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Biomedical subjects

J J Yu

Publications and source records attributed to J J Yu.

At least 37 records · Page 2Linked to original sources
PubMed1999

Isolation and confirmation of function of the Coccidioides immitis URA5 (orotate phosphoribosyl transferase) gene.

The OPRTase (URA5) gene of the human pathogenic fungus, Coccidioides immitis (Ci), was cloned, sequenced, chromosome-mapped and expressed both by transformation of Escherichia coli and by complementation of wdura5Delta, an auxotrophic strain of Wangiella dermatitidis (Wd) with a disrupted URA5 gene. A functional assay of the recombinant URA5 expressed by E. coli was conducted to ensure that the isolated Ci gene encodes the appropriate enzyme. In the absence of a transformation system for Ci, we also used a reported method of introduction of heterologous DNA into cells of the phylogenetically related fungus, Wangiella dermatitidis, to confirm the function of the Ci URA5 gene. Both the genomic and cDNA sequences of the Ci URA5 gene are presented. The transcription start point and two poly(A) addition sites were confirmed. The gene contains a 714-bp ORF that translates a 238-amino-acid (aa) protein of 25.5kDa and pI of 6.5. No introns are present. The translated protein contains a single, putative N-glycosylation site. The deduced Ci protein showed 55-63% aa sequence similarity to reported fungal OPRTases. The URA5 gene was mapped to chromosome IV of Ci, and was shown to be a single copy gene by Southern and Northern hybridizations. Transformation of the wdura5Delta mutant to prototrophy was accomplished by electroporation of Wd yeast cells with the Ci URA5 gene. Cellular uptake of the heterologous DNA was confirmed by Southern hybridization. The stable transformants were unable to grow on a medium containing 5-FOA. Expression of the Ci URA5 gene can be used as a selectable marker for a transformation system, and the latter is essential for molecular studies of this pathogenic fungus.

Amino Acid Sequence↗
PubMed1998

Reducing lipid peroxidation stress of erythrocyte membrane by alpha-tocopherol nicotinate plays an important role in improving blood rheological properties in type 2 diabetic patients with retinopathy.

The effects of alpha-tocopherol nicotinate on blood viscoelasticity and viscosity and on lipid peroxidation stress in erythrocyte membranes in patients with Type 2 DM were investigated. Thirteen Type 2 diabetic subjects with retinopathy were given alpha-tocopherol nicotinate 300 mg tds, after meals, for 3 months. The treatment resulted in significant reductions of blood viscosity at different shear rates (e.g. -2.23 +/- 2.82 p<0.015, gamma = 1.5 s(-1)) and viscoelasticity (p<0.004); resistance of erythrocyte deformation (p<0.001) and lipid peroxidation stress in red cell membrane (malondialdehyde or MDA reduced by 0.17 +/- 0.13 nmol l(-1) p<0.005). Plasma viscosity, red cell rigidity, and HbA1c were unchanged. There were negative linear correlations between the indices of red cell deformability and the levels of MDA of red cell membrane both pre- and post-treatment (e.g. R = -0.79, p<0.001; R = -0.78, p<0.002, n = 13; pre- and post-, respectively). We suggest that the improvements of rheological properties of blood and red cell deformability by alpha-tocopherol nicotinate are mainly attributed to reducing lipid peroxidation stress on membrane of red blood cells. The treatment may be useful in slowing deterioration of microangiopathy in Type 2 DM.

Aged↗
PubMed1998

Alternative splicing of ERCC1 and cisplatin-DNA adduct repair in human tumor cell lines.

Alternative splicing is a common natural tool for the inhibition of function of full length gene products. We explored whether there was evidence that alternative splicing of ERCC1 may serve such a function for nucleotide excision repair. The ratio of alternatively spliced species to full length species was assessed for the protein and/or for the mRNA, for a series of human cell lines and tissues. This ratio was plotted against the amount of cisplatin-DNA adduct repair in each cell line (n=9), as measured by atomic absorbance spectrometry. As the percentage of alternatively spliced protein and/or mRNA increased, the amount of cisplatin-DNA adduct that was repaired was reduced. This inverse relationship was associated with a substantial amount of scatter (r=0.635), particularly at low levels of repair. These data demonstrate an association between alternative splicing of ERCC1, and reduction in cellular capability to repair cisplatin-DNA adduct.

Alternative Splicing↗
PubMed1998

Cisplatin and phorbol ester independently induce ERCC-1 protein in human ovarian carcinoma cells.

Nucleotide excision repair (NER) is the DNA repair pathway by which cisplatin-induced damage is removed from DNA in human cells. ERCC-1 is one of the essential proteins in NER, and is essential for life. Enhanced ERCC-1 expression has been associated with clinical and cellular resistance to cisplatin. We therefore carried out this study to investigate the effect of cisplatin on ERCC-1 protein expression in A2780/CP70 human ovarian cancer cells. Western blot analysis showed that ERCC-1 protein levels were increased to more than 3 times control after a 1 h cisplatin exposure to A2780/CP70 cells in culture. This increase was time- and concentration-dependent. The effect of cisplatin was maximal at 40 mM and peaked 24-48 h after exposure to the drug. These results extend our previous observations that ERCC-1 mRNA expression is induced by cisplatin in this system. TPA, a known AP-1 activator and tumor-promoting phorbol ester, also induced ERCC-1 protein to the same extent as cisplatin, but did not synergize with cisplatin in this regard. These findings suggest that ERCC-1 gene up-regulation in these cells can result through a DNA damage-response pathway, or through the induction of AP-1 activity, independent of the occurrence of DNA damage.

Antineoplastic Agents↗
PubMed1997

The hsp60 gene of the human pathogenic fungus Coccidioides immitis encodes a T-cell reactive protein.

A heat shock protein-encoding gene (hsp60) from the human respiratory fungal pathogen, Coccidioides immitis (Ci), was cloned, sequenced, chromosome-mapped, expressed and immunolocalized in parasitic cells. Both the genomic and cDNA sequences are presented. The transcription start point and poly (A) addition site were confirmed. The hsp60 gene contains two introns and a 1782-bp ORF which translates a 594-amino acid (aa) protein of 62.4 kDa and pI of 5.6. The translated protein revealed two potential N-glycosylation sites. The deduced HSP60 showed 78-83% aa sequence similarity to reported fungal HSP60 proteins. The hsp60 gene was mapped to chromosome III of Ci and was shown to be a single copy gene by Southern and Northern hybridization. Expression of a 1737-bp cDNA fragment of the hsp60 gene in E. coli resulted in production of a recombinant protein. Amino acid sequence analysis of the recombinant protein confirmed that it was encoded by the Ci hsp60 gene. Antiserum raised in mice against the isolated recombinant protein immunolocalized HSP60 in the cytoplasm and wall of parasitic cells of Ci. The recombinant HSP60 was used to immunize BALB/c mice and was shown to induce proliferation of T cells isolated from lymph nodes of these animals. The hsp60 gene of Ci is the first reported heat-shock protein gene of this human pathogen.

Amino Acid Sequence↗
PubMed1997

Isolation and characterization of the urease gene (URE) from the pathogenic fungus Coccidioides immitis.

The urease (URE)-encoding gene from Coccidioides immitis (Ci), a respiratory fungal pathogen of humans, was cloned, sequenced, chromosome-mapped and expressed. Both the genomic and cDNA sequences are reported. The transcription start point and poly(A)-addition site were confirmed. The URE gene contains eight introns and a 2517-bp ORF that translates a 839-amino-acid (aa) protein of 91.5 kDa and pI of 5.5, as deduced by computer analysis of the nucleotide sequence. The translated protein revealed eight putative N-glycosylation sites. The deduced URE showed comparable levels of homology to reported URE of the jack bean plant (Canavalia ensiformis; 71.8%) and URE of several genera of bacteria (Bp, 71.7%; Hp, 68.3%; Ka, 71.6%; Pm, 71.9%). The URE gene was mapped to chromosome III of Ci and was shown to be a single copy gene by Southern hybridization. Expression of a 1687-bp fragment of the URE gene in E. coli resulted in the production of a 63-kDa recombinant protein that was recognized in an immunoblot by antiserum raised against the Ka URE homolog. This is the first report of a fungal URE gene.

Amino Acid Sequence↗
PubMed1997

A nucleotide polymorphism in ERCC1 in human ovarian cancer cell lines and tumor tissues.

We studied the DNA sequence of the entire coding region of ERCC1 gene, in five cell lines established from human ovarian cancer (A2780, A2780/CP70, MCAS, OVCAR-3, SK-OV-3), 29 human ovarian cancer tumor tissue specimens, one human T-lymphocyte cell line (H9), and non-malignant human ovary tissue (NHO). Samples were assayed by PCR-SSCP and DNA sequence analyses. A silent mutation at codon 118 (site for restriction endonuclease MaeII) in exon 4 of the gene was detected in MCAS, OVCAR-3 and SK-OV-3 cells, and NHO. This mutation was a C-->T transition, that codes for the same amino acid: asparagine. This transition converts a common codon usage (AAC) to an infrequent codon usage (AAT), whereas frequency of use is reduced two-fold. This base change was associated with a detectable band shift on SSCP analysis. For the 29 ovarian cancer specimens, the same base change was observed in 15 tumor samples and was associated with the same band shift in exon 4. Cells and tumor tissue specimens that did not contain the C-->T transition, did not show the band shift in exon 4. Our data suggest that this alteration at codon 118 within the ERCC1 gene, may exist in platinum-sensitive and platinum-resistant ovarian cancer tissues.

Antineoplastic Agents↗
PubMed1997

Status of the eradication of indigenous wild poliomyelitis in the People's Republic of China.

A large nationwide outbreak occurred in 1989-1990 in China, in which nearly 10,000 poliomyelitis cases were reported. After two rounds of oral poliovirus vaccine (OPV) supplemental activity in nearly every province in the 1992-1993 winter season, no wild poliovirus was detected in 1993 in 22 provinces in the middle of China that contained 86% of the population. During the first national immunization days (NIDs) conducted in December 1993 and January 1994, 83 million children 0-47 months of age were immunized. In 1994, wild poliovirus was identified in only 6 of 2397 children with stool specimens tested. After a second NID in December 1994 and January 1995, no wild poliovirus was detected in 1995 despite a very high level of virus surveillance. In summary, double-round mass supplemental OPV immunizations in children 0-3 years old in two consecutive winters eliminated wild poliovirus from 23% of the world's population (1.2 billion people).

Child, Preschool↗
PubMed1997

Effect of target age of supplemental immunization campaigns on poliomyelitis occurrence in China.

The World Health Organization recommends conducting supplemental immunization activities to eradicate poliomyelitis by the year 2000. Although effective in eliminating poliomyelitis from the Americas, supplemental campaigns require substantial resources. To assess differential campaign effectiveness in eliminating this disease, poliomyelitis occurrence was compared in counties in China that targeted children <3 versus <4 years of age. Counties that targeted children <3 years of age reported poliomyelitis more frequently after the campaigns. This association was observed even after accounting for the effects of previous poliomyelitis occurrence, urban versus rural setting, and population density. While several limitations emphasize the preliminary nature of these findings, these data support targeting the widest possible age group of susceptible children to ensure maximum effectiveness in eliminating poliomyelitis. Thus, while reducing the target age of these activities may result in considerable resource savings, such campaigns may not be as effective in eliminating poliomyelitis.

Age Factors↗
PubMed1996

Comparative analyses of relative ERCC3 and ERCC6 mRNA levels in gliomas and adjacent non-neoplastic brain.

Nucleotide excision repair (NER) is an ordered process in nonmalignant cells, in both human and nonhuman systems. We previously reported that in human brain there is discordant mRNA expression of excision repair cross-complementing (ERCC) 1 and ERCC2 in malignant tissues, concurrent with excellent concordance of these genes in nonmalignant tissues from the same patients. Here we have extended these studies to compare low-grade tumors to high-grade tumors and to include ERCC3 (which links DNA repair with DNA transcription) and ERCC6 (which is essential for gene-specific repair). Glial tumor and adjacent normal brain specimens from 19 individuals were studied. Paired malignant and nonmalignant tissues were obtained from 12 of these patients. For ERCC3, there was excellent concordance of mRNA expression between malignant and nonmalignant tissues from the same individuals (P = 0.003). For ERCC6, no concordance was observed (P = 0.314). Tumor tissue from patients with high-grade gliomas exhibited marked discordance of mRNA expression patterns in situations in which good concordance was observed in tumor tissue from low-grade gliomas. We previously established that malignant brain tumors show increased disorder of genes in the NER process, as compared with nonmalignant tissues. These data suggest that increasing disorder in the NER process may occur as cells move from low-grade to high-grade malignancy.

Astrocytoma↗
PubMed1995

Malignant and nonmalignant brain tissues differ in their messenger RNA expression patterns for ERCC1 and ERCC2.

Perturbation of the DNA repair process appears to be responsible for the occurrence of a number of human diseases, which are usually associated with a propensity to develop internal malignancies and/or disorders of the central nervous system. We have been interested in the possibility that a subtle abnormality in DNA repair competency might be associated with the transformation of nonmalignant cells to the malignant state. To study this question, we assayed malignant and nonmalignant brain tissues from 19 individuals for mRNA expression levels of the human DNA repair genes ERCC1, ERCC2, and XPAC and for differential splicing of the ERCC1 transcript. We separately compared expression levels of these genes in the following situations: concordance of expression within malignant tissues; concordance of expression within nonmalignant tissues; concordance between malignant and nonmalignant tissues within individuals of the cohort; and concordance of gene expression between two nonmalignant tissue sites within a single individual. Linear regression analyses of mRNA values obtained suggested orderly concordance of these three DNA repair genes in nonmalignant tissues within the patient cohort and an excellent concordance of these genes between two separate biopsy sites from the same individual. In contrast, malignant tissues showed disruption of concordance between the full-length ERCC1 transcript and ERCC2, which have excision and helicase functions, respectively. Furthermore, within the same individuals, malignant tissues were discordant with nonmalignant tissues for ERCC1 and ERCC2, although concordance for XPAC was preserved. These data suggest that one molecular characteristic of human malignancy may be the disruption of the normal relationship between the excision and the helicase functions of the nucleotide excision repair pathway.

Brain↗
PubMed1995

[Disseminated Strongyloides stercoralis infection mimicking pneumonia].

Strongyloides stercoralis is an intestinal nematode. In an immunocompetent host, Strongyloides infections usually produce only mild gastrointestinal symptoms. However, in an immunocompromised host, widespread dissemination of larvae to the extra-intestinal organs may occur. If unrecognized, the mortality rate is high. Here we report a case of disseminated strongyloidiasis in a chronic obstructive pulmonary disease (COPD) subject whose chest radiograph demonstrated multiple pneumonic patches and interstitial infiltrates. Strongyloides larvae were found in stool, sputum, and urine, and embryonated eggs were also found in sputum. The patient was treated successfully with mebendazole and alben albendazole. In conclusion, although high mortality rate is noted in disseminated strongyloidiasis, it is still a curable disease when early diagnosis and treatment could be made.

Animals↗
PubMed1994

Messenger RNA levels of XPAC and ERCC1 in ovarian cancer tissue correlate with response to platinum-based chemotherapy.

Nucleotide excision repair is a DNA repair pathway that is highly conserved in nature, with analogous repair systems described in Escherichia coli, yeast, and mammalian cells. The rate-limiting step, DNA damage recognition and excision, is effected by the protein products of the genes ERCC1 and XPAC. We therefore assessed mRNA levels of ERCC1 and XPAC in malignant ovarian cancer tissues from 28 patients that were harvested before the administration of platinum-based chemotherapy. Cancer tissues from patients whose tumors were clinically resistant to therapy (n = 13) showed greater levels of total ERCC1 mRNA (P = 0.059), full length transcript of ERCC1 mRNA (P = 0.026), and XPAC mRNA (P = 0.011), as compared with tumor tissues from those individuals clinically sensitive to therapy (n = 15). In 19 of these tissues, the percentage of alternative splicing of ERCC1 mRNA was assessed. ERCC1 splicing was highly variable, with no difference observed between responders and nonresponders. The alternatively spliced species constituted 2-58% of the total ERCC1 mRNA in responders (median = 18%) and 4-71% in nonresponders (median = 13%). These data suggest greater activity of the DNA excision repair genes ERCC1 and XPAC in ovarian cancer tissues of patients clinically resistant to platinum compounds. These data also indicate highly variable splicing of ERCC1 mRNA in ovarian cancer tissues in vivo, whether or not such tissues are sensitive to platinum-based therapy.

Carboplatin↗
PubMed1994

Rheological parameter alternations in blood may impair oxygen transported to the retina in type II diabetes (NIDDM).

Rheological properties of blood, retinal capillary blood flow velocity and oxygen transport efficiency (TE) were measured and calculated from 21 type II diabetic patients (NIDDM) and 19 normal subjects. The results showed that the diabetes group had increased plasma viscosity and blood viscosities (P < 0.05 or better) at all tested shear rates. For both shear rates (gamma) of 225 sec-1 and 450 sec-1, the diabetes group displayed increased internal viscosity of red cells (Tk) (P < 0.002 and P < 0.001, respectively) but lower values of TE (P < 0.001, both). A lower value of retinal capillary blood flow velocity (P < 0.005) was also noted. Upon further analysis of possible relations between Hct values and Tk of red cells, it was found that Hct values were linearly correlated with Tk for both shear rates of the diabetes group (gamma = 225 sec-1 and gamma = 450 sec-1; R = -0.63 and -0.65, n = 21 and 19, respectively, both P < 0.005); control group (gamma = 225 sec-1 and gamma = 450 sec-1; R = -0.84 and -0.82, respectively, n = 19, both P < 0.001) and all subjects (gamma = 225 sec-1 and gamma = 450 sec-1; R = -0.70 and -0.72, n = 40 and 38, respectively, both P < 0.001). The impaired rheological properties of blood and internal viscosity of red cells may result in both reduced capillary blood flow velocities and less oxygen transported to tissues. This may possibly contribute to the deterioration of retinopathy or microangiopathy in diabetic patients.

Blood Flow Velocity↗
PubMed1993

[Pulmonary siderosis: report of a case].

Pulmonary siderosis is one kind of pneumoconiosis caused by the long term inhalation of iron dust. It occurs in a number of occupations including steel rolling and grinding, welding, polishing, casting, boiler scaling, iron ore mining and emery working. Here we report a case of pulmonary siderosis. A 49-year-old male who had worked in an iron foundry for 30 years was admitted because diffuse micronodular lesions were seen in both lung fields on a routine chest radiographic study. A pulmonary function test disclosed a mildly restrictive ventilatory defect. Transbronchial lung biopsy revealed a significant amount of iron dust deposited within a fibrous nodule by which confirmed the diagnosis of pulmonary siderosis.

Humans↗
PubMed1992

Isolation and characterization of the cDNA encoding human DNA methyltransferase.

We have cloned a series of overlapping cDNA clones encoding a 5194 bp transcript for human DNA methyltransferase (DNA MTase). This sequence potentially codes for a protein of 1495 amino acids with a predicted molecular weight of 169 kDa. The human DNA MTase cDNA has eighty percent homology at the nucleotide level, and the predicted protein has seventy-four percent identity at the amino acid level, to the DNA MTase cDNA cloned from mouse cells. Like the murine DNA MTase, the amino terminal two-thirds of the human protein contains a cysteine-rich region suggestive of a metal-binding domain. The carboxy terminal one-third of the protein shows considerable similarity to prokaryotic (cytosine-5)-methyltransferases. The arrangement of multiple motifs conserved in the prokaryotic genes is preserved in the human DNA MTase, including the relative position of a proline-cysteine dipeptide thought to be an essential catalytic site in all (cytosine-5)-methyltransferases. A single 5.2 kb transcript was detected in all human tissues tested, with the highest levels of expression observed in RNA from placenta, brain, heart and lung. DNA MTase cDNA clones were used to screen a chromosome 19 genomic cosmid library. The DNA MTase-positive cosmids which are estimated to span a genomic distance of 93 kb have been localized to 19p13.2-p13.3 by fluorescence in situ hybridization. Isolation of the cDNA for human DNA MTase will allow further study of the regulation of DNA MTase expression, and of the role of this enzyme in establishing DNA methylation patterns in both normal and neoplastic cells.

Amino Acid Sequence↗