bovine leukemia virus is described. This test involves specific antibody-mediated inhibition of BLV-induced cytopathic effects in an indicator cell line. A total of 300 sera were screened commercially by agar gel immunodiffusion (AGID) and were then screened by Western blot and SIA. The new assay system provided results which were comparable to Western blot and AGID. The results obtained suggest that SIA may be more sensitive than either of the other two assay systems examined for the determination of the infection status of cattle.     

Spatio-temporally differential expression of genes for three members of fatty acid binding proteins in developing and mature rat brains

The chronological changes in the gene expression for three species of cytosolic fatty acid-binding proteins (FABPs) in the rat brain were examined by Northern and in situ hybridization analyses. The expression for heart(H)-FABP became evident after birth, with a gradual increase and confined to the gray matter, suggesting that the expression of H-FABP mRNA is neuron-specific in postnatal brain. The expression for brain(B)-FABP was very intense in the ventricular germinal zone, without expression in the cerebellar external granule cell layer, suggesting the dominant expression in the cells of glial lineage. B-FABP mRNA was transiently expressed in perinatal gray as well as white matter and the expression in glial cells persists only in the olfactory nerve fiber layer at the adult stage. On the other hand, the expression for skin type(S)-FABP was evident in the both ventricular germinal zone and cerebellar external granule cell layer, suggesting the expression in cells of neuronal lineage. The expression for S-FABP was evident in the prenatal gray matter and S-FABP mRNA was expressed in glial cells at early postnatal stage, whereafter the expression decreased to, but remained at weak levels in the adult brain. Discrete functions of the three FABPs were suggested in neurons and glia differentially at various developmental stages.     

重组猪戊型肝炎病毒ORF2区主要抗原域的原核表达及鉴定

目的：在大肠杆菌中表达猪戊型肝炎病毒ORF2区主要结构蛋白,并对其进行血清学鉴定。方法：通过RT-PCR技术从一份猪粪中扩增并克隆戊型肝炎病毒主要的结构基因片段,将该片段插入pET-32a表达载体,在原核系统中融合表达蛋白,Western blot和间接ELISA方法分析该蛋白的抗原性。结果：SDS-PAGE分析结果表明,获得45kD的目的蛋白,该重组蛋白可以与HEV阳性血清反应,而不与阴性血清反应,表明该蛋白具有良好的抗原性。结论：本研究获得了猪戊型肝炎病毒重组抗原,可与HEV阳性血清产生特异性的结合反应,可以作为诊断用的重组蛋白,为研制敏感、特异的HEV诊断试剂盒,行之有效的HEV基因工程疫苗及为HEV感染的预防和临床治疗提供资料。     

Real-time PCR method for the quantitative analysis of human T-cell receptor gamma and beta gene rearrangements

Analyzing the status of T-cell receptor (TCR) gene rearrangements has been an essential part of deciphering the stages of thymocyte development, understanding the β vs. γδ lineage decision, and characterizing T-cell leukemias. Methods such as PCR and quantitative Southern blotting provide useful information, but also have significant shortcomings such as lack of quantitation in the case of PCR and technical challenges in the case of Southern blotting. Here we describe a real-time PCR method that overcomes many of these shortcomings. This new method shows comparable results for the fraction of unrearranged TCRγ and TCRβ genes in human thymocytes and peripheral blood T cells as Southern blotting, and has the advantages of being simple to perform, highly quantitative, and requiring nanogram quantities of DNA. We also describe a real-time PCR method to quantitate T-cell receptor excision circles formed during TCRβ rearrangements.     

Production of a monoclonal antibody for evaluation of hard red winter wheat cultivars to wheat streak mosaic virus

The monoclonal antibody technology has provided a means to produce a supply of highly specific uniform antibody which is useful in the detection of plant viruses and which facilitates disease resistance screening. Because of the specificity of a monoclonal antibody to an epitope, a monoclonal antibody may not react to a partially degraded protein. Wheat streak mosaic virus (WSMV) is a member of the potyvirus group and is transmitted by the wheat curl mite Eriophyes tulipae Keifer. The capsid protein of WSMV, like many potyviruses, is degraded in planta. Monoclonal antibodies produced to WSMV reacted to native as well as trypsin treated virions. The antibodies were also useful for evaluation of hard red winter wheat cultivars inoculated with WSMV in the fall or in the spring under field conditions.     

Human papillomavirus types in anogenital warts of children

Tissue from anogenital warts of 25 children, 10 of whom were suspected of being victims of sexual abuse, was investigated by dot blot and Southern blot techniques for human papillomavirus (HPV) types. HPV DNA was detected in 22 children, two of whom had double infections. The genital HPV types 6 and/or 11 were detected in 20 children, and in three children other HPV types were found. One had HPV 18 (as well as 11); in a second child a possible skin type, HPV 2, was detected; and the third child was infected with an unidentified type. In three cases genital wart material was available from one of the parents, and in all three the HPV type was the same as that of the child. For nine other children one or both parents were reported to have genital warts. The source of infection appeared to be the adult genital tract, but sexual contact might not be the only means of transmission.