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Biomedical subjects

M Y Chou

Publications and source records attributed to M Y Chou.

At least 55 records · Page 3Linked to original sources

Cloning and expression of sialidase L, a NeuAcalpha2-->3Gal-specific sialidase from the leech, Macrobdella decora.

Sialidase L is a NeuAcalpha2-->3Gal linkage-specific sialidase that releases 2,7-anhydro-NeuAc instead of NeuAc from sialoglycoconjugates (Chou, M.-Y., Li, S.-C., Kiso, M., Hasegawa, A., and Li, Y.-T.(1994) J. Biol. Chem. 269, 18821-18826). A 2. 5-kilobase cDNA of sialidase L was cloned by a combination of methods based on polymerase chain reactions. The composite cDNA sequence reveals an open reading frame coding for 762 amino acids, including a putative 28-residue signal peptide at the N terminus that is similar to the signal sequence of the Clostridium septicum sialidase. The result suggests that sialidase L is a secretory enzyme. The coding sequence excluding the putative signal peptide of sialidase L was overexpressed in Escherichia coli. The purified recombinant enzyme was characterized to be as active as the enzyme isolated from the leech. It also possessed the strict NeuAcalpha2-->3Gal linkage specificity and released the unique cleavage product, 2,7-anhydro-NeuAc from sialoglycoconjugates. The deduced amino acid sequence of sialidase L exhibits little similarity with other reported sialidases. However, sialidase L contains a conserved "FRIP region" and four repeating "Asp box" motifs that align well with the corresponding positions of bacterial sialidases. The predicted beta-strand structures near the conserved motifs of sialidase L are similar to those of Salmonella typhimurium sialidase. Several conserved single amino acid residues of bacterial sialidases, including those known to be involved in the active site of Salmonella enzyme, are conserved in the deduced amino acid sequence of sialidase L. This observation suggests that part of the catalytic mechanism of sialidase L may be similar to the ordinary sialidase.

Amino Acid Sequence↗

Suppressive effects of methyl methacrylate on the mutagenicity and DNA adduct formation induced by 1-nitropyrene and benzo[a]pyrene.

Methyl methacrylate (MMA) is widely used as a cement in dentistry, orthopaedic surgery and ophthalmology. Studies based on short-term genotoxicity tests have produced conflicting results in the last two decades. In the present study, the effects of MMA on the mutagenicity of 1-nitropyrene (1-NP) and benzo[a]pyrene (B[a]P) were evaluated with the Salmonella typhimurium TA98 strain in the absence and presence of S9 mix. The direct-acting mutagenicity of 1-NP was markedly decreased by MMA in a dose-dependent manner. However, a low inhibitory effect of MMA on the metabolic-acting mutagenicity of B[a]P was observed. MMA did not show mutagenicity within the concentrations of 4.7-37.6 microM either with or without S9 mix. The inhibitory effect of MMA was not due to its cytotoxicity because very low and/or no cytotoxicity of MMA to S. typhimurium TA98 was observed. To confirm the antimutagenicity of MMA against 1-NP and B[a]P, a 32P-postlabelling method was used to determine whether MMA modified DNA adduct formation produced by both compounds in calf thymus DNA. MMA inhibits the formation of 1-NP- and B[a]P-DNA adducts in a dose-dependent manner. The DNA adduct of 1-NP reduced by MMA was greater than that of B[a]P. Thus, we suggested that MMA was possibly acting as an inhibitor of chemical carcinogenesis.

Animals↗

Deleted in oral cancer-1 (doc-1), a novel oral tumor suppressor gene.

We have identified, isolated, and partially characterized doc-1, a novel cDNA sequence whose activity is consistent with a suppressor of hamster oral carcinogenesis. Doc-1 is an evolutionarily conserved gene exhibiting loss of heterozygosity and marked reduction in expression in malignant hamster oral keratinocytes. The full-length doc-1 cDNA encodes an 87 amino acid product that shows a significant homology to one of the seven novel genes induced in mouse fibroblasts by TNF-alpha. Transfection of the full-length doc-1 cDNA into malignant hamster oral keratinocytes alters the behavior of the recipients in terms of morphology, growth rate, and anchorage-independent growth, suggesting reversion of transformation phenotypes. We propose that doc-1 is a novel tumor suppressor gene in oral cancer development.

Amino Acid Sequence↗

Comparison of pyogenic liver abscesses caused by Klebsiella pneumoniae and non-K. pneumoniae pathogens.

A retrospective study on patients with pyogenic liver abscesses was conducted to clarify the different clinical presentations among patients with Klebsiella pneumoniae abscesses and those with non-K. pneumoniae abscesses. From 1981 to 1993, the medical records of 146 adults with culture-confirmed pyogenic liver abscesses who attended Tri-Service General Hospital in Taipei were studied. Abscesses due to K. pneumoniae accounted for 114 (78%) of pyogenic liver abscesses. When compared to patients with non-K. pneumoniae abscesses, patients with K. pneumoniae liver abscesses had significantly higher proportions of monomicrobial infections, unknown sources of infection and solitary abscesses. Patients with K. pneumoniae liver abscesses were found to have diabetes mellitus more often than patients with non-K. pneumoniae liver abscesses (66% vs 19%). Septicemia was found more frequently in patients with K. pneumoniae liver abscesses than in patients with non-K. pneumoniae liver abscesses (50% vs 27%). The clinical presentations among the two groups were, otherwise, not significantly different. Regardless of the microbial etiology, patients with diabetes mellitus had longer periods of fever after treatment and hospitalization than patients without diabetes. The reason for the high relative frequency of liver abscesses in Taiwan and its more frequent occurrence in diabetes mellitus remains unclear.

Adult↗

Purification and characterization of sialidase L, a NeuAc alpha 2-->3Gal-specific sialidase.

Sialidase L releases 2,7-anhydro-NeuAc from sialoglycoconjugates (Li, Y.-T., Nakagawa, H., Ross, S. A., Hansson, G., and Li, S.-C. (1990) J. Biol. Chem. 265, 21629-21633). This enzyme has been purified more than 10,000-fold from Macrobdella leech. The final preparation gives a single protein band on SDS-polyacrylamide gel electrophoresis with the molecular mass of 84 kDa. The pI is determined to be 6.0 using isoelectric focusing. With 4-methylumbelliferyl-alpha-NeuAc as substrate, the pH optimum is between pH 5.5-7.0. Unlike regular sialidases, sialidase L is not inhibited by 2-deoxy-2,3-dehydro-NeuAc. Two of the seven tryptic peptides derived from sialidase L contain the consensus repeat S-X-D-X-G-X-T-W that has been found in the regular sialidases. Among various sialoglycoconjugates tested, sialidase L cleaves only the NeuAc alpha 2-->3Gal linkage. NeuAc alpha 2-->6Gal, NeuAc alpha 2-->6GalNAc, NeuAc alpha 2-->6GlcNAc, NeuAc alpha 2-->8-NeuAc, and NeuAc alpha 2-->9NeuAc linkages are not hydrolyzed. At pH 7.0, sialidase L and Clostridial sialidase release 46 and 92% of sialic acid, respectively, from bovine fetuin, indicating that sialidase L selectively cleaves NeuAc alpha 2-->3Gal linkages in fetuin. Sialidase L is the first sialidase found to exhibit a strict specificity toward the hydrolysis of the NeuAc alpha 2-->3Gal linkage, and it should become useful for the selective cleavage of NeuAc alpha 2-->3Gal linkages in sialoglycoconjugates without destroying other sialosyl linkages.

Amino Acid Sequence↗

Sequential cytogenetic alterations in hamster oral keratinocytes during DMBA-induced oral carcinogenesis.

Using the hamster cheek pouch oral cancer model, we have performed a comprehensive analysis of the cytogenetic changes in hamster oral keratinocytes during 7,12-dimethylbenz[a]anthracene (DMBA)-induced carcinogenesis. Tumour induction in the hamster cheek pouch required repeated application of the carcinogen for 14 weeks. We have found that this hamster oral cancer model to be suitable for cytogenetic studies. Unlike human oral cancers where chromosome breaks have been shown, this is only infrequently observed in DMBA-treated hamster oral keratinocytes. Of importance is the finding that at the beginning of the second week of DMBA treatment, there is a significant increase of karyotypes demonstrating tetraploid or near-tetraploidy. We propose that the significant increase in hamster oral keratinocytes exhibiting tetraploidy be further evaluated as a marker of premalignancy/malignancy.

9,10-Dimethyl-1,2-benzanthracene↗

Enterococcal bacteremia in a medical center.

BACKGROUND: The enterococci have become important nosocomial pathogens. They can cause multiple site infections and enterococcal bacteremia becomes more frequently associated with a high mortality rate. Previous studies of enterococcal bacteremia showed a variety of results. To establish the significance and importance of enterococci as nosocomial pathogens in this hospital, to characterize their clinical pictures and to search for the risk factors for mortality, this retrospective study was performed. METHODS: There were 208 cases of enterococcal bacteremia which occurred from 1988 to 1992. Twenty-seven cases had no medical charts, dismissing possibility of evaluation. Finally, 181 cases of enterococcal bacteremia were analysed. RESULTS: One hundred and eighteen episodes were nosocomial infections. Polymicrobial bacteremia occurred in 68.5% of the patients and the most common co-isolate was Pseudomonas aeruginosa. Those patients (78.5%) with underlying diseases and malignancies were the most common underlying problems. The portal of entry could be found in 69.6 percent of patients, with the gastrointestinal tract the most common sources. Antimicrobial susceptibility testing showed high gentamicin resistance rate (89.5%), and ampicillin still had about 80 percent sensitivity rate. The group who received specific antibiotic therapy for enterococcus showed lower mortality (36.4% versus 47.6%). Only one case had infective endocarditis. Forty-nine patients suffered from septic shock, the cause of 30 deaths. Totally 75 patients died during hospitalization. Besides sepsis, another major cause of death was their underlying diseases itself. CONCLUSIONS: Enterococci have no doubt become important nosocomial pathogens and enterococcal bacteremia were associated with high mortality, especially in elderly patients with underlying diseases such as malignancy or diabetes. When clinically dealing with sepsis from the gastrointestinal or biliary tract, especially when previous cephalosporins therapy showed no response, the possibility of enterococcal bacteremia should always be considered.

Adolescent↗

Serodiagnosis of tuberculosis using the A60 IgG ELISA.

The serodiagnosis of tuberculosis (TB) by using an enzyme-linked immunosorbent assay (ELISA) to detect IgG against mycobacterial antigen 60 (A60) was evaluated. Forty-three controls (10 healthy adults, 19 patients with non-tuberculous pulmonary diseases and 14 patients with non-pulmonary infectious diseases) and 63 patients with TB in different disease activity (50 patients with active TB and 13 patients with old pulmonary TB) were studied. When a cut-off value of 200 units was selected, the results obtained with controls were: 100% negative in IgG titers in healthy adults and patients with non-pulmonary infectious diseases, 89% negative in IgG titers in patients with non-tuberculous pulmonary diseases. The overall "false positive" rate was 5% in the control groups. The percentage of serologically positive cases of active TB was: 82% in smear-positive, 67% in smear-negative/culture-positive and 38% in culture-negative. Of the 13 patients with calcified pulmonary TB, four had a positive serological response (31%). Four out of eight patients with active TB showed an elevation in IgG titer 1 to 3 months after diagnosis and anti-TB chemotherapy. The positive and negative predictive values of the test may provide valuable information according to the prevalence of TB in different populations. When the TB prevalence is high, a positive IgG ELISA test may support the diagnosis of active TB.

Antibodies, Bacterial↗

Gram-negative bacillary meningitis in adults.

To evaluate the clinical aspects of gram-negative bacillary meningitis (GNBM), we reviewed 41 adult patients with bacteriologically proven gram-negative bacillary meningitis, seen from 1985 to 1990. Thirty-two patients had post-neurosurgical GNBM and nine patients had spontaneous GNBM. Spontaneous GNBM appeared to have a sudden onset, a relatively fulminant course, and was caused most often by Escherichia coli. Post-neurosurgical GNBM, however, had a more insidious onset, a more protracted course, and was more often caused by nosocomial organisms which were resistant to multiple antibiotics. The overall mortality was 39%. Patients treated with combined aminoglycoside therapy had a lower mortality rate than those treated with intravenous aminoglycoside (17% vs 48%). The use of third-generation cephalosporins has made a significant therapeutic advance in the treatment of GNBM, with a lower mortality of 21%. We recommend treatment of GNBM with third-generation cephalosporins and aminoglycosides. If aminoglycosides are to be employed, it is suggested that they be administered both intravenously and directly into the central nervous system.

Adult↗

Acute renal failure associated with scrub typhus: report of a case.

We report on a 25-year-old man who had scrub typhus with the unusual complication of acute renal failure. The clinical features of fever, headache, high Weil-Felix OX-K agglutination and Rickettsia tsutsugamushi immunofluorescence titers confirmed the diagnosis of scrub typhus. Acute renal failure was proven by oliguria, urinary diagnostic indices and renal biopsy. The patient had a complete recovery after adequate medical treatment.

Acute Kidney Injury↗

Septicemia caused by Vibrio parahemolyticus: a case report.

Vibrio parahemolyticus is a halophilic marine vibrio commonly associated with outbreaks of acute gastroenteritis which also sometimes causes serious wound infection. It is an uncommon cause of septicemia. A few reports suggest that patients with chronic liver disease and leukemia are more susceptible. A case of liver cirrhosis with septicemia caused by this organism is discussed. The patient's condition rapidly deteriorated, and he died 12 hours after admission.

Adult↗

[Infected common iliac aneurysm due to Salmonella: a case report].

A 62-year-old male, presenting with chills and fever and RLQ abdominal pain, was found to have an infected aneurysm of the right common iliac artery infected by Salmonella group C1. CT scan of abdomen and arteriography definitely identified the lesions. Antibiotic treatment and surgical intervention with aneurysmectomy and suprapubic crossover femoro-femoral artery graft resulted in a good recovery and outcome.

Humans↗

Acute fatty liver of pregnancy: a report of two cases.

In the past two years, two women with acute fatty liver of pregnancy have presented at this hospital. Both patients survived, but their babies died; published rates for both mother and fetus are up to 85% mortality with this condition. Both patients presented with symptoms and signs of impending hepatic failure including jaundice, elevated aminotransferases, leukocytosis, hepatic encephalopathy and disseminated intravascular coagulation during the third trimester. Diagnosis was established by liver biopsy in both cases. The unusual good outcome for both women can be attributed to early recognition and prompt delivery, once diagnosis was considered.

Acute Disease↗

Eosinophils from patients with blood eosinophilia express transforming growth factor beta 1.

The infiltration of eosinophils into tissues during pathologic responses is often associated with extracellular matrix alterations such as fibrosis. Transforming growth factor-beta 1 (TGF-beta 1) is a well-characterized multifunctional cytokine known to exert potent effects on the extracellular matrix. In this report, we showed the production of TGF-beta 1 by human eosinophils from patients with blood eosinophilia. Northern blot analysis using RNA isolated from eosinophils purified from a patient with the idiopathic hypereosinophilic syndrome (HES) detected the 2.5-kb TGF-beta 1 transcript. In situ hybridization and immunohistochemistry of leukocytes from two patients with HES and two patients with blood eosinophilia localized TGF-beta 1 messenger RNA (mRNA) and protein to eosinophils. No other cell type contained TGF-beta 1 mRNA by in situ hybridization, whereas other leukocytes contained detectable TGF-beta 1 protein by immunohistochemistry. Eosinophils from four normal donors contained little or no detectable TGF-beta 1 protein by immunohistochemistry, whereas eosinophils from two of these four normal donors labeled weakly for TGF-beta 1 mRNA by in situ hybridization. These results show that eosinophils in the peripheral blood of patients with blood eosinophilia can express TGF-beta 1, but that eosinophils in the blood of normal donors contained little or no TGF-beta 1.

Adult↗

Neuropsychopharmacological studies of a Ca2+ channel blocker on the modulation of brain Ca2+ mobilization of spontaneously hypertensive rats under mild stress.

The psychotropic effects of a calcium channel blocker (Ca antagonist) were examined in behavioral studies following changes in 45Ca2+ influx in synaptosomal fractions of brain tissues using spontaneously hypertensive rats (SHR). Under a novel circumstance utilizing 85-dB noise, SHR demonstrated hyperactivity and a significant increase in 45Ca2+ uptake into synaptosomal fractions of frontal cortex (FC) and hippocampus. Such hyperactivity may be caused not only be seeking behavior but also by stress-induced anxiety. Such hyperactivity was significantly blocked after 10 days of repeated administration of diazepam (DZP), tandospirone (SM-3997; SM), a 5-HT1A anxiolytic, and nitrendipene (Nit), a Ca antagonist. Moreover, repeated administration of DZP, SM and Nit reduced the maximum binding density of 3H-PN200-110 and reduced the 45Ca2+ uptake in FC of SHR. In hippocampus, midbrain, hypothalamus and striatum, the increased ratio of 45Ca2+ uptake was reduced after repeated administration of Nit or SM. These results suggest that the hyperactivity induced by this novel circumstances was reduced by DZP, SM and Nit and may be attributed to inhibition of voltage-dependent Ca channel activities in FC. In addition, Nit may induce anti-anxiety through the modulation of Ca2+ mobilization in the central nervous system.

Acoustic Stimulation↗

Molecular cloning of the complementary DNA encoding for the hamster TGF-alpha mature peptide.

The expression of transforming growth factor alpha (TGF-alpha) is consistently associated with the malignant transformation of oral mucosal tissues in both hamster and human. The cheek pouch of the Syrian hamster represents an ideal model to elucidate the role of TGF-alpha in epithelial cancer development. A prerequisite for such investigations at the molecular level is to obtain hamster-specific TGF-alpha molecular probes. Here we report the successful cloning of the hamster TGF-alpha cDNA from a hamster oral cancer cell line (HCPC-1) by the polymerase chain reaction technique using synthetic oligonucleotide primers based on human TGF-alpha cDNA sequence. Analysis of the nucleotide sequence of the newly isolated hamster cDNA encoding the portion of the mature TGF-alpha peptide revealed that it is 92.6% (139/150) homologous to that of the rat and 93.3% (140/150) homologous to that of the human sequences. The predicted hamster TGF-alpha amino acid sequence is 96% (48/50) similar to that of human, while 94% (47/50) similar to that of rat. Using this hamster TGF-alpha cDNA as a probe, molecular hybridization experiments revealed that it detects hamster TGF-alpha mRNA with approximately 40 times and approximately 5 times greater sensitivity than similar probes from human and rat origin respectively. This hamster TGF-alpha cDNA should be of great value as a probe to evaluate the role of TGF-alpha in the normal and pathological processes using the hamster as an experimental model.

Amino Acid Sequence↗

Cellular sources of transforming growth factor-alpha in human oral cancer.

Aberrant expression of TGF-alpha is associated with human malignant oral epithelium. Experiments were initiated to determine the cellular sources of transforming growth factor-alpha (TGF-alpha) in human oral cancer. Ten freshly resected human oral cancers and four specimens of normal human oral epithelium were studied by in situ hybridization and immunohistochemistry. Tissues were probed with 35S-labeled sense and antisense riboprobes to (i) human TGF-alpha (hTGF-alpha), (ii) human epidermal growth factor receptor (EGFR) to determine the distribution of TGF-alpha responsive cells, and (iii) histone H3 to examine TGF-alpha and/or EGFR's possible contribution to altered proliferation in transformed epithelium. Results of our experiments showed that TGF-alpha mRNA could be detected in normal and transformed human oral epithelium. More surprising, we have identified the major source of TGF-alpha mRNA to be the infiltrating eosinophils. A monoclonal antibody to the mature human TGF-alpha peptide stained similar areas in normal and malignant specimens. Eosinophils associated with tumors exhibited positive cytoplasmic immunostaining for TGF-alpha protein. Labeling of EGFR mRNA in human oral epithelium demonstrated uniform labeling of basal layers in normal, hyperplastic, and mildly dysplastic epithelium. In severely dysplastic epithelium and carcinomas (particularly moderate to poorly differentiated types), cellular levels of EGFR mRNA were significantly higher. The profile of altered cellular levels of EGFR mRNA correlated well with the profile of altered proliferation as indicated by H3 mRNA labeling. We hypothesize that the overproduction of EGFR mRNA in tumor epithelium--together with the localized delivery of high amounts of TGF-alpha by eosinophils at tumor-developing sites--is responsible for the increased proliferation of the tumor epithelium.

Antibodies, Monoclonal↗

Localization of transforming growth factor-alpha in adult Syrian hamster tissues.

Using the cheek pouch of the Syrian hamster as an experimental model for oral carcinogenesis, it has been shown that the expression of transforming growth factor-alpha (TGF-alpha) is consistently associated with the malignant transformation process. We have recently shown that production of TGF-alpha has been localized to normal hamster oral epithelium and bone marrow eosinophils. In this study we investigated the production of this cytokine in other normal hamster adult tissues. By using an EGF-radioreceptor assay, immunohistochemistry, Northern blot analysis, and in situ hybridization we have now further detected the presence of TGF-alpha mRNA and/or protein in the kidney, stomach, and pancreas of normal adult hamster. Together with the previously reported detection of TGF-alpha in oral mucosa and bone marrow eosinophils, these adult normal tissue/cellular sources can serve as sites of TGF-alpha production. The availability of hamster specific reagents (cDNA and monoclonal antibodies) and the delineation of the various adult tissues that could produce TGF-alpha make the Syrian hamster a suitable model for the study of how this multifunctional cytokine can influence normal and pathological processes.

Animals↗