Genetic modification of the spontaneous rho − mutability in Saccharomyces cerevisiaee

Summary The phenotypic trait starry colony in Saccharomyces is associated with a high spontaneous rho ^– petite mutability. Genetic analysis of this trait has shown the high rho ^– mutability to be caused by several modifying genes present together in the strains studied. Every single modifying gene produces only a relatively small enhancement of the rho ^– mutability.     

SAGE遗传分析系统的功能及应用

SAGE是集多功能于一体的医学遗传学群体与家系资料计算机分析系统。本文概述SAGE系统的主要功能及应用环境。重点介绍了FCOR2和TDTEX两个功能模块的数学原理和使用方法。应用TDTEX模块 ,我们发现微卫星标记 85ca与小儿失神症存在连锁不平衡 ,提示在该位点附近存在小儿失神症的易感基因     

A Complementary Method for the Detection of Osteoblastic Metastases on Digitized Radiographs

Purpose This study was conducted to evaluate the diagnostic usefulness of gray level parameters in order to distinguish healthy bone from osteoblastic metastases on digitized radiographs. Materials and methods Skeletal radiographs of healthy bone (n = 144) and osteoblastic metastases (n = 35) were digitized using pixels 0.175 mm in size and 4,096 gray levels. We obtained an optimized healthy bone classification to compare with pathological bone: cortical, trabecular, and flat bone. The osteoblastic metastases (OM) were classified in nonflat and flat bone. These radiological images were analyzed by using a computerized method. The parameters (gray scale) calculated were: mean, standard deviation, and coefficient of variation (MGL, SDGL, and CVGL, respectively) based on gray level histogram analysis. Diagnostic utility was quantified by measurement of parameters on healthy and pathological bone, yielding quantification of area under the receiver operating characteristic (ROC) curve, AUC. Results All three image parameters showed high and significant values of AUC when comparing healthy trabecular bone and nonflat bone OM, showing MGL the best discriminatory ability (0.97). As for flat bones, MGL showed no ability to distinguish between healthy and flat bone OM (0.50). This could be achieved by using SDGL or CVGL, with both showing a similar diagnostic ability (0.85 and 0.83, respectively). Conclusion Our results show that the use of gray level parameters quantify healthy bone and osteoblastic metastases zones on digitized radiographs. This may be helpful as a complementary method for differential diagnosis. Moreover, our method will allow us to study the evolution of osteoblastic metastases under medical treatment.     

Cell-free fetal DNA and intact fetal cells in maternal blood circulation: implications for first and second trimester non-invasive prenatal diagnosis

Both intact fetal cells as well as cell-free fetal DNA are present in the maternal circulation and can be recovered for non-invasive prenatal genetic diagnosis. Although methods for enrichment and isolation of rare intact fetal cells have been challenging, diagnosis of fetal chromosomal aneuploidy including trisomy 21 in first- and second-trimester pregnancies has been achieved with a 50-75% detection rate. Similarly, cell-free fetal DNA can be reliably recovered from maternal plasma and assessed by quantitative PCR to detect fetal trisomy 21 and paternally derived single gene mutations. Real-time PCR assays are robust in detecting low-level fetal DNA concentrations, with sensitivity of approximately 95-100% and specificity near 100%. Comparing intact fetal cell versus cell-free fetal DNA methods for non-invasive prenatal screening for fetal chromosomal aneuploidy reveals that the latter is at least four times more sensitive. These preliminary results do not support a relationship between frequency of intact fetal cells and concentration of cell-free fetal DNA. The above results imply that the concentration of fetal DNA in maternal plasma may not be dependent on circulating intact fetal cells but rather be a product of growth and cellular turnover during embryonic or fetal development.     

中国庚型肝炎病毒河南株非结构基因（NS）3区cDNA的克隆 …

目的 为了研究中国庚型肝炎病毒（ＨＧＶ）非结构（ＮＳ３）区基因结构特征。方法 利用逆转录－半巢式－聚合酶链反应从河南１份ＨＧＶ ＲＮＡ阳性血清获得覆盖ＨＢＶ ＮＳ３全长ｃＤＮＡ的４个片段，并克隆到ｐｃＤＮＡⅡ载体中，采用Ｓａｎｇｅｒ双脱氧末端终止法测定全部ｃＤＮＡ序列。结果 发现克隆到的包括ＨＢＶ ＮＳ３区在内的ｃＤＮＡ序列和度为２１３７个核苷酸，编码７１１个氨基酸。与国内外已测定的５株全序列的相     

Identification of the novel allele HLA-A*6813 in two members of a family of Syrian origin: implications for bone marrow transplantation

The identification of the new allele HLA-A*6813, which was found in a woman of Syrian origin and her son, is described. In the sequence analysis the new allele differs from A*68011 by positions 259 (A>G) and 261 (C>G) in exon 2. As the structure is thus identical to the HLA-A consensus sequence it is likely that the new allele originated by gene conversion. At the protein level, the new allele has one amino acid difference from A*6801 (Asn63Glu), which results in a distinct banding pattern in one dimensional-isoelectric focusing. Amino acid residue 63 contributes to the formation of pocket A and B and is thus important for peptide binding. A*6813 was serologically detectable only by two of six polyclonal, but by three monoclonal antisera. The restricted serological A68 activity may be explained by altered peptide binding as presented peptides can affect the serological recognition of major histocompatibility complex (MHC) class I molecules. Moreover, our findings suggest that a possible mismatch with the other known A*68 variants may impair clinical outcome of bone marrow transplantation.     

Possible causal relationships between cerebellar patterns of foliation and hindlimb coordination in laboratory mice: a quantitative trait locus analysis

The cerebellum is involved in a large set of integrative functions including memory, affect, and motricity. The cerebellar patterns of foliation and their causal relationships with motricity were investigated via a wide genome scan approach and quantitative trait locus (QTL) strategy. QTLs were mapped in an F₂ population derived from NZB/B1NJ and C57BL/6By inbred strains of mice for cerebellar fissures in the four vermal lobules (intraculminate, uvula, declival, and intracentral) and for hindpaw slips in a bar crossing test. No linkage was detected for uvula and intracentral fissures. We found five QTLs linked to declival fissure: Cpfd-1q and Cpfd-2q (chromosome 1), Cpfd-3q (chromosome 5), Cpfd-4q (chromosome 9), and Cpfd-5q (chromosome 13). Two QTLs were also mapped for intraculminate fissure Cpfi-1q (chromosome 4) and Cpfi-2q (chromosome 1). Most of the confidence intervals of these QTLs included genes that were previously identified for their implication in the physiological mechanisms underlying cerebellar patterns of foliation. Only one significant QTL was found for the measure of hindpaw coordination (Tne-1q). It was linked with Cpfd-1q and Cpfd-2q on the telomeric part of chromosome 1.     

Large Envelope Glycoprotein and Nucleocapsid Protein of Equine Arteritis Virus (EAV) Induce an Immune Response in Balb/c Mice by DNA Vaccination; Strategy for Developing a DNA-Vaccine Against EAV-Infection

Equine arteritis virus (EAV) is a member of the Arteriviridae family, that includes lactate dehydrogenase-elevating virus (LDV), porcine reproductive and respiratory syndrome virus (PRRSV), and simian haemorrhagic fever virus (SHFV). Equine arteritis is a contagious disease of horses and is spread via respiratory or reproductive tract. The objective of the present study is to evaluate the possibility for developing a model system for prevention horses against an EAV infection by DNA vaccination. A cDNA bank from the RNA of EAV was established. This gene library contains the translation unit of the EAV open reading frames (ORF) 1 to 7. The identity of the cDNA was confirmed by nucleotide sequence analysis. Using this defined EAV cDNA gene library the cDNA sequence of the viral ORFs were molecularly cloned into the corresponding sites of well characterized and powerful expression vectors (pCR3.1, pDisplay, and/or pcDNA3.1/HisC).The capability of these recombinant plasmids expressing the gene products of the individual viral ORFs 3 to 5, and 7 in induction of an immune response in mouse system was investigated. The Balb/c mice (ten mice per assay) were inoculated with the DNA of the constructed expression vectors harboring and expressing the EAV cDNA of the viral ORFs. The Balb/c mice were injected with about 100 g DNA diluted in 100 l PBS. The DNA was injected subcutaneously and into the tibialis cranialis muscle (Musculus gastrocnemius). The mice were boosted 3 to 5 times with the same quantities of DNA and under the same conditions at about two week intervals. Control mice received the same amount of parental expression vectors via an identical route and frequency.The pre- and post-vaccinated sera of the individual animals were screened by neutralization tests (NT). Neutralizing antibodies against EAV were detected when the animals were inoculated with the DNA of the expression vectors harboring cDNA of the EAV ORFs 5 and 7. Highest NT-titers were observed when the animals were administered with the cDNA of ORF 5 and/or with the cDNA of the neutralization determinants of EAV that is located on the N-terminal ectodomain of the gene product of ORF 5 between the amino acid positions 1–121. These results obtained from these studies justified proofing the capability of the EAV cDNA sequences of the viral genes including ORFs 5 and 7 in the autologous animal system horse.     

Hepatitis B virus DNA in serum of healthy black African adults positive for hepatitis B surface antibody alone: possible association with recombination between genotypes A and D

In some patients with chronic liver disease induced by hepatitis B virus, viral DNA is known to persist in low concentration in serum after seroconversion to hepatitis B surface antibody-positivity. This phenomenon has, however, not been documented in asymptomatic black African carriers of hepatitis B virus. Using nested amplification by the polymerase chain reaction, we detected low concentrations of hepatitis B virus DNA in the serum of 6 of 23 (26%) healthy black African adults with normal liver function and with hepatitis B virus surface antibody as the only serological marker of the virus. This finding offers one explanation for the earlier observation of integrated hepatitis B virus DNA in hepatocellular carcinomas in black Africans whose serum was positive for surface antibody alone. A number of genetic changes were found in the six isolates that might be responsible for evasion of the immune response and persistence of the virus. Isolated mutations were detected in the "a" determinant of the surface gene and in the encapsidation signal. In all five isolates sequenced in the core promoter, mutations were present in the upstream regulatory region. Recombination between genotypes A and D was present in three of the isolates, including both of those in which the entire genome was sequenced. This change in genotype also overlapped the amino end of the polymerase domain and may result in sufficiently low levels of replication to allow viral persistence. Topoisomerase 1 specific trinucleotides were concentrated in the vicinity of the recombination breakpoints.     

重庆市2004—2007年突发公共卫生事件分析

目的对重庆市2004—2007年报告的突发公共卫生事件进行分析，以加强对突发公共卫生事件的预防和应急处理。方法应用描述流行病学方法，分析事件特征。结果全市4年报告突发公共卫生事件478起，发病19884例，死亡56例；渝东南地区多发，时间分布基本呈双高峰：传染病事件占80．98％．学校事件占78．87％；事件平均报告时间3．44d，平均处理时间46．72d。结论突发公共卫生事件的发生与地区经济文化水平密切相关；应提高对乙、丙类传染病和非法定传染病事件的重视程度；学校卫生工作有待进一步加强：突发公共卫生事件报告管理体系有待进一步理顺。