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81.
By developing an appropriate immunization protocol for SCID (hu-PBL-SCID) mice engrafted with human peripheral blood lymphocytes in combination with scFv phage display library, we were able to establish an efficient strategy to obtain human scFv clones against a human self-antigen, TNF-alpha. The mice pretreated with gamma-radiation (3Gy) and anti-asialo GM1 antibody were immunized with a mixture of human TNF-alpha-keyhole limpet hemocyanin and Freund's adjuvant. Human antibody maturation was suggested to be induced in the mice with the immunization protocol. The scFv clones obtained from the mice were shown to exhibit binding affinities in the range of 10(7)-10(8) M(-1). Together with our previously published work on the isolation of respiratory syncytial virus neutralizing scFvs, the results of this study have implicated that this combined approach is one of the effective alternatives for the cloning of human monoclonal antibodies specific for a wide range of antigens of interest. 相似文献
82.
83.
We have recently identified and sequenced a molecular clone of the serogroup 2 simian retrovirus (SRV), D2/RHE/OR/V1, that retains an enhanced ability to infect specific T cell lines. In this report, using deletion mutagenesis, we localized the psi packaging signal, necessary for packaging of D2/RHE/OR/V1 particles, to the genomic region 345-650, which comprises the 5' intergenic region (IR) and the extreme 5' portion of the gag gene. To build an SRV-based gene transfer system and to reduce the possibility of recombination and regeneration of replication-competent viruses, we constructed split-genome D2/RHE/OR/V1 plasmid recombinants containing distinct and non-overlapping retroviral gene regions and several replacement components. For the retrovirus gene transfer vehicle, we deleted the D2/RHE/OR/V1 structural genes and substituted a cassette including the psi-packaging region, the beta-galactosidase reporter gene, and the 3' IR. Both packaging cell recombinants were used to generate stable monkey packaging cell lines; the gene transfer vehicle was subsequently transfected into the packaging cell lines, and replication-defective viruses were recovered for subsequent infection into fresh monkey cells. Successful infection by the recovered viruses verifies the potential efficacy of the SRV-based system as a research tool for gene transfer of heterologous genes into nonhuman primate cells. 相似文献
84.
Timo D Müller Anke Hinney André Scherag Thuy T Nguyen Felix Schreiner Helmut Schäfer Johannes Hebebrand Christian L Roth Thomas Reinehr 《BMC medical genetics》2008,9(1):85
Background
We have previously identified strong association of six single nucleotide polymorphisms (SNPs) in FTO (fat mass and obesity associated gene) to early onset extreme obesity within the first genome wide association study (GWA) for this phenotype. The aim of this study was to investigate whether the obesity risk allele of one of these SNPs (rs9939609) is associated with weight loss in a lifestyle intervention program. Additionally, we tested for association of rs9939609 alleles with fasting blood parameters indicative of glucose and lipid metabolism. 相似文献85.
Lai PS Takeshima Y Adachi K Van Tran K Nguyen HT Low PS Matsuo M 《Journal of human genetics》2002,47(10):0552-0555
The frequency and distribution of deletions of 19 deletion-prone exons clustered in two hot spots in the proximal and central
regions of the dystrophin gene were compared in three populations from Singaporean, Japan, and Vietnam. DNA samples obtained
from 105 Singaporean, 86 Japanese, and 34 Vietnamese Duchenne muscular dystrophy patients were examined by polymerase chain
reaction amplification. Deletions of the examined exons were found in 51.2% of Japanese patients but in 40.0% or less of the
Singaporeans and Vietnamese. About two thirds of the deletions were localized in the central region and the remaining deletions
were clustered at the proximal region. The most commonly deleted exons at the central deletion hot spot were exon 50 in the
Singaporean, exons 49 and 50 in the Japanese, and exon 51 in the Vietnamese population. At the proximal deletion hot spot,
the most commonly deleted exons were exons 6 and 8 in the Singaporeans, exons 12 and 17 in the Japanese, and exons 8 and 12
in the Vietnamese. Two cases each from Singapore and Japan had large-scale gross mutations spanning both deletion hot spots.
Our results suggest that, although the presence and frequency of the two deletion hot spots may be similar in the three Asian
populations analyzed, the distribution and frequency of deletions among the different exons can vary as a result of population-specific
intronic sequences that predispose individuals to preferential deletion breakpoints.
Received: May 20, 2002 / Accepted: July 1, 2002 相似文献
86.
Lin H Lei J Wininger D Nguyen MT Khanna R Hartmann C Yan WL Huang SC 《Stem cells (Dayton, Ohio)》2003,21(2):152-161
Human stem cells derived from human fertilized oocytes, fetal primordial germ cells, umbilical cord blood, and adult tissues provide potential cell-based therapies for repair of degenerating or damaged tissues. However, the diversity of major histocompatibility complex (MHC) antigens in the general population and the resultant risk of immune-mediated rejection complicates the allogenic use of established stem cells. We assessed an alternative approach, employing chemical activation of nonfertilized metaphase II oocytes for producing stem cells homozygous for MHC. By using F1 hybrid mice (H-2-B/D), we established stem cell lines homozygous for H-2-B and H-2-D, respectively. The undifferentiated cells retained a normal karyotype, expressed stage-specific embryonic antigen-1 and Oct4, and were positive for alkaline phosphatase and telomerase. Teratomatous growth of these cells displayed the development of a variety of tissue types encompassing all three germ layers. In addition, these cells demonstrated the potential for in vitro differentiation into endoderm, neuronal, and hematopoietic lineages. We also evaluated this homozygous stem cell approach in human tissue. Five unfertilized blastocysts were derived from a total of 25 human oocytes, and cells from one of the five hatched blastocysts proliferated and survived beyond two passages. Our studies demonstrate a plausible "homozygous stem cell" approach for deriving pluripotent stem cells that can overcome the immune-mediated rejection response common in allotransplantation, while decreasing the ethical concerns surrounding human embryonic stem cell research. 相似文献
87.
K. V. I. Rolston M. Messer H. Nguyen D. Ho B. LeBlanc G. P. Bodey 《European journal of clinical microbiology & infectious diseases》1991,10(7):581-585
The in vitro activity of cefpodoxime, an oral cephalosporin ester, against 792 bacterial isolates representing 36 species was evaluated in comparison to that of ciprofloxacin and trimethoprim/sulfamethoxazole (TMP/SMX). Cefpodoxime inhibited the majority ofStreptococcus spp.,Haemophilus influenzae andProteus mirabilis at a concentration of 0.12 µg/ml. It was also active againstCitrobacter diversus, Escherichia coli, Klebsiella spp.,Proteus vulgaris, Serratia marcescens and methicillin-susceptibleStaphylococcus aureus isolates. Overall, cefpodoxime appeared to be less active than ciprofloxacin and TMP/SMX against many pathogens common in cancer patients. 相似文献
88.
89.
de Jong MD Bach VC Phan TQ Vo MH Tran TT Nguyen BH Beld M Le TP Truong HK Nguyen VV Tran TH Do QH Farrar J 《The New England journal of medicine》2005,352(7):686-691
In southern Vietnam, a four-year-old boy presented with severe diarrhea, followed by seizures, coma, and death. The cerebrospinal fluid contained 1 white cell per cubic millimeter, normal glucose levels, and increased levels of protein (0.81 g per liter). The diagnosis of avian influenza A (H5N1) was established by isolation of the virus from cerebrospinal fluid, fecal, throat, and serum specimens. The patient's nine-year-old sister had died from a similar syndrome two weeks earlier. In both siblings, the clinical diagnosis was acute encephalitis. Neither patient had respiratory symptoms at presentation. These cases suggest that the spectrum of influenza H5N1 is wider than previously thought. 相似文献
90.
The 11q13 amplicon of a mammary carcinoma cell line 总被引:1,自引:0,他引:1
M Lafage C Nguyen P Szepetowski M J Pébusque J Simonetti G Courtois P Gaudray O deLapeyriere B Jordan D Birnbaum 《Genes, chromosomes & cancer》1990,2(3):171-181
Fifteen to 20% of breast carcinomas show amplification of genes located at 11q13. The HST/FGFK and INT2 fibroblast growth factor (FGF)-related genes and the BCL1 locus are usually present in the amplification units. We have investigated the structure and chromosomal location of the 11q13 amplicon of the MDA-MB-134 mammary carcinoma cell line by using in situ chromosomal and pulsed field gel hybridizations. The results indicate that a limited number of amplification units are involved in the constitution of an extended chromosomal region located on 11q. These units do not show any important rearrangement over rather large distances around the HST/INT2 and BCL1 loci. 相似文献