Biomedical subjects

T Z Liu

Publications and source records attributed to T Z Liu.

At least 37 records · Page 2Linked to original sources

PubMed1996

Purification and characterization of endogenous digoxin-like immunoreactive factors in chicken blood.

Studies have been performed to determine whether an endogenous material capable of binding to digoxin antibodies is present in the chicken plasma. In the blood of 12 chickens without feed control, endogenous digoxin-like immunoreactive factors (DLIF) binding of digoxin antibodies in enzyme immunoassays amounted to 866 / 302 pg digoxin equivalents/mL of plasma (mean +/- SEM). Immunoreactivity of DLIF increased to 1848***331 pg/mL with a double value of control after boiling and acid pretreating the plasma. The major purification steps employed in this report were gel filtration column chromatography, high performance liquid chromatography (HPLC) and isoelectric focusing (IEF). Using HPLC for the separation, at least 10 chicken DLIFs with different molecular weight (MW) have been found. The MW of the smallest is 300 daltons (Da) while the largest is 100 kDa. The value of the isoelectric point of the most abundant type of DLIF from untreated chicken plasma is 6.3 as determined by IEF. The partially purified DLIF inhibits Na+, K(+)-ATPase from a porcine cerebral cortex as well as three human red blood cell membrane preparations in a dose-response fashion.

Animals↗

PubMed1996

Correlation of membrane lipid peroxidation with oxidation of hemoglobin variants: possibly related to the rates of hemin release.

Experiments were performed to delineate the biochemical mechanism of hemoglobin (Hb)-catalyzed lipid peroxidation in human red blood cells (RBCs). Using a modified Langmuir trough lipid monolayer technique, we found that oxidized Hb induced an increase in lipid monolayer surface pressure, suggesting that oxidized Hb readily releases its heme moiety into the lipid monolayer. To confirm our interpretation that oxidized Hb readily releases its heme moiety, we monitored the fluorescence of Hb tryptophan upon oxidation of Hb. We found an increase in Hb fluorescence in the aqueous phase of our monolayer system after the addition of H2O2. The increase in fluorescence should reflect the departure of heme from globin due to a decrease in fluorescent quenching effect by the heme moiety. The rate of increase in lipid monolayer surface pressure upon Hb oxidation differed from Hb to Hb with an order of Hb E > F > S > A. The ability of various Hbs to affect lipid peroxidation in the RBC membrane, as monitored by the parinaric acid oxidation technique, followed this same order. In addition, hemin was shown to be a more potent catalyst of lipid peroxidation in RBC membrane than nonheme irons.

Adult↗

PubMed1996

Improved fluorometric quantification of urinary xanthurenic acid.

Measurement of urinary xanthurenic acid (XA) has been used clinically to study a variety of disorders caused by vitamin B6 deficiency. To obviate some cumbersome steps of current methods for measuring XA in human urine, we have developed a simple fluorometric method. We apply the urine sample to a solid-phase extraction column containing trimethylaminopropyl group bound to silica, which enables us to purify and concentrate the XA from the urine without contamination from various tryptophan metabolites. The XA in the acidic eluate can then be quantified fluorometrically. The linearity of the proposed method extends from 0.2 to 10.0 mg/L. The method is precise, yielding day-to-day CVs for two pooled control specimens (1.08 and 1.90 mg/L) of 1.2% and 2.6%, respectively. Correlation studies with an established HPLC method and with a spectrophotometric procedure showed correlation coefficients of 0.99 and 0.98, respectively. Interference from vitamin C, uric acid, salicylate, acetaminophen, vanillylmandelic acid, and homovanillic acid was insignificant. The proposed method for urinary XA is rapid, simple, and suitable for routine use in the clinical laboratory.

Chromatography, High Pressure Liquid↗

PubMed1996

Induction of superoxide dismutase isozymes by tumor necrosis factor-alpha and lipopolysaccharide in cultured normal and hyperplastic gingival fibroblasts.

To determine whether lipopolysaccharide (LPS) and tumor necrosis factor-alpha (TNF-alpha) are involved in the induction of superoxide dismutase (SOD) in gingival tissue, we examined their effect on induction of SOD isozymes in cultured normal (NGF) and phenytoin-induced hyperplastic (PHF) gingival fibroblasts. Treatment of both NGFs and PHFs with 10 to 50 ng/mL TNF-alpha for 24 hours increased the level of manganese SOD (MnSOD) to as much as four times the level of untreated cultures. PHFs, but not NGFs, were shown to be responsive to TNF-alpha in eliciting a significant increase in copper-zinc SOD (Cu/ZnSOD), albeit in a lesser amount than MnSOD. Additionally, treatment of both types of cells with 5 to 50 mg/mL of LPS for 24 hours also elicited an increase in the levels of MnSOD. Again, an LPS-induced increase in Cu/ZnSOD levels could only be demonstrated in PHFs, but not in NGFs. These observations were further confirmed by comparing the achromatic bands associated with SOD isozymes exhibited in the electrophoretogram using a nondenaturing polyacrylamide electrophoresis technique. These results indicate that TNF-alpha and LPS were capable of inducing both MnSOD and Cu/ZnSOD simultaneously in PHF fibroblasts. PHFs may be inherently more capable than NGFs in combating oxidative stress.

Anticonvulsants↗

PubMed1994

Detection of a novel lactate dehydrogenase isozyme and an apparent differentiation-associated shift in isozyme profile in hepatoma cell lines.

A hitherto unreported lactate dehydrogenase (LD) isoenzyme, which migrates electrophoretically to the relative position between LD2 and LD3 has been identified in the electropheratogram in 7 of 7 (100%) cultured hepatoma cell lines with various degrees of differentiation and is thus given the name LD2-3. LD2-3 seems to be specific for hepatoma cells because this atypical isoenzyme can not be detected in other tumor cell lines. In addition, the hepatoma cell lines also show a distinct pattern of LD isoenzyme and the isoenzyme pattern varies with the degree of differentiation. Hence, the expression pattern of LD isoenzyme phenotypes may provide a good marker for the investigation of human hepatoma cell differentiation.

Carcinoma, Hepatocellular↗

PubMed1994

Enhanced susceptibility of erythrocytes deficient in glucose-6-phosphate dehydrogenase to alloxan/glutathione-induced decrease in red cell deformability.

It has been hypothesized that enhanced oxidant sensitivity of glucose-6-phosphate dehydrogenase (G6PD) deficient red cells(RBCs) is the underlying mechanism for drug- or chemical-induced hemolytic crises in G6PD-deficiency. To further test this hypothesis, we used an alloxan-glutathione system to mimic oxidative stress and see how oxidative damage might affect RBC deformability. RBC deformability, a major determinant of RBC survival in vivo, was monitored by a laser viscodiffractometer. Under our experimental conditions, GSH alone had very little effect on the deformability of either normal or G6PD-deficient RBCs. In contrast, alloxan alone induced a small but significant decrease in the deformability of either normal or G6PD-deficient RBCs. Interestingly, alloxan and GSH together induced a further decrease in the deformability of either normal or G6PD-deficient RBCs. The decrease in deformability in G6PD-deficient RBCs was much more profound than in normal RBCs. In addition, an alloxan-vitamin C system produced a similar deleterious effect on RBC deformability as that produced by the alloxan-GSH system. Appreciable amount of hydroxyl radicals was generated by both alloxan-GSH and alloxan-vitamin C systems as evidenced by the production of hydroxylated products of salicylate which was used as a radical trap. Moreover, salicylate could ameliorate the deleterious effect of the alloxan system on the deformability of RBCs. Taken together, our results demonstrated that G6PD-deficient RBCs were particularly susceptible to oxidant-induced damage leading to a dramatic decrease in their deformability and thus provided strong support for the hypothesis that enhanced oxidant sensitivity of G6PD-deficient RBCs is the underlying mechanism for accelerated destruction of these RBCs in vivo.

Alloxan↗

PubMed1993

Molecular characterization of glucose-6-phosphate dehydrogenase (G6PD) deficiency in patients of Chinese descent and identification of new base substitutions in the human G6PD gene.

The underlying DNA changes associated with glucose-6-phosphate dehydrogenase (G6PD)-deficient Asians have not been extensively investigated. To fill this gap, we sequenced the G6PD gene of 43 G6PD-deficient Chinese whose G6PD was well characterized biochemically. DNA samples were obtained from peripheral blood of these individuals for sequencing using a direct polymerase chain reaction (PCR) sequencing procedure. From these 43 samples, we have identified five different types of nucleotide substitutions in the G6PD gene: at cDNA 1388 from G to A (Arg to His); at cDNA 1376 from G to T (Arg to Leu); at cDNA 1024 from C to T (Leu to Phe); at cDNA 392 from G to T (Gly to Val); at cDNA 95 from A to G (His to Arg). These five nucleotide substitutions account for over 83% of our 43 G6PD-deficient samples and these substitutions have not been reported in non-Asians. The substitutions found at cDNA 392 and cDNA 1024 are new findings. The substitutions at cDNA 1376 and 1388 account for over 50% of the 43 samples examined indicating a high prevalence of these two alleles among G6PD-deficient Chinese. Our findings add support to the notion that diverse point mutations may account largely for much of the phenotypic heterogeneity of G6PD deficiency.

Asian↗

PubMed1993

Direct alteration of erythrocyte membrane properties by 1-chloro-2,4-dinitrobenzene without oxidant challenge.

The electrophilic agent, 1-chloro-2,4-dinitrobenzene (CDNB), has been widely used as an intracellular glutathione-depleting agent. However, its possible effect on the functional integrity of cell membrane has largely been neglected. Incubation of human erythrocytes (RBC) with various concentrations of CDNB (0.5 to 5.0 mM) in potassium-free, phosphate buffered saline containing ouabain resulted in a drastic depletion of cellular glutathione as well as a dose-dependent increase in passive potassium leakage. Further, an osmotic gradient ektacytometry profile indicated that the deformability index (DI) of CDNB-treated RBC was substantially lower than the DI value of the control. Also, CDNB caused a dose-dependent increase in the rate of shear-induced fragmentation of resealed ghost prepared from treated, intact erythrocytes. These CDNB-induced changes were accompanied by stomatocytic transformations as evidenced by scanning electron micrographs. Additional study indicated that CDNB caused a dose-dependent decrease in thiol concentrations of RBC membrane. SDS-PAGE analysis of membrane proteins revealed new Coomassie blue stainable bands, most noticeable below band-7 (M.W. 20,000). The effects of CDNB on RBC deformability and membrane proteins were also investigated under an atmosphere without oxygen (under nitrogen) and similar effects were observed between that under room air and that under nitrogen. Taken together, these data strongly indicate that CDNB has an adverse effect on the RBC membrane integrity in addition to its ability to deplete intracellular glutathione, possibly through its interaction with membrane sulfhydryl groups.

Dinitrochlorobenzene↗

PubMed1993

High-molecular-mass alkaline phosphatase as a tumor marker for colorectal cancer: comparison of two test methods.

We measured high-molecular-mass alkaline phosphatase (HiMwALP) in serum samples from patients with colorectal cancer by polyacrylamide gel electrophoresis (PAGE) and by column chromatography on diethylaminoethyl (DEAE)-cellulose. Determination of carcinoembryonic antigen (CEA) by an enzyme immunoassay, a frequently used cancer assessment method, was used for comparison. We studied patients with primary colorectal cancer (n = 72), using others with hemorrhoids (n = 38) for a comparison group. HiMwALP activities twice those of pooled normal sera were adopted as cutoff values. The diagnostic sensitivity of the PAGE method for 72 colorectal cancer patients was 63% vs 36% for the DEAE method and 50% for the CEA method. The diagnostic specificities of the PAGE, DEAE, and CEA methods were 89%, 79%, and 95%, respectively. Using both HiMwALP (PAGE method) and CEA for the detection of primary colorectal cancer increased the sensitivity to 72% but decreased specificity to 87%.

Adult↗

PubMed1992

Simultaneous presence of three or more creatine kinase isoenzymes in patient serum.

Samples of sera from 1,508 consecutive patients, which were sent to the clinical laboratories of Chang Gung Medical College for creatine kinase (CK) isoenzyme determination, were examined for the simultaneous presence of various patterns of three or more usual or atypical isoenzymes. Of these samples, a total of 29 cases (1.9%) exhibited this abnormal isoenzyme condition. Nineteen (66%) of these 29 cases were patients with cancer of various types. Furthermore, five (26%) of these 19 cases had the same pattern of four isoenzymes, and 17 (89%) of these 19 cases showed the cathodic CK variant (macro CK type II). Eleven (58%) of these 19 cancerous cases also showed a CK-MB variant, evidently of nonmyocardial origin.

Creatine Kinase↗

PubMed1992

A new mutation responsible for severe G6PD deficiency in two ethnic Chinese with different clinical presentations: determination by a direct PCR sequencing technique.

To test the hypothesis that clinical manifestation in G6PD deficiency correlates with a molecular lesion, we investigated the G6PD gene of two Chinese Americans both of whom had G6PD deficiency, but who manifested different clinical presentations. In this study, we have developed a direct PCR sequencing protocol to examine the human G6PD gene. By using optimized PCR conditions with internal primers, we were able to amplify a 4.2 kb DNA fragment (covering exon 3 through 13 of the G6PD gene) of consistently high quality. From this we were then able to generate high quality single-stranded DNA templates by asymmetric PCR for subsequent sequencing. We also overcame the crossband problem by using internal primers, high temperature reaction with Taq I DNA polymerase, and/or sequencing with gene 32 protein. We could consistently amplify exons 1 and 2 despite their high G/C content by substituting 75% of dGTP with deoxy-7-deaza-guanosine triphosphate. By using this novel approach, we have identified a new mutation at cDNA position 1376 from G to T, which causes substitution of Leu for Arg at amino acid position 459. This mutation has not been reported in other ethnic groups. It is the only genetic defect in the coding regions of the G6PD gene of these two G6PD deficient individuals. We speculate that in addition to a defect in the G6PD gene, other factors also play a role in the clinical manifestation of G6PD deficiency.

Base Sequence↗

PubMed1991

Two commonly occurring nucleotide base substitutions in Chinese G6PD variants.

Using a direct PCR sequencing technique, we have identified two DNA base substitutions in 8 different biochemical G6PD variants of Chinese origin. Neither one of these abnormalities has been reported in other ethnic groups. An abnormality (C1) of G to T substitution at cDNA 1376 causing an amino acid change from Arg to Leu has been found in 3 variants. Another abnormality (C2) of G to A substitution at cDNA 1388 causing an amino acid change from Arg to His has been found in 5 variants. Both C1 and C2 are located in exon 12 of the G6PD gene and are only 12 base pairs apart. However, C1 is associated with a significant increase in the deamino-NADP utilization rate, whereas C2 is not. Taken together, our data suggest that C1 and C2 are very common among Chinese with a G6PD deficiency and exon 12 may define an important functional domain of the human G6PD.

Base Sequence↗

PubMed1991

Rapid laser nephelometric determination of amylase activity in serum and urine.

We describe herein a rapid and sensitive laser nephelometric method for the determination of serum and urinary amylase activities. Our data showed that the change in relative light scattering (RLS) of an amylopectin substrate measured by a laser nephelometer related directly with amylolytic activity of amylase from 50 to 600 IU/L. Within-run variations at 293 and 769 IU/L sera showed CV's of 5.0% and 3.1%, respectively. Day-to-day variation for the same sera showed CV's of 7.2% and 4.7%, respectively. Correlation studies using the manual Phadebas dye-starch complex method and with the Roche amylochrome method showed correlation coefficients of 0.99 and 0.95, respectively. Using urine specimens, the correlation studies also showed a correlation coefficient of 0.98. These studies indicated that the proposed method was sensitive, fast, economical and easily adaptable to emergency and routine applications.

Amylases↗

PubMed1990

Creatine kinase and lactate dehydrogenase isoenzyme patterns in cultured normal and pathological gingival fibroblasts.

The isoenzyme patterns of creatine kinase (CK) and lactate dehydrogenase (LDH) were examined in cultured fibroblasts derived from the gingival tissue of a healthy person and patients with periodontitis and gingival fibromatosis. Human foreskin fibroblasts were also included for comparison. Cytosol fractions were prepared by sonication and centrifugation. Gingival fibroblasts from normal subjects (A), and from patients with periodontitis (B) or fibromatosis (C), exhibited CK-MM (muscle form) as a predominant isoenzyme and CK-BB (brain form) as a minor fraction. There was no significant difference in specific activities of CK-BB fraction in all 3 types of fibroblasts. However, B fibroblasts contained a higher proportion of MM type isoenzyme than A fibroblasts. C fibroblasts showed the lowest amount of CK-MM isoenzyme as compared to either A or B fibroblasts. In contrast, fibroblasts derived from human foreskin (D) contained only a small amount of CK-BB isoenzyme, but CK-MM was not present. In the studies of LDH isoenzymes, A, B, and C fibroblasts possessed only 3 isoenzymes, namely, LDH-3, LDH-4 and LDH-5. In terms of the relative abundance of M vs H subunits, A, B, and C fibroblasts were shown to have a preponderance of M subunits (85-87% M vs 13-15% H subunits). In comparison, D fibroblasts contained 4 isoenzymes (LDH-2, LDH-3, LDH4, LDH-5) with 73% M and 27% H subunits. The ratio of LDH-5 to LDH-4 was calculated for all fibroblasts tested, and the values were 1.11 in A, 1.39 in B, 1.16 in C and 0.64 in D.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1990

Diagnostic efficacy of a monoclonal antibody-based immunoenzymetric technique for quantitation of creatine kinase MB isoenzyme in serum.

The clinical usefulness of the Tandem-E CK-MB method (A) using two monoclonal antibodies to measure the intact cardiac specific creatine kinase isoenzyme (CK-MB) was evaluated against results obtained by Corning's electrophoretic fluorescence scanning method (B) and the DuPont aca mechanized column chromatographic method (C). In 95 patients suspected to have myocardial infarction, a total of 127 CK's with MB's were simultaneously determined by methods "A", "B", and "C". "C" produced a total of 16.5% (21 out of 127) false-negative or false-positive results. In contrast, CK-MB results obtained by "A" correlated very well with those determined by "B" (r = 0.97; N = 64). In addition, the clinical course of myocardial infarction as monitored by the measurement of CK-MB at various time intervals by both methods showed excellent parallelism. Furthermore, "A" was shown to be free of interference from the presence of macro CK type I and II. Also, the presence of increasing concentrations of CK-BB (up to 29 IU/L) did not alter the assay response of CK-MB in these serum samples. In conclusion, we found the Tandem-E CK-MB method to be sensitive (sensitivity = 94%) and specific (specificity = 96%).

Antibodies, Monoclonal↗

PubMed1990

Laser nephelometric quantitation of protein in urine and cerebrospinal fluid.

We describe a rapid method for the routine determination of protein in cerebrospinal fluid (CSF) and in urine using laser nephelometer. The relative light scattering (RLS) resulting from the interaction of trichloroacetic acid (TCA) and protein is linearly related to the concentration of protein from 0.01 to 6.0 g/L. Precision studies indicate that the day-to-day coefficient of variation (CV) for two CSF controls (0.32 and 0.64 g/L) were 6.4% and 4.9%, respectively. The day-to-day CV's for two urine controls (0.39 and 0.84 g/L) were 5.8% and 2.9%, respectively. Correlation studies with a manual spectroturbidimetric method revealed a correlation coefficient of 0.99 for both CSF and urine specimens. The proposed method, which uses a smaller sample size, is fast and provides greater sensitivity at low ranges due to the ability to increase the sensitivity by the laser nephelometer.

Cerebrospinal Fluid Proteins↗

PubMed1990

Histologic, morphometric, and biochemical evolution of vein bypass grafts in a nonhuman primate model. II. Modification of early changes by platelet inhibition with aspirin and dipyridamole.

The objective of this study was to determine the early influence of platelet inhibition on the histologic, morphometric, and biochemical evolution of vein bypass grafts in a nonhuman primate model. Cephalic vein grafts were interposed bilaterally in the femoral arteries of 15 stump-tailed macaque monkeys fed a diet that sustains plasma cholesterol levels of approximately 225 mg/dl. All animals received in combination aspirin, 80 mg/day, and dipyridamole, 50 mg/day. Grafts were excised from five animals for analysis on each of postoperative days 3, 7, 14, 30, 60, and 90. In animals subjected to platelet inhibition, cholesterol content in the graft was 170 +/- 52 micrograms/100 mg at 90 days, 205% of the level in ungrafted vein (p less than 0.01). This change was small in comparison with the increase to 686% of ungrafted vein observed in our study of control animals. In stepwise regression analysis, cholesterol content of grafts was best predicted by prevalence of foam cells (r2 = 0.82), and the proportion of intima as a fraction of total wall area was best predicted by the presence of macrophages (r2 = 0.69). Platelet inhibition did not decrease the extent of intimal hyperplasia. The prevalence of adherent platelets (r = -0.72) and the amount of fibrin (r = -0.78) correlated inversely with the amount of endothelium present during the first 14 days. The strength of these correlations declined with time, despite persistent lack of endothelium in some areas. Medial fibrosis occurred to the same extent as in control grafts, as did the early appearance of platelet factor VIII and fibronectin and the lack of vasa vasorum at 3 days followed by reappearance at 7 days. These data demonstrate that platelet inhibition dramatically reduces lipid uptake by grafts in the first 90 days but has less influence over histologic or morphometric changes.

Animals↗

PubMed1990

Histologic, morphometric, and biochemical evolution of vein bypass grafts in a nonhuman primate model. I. Sequential changes within the first three months.

The objective of this study was to define the histologic and morphometric evolution that accompanies the increase in cholesterol content of vein bypass grafts in a nonhuman primate model. Cephalic vein grafts were interposed bilaterally in the femoral arteries of 15 stump-tailed macaque monkeys (Macaca arctoides), which were fed a diet that sustains plasma cholesterol levels of approximately 225 mg/dl. Grafts were excised from five animals for analysis on each of postoperative days 3, 7, 14, 30, 60, and 90. Cholesterol content increased from 69 +/- 24 micrograms/100 mg (mean +/- standard deviation) in ungrafted vein to 473 +/- 122 micrograms/100 mg in grafts 90 days after implantation (p less than 0.05). By stepwise regression analysis, cholesterol content was best predicted by abundance of foam cells (r2 = 0.82). Intima comprised 13% +/- 5% of the total cross-sectional area of the wall in ungrafted vein and 59% +/- 11% at day 90 (p less than 0.001). With cholesterol content excluded from the stepwise regression, intimal area was best predicted by the presence of foam cells (r2 = 0.39). There was consistently an increase in the prevalence of polymorphonuclear leukocytes on the luminal surface and in both the intima and media during the first 14 days after grafting. Vasa vasorum, which were always present in ungrafted vein, were sparse at 3 days but reappeared by day 7. Medial fibrosis occurred in grafts, and in the 30- to 90-day interval it was directly correlated with the number of adventitial vasa vasorum present (r = 0.64, p less than 0.05). Immunohistochemistry revealed prominent staining for both platelet factor VIII and fibronectin during the first month, with a gradual decline in staining intensity thereafter. The evolution of changes in vein bypass grafts documented in this report are in general agreement with graft changes observed in humans and support the validity of our model in evaluating the histologic correlates of increased graft cholesterol content.

Animals↗