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Biomedical subjects

Y H Yang

Publications and source records attributed to Y H Yang.

At least 19 recordsLinked to original sources

Prolactin regulation of mitochondrial aspartate aminotransferase and protein kinase C in human prostate cancer cells.

Citrate production is a major physiological function of the prostate that is regulated by testosterone and prolactin. Mitochondrial aspartate aminotransferase (mAAT) is a key enzyme in the metabolic pathway of prostate citrate production. In addition, prolactin stimulates expression of mAAT in the rat lateral prostate. In this report we establish the role of prolactin in the regulation of mAAT in two prostate cancer cell lines, LNCaP and PC-3. LNCaP cells respond to hormonal stimulation with increased secretion of prostate specific products. PC-3 cells, on the other hand, are testosterone independent and apparently do not respond to other growth factors either. Results showed that both LNCaP and PC-3 cells responded to prolactin with increased mAAT activity and an increased steady state level of mAAT mRNA. Prolactin also increased protein kinase C (PKC) activity in both these cell lines. Treatment of LNCaP and PC-3 cells with the phorbol ester 12-O-tetradecanoylphorbol (TPA) caused the same effect on mAAT activity and mRNA level as prolactin. The results suggest that the diacylglycerol-PKC signal transduction system mediates the prolactin effect on mAAT. In addition, these results also show that the prolactin effect on mAAT is independent of androgens since PC-3 cells reportedly lack androgen receptor expression. Thus, these results provide evidence that prolactin is a physiological regulator of prostate function in human as well as rat prostate. In addition, the results also show that though prostate cancer cells are androgen independent, they remain responsive to prolactin. This could have important implications for the treatment and management of prostate cancer.

Animals

The genomic structure of a mouse seminal vesicle autoantigen.

The genomic structure of an androgen-stimulated mouse seminal vesicle autoantigen was determined. Analysis of the nucleotide sequence established 2135 bp of the 5'-flanking region, four exons of 123, 136, 112, 227 bp, three introns of 1555, 1931, 316 bp, and 185 bp of the 3'-flanking region of this gene. Ten DNA segments, five in the 5'-flanking region, two in the first intron, and three in the second intron were identified to have more than 50% homology with the consensus sequence of the androgen response element (ARE). Two sets of adjacent DNA segments, one including -213 to -199 bp and -124 to -110 bp in the 5'-flanking region and the other including 2532 to 2546 bp and 2582 to 2596 bp in the second intron, are noticeable for their high degree of homology with ARE.

Androgens

Heterogeneity of the human H blood group alpha(1,2)fucosyltransferase gene among para-Bombay individuals.

BACKGROUND AND OBJECTIVES: The para-Bombay phenotype has a relatively high frequency of about 1 in 8,000 Taiwanese. Studies were carried out on eight healthy and unrelated Taiwanese with the para-Bombay phenotype to cast light on its immunogenetic basis. MATERIALS AND METHODS: Blood and saliva samples were tested with standard hemagglutination techniques. Salivary ABH substances were determined by hemagglutination inhibition. PCR techniques were used to amplify the coding region of the H genes. RESULTS: Five different h alleles, designated as h1, h2, h3, h4 and h5, were identified in the Taiwanese with the para-Bombay phenotype. The h1 allele loses one of the three AG repeats located at the nucleotides 547-552 of the H gene, whereas two of the three T repeats located at the nucleotides 880-882 are deleted in the h2 allele. The h3 allele contains a C658 to T missense mutation, whereas two missense mutations, C35 to T and A980 to C were identified in the h4 allele. A T460 to C missense is present in the h5 allele. The h5 allele was identified in an individual whose red blood cells contain blood group A antigen but not H antigen, and thus may be considered a weak variant of the H gene. CONCLUSIONS: So far no biologic relevance of the H antigen has been discovered, and its deficiency does not seem to produce any deleterious effects. There may be better understanding of the evolutionary basis for the polymorphisms at these loci after systematic study of different ethnic populations.

ABO Blood-Group System

Rapid polymerase chain reaction analysis of St14 (DXS52) VNTR: carrier detection of hemophilia A.

OBJECTIVE: To determine the frequency of St14 VNTR allele in Koreans as a marker of the hemophilia A and to evaluate the efficacy of this marker for carrier detection of hemophilia A METHODS: PCR amplified RFLP analysis of St14 VNTR was done in 312 X-chromosomes of 122 unrelated Korean males and 95 females and the same method was applied to carrier detection in the 2 hemophilia A families. RESULTS: There were 13 alleles of different sizes of St14 VNTR locus appeared in 312 X-chromosomes of unrelated Koreans. For carrier detection of hemophilia A, in the family A, the mother showed 1390/ 1330 bp alleles and the father showed 700 bp allele. The affected son has inherited 1390 bp allele from his mother. The daughter at risk showed 1330/700 bp alleles. In family B, the mother showed 1280/700 bp alleles and the stepfather showed 1390 bp allele. The affected son has inherited 1280 bp allele. The daughter at risk showed 1390/700 bp alleles. And so the daughters of the 2 families were not carriers for hemophilia A. CONCLUSION: PCR analysis of St14 VNTR was a useful tool for carrier detection of hemophilia A.

Alleles

Identification and characterization of acoK, a regulatory gene of the Klebsiella pneumoniae acoABCD operon.

By using transposon insertional mutagenesis and deletion analyses, a recombinant clone containing the region upstream of the acoABCD operon of Klebsiella pneumoniae was found to be required for acetoin-inducible expression of the operon in Escherichia coli. The nucleotide sequence of the region was determined, and it displayed an open reading frame of 2,763 bp that is transcribed divergently to the acoABCD operon. This gene, designated acoK, is capable of encoding a protein with an overall 58.4% amino acid identity with MalT, the transcriptional activator of the E. coli maltose regulon. A conserved sequence for nucleotide binding at the N-terminal region, as well as a helix-turn-helix motif belonging to the LuxR family of transcriptional regulators at the C terminus, was also identified. Primer extension analysis identified two transcription initiation sites, S1 and S2, located 319 and 267 bp, respectively, upstream of the putative start codon of acoK. Several copies of NtrC recognition sequence [CAC-(N11 to N18)-GTG] were found in the promoter regions of both the acoK gene and the acoABCD operon. Acetoin-dependent expression of the acoABCD operon could be restored in the E. coli acoK mutants by supplying a plasmid carrying an intact acoK, suggesting a transactivating function of the gene product. The AcoK protein overproduced in E. coli was approximately 100 kDa, which is in good agreement with the molecular mass deduced from the nucleotide sequence. A specific DNA binding property and an ATPase activity of the purified AcoK were also demonstrated.

Acetoin

Exacerbation of adjuvant arthritis by adrenalectomy is associated with reduced leukocyte lipocortin 1.

OBJECTIVE: Lipocortin 1 is a mediator of the antiinflammatory actions of therapeutic glucocorticoids. ENdogenous glucocorticoids modulate inflammatory arthritides including rheumatoid and adjuvant arthritis (AA), but the role of lipocortin 1 in this phenomenon is not known. We studied the effects of endogenous glucocorticoids on adjuvant arthritis and leukocyte lipocortin 1 content. METHODS: Adrenalectomy of sham adrenalectomy was performed 2 days before adjuvant injection in 170 g inbred Sprague-Dawley rats. Peripheral blood was obtained and disease severity assessed by delta paw volume and clinical score 14 days later. Leukocyte subset lipocortin 1 content was determined by double labeling permeabilization flow cytometry using specific monoclonal antibodies. RESULTS: Lipocortin 1 fluorescence was readily detected in control rat peripheral blood cells labeled with OX-1 (pan-leukocyte), OX-19 (CD5), W3/25 (CD4), and OX-8 (CD8). Lipocortin 1 fluorescence was significantly greater in polymorphonuclear leukocytes (PMN) (RP3; p < 0.01). Induction of AA was accompanied by significant increases in lipocortin 1 (p < 0.001) in all subsets. Sham adrenalectomy induced no significant change in AA rat leukocyte lipocortin 1. Adrenalectomy induced significant exacerbation of AA disease severity compared to sham operation (delta paw volume 1.43 +/- 0.1 vs 1.13 +/- 0 ml; p < 0.05). Adrenalectomy was also associated with significant reduction in lipocortin 1 content in all leukocyte subsets except PMN. Leukocyte lipocortin 1 content exhibited significant negative correlation with clinical disease severity (R2 = -0.55; p < 0.05). CONCLUSION: Endogenous glucocorticoids modulate leukocyte expression of lipocortin 1 in inflammatory disease, and reduced lipocortin 1 may be involved in the exacerbation of AA by adrenalectomy.

Adrenalectomy

Heterogeneity of the human Secretor alpha(1,2)fucosyltransferase gene among Lewis(a+b-) non-secretors.

The human Secretor alpha (1,2)fucosyltransferase gene determines the ABH secretor status and influences the Lewis phenotype of an individual. Two different se alleles with point mutations, C571 to T and G849 to A respectively, in the coding region were identified in Le(a+b-) non-secretors from one of the Taiwanese indigenous groups. The base substitutions predict the alteration of Arg191 and Trp283 to stop codons respectively, resulting in deletion of the Secretor enzyme's C-terminal segment. Both alleles of the Secretor locus in all Le(a+b-) non-secretors, but not in Le(a+b-) secretors, were further demonstrated to be either one of these two se alleles with nonsense mutations. These results suggest two new molecular bases for the null se allele responsible for the formation of the non-secretor phenotype.

ABO Blood-Group System

Identification and characterization of the acoD gene encoding a dihydrolipoamide dehydrogenase of the Klebsiella pneumoniae acetoin dehydrogenase system.

The acoD gene, which encodes a dihydrolipoamide dehydrogenase component of the acetoin dehydrogenase enzyme system of Klebsiella pneumoniae was isolated and the nucleotide sequence determined. The gene is capable of encoding a protein of 465 amino acid residues with conserved binding domains for NAD and FAD, and two redox-active cysteine residues. The acoD gene product exhibited a Michaelis constant of 170 microM for NAD, while NADP can not be used as a substrate. The purified enzyme appeared to be a dimer of the acoD gene product. It did not associate tightly with the E1 and E2 components of either acetoin dehydrogenase or 2-oxoglutarate dehydrogenase to form an active multi-enzyme complex.

Acetoin Dehydrogenase

Chorionic villus sampling: clinical experience of the initial 750 cases.

OBJECTIVE: To assess the factors that might influence the success rate, safety and reliability of chorionic villus sampling (CVS) and to evaluate the relationship between CVS and other congenital anomalies. DESIGN: Analysis of the outcome of 750 cases of CVS (730 cases with transcervical and 20 cases with transabdominal). SETTING: The outpatient prenatal genetic clinic of a university tertiary care center. SUBJECT: Seven hundred and fifty pregnant women that underwent CVS for prenatal genetic diagnosis from 7 to 12 weeks of gestation. RESULTS: Advanced maternal age was the most common indication for CVS (32.8%). The overall sampling success rate was 98.0% (735/750), representing 93.9% at 7 to 8 weeks, 98.1% at 9 to 10 weeks, and 98.3% at 11 to 12 weeks of gestation. The majority of cases (93.1%) required one or two aspirations. Cytogenetic analysis routinely included direct overnight and long-term culture methods which revealed 16 abnormalities (2.2%). Of 735 cases in which CVS was successful, 700 advanced to normal offspring, 17 had therapeutic termination, and 18 resulted in spontaneous abortions; there was an overall fetal loss rate of 2.4% (18/750). CONCLUSION: CVS in early pregnancy is a relatively safe and reliable method of prenatal genetic diagnosis capable of replacing genetic amniocentesis. However, it must be done by experienced personnel. No congenital anomalies were found to be related to CVS in this series.

Adult

[Vaginal colonization of group B Streptococcus: a study in 267 cases of factory women].

An epidemiologic study on vaginal colonization of group B streptococcus (GBS) from non-pregnancy women was carried out. Two hundred sixty seven female workers were studied. The carrier rate of GBS in vaginal specimens was 10.86%. Women aged 45 years old and above had more cases with genital tract GBS colonization. Women with vaginal colonization had more history of miscarriage and using IUD. We did not find the positive correlations between vaginal colonization and oral contraceptive, ovarian cyst, hysteromyoma in our study group. Women with gynecologic inflammation had more cases with vaginal GBS colonization. There is a significant increase for women with vaginitis and cervicitis. Serotyping study showed that types III and II were the most frequent GBS types isolated from the carriers. Antibiotic sensitivity test showed that more than half GBS strains were resistant to oxcillin and amikacin.

Adult

Correlation of a missense mutation in the human Secretor alpha 1,2-fucosyltransferase gene with the Lewis(a+b+) phenotype: a potential molecular basis for the weak Secretor allele (Sew).

A missense mutation (A385 to T), predicting an Ile129 to Phe substitution, in the human Secretor alpha 1,2-fucosyltransferase gene was present in double dose in Lewis(a+b+) individuals, but not in Lewis(a-b+) individuals. Co-segregation of the Lewis(a+b+) phenotype with homozygosity for the mutation was also verified. These results yield a potential molecular basis for the weak Secretor allele (Sew) accounting for the Lewis(a+b+) phenotype.

Alleles

Rapid chromosome analysis and prenatal diagnosis using fluid from the cystic hygroma, hydrothorax and isolated ascites: new source for chromosome analysis.

OBJECTIVE: To determine whether abnormally collected fetal body fluid could be used as alternative cell sources for cytogenetic assessment. METHOD: Using modified lymphocyte culture, the abnormal fetal body fluids obtained from the five fetal anomaly cases (3 cystic hygromas, 1 hydrothorax, and 1 isolated ascites) were analyzed in order to determine their karyotypes. RESULTS: Two cases showed 45, X and 46, XX/45, X and the other three showed a normal karyotype. CONCLUSION: The cytogenetic analysis from body fluid aspiration yields a rapid and accurate result within 96 hours. This method could be helpful when cordocentesis or placental biopsy is not feasible or when time requirement for amniotic fluid cell culture is burdensome to patient management.

Adult

CDEBP, a site-specific DNA-binding protein of the 'APP-like' family, is required during the early development of the mouse.

A murine protein, termed CDEBP, was previously shown to bind the double-stranded DNA motif GTCACATG, identical to the yeast centromeric element CDEI. The cDNA sequence showed three domains with extensive similarities to the amyloid beta precursor protein (APP). The protein is homologous over its entire length to the human protein designated APPH. In situ immunofluorescence assays using antibodies raised against distinct parts of CDEBP detected discrete sites of accumulation inside the interphase nucleus, and the bulk of the protein was not associated with mitotic chromosomes. One of the complexes with double-stranded CDEI oligonucleotides detected by gel shift assay was not present when the protein had been selectively removed from nuclear extracts by immunoprecipitation. We reported previously that microinjection into one-cell mouse embryos of DNA fragments including the CDEI sequence results in an early arrest of development with abnormal nuclei containing variable amounts of DNA. The same characteristic figures were observed when embryos were treated with antisense oligonucleotides complementary to parts of the CDEBP coding region. Complexes between the CDEBP protein and CDEI sites in the mouse genome thus appear to play a critical role in the replication/segregation of the embryonic genome.

3T3 Cells

Prenatal fetal sex determination from maternal peripheral blood using polymerase chain reaction.

We have investigated the use of a nested polymerase chain reaction(PCR) assay with Y-specific sequence from the DYS 14 locus on the short arm of Y-chromosome for prenatal sex determination in the peripheral blood of 22 pregnant women who participated in the antenatal genetic diagnosis program. The sensitivity and specificity of the nested PCR using DYS 14 locus primers(Y1.5,Y1.6, and Y1.7,Y1.8) were 76.4% and 55.5%, respectively. In terms of gestational age, positive predictive values of 66.6%, 66.6%, and 80% were obtained for the first, second, and third trimester respectively. The corresponding negative predictive values were 50%, 50%, and 100% respectively. Male specific band was positive in three of the six cases of female bearing women and male specific band was negative in three of the seven cases of male bearing women during 9-16 gestational weeks showing low sensitivity. But all cases except one show the male specific band during the male fetus and all female fetuses did not show the male specific 198 base pair band during 18 approximately 40 gestational weeks. This study suggests that prenatal sex determination by PCR employing maternal peripheral blood was usually possible in late pregnancy but less reliable in early pregnancy. It seems that if we used a method separating fetal cells from maternal blood and then run PCR on these cells with DYS 14 locus primers we could make a fairly accurate fetal sex determination.

Base Sequence