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Biomedical subjects

H Y Hu

Publications and source records attributed to H Y Hu.

At least 37 records · Page 2Linked to original sources
PubMed2001

Effects of temperature on biodegradation characteristics of organic pollutants and microbial community in a solid phase aerobic bioreactor treating high strength organic wastewater.

The BOD removal rate and microbial community structure in a solid phase aerobic bioreactor using polyvinyl alcohol gel particles as packing material for the treatment of high strength organic wastewater were investigated at various temperatures. The BOD removal rate in the bioreactor increased when the temperature increased from 20 degrees C to 30 degrees C, 40 degrees C, and 50 degrees C, but it decreased when the temperature increased from 50 degrees C to 60 degrees C. Higher temperature enhanced the endogenous respiration of microbes in the bioreactor. The microbial community structure in the bioreactor was analyzed with quinone profile. The experimental results showed that the microbial community structure in the bioreactor was significantly affected by temperature. The dominant quinone of the microbes inhabiting the bioreactor was ubiquinone-8 at 30 degrees C, but that at 50 degrees C and 60 degrees C was menaquinone-7. It was estimated that the thermophilic Bacillus having menaquinone-7 dominated in the bioreactor at higher temperature. The microbial diversity in the bioreactor varied with temperature.

Bacillus↗
PubMed2001

Characterization of microbial community in an activated sludge process treating domestic wastewater using quinone profiles.

The dynamics of microbial community structure of activated sludges in a small-scale domestic wastewater treatment process were examined using a novel approach of quinone profiles. The composition and content of quinones in the activated sludges were analyzed monthly over a period of one year. More than 4 types of ubiquinones and 12 types of menaquinones were observed in the activated sludges, with the dominant quinones being ubiquinone (UQ)-8, menaquinone (MK)-7, followed by UQ-10, MK-8 and MK-6. The total quinone contents in the activated sludges varied from 0.93 to 2.68 mumol per gram of particle organic carbon. The molar ratio of ubiquinones to menaquinones (UK/MK) changed from 0.38 to 0.98, indicating that anaerobic bacteria dominated the microbial community of the activated sludges examined. The ratio of UQ/MK varied similar to that of dissolved oxygen in the bulk. The microbial diversity of the activated sludges calculated from the quinone compositions was 13.4-16.8. The diversity of menaquinones was much higher than that of ubiquinones, and increased slightly with increasing temperature. The microorganisms containing menaquinones appear to be sensitive to the change in temperature than those containing ubiquinones.

Bacteria↗
PubMed2001

Influence of gamma irradiation and storage on apheresis platelets.

BACKGROUND AND PURPOSE: Gamma irradiation of platelet concentrates to prevent graft-versus-host disease may inactivate contaminated lymphocytes and subsequently inhibit the synthesis of cytokines in the apheresis platelets during storage. We investigated the influence of irradiation and storage on apheresis platelets collected with the COBE Spectra or Fenwal CS-3000 Plus systems. METHODS: Eleven units of apheresis platelets were collected with a COBE Spectra cell separator and another 11 units with a Fenwal CS-3000 Plus system. Each unit of apheresis platelets was divided into two equal parts: one was irradiated with 3000 cGy directly after blood donation, and the other served as a control. Cell counts, platelet activation marker CD62 antigen, blood gas values, and supernatant concentrations of K+, Na+, lactate, glucose, interleukin-1 beta (IL-1 beta), IL-8, and tumor necrosis factor-alpha (TNF-alpha) were determined in paired samples on the day of collection (day 0) and after 5 days of storage (day 5). RESULTS: No significant differences in white cell counts or TNF-alpha concentrations were noted between the irradiated and control platelets on day 0 or day 5, whereas the mean proportion of platelets expressing CD62P (22.65% vs 25%, p = 0.014) and the mean IL-1 beta (45.55 pg/mL vs 52.75 pg/mL, p = 0.004) and IL-8 concentrations (10.68 pg/mL vs 13.07 pg/mL, p = 0.015) were significantly lower in irradiated than control platelets on day 5. The 5-day storage significantly increased the mean proportion of platelets expressing CD62P (25.00% vs 15.02%, p = 0.008), mean PO2 (116.34 mm Hg vs 98.07 mm Hg, p = 0.002), and mean concentrations of K+ (3.30 mmol/L vs 3.06 mmol/L, p < 0.001), lactate (15.12 mmol/L vs 3.23 mmol/L, p < 0.001), IL-1 beta (52.75 pg/mL vs 29.73 pg/mL, p = 0.001), and IL-8 (13.07 pg/mL vs 3.62 pg/mL, p < 0.001). Five-day storage also significantly decreased white cell count (0.18 x 10(8) vs 0.74 x 10(8), p < 0.001), PCO2 (19.38 mm Hg vs 50.51 mm Hg, p < 0.001), and concentrations of HCO3- (10.36 mmol/L vs 21.34 mmol/L, p < 0.001) and glucose (193.37 mg/dL vs 309.18 mg/dL, p < 0.001). Platelet counts and concentrations of IL-1 beta, IL-8, and TNF-alpha on day 0 did not differ significantly between control apheresis platelets collected with the Fenwal CS-3000 Plus and those collected with COBE Spectra. The mean white cell count (1.29 x 10(8) vs 0.19 x 10(8), p = 0.002) and the proportion of platelets expressing CD62P (24.71% vs 7.09%, p < 0.001) on day 0, however, were significantly higher in the platelets collected with the Fenwal CS3000-Plus than in those collected with the COBE Spectra. CONCLUSIONS: Gamma irradiation of apheresis platelets inhibits the expression of platelet CD62P and the secretion of IL-1 beta and IL-8 after 5 days' storage.

Blood Component Removal↗
PubMed2001

[High expression of rabbit sperm membrane protein rSP10 in Escherichia coli and preparation of its specific antisera].

To study the position and immunogenicity of rSP10, the rabbit rsp10 gene which did not include sequences coding for signal peptides was inserted into expression vector pET30a. An in-frame fusion protein was made such that a His6 stretch was produced at the N terminus of re-rSP10. High expression was obtained, the amount of re-rSP10 up to 67% in the total bacterial protein. The re-rSP10 was purified by DEAE chromatography and the yield of purified re-rSP10 was approximately 50 micrograms/mL of culture. Western blotting analysis of re-rSP10 with rabbit polyclonal sera raised against rabbit sperm membrane protein showed that the synthesized antigen possessed immunogenicity of rSP10. Specific antisera against re-rSP10 was induced using purified re-rSP10 as an antigen. The motility of capacitated sperms were affected but no aggregation was observed.

Animals↗
PubMed2001

[Genetic studies of 13 loci in Guangdong Han population by two multiplex PCR systems and its forensic application].

OBJECTIVE: The genetic studies of 13 short tandem repeats(STRs) loci in two multiplex amplification systems were carried out on Chinese Han population in Guangdong. METHODS: DNA samples from 328 unrelated individuals were screened. The 13 loci were D3S1358, vWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317, D7S820, D16S539, TH01, TPOX and CSF1PO. The PCR products were analyzed and genotyped by ABI 377-96 Sequencer. RESULTS: The combined power of discrimination (DP) was 0.999999999999993 and the combined paternity of exclusion(PE) was 99.999%. These 13 loci met the Hardy-Weinberg expectations. CONCLUSION: The two multiplex amplification systems were very useful in forensic case investigation.

Asian People↗
PubMed2000

Alpha-to-beta structural transformation of ovalbumin: heat and pH effects.

Ovalbumin is an important member of the serpin superfamily without inhibitory activity. The heat- and pH-induced alpha-to-beta structural transformations of ovalbumin were investigated by means of circular dichroism and binding of ANS and Congo red dyes. The native ovalbumin shows a mixture of alpha-helix and beta-sheet, while both the heat and alkali treatments are able to transform the native protein into a predominance of beta-sheet secondary structure. The free energy changes during transitions to the unfolded state are 5.19 kcal/mol from the native state and 4.00 kcal/mol from the heat-treated one. The binding abilities of the heat-treated and the alkali-treated forms to ANS and Congo red suggest that the altered forms exhibit hydrophobic exposure and intermolecular interaction. The results substantiate that the altered protein forms bearing increased beta-sheet structures are prone to aggregation, which is implicated in the pathogenesis of some conformational diseases.

Animals↗
PubMed2000

Determination of sibship by PCR-amplified short tandem repeat analysis in Taiwan.

BACKGROUND: Sibship determination for any two persons whose parents have died is one of the most fundamental issues of personal identification, second only to those of a parent-child relationship. STUDY DESIGN AND METHODS: By automated fluorescence analysis of a PCR-amplified short tandem repeat (STR) system in conjunction with capillary electrophoresis, a panel of up to 15 polymorphic, autosomal, unlinked STR loci was used to investigate sibship index (SI) values in a cohort of 126 true sibling pairs. These SI values were then compared with those of 126 random pairs. RESULTS: The 15-loci STR panel provides a cumulative power of exclusion of 0. 9999997. Of the 126 random pairs, 124 (98.4%) had cumulative sibship indices (CSIs) of <1.0, and none had a CSI of >3.0 (median, 0.0101; range, 0.0000003-2.5376). In contrast, 107 (85%) of the 126 sibling pairs had a CSI of >100 (median, 5,579.9853; range, 0.0747-9,406,829, 249.8461). However, five pairs (4%) of the sibling group had a CSI of <3.0. True sibship was confirmed for this particular group by additional paternity testing and mitochondrial DNA sequence analysis. Among a total of 1890 observations (15 loci x 126 pairs), two alleles per locus were shared 760 times (40.21%) (mean, 6.03 loci; range, 1-10) in the sibling group, but only 192 times (10.16%) in the random group (mean, 1.52 loci; range, 1-5) (p<0.001). No alleles were shared 696 times (36.83%) in the unrelated pairs, as compared to 176 times (9.31%) in the sibling group (p<0.001). A polarized distribution was not noted in the sharing of single alleles in either the random or the sibling group: 1002 observations (53.02%) and 954 observations (50.48%), respectively. CONCLUSION: Highly polymorphic STR analysis can be discriminative in most sibship determinations, and the sharing of two alleles per locus is most informative in indicating sibship. Complementary mitochondrial DNA sequence analysis is mandatory in a few cases to exclude or establish true sibship when CSIs are equivocal and neither parent is available.

Alleles↗
PubMed1999

Acute and chronic phase platelet aggregability studies in Chinese patients after implantation of a permanent transvenous pacemaker.

It has been suggested that the incidence of thromboembolic events always increases in patients after insertion of a transvenous pacemaker. Blood samples from twenty consecutive patients (fifteen males and five females) before and after pacemaker implantation was retained for platelet aggregability studies which were analyzed separately with ADP, collagen, epinephrine and arachidonic acid. The maximal amplitude of platelet aggregatory curve was detected by an aggregometer. The samples collected the day before pacemaker implantation (day 0) were used as self-control. Day 1 and day 3 after pacemaker implantation were defined as the acute phase, while day 30 was defined as the chronic phase. The maximal amplitude of platelet aggregatory curve was observed to be lowest on day 1 and then return to normal on day 3 and day 30. The results of platelet aggregability, however, showed no significant difference (P>0.05) between self-control and post-implantation samples. In conclusion, there was no significant change in platelet aggregability for either acute or chronic phases after pacemaker implantation. Antiplatelet medications may not be necessary for the prevention of thromboembolic events after the implantation of a pacemaker.

Aged↗
PubMed1999

Development of a novel solid phase extraction method for the analysis of bacterial quinones in activated sludge with a higher reliability.

A novel analytical method for analysis of microbial quinones in activated sludge sample was developed with improved reliability compared to the conventional method. The operating conditions for the extraction of quinones from activated sludge sample with a methanol-chloroform mixture and hexane were optimized. A solid phase extraction method using the Sep-Pak Plus Silica, a small column packed with silica gel, was employed for the purification and separation of quinones instead of column chromatography and thin layer column chromatography used in the conventional method. The analytical operation was simplified and the analytical time was shortened by a half or two-thirds of that required for the conventional method. The recoveries of quinones were markedly increased from 30-60% with the conventional method to as high as 90-101% with the improved method, demonstrating that the reliability of the analytical results of the improved method is significantly higher than that of the conventional method. The quinone content value obtained using the improved method was twice as high as that obtained using the conventional method for the same activated sludge sample, but the quinone profiles expressed as the mole fraction of each quinone type were the same for each method.

Journal Article↗
PubMed1999

Study of the redox properties of metallothionein in vitro by reacting with DsbA protein.

Mammalian metallothionein (MT) contains 20 cysteine residues involved in the two metal clusters without a disulfide bond. The redox reaction of the Cys thiols was proposed to be associated with the metal distribution of MT. The E. coli DsbA protein is extremely active in facilitating thiol/disulfide exchange both in vivo and in vitro. To further investigate the redox properties of MT, reaction between MT and DsbA was carried out in vitro by fluorescence detection. Equilibrium characterization indicates that the reaction is stoichiometric (1:1) under certain conditions. Kinetic study gives a rate constant of the redox reaction of 4.42 x 10(5) sec(-1) M(-1), which is 10(3)-fold larger than that of glutathione reacting with DsbA. Metal-free MT (apo-MT) shows a higher equilibrium reduction potential than MT, but exhibits an indistinguishable kinetic rate. Oxidation of MT by DsbA leads to metal release from the clusters. The characteristic fluorescence increase during reduction of DsbA may provide a sensitive probe for exploring the redox properties of some reductants of biological interest. The result also implies that oxidation of Cys thiols may influence the metal release or delivery from MT.

Animals↗
PubMed1998

The effects of helix breaking mutations in the diphtheria toxin transmembrane domain helix layers of the fusion toxin DAB389IL-2.

The fusion protein toxin DAB389IL-2 is composed of the catalytic and transmembrane domains of diphtheria toxin genetically linked to human interleukin 2 (IL-2). This fusion toxin is selectively toxic for eukaryotic cells which express the high-affinity form of the IL-2 receptor and the mechanism of intoxication parallels that of native diphtheria toxin. We used site-directed mutagenesis to introduce Pro residues into each of the three helical layers of the transmembrane domain. Although each of the mutations results in the complete loss of cytotoxic activity, individual mutants were found to vary with respect to channel formation in planar lipid bilayers, binding affinity and melting temperature. We propose that each of the three helix layers plays a critical role in the productive delivery of the catalytic domain to the cell cytosol.

Amino Acid Sequence↗
PubMed1998

Allogeneic peripheral blood stem cell transplantation and early detection of donor engraftment by polymerase chain reaction.

Patients with hematologic malignancy or severe aplastic anemia after myeloablative chemo- and radiotherapy were given granulocyte colony-stimulating factor (G-CSF)-mobilized, cryopreserved allogeneic peripheral blood stem cells (PBSCs) from 15 healthy donors who were either human leukocyte antigen (HLA)-matched siblings (n = 13) or haploidentical offspring (2). Polymerase chain reaction-amplified short tandem repeat genotyping was used for early confirmation of donor engraftment after PBSC transplantation (PBSCT). A standard cyclosporine A/methotrexate combination was used to prevent acute graft-versus-host disease (GVHD). All donors, including one in the third trimester of pregnancy, tolerated G-CSF administration and 3-day PBSC harvesting procedures well. Engraftment was prompt for all patients; it was verified using a panel of 12 human polymorphic short tandem repeat loci from bone marrow as early as 7 days posttransplantation. This status was maintained until relapse, when mixed chimerism was detected using the polymerase chain reaction. A minimum resurgence of recipient cells to 1% of the population was required to detect chimerism. The median times to recovery of the absolute neutrophil count to greater than 0.5 x 10(9)/L and the sustained platelet count to greater than 20 x 10(9)/L without transfusion were 10 and 12 days after PBSCT, respectively. Six patients experienced acute GVHD, Grade I in two patients and Grade II in four, including two HLA-haploidentical recipients. Chronic GVHD was noticed in three of the 11 patients who were followed for at least 100 days after PBSCT. Ten patients were still alive at the latest follow-up and have been disease free for a median of 278 days (range 60-671). Five patients died from causes other than graft failure: three from leukemia relapse and two from transplant-related complications. The results confirm that G-CSF can be safely administered to healthy donors and that engraftment after allogeneic PBSCT is fast and durable. Complete chimerism can be detected early by genomic analysis. PBSCT may offer an alternative to bone marrow transplantation.

Adolescent↗
PubMed1998

Polymorphisms of twelve short tandem repeat loci in a Taiwanese population and their application in parentage testing.

With the advancement of techniques in molecular biology, rapid, sensitive, and reliable methods of DNA typing for parentage testing have become available. In this study, we evaluated the usefulness of multiplex polymerase chain reaction (PCR) with 12 unlinked short tandem repeat (STR) loci for paternity testing in Taiwan. The genetic informativeness of this test was then compared with that of conventional human leukocyte antigen (HLA) analysis in 167 parentage studies. The 12 STR loci alone provided a cumulative power of exclusion of up to 0.9998. Paternity was excluded in 59 (35.3%) cases, including 40 of 112 paternity trios and 19 of 55 paternity duos. In the 40 trios in which paternity was excluded, a mean of 6 (range, 3-9) incompatible STR markers were in the 19 duos in which paternity was excluded, a mean of 4 (range, 1-8) incompatible STR markers were noted. In the 72 trios in which the alleged paternity could not be excluded, the mean probabilities of paternity (PP) were 90.6863% with HLA testing alone, 99.9847% with STR analysis alone, and 99.9972% with combined HLA and STR analysis. In the 36 duos in which the alleged paternity could not be excluded, the mean PPs were 81.4768% with HLA testing alone, 99.6124% with STR analysis alone, and 99.9145% with combined HLA and STR analysis. These results suggest that STR analysis is very powerful when used alone for paternity trio testing and when combined with conventional serologic HLA typing for duo parentage testing in the Taiwan population.

Chromosome Mapping↗
PubMed1998

Factors affecting progenitor cell yields using three tandem leukaphereses in previously treated malignancies.

BACKGROUND: Mobilized peripheral blood progenitor cells (PBPCs) have increasingly been used to replace autologous bone marrow to allow faster hematopoietic reconstitution after myeloablative therapy in various malignancies. There is a paucity of data concerning factors that affect the total yield of three tandem leukaphereses. METHODS: Factors affecting the yield of PBPCs were analyzed in a series of 121 consecutive patients including 36 with non-Hodgkin's lymphoma, two with Hodgkin's disease, four with multiple myeloma, 44 with acute leukemia, 20 with breast cancer and 15 with other solid tumors. PBPCs were mobilized using granulocyte-colony-stimulating factor (G-CSF) alone (group I, n = 15), or after conventional-dose (group II, n = 70) or high-dose (group III, n = 36) chemotherapy followed by G-CSF. The total yield of three tandem PBPC collections for each patient was assessed by the number of mononuclear cells (MNCs), CD34+ cells and colony-forming units of granulocyte macrophages (CFU-GM). The factors evaluated included age, sex, diagnosis, history of marrow involvement, previous radiotherapy, the number of prior chemotherapy cycles and mobilization method. The two -sample t-test and logistic regression analysis were performed for univariate and multivariate analysis, respectively. RESULTS: With univariate analysis, a diagnosis of acute leukemia, positive history of bone marrow involvement, more chemotherapy cycles and mobilization with high-dose chemotherapy adversely affected the yields of CD34+ cells. By multivariate analysis, Group II had higher yields of MNCs (p = 0.039), CFU-GM (p = 0.002) and CD34+ cells (p = 0.011) than Group III. Fewer cycles of prior chemotherapy is the common favorable factor for the yields of both CD34+ cells (p = 0.016) and CFU-GM (p = 0.017). CONCLUSIONS: The number of prior chemotherapy cycles adversely affects progenitor cell yield. Conventional-dose chemotherapy followed by G-CSF seems to be the mobilization methods of choice for heavily pretreated cancer patients with limited bone marrow reserve. PBPCs should be harvested early, when the tumor burden is less, to avoid cumulative marrow toxicity from chemotherapy.

Adolescent↗
PubMed1997

Only snake curaremimetic toxins with a fifth disulfide bond have high affinity for the neuronal alpha7 nicotinic receptor.

Long chain and short chain curaremimetic toxins from snakes possess 66-74 residues with five disulfide bonds and 60-62 residues with four disulfide bonds, respectively. Despite their structural differences all of these toxins bind with high affinity to the peripheral nicotinic acetylcholine receptors (AChR). Binding experiments have now revealed that long chain toxins only, like the neuronal kappa-bungarotoxin, have a high affinity for a chimeric form of the neuronal alpha7 receptor, with Kd values ranging from about 1 to 12 nM. In contrast, all other toxins bind to the chimeric alpha7 receptor with a low affinity, with Kd values ranging between 3 and 22 microM. These results are supported by electrophysiological recordings on both the wild-type and chimeric alpha7 receptors. Amino acid sequence analyses have suggested that high affinities for the neuronal receptor are associated with the presence of the fifth disulfide at the tip of the toxin second loop. In agreement with this conclusion, we show that a long chain toxin whose fifth disulfide is reduced and then dithiopyridylated has a low affinity (Kd = 12 microM) for the neuronal alpha7 receptor, whereas it retains a high affinity (Kd = 0.35 nM) for the peripheral AChR. Thus, a long chain curaremimetic toxin having a reduced fifth disulfide bond behaves like a short chain toxin toward both the peripheral and neuronal AChR. Therefore, functional classification of toxins that bind to AChRs should probably be done by considering their activities on both peripheral and neuronal receptors.

Amino Acid Sequence↗
PubMed1997

Solution conformation of N-terminal fragments of trichosanthin small domain (TCS 182-200). Circular dichroic studies.

Three peptides, T14, T18 and TDK, derived from the N-terminus of trichosanthin small domain (TCS 182-200) have been investigated by circular dichroism. Secondary structure and structural transitions of the above peptides under different conditions were studied. Alcohol prompts a transition of the T18 peptide from a beta-sheet to an alpha-helical structure. It also increases the alpha-helicities of T14 and TDK. The beta-sheet of T18 peptide appears more hydrophobic than the alpha-helix of T14 or TDK. The effects of polypeptide sequence and solvent on secondary structure formation of these model peptides are discussed.

Amino Acid Sequence↗
PubMed1996

Analytic performance and contamination control methods of a ligase chain reaction DNA amplification assay for detection of Chlamydia trachomatis in urogenital specimens.

A ligase chain reaction (LCR) DNA amplification assay that targeted the cryptic plasmid of Chlamydia trachomatis was developed to detect C. trachomatis urogenital tract infection. The objectives of this study were to determine the cutoff and analytic performance of the LCR assay and to characterize the effectiveness of its postdetection contamination control method. The assay's cutoff was determined after receiver-operator characteristic (ROC) analysis of 4660 clinical data points. The assay detected one infectious unit per reaction of each of the 15 C. trachomatis serovars and did not cross-react with 13 Chlamydia pneumoniae strains, 13 Chlamydia psittaci strains, and 87 other bacteria, fungi, parasites, or viruses. In addition, the assay did not detect 77 processed urine specimens collected from patients with urinary tract infections caused by yeast or bacteria other than C. trachomatis. The assay was sufficiently precise to detect consistently two infectious units of C. trachomatis per reaction. False-positive assay results attributable to contamination with amplified product were minimized by the use of standard procedures as well as by a postdetection chemical inactivation method that could reduce the amount of amplified LCR product by a factor of > or = 10(7).

Chlamydia trachomatis↗