Lack of activated protein C resistance in healthy Hong Kong Chinese blood donors--correlation with absence of Arg506-Gln mutation of factor V gene.
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Adolescent↗
Biomedical subjects
H W Liu
Publications and source records attributed to H W Liu.
[Progress in the studies on traditional Chinese medicine syndrome differentiation of chronic glomerulonephritis with laboratory findings].
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Chronic Disease↗
Modulatory effects of nitric oxide on baroreflex activation in the brainstem nuclei of rats.
It has been shown that nitric-oxide (NO) was synthesized in the central nervous system as well as in vascular endothelial cells. We reported recently that nitric oxide was involved in central cardiovascular regulation of rat brainstem nuclei. In the present study, we evaluated further the baroreflex response of NO in the nucleus tractus solitarii (NTS) of rats. Male Sprague-Dawley rats were anesthetized with urethane, blood pressure and heart rate was monitored intra-arterially. Intramedullary microinjection (60 nl) of NO synthase inhibitor NG-methyl-L-arginine (L-NMMA), NG-nitro-L-arginine methyl ester(L-NAME) or normal saline was made into the NTS. Baroreflex response were elicited by increasing doses of phenylephrine (10-30 micrograms/Kg i.v.) before and after intra-NTS administration of L-NMMA, L-NAME or normal saline. The reflex bradycardia elicited by phenylephrine was significantly inhibited by the pretreatment of L-NMMA or L-NAME. Furthermore, the inhibitory effects on baroreflex activation by L-NMMA were significantly reversed by pretreatment with L-arginine. These results suggested that central endogenous NO was involved in the medullary regulation of blood pressure and that NO synthase inhibitor attenuated baroreflex activation.
Animals↗
Decreased bone mineral density in premenopausal patients with systemic lupus erythematosus.
To study bone mineral density (BMD) in premenopausal adult female patients with systemic lupus erythematosus (SLE) and its relation with clinical parameters, 56 SLE patients (mean age 31 years, mean disease duration 6.3 years) and 15 normal controls were studied. BMD at the lumbar vertebrae (L2-L4) was measured by dual energy X-ray absorptiometry (DEXA). Classification of BMD was made according to the WHO criteria in 1994. Correlation between BMD and clinical parameters was calculated. It was found BMD in the SLE patients (0.942 +/- 0.136 g/cm2) was lower than in the control group (1.055 +/- 0.080 g/cm2) (P < 0.01). According to the WHO criteria, 17 patients (30%) had normal BMD, 22 patients (40%) had osteopenia and 17 patients (30%) had osteoporosis. BMD was inversely correlated with disease duration in SLE patients (p < 0.005). The minimal disease duration for a female SLE patient to develop osteopenia was 3.5 years. In conclusion, SLE patients have lower lumbar BMD than normal controls. SLE patients with longer disease duration have lower BMD. In order to achieve early prevention of osteoporosis, we suggest that female SLE patients with disease duration for more than 3.5 years should take a BMD examination.
Adult↗
Mechanistic studies on CDP-6-deoxy-L-threo-D-glycero-4-hexulose 3-dehydrase identification of His-220 as the active-site base by chemical modification and site-directed mutagenesis.
CDP-6-deoxy-L-threo-D-glycero-4-hexulose 3-dehydrase (E1) purified from Yersinia pseudotuberculosis is a pyridoxamine 5'-phosphate (PMP) dependent iron-sulfur-containing enzyme which catalyzes the C-O bond cleavage at C-3 of its substrate leading to the formation of 3,6-dideoxyhexose. This enzyme is rapidly inactivated by diethyl pyrocarbonate (DEP) at pH 6.0 and 25 degrees C. The inactivation of E1 by DEP, which is reversible upon treatment of hydroxylamine, appears to be attributable solely to the modification of histidine residues. The fact that coincubation of E1 with its substrate gave almost total protection against DEP inactivation and that only one less histidine residue was modified in the presence of substrate strongly suggested that inactivation is due to the modification of only one reactive histidine residue which resides in or near the active site of E1 and is critical for E1's activity. Sequence alignment between the translated ascC (E1) gene and several representative pyridoxal 5'-phosphate (PLP)/PMP dependent enzymes revealed that three of the four invariant residues, glycine, aspartate, and arginine found in all other aminotransferases, are conserved in the E1 sequence (G169, D191, and R403). However, the highly conserved lysine is replaced by a histidine residue (H220) in E1. In order to test whether H220 plays an essential role in E1 catalysis, H220N mutant was constructed and the encoding protein was found to exhibit nearly identical physical characteristics as the wild-type E1. Interestingly, the mutant protein had lost most of its catalytic activity, and one less histidine residue was modified upon treatment of H220N-mutated protein with DEP. Such a single-point mutation also impaired E1's capability of catalyzing the solvent hydrogen exchange at C-4' position of the PMP coenzyme. Our findings strongly suggested that H220 is most likely the active-site base which abstracts the C-4' proton from the PMP-substrate adduct and initiates the catalysis. Furthermore, E1's preservation of other invariant residues found in many PLP/PMP dependent enzymes allowed a speculation of their roles in E1 catalysis.(ABSTRACT TRUNCATED AT 400 WORDS)
Amino Acid Sequence↗
Mechanistic studies on CDP-6-deoxy-delta 3,4-glucoseen reductase: the role of cysteine residues in catalysis as probed by chemical modification and site-directed mutagenesis.
CDP-6-deoxy-delta 3,4-glucoseen reductase (E3), which catalyzes the reduction of the C-3 deoxygenation step during the formation of CDP-ascarylose, a 3,6-dideoxyhexose found in the lipopolysaccharide of Yersinia pseudotuberculosis, has been expressed at high level in Escherichia coli (670 times over the wild-type strain). This flavoenzyme, which also contains one plant ferredoxin type [2Fe-2S] cluster, was inactivated by 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) and N-ethylmaleimide. In both cases the inactivation followed a pseudo first order kinetics. The second order rate constant for the reaction of DTNB with E3 was 0.25 mM-1 min-1 at 20 degrees C, pH 8.0. Detailed characterization of the inactivated enzyme showed that neither the flavin nor the [2Fe-2S] cluster was altered during inactivation. Since this inactivation was reversible by treating the inactivated enzyme with 1 mM D,L-dithiothreitol (DTT), it was concluded that only cysteine residues were modified during inactivation. Analysis of the inactivation using the method developed by Tsou revealed that two cysteines react with DTNB at similar rates and modification of either one is enough to impair E3's activity. Tryptic digestion of E3 labeled with N-ethyl[2,3-14C]maleimide, followed by fractionation of the digest by high performance liquid chromatography, gave two labeled peptides, both of which were separately isolated as a pair of interconvertible diastereoisomers. Sequence analysis of these labeled peptides allowed the identification of Cys-75 and Cys-296 as the reactive cysteine residues. Interestingly, the C75S and C296S mutant proteins exhibit identical physical and comparable catalytic properties as the wild-type enzyme. Since Cys-296 is a conserved residue in the NAD(P) binding domain of enzymes belonging to the same class, this residue may be involved in stabilizing the charge-transfer complex between E3 and NADH, thus facilitating hydride transfer from the nicotinamide nucleotide to flavin. A chemically modified Cys-75 which is immediately adjacent to the [2Fe-2S] center in E3 may prevent the proper juxtaposition of the redox centers and thus impede electron transfer leading to enzyme inactivation. These results may be useful for placing constraints on the peptide folding comprising the active site of E3 for electron transfer between NADH, FAD, and the [2Fe-2S] center.
Amino Acid Sequence↗
Relationship of mitochondrial function and cellular adenosine triphosphate levels to pMC540 and merodantoin cytotoxicity in MCF-7 human breast cancer cells.
In previous studies we have reported that preactivated merocyanine 540 (pMC540) and its chemically synthesized isolates merocil and merodantoin mediate their preferential cytotoxicity towards certain types of malignant cells including human breast cancer cells in vitro and in vivo. The mechanism of cytotoxic action appears to be, in part, via initial interaction with topoisomerase II leading to apoptosis. To further build upon these findings we now show that pMC540 and merodantoin disrupt mitochondrial morphology and function in intact MCF-7 human breast cancer cells as seen by their causing the release of rhodamine 123 from prestained cells, a rapid reduction in ATP levels, inhibition of succinate dehydrogenase activity and oxygen consumption. These data suggest that mitochondria may also be an important target for the cytotoxic action of pMC540 and merodantoin mediated through disruption of the energy balance.
Adenosine Triphosphate↗
Synthesis and antiplatelet effects of omega-aminoalkoxylxanthones.
A series of omega-aminoalkoxylxanthones were synthesized and tested in-vitro for their ability to inhibit aggregation of rabbit washed platelets and human platelet-rich plasma (PRP) induced by various inducers. Nine of these compounds showed more potent antiplatelet effects than natural norathyriol tetraacetate on collagen-induced aggregation. The various omega-aminoalkoxyl side chains of the synthesized compounds modified the antiplatelet effects. All the compounds tested in human PRP showed significant inhibition of secondary aggregation induced by adrenaline, suggesting that the antiplatelet effects of these compounds is mainly due to an inhibitory effect on thromboxane formation. These compounds at high concentration also cause vasorelaxing action in rat thoracic aorta.
Animals↗
Impact of alcohol consumption and cigarette smoking on stroke among the elderly in Taiwan.
BACKGROUND AND PURPOSE: We investigated the influence of alcohol consumption and cigarette smoking on all types of stroke and cerebral infarction, in particular among a representative sample of elderly residents in Taiwan. METHODS: This study was a component of a nationwide survey of health and living status of residents aged 65 years or older in Taiwan in which subjects received detailed physical, neurological, and laboratory examinations. Inquiries were made about medical history, and information on the amount and duration of drinking and smoking was obtained. Diagnoses of stroke were made according to the results of brain computed tomography at the onset of disease or were based on criteria established by the World Health Organization. RESULTS: Of the 2600 subjects, there were 155 elderly persons with stroke (prevalence, 6%). Excessive drinking of more than 367.6 g/wk of alcohol was associated with a high prevalence of cerebral infarction. Consumption of < or = 367.5 g/wk of alcohol did not have an influence on stroke prevalence. The relationship between duration of alcohol drinking and stroke was equivocal. More than 30 pack-years of cigarette smoking was a significant risk factor for all types of stroke and cerebral infarction in particular. Using multiple logistic regression to control for possible confounders, it was found that smoking was an independent risk factors for all stroke and was of borderline significance for cerebral infarction. Although excessive drinking was a significant risk factor for cerebral infarction in univariate analysis, this effect was lost after adjustment for other confounders. CONCLUSIONS: Cigarette smoking was a more important risk factor for stroke and cerebral infarction than excessive drinking of alcohol.
Aged↗
Chronic lead intoxication at an indoor firing range in Taiwan.
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Adolescent↗
HLA-DRB1 genotyping in patients with rheumatoid arthritis in Taiwan.
OBJECTIVE: To investigate the relation of HLA-DR alleles to the development of rheumatoid arthritis (RA) and the correlations between HLA-DR alleles and clinical manifestations of patients with RA in Taiwan. METHODS: The HLA-DRB1 alleles were studied in 144 patients with RA and 154 healthy controls using polymerase chain reaction/sequence specific oligonucleotide probe methods. The subtypes of HLA-DR4 were detected by cloning sequencing. RESULTS: The prevalence of HLA-DR4 was significantly higher in patients with RA (46.53%) than in healthy controls (25.97%). HLA-DRB1*0405 was more common in patients than in controls (83.58 vs 50%), while the prevalence of DRB1*0403 in patients (4.48%) was significantly lower than in controls (27.5%). There was no significant difference in frequencies of HLA-DR alleles between DRB1*0405 negative patients and DRB1*0.05 negative controls. We also noted a positive correlation in patients between HLA-DR4 and bone erosion, but not HLA-DR4 and age of onset, seropositivity, or extraarticular involvement. There was no significant difference in seropositivity, extraarticular involvement, and bone erosion between homozygous and heterozygous DR4 patients. CONCLUSION: HLA-DRB1*0405 is related to the development of RA in Taiwan, while DRB1*0403 was a negative risk factor. HLA-DR4 was associated with bone erosion in patients.
Alleles↗
Application of ABO genotyping using polymerase chain reaction method to assess engraftment in bone marrow transplantation--a case report.
We described a case of acute promyelocytic leukemia with O blood type who received bone marrow transplantation during a second remission from his HLA identical, B blood type sibling. Using PCR genotyping of the ABO group and analysis of the variable number of tandem repeats of the D1S80 locus, we could quickly document the successful early engraftment in this patient. We emphasize that the use of PCR ABO genotyping is a quick and valuable method which could be applied to assess the engraftment and its follow up in those transplant cases with donors of mismatched blood groups.
ABO Blood-Group System↗
MDR1 gene expression in acute myeloid leukemia: clinical correlation.
Acute myeloid leukemia (AML) has a poor long-term prognosis, with a disease-free survival rate of only 20%. Drug resistance remains the major obstacle in this disease, although relatively little data are available on the correlation of multi-drug resistance (MDR) and clinical outcome. We examined 28 bone marrow samples from 24 patients with AML. The MDR1 RNA levels were determined by slot blot analysis. The incidence of positive expression of MDR1 mRNA in patients was 64.7% in samples from newly-diagnosed patients and 85.7% in relapse samples. The overall complete remission rate was 45%. However, the complete remission rate was lower for patients in the MDR1 RNA positive group than in the negative group. In three cases, the MDR1 RNA expression steadily rose during the period of disease progression and after several courses of chemotherapy. We suggest that a high frequency of MDR1 gene expression in patients with AML and higher MDR1 RNA levels are associated with a lower incidence of complete remission as well as a shorter disease-free time. The expression of MDR1 is a poor prognostic factor for AML treatment and it may play an important role in resistance to chemotherapy.
ATP Binding Cassette Transporter, Subfamily B, Mem↗
HLA-DPB1 polymorphism in patients with rheumatoid arthritis in Taiwan.
OBJECTIVE: To investigate the role of HLA-DP alleles in the pathogenesis of rheumatoid arthritis (RA). METHODS: HLA-DPB1 alleles were studied in 144 patients with RA and 144 healthy controls using the polymerase chain reaction/sequence specific oligonucleotide probe method. RESULTS: HLA-DPB1*0510 was the the most common allele in patients with RA and healthy controls in Taiwan. There was no significant difference in frequencies of HLA-DPB1 alleles between patients and controls. The HLA-DPB1 alleles were not associated with seropositivity for rheumatoid factor, extraarticular involvement, or bone erosion in patients. There was also no significant difference in frequencies of HLA-DPB1 alleles among DR4-positive and DR4-negative patients and controls. CONCLUSION: There was no significant difference in the prevalences of HLA-DPB1 alleles between Taiwanese patients with RA and healthy controls. HLA-DPB1 alleles were not related to the clinical manifestations of patients with RA in Taiwan.
Alleles↗
Inhibition of the human methylmalonyl-CoA mutase by various CoA-esters.
Human methylmalonyl-CoA mutase is inhibited by ethylmalonyl-CoA, cyclopropylcarbonyl-CoA carboxylate, and methylenecyclopropylacetyl-CoA, which are substrate, intermediate, and product analogs, respectively. The mode of inhibition by each analog is reversible and mixed with respect to the substrate, methylmalonyl-CoA. This implies that the inhibitors are able to bind to both free enzyme and to the enzyme-substrate complex, although with affinities that are 4.5- to 10-fold different for the two species. The Ki1 for the cyclopropylcarbonyl-CoA carboxylate (0.26 +/- 0.07 mM), is 4-fold greater than the Km(app) measured for the substrate, methylmalonyl-CoA. Additionally, ethylmalonyl-CoA functions as an alternate substrate and is metabolized to methylsuccinyl-CoA. The human mutase is a homodimer that binds 1 mol of cobalamin per subunit. So, the observed mixed inhibition kinetics by substrate analogs is curious. Our finding that methylenecyclopropylacetyl-CoA, the causative agent of Jamaican "vomiting sickness," inhibits methylmalonyl-CoA mutase, while interesting, is probably not physiologically important because of the relatively high inhibition constants (Ki1 = 0.47 +/- 0.12 mM and Ki2 = 2 +/- 0.34 mM) observed with this compound.
Acyl Coenzyme A↗
Trisomy 12 in chronic lymphocytic leukemia. An interphase cytogenetic study by fluorescence in situ hybridization.
B-cell chronic lymphocytic leukemia (CLL) is a rare disorder in the Chinese population. We evaluated the use of fluorescence in situ hybridization (FISH) with a chromosome 12-specific probe in the detection of trisomy 12 in interphase cell of 19 Chinese CLL patients. FISH successfully detected trisomy 12 in two cases, one of which had normal conventional cytogenetic findings, giving an incidence of 10%. The low incidence of trisomy 12 in our CLL patients may also reflect a biologic difference of this rare disorder in our population, compared to that of the West.
Adult↗
Diagnosis of thalassaemia by non-isotope detection of alpha/beta and zeta/alpha mRNA ratios.
The alpha/beta and zeta/alpha messenger RNA (mRNA) ratios in the thalassaemia syndromes were investigated by polymerase chain reaction (PCR) with silver staining of the PCR products. In this study we used the PCR to amplify cDNA copies of circulating erythroid cell mRNA in order to measure the relative amounts of alpha-, beta- and zeta-globin contained within. Quantitation was performed by scanning the silver stain of specific globin cDNA bands. We found that there were significant differences of alpha/beta-mRNA and zeta/alpha-mRNA in patients with Hb H disease and alpha-thalassaemia-1 compared to normal subjects. There was a marked increase in the alpha/beta-mRNA ratio but not in the zeta/alpha-mRNA ratio in patients with beta-thalassaemia. In two beta-thalassaemia cases abnormal increases of zeta-globin bands were noted and they were confirmed through DNA analysis to be combined with alpha-thalassaemia-1. This method provides a simple, rapid and non-radioactive approach to detect thalassaemia syndromes, and can help to screen cases of beta-thalassaemia with alpha-thalassaemia-1.
Adult↗
Studies on the synthesis of some xanthonoid derivatives possessing antiplatelet effects.
2,3- and 3,4-Dihydroxyxanthone react with ethyl 2,3-dibromopropanoate to form the new, substituted 1,4-benzodioxanes 3 and 4, respectively. The regioisomers 3a and 3b; 4a and 4b were separated by column chromatography and characterized for evaluation of the antiplatelet effects in rabbit washed platelets and human platelet-rich plasma. The ethoxycarbonyl derivatives 3a (20 microM) and 3b (20 microM) strongly inhibited the aggregation of rabbit washed platelets induced by arachidonic acid and collagen. The compound 4b showed the most potent inhibition of rabbit washed-platelet aggregation induced by arachidonic acid (IC50 = 8.3 microM). Of the compounds tested in human platelet-rich plasma, compound 4b exhibited the most potent inhibition of primary and secondary aggregation induced by adrenaline (IC50 = 8.6 microM). We conclude that the antiplatelet effects of these four ethoxycarbonyl derivatives are mainly due to an inhibitory effect on thromboxane formation and interference in the adrenaline-receptor interaction.
Animals↗