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Biomedical subjects

J Y Huang

Publications and source records attributed to J Y Huang.

At least 19 recordsLinked to original sources

The EMOTIF database.

The EMOTIF database is a collection of more than 170 000 highly specific and sensitive protein sequence motifs representing conserved biochemical properties and biological functions. These protein motifs are derived from 7697 sequence alignments in the BLOCKS+ database (released on June 23, 2000) and all 8244 protein sequence alignments in the PRINTS database (version 27.0) using the emotif-maker algorithm developed by Nevill-Manning et al. (Nevill-Manning,C.G., Wu,T.D. and Brutlag,D.L. (1998) Proc. Natl Acad. Sci. USA, 95, 5865-5871; Nevill-Manning,C.G., Sethi,K.S., Wu,T. D. and Brutlag,D.L. (1997) ISMB-97, 5, 202-209). Since the amino acids and the groups of amino acids in these sequence motifs represent critical positions conserved in evolution, search algorithms employing the EMOTIF patterns can identify and classify more widely divergent sequences than methods based on global sequence similarity. The emotif protein pattern database is available at http://motif.stanford.edu/emotif/.

Amino Acid Motifs↗

Serum interleukin-6 (IL-6), IL-10, tumor necrosis factor (TNF) alpha, soluble type II TNF receptor, and transforming growth factor beta levels in human immunodeficiency virus type 1-infected individuals with Mycobacterium avium complex disease.

To characterize changes in serum cytokine levels in human immunodeficiency virus type 1 (HIV-1)-infected persons with Mycobacterium avium complex (MAC) bacteremia, the levels of IL-1alpha (interleukin-1alpha), IL-6, IL-10, tumor necrosis factor alpha (TNF-alpha), soluble type II TNF receptor (sTNF-RII), and transforming growth factor beta (TGF-beta) in serum were measured in two cohorts of HIV-1-infected persons with MAC bacteremia. The first cohort was part of a MAC prophylaxis study. Patients with bacteremia were matched with controls without bacteremia. Elevated IL-6, IL-10, TNF-alpha, sTNF-RII, and TGF-beta levels were noted at baseline for all subjects, a result consistent with advanced HIV-1 disease. IL-1alpha was not detected. No differences in cytokine levels in serum were noted at baseline and at the time of bacteremia between patients with MAC and controls. In the second cohort, subjects had serum samples collected at the time of MAC bacteremia and thereafter while on macrolide therapy. Serum samples at time of bacteremia were collected from HIV-1-infected persons at a time when neither highly active antiretroviral therapy (HAART) nor MAC prophylaxis was used routinely. MAC treatment resulted in decreased levels of IL-6 and TNF-alpha in serum, which were evident for IL-6 by 4 to 6 weeks and for TNF-alpha by 8 to 16 weeks. Thus, antibiotic treatment for MAC results in decreased levels of IL-6 and TNF-alpha in serum in HIV-1-infected persons who are not on HAART.

AIDS-Related Opportunistic Infections↗

Centrosomes have a role in regulating the destruction of cyclin B in early Drosophila embryos.

We reported previously that the disappearance of cyclin B at the end of mitosis in early Drosophila embryos starts at centrosomes and spreads into the spindle [1]. Here, we used a novel mutation, centrosome fall off (cfo), to investigate whether centrosomes are required to initiate the disappearance of cyclin B from the spindle. In embryos laid by homozygous cfo mutant mothers, the centrosomes co-ordinately detached from the mitotic spindle during mitosis, and the centrosomeless spindles arrested at anaphase. Cyclin B levels decreased on the detached centrosomes, but not on the arrested centrosomeless spindles, presumably explaining why the spindles arrest in anaphase in these embryos. We found that the expression of a non-degradable cyclin B in embryos also caused an anaphase arrest, but most centrosomes remained attached to the arrested spindles, and non-degradable cyclin B levels remained high on both the centrosomes and spindles. These findings suggest that the disappearance of cyclin B from centrosomes and spindles is closely linked to its destruction, and that a connection between centrosomes and spindles is required for the proper destruction of the spindle-associated cyclin B in early Drosophila embryos. These results may have important implications for the mechanism of the spindle-assembly checkpoint, as they suggest that unattached kinetochores may arrest cells in mitosis, at least in part, by signalling to centrosomes to block the initiation of cyclin B destruction.

Anaphase↗

A study of secondary cutaneous amyloidosis in basal cell carcinoma in Chinese patients: lack of correlation with bcl-2 or p53 protein expression.

Pigmented basal cell carcinoma (BCC) is an uncommon form of BCC among Caucasians. In contrast, the majority of BCC in Orientals are pigmented BCC. To assess the occurrence of secondary amyloid deposits in BCC among Chinese patients, a retrospective study was conducted on 53 BCC specimens. We used a terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling method for detecting the apoptotic cells. In addition, immunohistochemical staining was performed to examine the expression of the B-cell leukemia/lymphoma-2 gene (bcl-2) and p53 protein. Of the 53 BCC specimens, 37 (69.8%) were pigmented BCC and 31 (58.5%) showed amyloid deposits in the stroma of the tumor cells. The mean percentage of apoptotic tumor cells was 0.29%. Immunostaining of bcl-2 and p53 was detected in 33 BCCs (62.3%) and 19 BCCs (35.8%), respectively. No relationship between amyloid deposition and the number of apoptotic cells was found. In addition, there was no correlation between amyloid deposition and bcl-2 protein expression or between amyloid deposition and p53 protein expression. Our results indicate that the frequency of secondary amyloidosis in BCC among Chinese patients is not higher than that reported in the West. Although no correlation could be detected between amyloid deposition and bcl-2/p53 protein expression, the possible role of apoptosis in the pathogenesis of amyloid deposition in BCC still needs further investigation.

Adult↗

Total creatinine appearance as indicator of risk of infectious complication in peritoneal dialysis.

Peritonitis and exit-site infections are the main causes of complications in peritoneal dialysis. Death due to infectious complication is also one of the major causes of drop-out. The underlying cause of infection may include malnutrition. Total creatinine appearance (TCA) may reflect overall nutritional status. We determined TCA from the daily dialysate, urine, and estimated gut creatinine of patients and normalized it to actual body weight (nTCA). We examined the relationship between nTCA and the incidence of infection, and between nTCA and infection-related survival. The study included 323 adult patients in a single dialysis center. The mean nTCA of all patients was 19.73 +/- 4.75 mg/kg/day. The patients with an nTCA below 1 standard deviation from the mean (nTCA < 14.98 mg/kg/day) had a significantly higher peritonitis and exit-site infection rate (p < 0.01) and a higher chance of drop-out owing to infection-related complications (p < 0.0001). Our study concluded that the adult patient with malnutrition (nTCA < 14.98 mg/kg/day) has higher risk of infection.

Bacterial Infections↗

Beta2-integrin, LFA-1, and TCR/CD3 synergistically induce tyrosine phosphorylation of focal adhesion kinase (pp125(FAK)) in PHA-activated T cells.

Complete T cell activation requires not only a first signal via TCR/CD3 engagement but also a costimulatory signal through accessory receptors such as CD2, CD28, or integrins. Focal adhesion kinase, pp125(FAK) (FAK), was previously shown to be localized in focal adhesions in fibroblasts and to be involved in integrin-mediated cellular activation. Although signaling through beta1- or beta3-integrins induces tyrosine phosphorylation of FAK, there has been no evidence that activation of T cells through the beta2-integrin, LFA-1, involves FAK. We report here that crosslinking of LFA-1 induces tyrosine phosphorylation of FAK in PHA-activated T cells. Moreover, cocrosslinking with anti-LFA-1 mAb and suboptimal concentration of anti-CD3 mAb markedly increases tyrosine phosphorylation of FAK in an antibody-concentration-dependent and time-kinetics-dependent manner compared with stimulation through CD3 alone, which correlates well with enhanced proliferation of PHA-activated T cells. Furthermore, LFA-1beta costimulation with CD3 induces tyrosine phosphorylation of Syk associated with FAK. These results indicate, for the first time, that signals mediated by LFA-1 can regulate FAK, suggesting that LFA-1-mediated T cell costimulation may be involved in T cell activation at least partially through FAK.

CD18 Antigens↗

The crystal structure of a bacterial, bifunctional 5,10 methylene-tetrahydrofolate dehydrogenase/cyclohydrolase.

The structure of a bifunctional 5,10-methylene-tetrahydrofolate dehydrogenase/cyclohydrolase from Escherichia coli has been determined at 2.5 A resolution in the absence of bound substrates and compared to the NADP-bound structure of the homologous enzyme domains from a trifunctional human synthetase enzyme. Superposition of these structures allows the identification of a highly conserved cluster of basic residues that are appropriately positioned to serve as a binding site for the poly-gamma-glutamyl tail of the tetrahydrofolate substrate. Modeling studies and molecular dynamic simulations of bound methylene-tetrahydrofolate and NADP shows that this binding site would allow interaction of the nicotinamide and pterin rings in the dehydrogenase active site. Comparison of these enzymes also indicates differences between their active sites that might allow the development of inhibitors specific to the bacterial target.

Aminohydrolases↗

Human Nopp140, which interacts with RNA polymerase I: implications for rRNA gene transcription and nucleolar structural organization.

Nopp140 is thought to shuttle between nucleolus and cytoplasm. However, the predominant nucleolar localization of Nopp140 homologues from different species suggests that Nopp140 is also involved in events occurring within the nucleolus. In this study, we demonstrated that the largest subunit of RNA polymerase I, RPA194, was coimmunoprecipitated with the human Nopp140 (hNopp140). Such an interaction is mediated through amino acids 204 to 382 of hNopp140. By double immunofluorescence, hNopp140 was colocalized with RNA polymerase I at the rDNA (rRNA genes) transcription active foci in the nucleolus. These results suggest that Nopp140 can interact with RNA polymerase I in vivo. Transfected cells expressing the amino-terminal half of hNopp140, hNopp140N382 (amino acids 1 to 382), displayed altered nucleoli with crescent-shaped structures. This phenotype is reminiscent of the segregated nucleoli induced by actinomycin D treatment, which is known to inhibit rRNA synthesis. Consistently, the hNopp140N382 protein mislocalized the endogenous RNA polymerase I and shut off cellular rRNA gene transcription as revealed by an in situ run-on assay. These dominant negative effects of the mutant hNopp140N382 suggest that Nopp140 plays an essential role in rDNA transcription. Interestingly, ectopic expression of hNopp140 to a very high level caused the formation of a transcriptionally inactive spherical structure occupying the entire nucleolar area which trapped the RNA polymerase I, fibrillarin, and hNopp140 but excluded the nucleolin. The mislocalizations of these nucleolar proteins after hNopp140 overexpression imply that Nopp140 may also play roles in maintenance of nucleolar integrity.

Animals↗

Insulin-like growth factor I plays a role in regulating erythropoiesis in patients with end-stage renal disease and erythrocytosis.

Erythroid progenitor growth, the serum hormones that regulate erythropoiesis, and the effect of patient's serum on the growth of normal erythroid progenitors were assessed in eight patients with end-stage renal disease (ESRD) and erythrocytosis. All patients were male and had been on maintenance dialysis, they had a hematocrit >50% and/or a red blood cell count >6 x 10(12)/L and an arterial oxygen saturation >95%. Four had acquired cystic disease of the kidney (ACDK), and four other non-ACDK patients did not have known causes of secondary erythrocytosis after appropriate investigations and long-term follow-up. The methylcellulose culture technique was used to assay the erythroid progenitor (BFU-E/CFU-E) growth. Serum erythropoietin (EPO) and insulin-like growth factor I (IGF-I) levels were measured by RIA. Paired experiments were performed to determine the effects of 10% sera from ESRD patients and control subjects on normal marrow CFU-E growth. The numbers of EPO-dependent BFU-E in marrow and/or blood of patients with ESRD and erythrocytosis were higher than those of normal controls. No EPO-independent erythroid colonies were found. Serum EPO levels were constantly normal in one patient and elevated in three patients with ACDK; for non-ACDK patients, EPO levels were normal or low in two patients and persistently increased in one, but fluctuated in the remaining one on serial assays. There was no correlation between serum EPO levels and hematocrit values. The serum IGF-I levels in patients with ESRD and erythrocytosis were significantly increased compared with normal subjects or ESRD patients with anemia. We found an inverse correlation between serum EPO and IGF-I levels. Sera from patients with ESRD and erythrocytosis exhibited a stimulating effect on normal marrow CFU-E growth. The stimulating effect of sera from patients who had a normal serum EPO level and an elevated IGF-I level could be partially blocked by anti-IGF-I. The present study suggests that IGF-I plays an important role in the regulation of erythropoiesis in patients with ESRD and erythrocytosis who did not have an increased EPO production.

Adult↗

[The study of family structure, illness symptom, and stress adaptation of psychotic patients].

The major purposes of this paper are to explore the phenomena of family structure, illness symptoms, family coping and adaptation for patients with schizophrenia or manic-depression psychosis. This paper tries to provide a good reference instrument for application by nurses in home care, in order to understand and evaluate family needs. Subjects are schizophrenic or manic-depression outpatients from 3 hospitals located in northern Taiwan. Data were collected through home interview with primary caregivers and observations. There were fifty subjects from each of the 3 hospitals, and 151 subjects in total. Descriptive statistics, t-test, one way ANOVA, Pearson correlation and multiple stepwise correlation were used to analyze data. Research indicates that most patients are aged between 31 to 40, with over 10 years elapsed since onset, and are not married. Most primary caregivers are parents over 60 years old. Most family development was at the stage with young adult offspring. The manic-depressive patients have more working opportunities than schizophrenic patients. For schizophrenic patients, paranoia was the most serious in active symptoms; lack of interpersonal interaction was the most serious in negative symptoms; the other major problem was sleep disturbance in emotion-behavior assessment. Patient's disposition was the most concerning issue for families and the worst coping efficiency occurred with lazy behavior and sleep disturbance. For manic-depressive patients, aggressive behavior was the most serious active symptom, lack of energy was the most serious in negative symptom, and sleep disturbance was the most concerning problem in emotion-behavior assessment. The patient's symptom was the most concerning issue for families and the worst coping efficiency was found in drug side effect. The result also indicates that active and negative symptoms are both related to coping efficiency.

Adaptation, Psychological↗

Profiling expression patterns and isolating differentially expressed genes by cDNA microarray system with colorimetry detection.

A high-density cDNA microarray with colorimetry detection system to simultaneously monitor the expression of many genes on nylon membrane is described and characterized. To quantify the expression of genes and to isolate differentially expressed genes, the southern hybridization process on filter membranes was employed. The levels of gene expression were represented by color intensities generated by colorimetric reactions in place of hazardous radioisotopes or costly laser-induced fluorescence detection. The gene expression patterns on nylon membranes were digitized by devices such as an economical flatbed scanner or a digital camera. The quantitative information of gene expression was retrieved by image analysis software. Quantitative comparison of the northern dot-blotting method with the microarray system is described. Applications employing single-color detection as well as dual-color detection to isolate differentially expressed genes among thousands of genes are demonstrated.

Blotting, Northern↗

Microstructure investigations of ball milled materials.

HREM and FEG TEM were emphasized and extensively used to follow the most subtle changes in the structure and composition of ball-milled Cu, Fe-Cu, and thermally decomposed Fe60Cu40. Some significant results are obtained and summarized as follows: HREM shows that the deformation of ball-milled copper proceeds mainly by twinning and shear bands (SBs) formation. The nano-grains formed during ball milling (BM) contain a high density of dislocations. The grain boundaries (GBs) of nanocrystalline (NC) Cu prepared by BM are ordered, curved, and strained, but disordering, lattice distortion, and nanovoids in local regions were frequently observed. Nanoscale composition analysis on mechanically alloyed Fe16Cu84 shows that the average Fe content in both the interior of grains and the GBs is close to the designed composition, which proves that a supersaturated solid solution has really formed. However, the Fe content is rather inhomogeneous between the larger and smaller grains, which infers the inhomogeneous mixing of Fe and Cu during mechanical alloying (MA). NC structure and the mechanical force-enhanced fast diffusion are the reasons of the formation of supersaturated solid solutions in immiscible systems with positive enthalpy of mixing. HREM observations carried out with the thermally decomposed Fe60Cu40 solid solution show that the Nishiyama (N-W) or Kurdyumov-Sachs (K-S) orientation relationships exist between alpha-Fe and Cu. Energy dispersive X-ray spectra (EDXS) results show that the Cu content in these alpha-Fe grains reaches as high as 9.5 at.% even after heating to 1,400 degrees C, which is even higher than the maximum solubility of Cu in gamma-Fe at 1,094 degrees C.

Alloys↗

Co-stimulation of T cells with CD2 augments TCR-CD3-mediated activation of protein tyrosine kinase p72syk, resulting in increased tyrosine phosphorylation of adapter proteins, Shc and Cbl.

Complete T cell activation requires not only the first signal via TCR-CD3 engagement, but also a co-stimulatory signal through accessory receptors such as CD2, LFA-1 and CD28. However, the pathway of co-stimulatory signaling through accessory receptors is incompletely understood. We report here that CD2 provides a co-stimulus for activation of CD3-mediated syk/ZAP-70 family kinase, p72Syk (Syk), in Jurkat T cells. Although cross-linking of CD2 alone or any combination of CD2 with LFA-1alpha, LFA-1beta or CD28 did not induce tyrosine phosphorylation of Syk, co-cross-linking of CD2 with CD3 enhanced CD3-mediated tyrosine phosphorylation of Syk. Enhancement of tyrosine phosphorylation of Syk by CD2 co-stimulation was CD2 antibody concentration-dependent, and time course studies showed that CD2 co-stimulation enhanced Syk tyrosine phosphorylation by 30 s and through 5 min stimulation compared with the control. In vitro kinase assay revealed that co-cross-linking of CD2 with CD3 augmented Syk kinase activity using myelin basic protein as a substrate. Furthermore, CD2 co-stimulation with CD3 resulted in enhanced tyrosine phosphorylation of adapter proteins, such as Shc and Cbl, in an antibody concentration-dependent manner. Finally, CD2 provided a co-stimulatory signals for synthesis of IL-2 in Jurkat cells and phytohemagglutinin (PHA)-activated T cells and for proliferation of PHA-activated T cells. Taken together, these results indicate that CD2 is an important co-stimulatory receptor for CD3-mediated T cell activation and functions in concert with CD3.

Adaptor Proteins, Signal Transducing↗

Cyclosporin inhibits nitric oxide production in medullary ascending limb cultured cells.

BACKGROUND: Nitric oxide (NO) has been shown to play a role in cyclosporin (CsA) nephrotoxicity, but its mechanism of action is still unclear. As inducible NO synthase (iNOS) mRNA has been found to be expressed in rat medullary thick ascending limb (mTAL) cells, we investigated the effects of CsA on NO production in a model of mouse cultured mTAL cells. MATERIALS AND METHODS: The experiments were carried out on sub-cultured cells derived from isolated mTAL microdissected from the kidney of C57BL/6 mice. The identification of the iNOS mRNA in mTAL microdissected segment and cultured cell was confirmed by RT PCR and RsaI digestion. Nitrite (NO2) released by mTAL cells was determined using the modified Griess reagent method and taken as an index of nitric oxide production. The cultured cells were treated with various concentrations of CsA and different signal transduction regulators to assess the effect and possible pathway(s) of action of CsA on NO production in mTAL cells. RESULTS: The basal production of NO by mTAL cells increased by 1.8-fold following incubation with bacterial lipopolysaccaride (LPS). Both aminoguanidine and L-NAME inhibited NO production. CsA (10-300 ng/ml) also inhibited NO production in a dose-dependent manner and prevented its increase induced by LPS. Phorbol 12-myristate 13-acetate (PMA), a PKC stimulator, enhanced slightly the production of NO under basal conditions and prevented the inhibitory action of CsA on NO production. These results suggest that the NO secreted by mouse cultured mTAL cells is dependent on the PKC pathway. CONCLUSION: These results show that CsA may down-regulate the production of NO by cultured mTAL cells expressing iNOS mRNA and that the PKC pathway is involved in this process.

Animals↗

End-stage renal disease following polycythemia vera: in vitro and in vivo response of erythroid progenitors to erythropoietin and effects of sera on normal erythropoiesis.

We investigated the in vitro erythroid progenitor growth and the effects of sera on normal-marrow CFU-E (colony-forming units - erythroid) growth in 2 patients with renal failure on regular hemodialysis following a prior history of polycythemia vera (PV). PV was diagnosed 3 and 11 years, respectively, before the development of terminal renal failure. One of the patients had entered a spent phase of PV as characterized by diffuse extensive myelofibrosis and anemia; the other also had mild myelofibrosis. The serum erythropoietin (EPO) levels were low or normal on serial measurements by radioimmunoassay. There was no correlation between the hematocrit values and serum EPO levels. EPO-independent erythroid colonies were present in the cultures of bone marrow and peripheral blood cells from both patients after renal failure in the anemic state. With the addition of various concentrations of EPO, the number of erythroid colonies increased as the concentrations of EPO increased which was in accordance with the clinical observation that 1 patient with postpolycythemic myeloid metaplasia partially responded to recombinant human EPO therapy. In the EPO-dependent CFU-E assay, normal-marrow CFU-E numbers supported by 10% of the patient sera were less than those by normal sera. In the absence of EPO in cultures, no erythropoietic activity was found in the patients' sera. Our study on uremic patients with underlying PV showed that the biologic characteristics of autonomous erythroid progenitor growth for PV persisted during the spent phase and after the development of terminal renal failure with anemia. The erythroid progenitors responded to EPO both in vitro and in vivo. Their sera exhibited an inhibiting effect on the growth of normal-marrow erythroid progenitors.

Adult↗

Development of a dual-probe detection system for positron emission radiotracer detection.

BACKGROUND: Positron emission radiotracers are very useful in biochemical and pharmacological research. The purpose of this work was to develop and build a simple and inexpensive dual-probe detector system (DPDS) for detecting the activity of positron emission radiotracers. MATERIALS AND METHODS: The design of the DPDS was based on the coincident detection of two 511 KeV annihilation gamma-rays, which determined the positron activity in the body. A pair of sodium iodide (NaI) scintillators coupled to a photomultiplier tube (PMT) were positioned face-to-face as in a dual-probe system. The signals from each probe were then sent to a preamplifier, a linear amplifier, a single channel analyzer (SCA), and the primary and secondary coincidence units, respectively, to determine the number of total coincidence events and random events. Validation of basic characteristics, including stability, sensitivity, linearity and geometric counting efficiency, were tested. RESULTS: The sensitivity and linearity of the DPDS worked well in detecting radioactivity within 100 microCi. The DPDS was very stable in continuous counting for more than 2 hours. The geometric counting efficiency changed less than 5% when the source was placed 2 cm from the central coincidence line. CONCLUSION: This DPDS is a promising tool for positron emission radiotracer detection. It may be helpful in future in both clinical and basic biomedical research, such as neuroreceptor occupancy or glucose metabolic studies. Eventually, we will be able to use the DPDS in the human or animal body for evaluating position emission radiotracer metabolism and distribution.

Humans↗

Involvement of protein tyrosine kinase p72syk and phosphatidylinositol 3-kinase in CD2-mediated granular exocytosis in the natural killer cell line, NK3.3.

The granular exocytosis pathway is one mechanism by which NK cells and CTLs induce cytolysis of target cells. Triggering through adhesion molecules such as CD2 and LFA-1 as well as Fc gammaRIII (CD16) can invoke this pathway. CD2 is a cell surface glycoprotein present on CTLs and NK cells that plays an important role in both cellular adhesion and signal transduction. Here we report that cross-linking of CD2 as well as CD16 by immobilized Abs enhances granular exocytosis in an NK cell line, NK3.3. Herbimycin, a protein tyrosine kinase (PTK) inhibitor, or wortmannin, a specific inhibitor of phosphatidylinositol 3-kinase (PI 3-K), inhibited completely or almost completely CD2- or CD16-mediated granular exocytosis, suggesting the involvement of protein tyrosine kinases and PI 3-K in both CD2- and CD16-mediated granular exocytosis. We also observed that cross-linking of CD2 as well as CD16 enhances p72syk tyrosine kinase activity, and this enhancement correlated well with the increased tyrosine phosphorylation of several cellular proteins, including the adapter protein Shc. Furthermore, we have observed that cross-linking of CD2 as well as CD16 enhances the PI 3-K activity associated with the tyrosine-phosphorylated cellular proteins and Shc. These results provide insight into the signaling pathways by which triggering of CD2 and CD16 on NK cells leads to cytolysis of target cells.

Androstadienes↗