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Biomedical subjects

X Chen

Publications and source records attributed to X Chen.

At least 469 records · Page 26Linked to original sources
PubMed2000

L-type Ca2+ channel alpha 1c subunit isoform switching in failing human ventricular myocardium.

UNLABELLED: The objectives of this study were to determine the relative abundance of the L-type Ca channel alpha 1c IVS3 isoforms that result from alternative splicing in normal human ventricular myocytes and to measure the changes in isoform expression in end stage heart failure. METHODS: mRNA was isolated from left ventricular tissue and myocytes from non-failing and failing human hearts. RT-PCR with isoform-specific primers was used to obtain cDNAs that were then mutated for use in competitive PCR reactions. An RNase protection assay was also used to confirm the presence of one of the novel isoforms. RESULTS: Four different alpha 1c IVS3 isoforms were found in non-failing human ventricular myocytes using RT-PCR. Two isoforms contained exon 31 (termed IVS3A isoforms) and two isoforms contained exon 32 (termed IVS3B isoforms). One of these isoforms has not been observed previously and contains exon 31 and all but the last six base pairs of exon 32. In non-failing human ventricular myocytes the IVS3A isoform is 2.5 times more abundant than the IVS3B isoform. There were significant changes in the relative abundance of these isoforms in failing hearts, with the IVS3B isoform being twice as abundant as the IVS3A isoform. All isoforms were confirmed by RNase protection analysis. CONCLUSIONS: These experiments show that there are at least four L-type Ca channel mRNA isoforms in the normal human heart and that the relative abundance of these isoforms changes significantly in heart failure. These alpha 1c isoform changes in heart failure are associated with dysfunctional electromechanical disturbances, but the specific physiological role of each L-type Ca channel isoform in normal and failing hearts needs to be defined.

Adult↗
PubMed2000

Drug injection rates and needle-exchange use in New York City, 1991-1996.

Objectives included (1) to develop methods for identifying injection drug users with accelerating injection habits so they might be referred to counseling and treatment and (2) to investigate behavioral correlates of accelerating injection habits, including syringe-exchange program utilization. Data on drug use, enrollment in methadone maintenance, and demographic variables were obtained from 328 subjects who were seronegative for human immunodeficiency virus (HIV) who attended anywhere from 4 to 11 quarterly study visits for interview, HIV pretest counseling and risk reduction counseling, and blood donation for HIV antibody testing. Subjects were recalled 2 weeks after each study visit to receive their results and post-test counseling. We characterized subjects according to their patterns of drug injection as accelerating, decelerating, or stable, using intraindividual regression analyses and categorization rules, and by syringe-exchange use as consistent users, sporadic users, or nonusers. The present subjects included 52% with decelerating, 29% with stable, and 19% with accelerating rates of drug injection. There were 128 subjects (39%) who were categorized as consistent users of syringe-exchange programs, 84 (25%) were categorized as sporadic users, and 116 (35%) were categorized as nonusers. All syringe-exchange groups showed significantly decelerating drug injection. Rates of decline were significantly less, however, among consistent syringe-exchange users than sporadic or nonusers of syringe exchanges. Categorical analysis also showed significant differences among groups, with 30% of consistent syringe-exchange program users having accelerating rates of drug injection compared to 9% of nonusers and 17% of sporadic users. That consistent syringe-exchange users included a larger proportion of individuals whose drug habits were accelerating than did sporadic users or nonusers of syringe exchanges suggests a need for improved identification and counseling of such subjects by syringe-exchange program staff. The present statistical approaches may be of value in targeting such efforts. The ability of a syringe-exchange program to attract a disproportionate share of drug users with accelerating rates of drug injection underscores the importance of these programs to HIV prevention efforts.

Adult↗
PubMed2000

Flow cytometric analysis of DNA content in parotid tumor and its contiguous acini.

To investigate the relationship between proliferative capacity of salivary gland cells in contiguous acini of parotid tumors and recurrent neoplasma, DNA contents of 30 fresh specimens of parotid were studied by using cytometry in tumors, normal and shallow or deep lobe acini of the masses. The results showed that the DI was 1.369, S% 16.95, PI 26.18 in malignant tumors; DI was 1.171, S% 12.41, PI 15.54 in recurrent pleomorphic adenoma; DI was 1.141, S% 12. 74, PI 13.07 in pleomorphic adenoma, DI was 0.999, S% 5.10, PI 8.00 in normal acini. Analysis of variance showed there was a significant difference (P < 0.01). The average DNA contents of shallow on deep lobe of contiguous tumors was 1.08 in DI, 10.65 in S%, 13.49 in PI in malignant tumor, 1.06 in DI, 8.96 in S% and 9.85 in PI in pleomorphic adenoma, which were all higher than in normal acini (P > 0.05). It was concluded that the levels of DI and S% of parotid tumor and its contiguous acini are related to degree of malignancy or recurrent condition of the tumors, suggesting contiguous acini of parotid tumors had the strong capacity of proliferation, which might play an important role in recurrent or malignant change of the parotid tumors.

Adenoma, Pleomorphic↗
PubMed2000

Endotoxin exacerbates immunologically induced liver injury in cooperation with interferon-gamma.

OBJECTIVE AND DESIGN: To investigate the role of endotoxin in the development of immunologically induced liver injury. MATERIALS AND METHODS: A new model of liver injury induced in BALB/c mice by delayed-type hypersensitivity to picryl chloride and its in vitro assay for the interaction between liver nonparenchymal and parenchymal cells were used. RESULTS: Plasma endotoxin in the injured liver correlated well with serum alanine transaminase (ALT) activity (r = 0.601). Tolerance to lipopolysaccharide led to a significant inhibition of serum ALT elevation. However, lipopolysaccharide in vitro increased the ALT release from hepatocytes caused by nonparenchymal cells only in the presence of IFN-gamma. When nonparenchymal cells were separated into Kupffer and non-Kupffer cell populations, the synergistic hepatotoxicity of lipopolysaccharide and IFN-gamma was still observed in the former but not in the latter cell type. Lipopolysaccharide with IFN-gamma also enhanced TNF-alpha levels. Anti-TNF-alpha almost completely inhibited the ALT release. Combined use of TNF-alpha and IFN-gamma caused marked hepatocyte damage, while these cytokines alone did not. CONCLUSIONS: We suggest that elevated endotoxin levels accompanying the development of liver injury may activate Kupffer cells to release TNF-alpha leading to exacerbation of hepatocyte damage in cooperation with IFN-gamma produced during liver injury.

Alanine Transaminase↗
PubMed2000

Characterization of chenodeoxycholic acid as an endogenous antagonist of the G-coupled formyl peptide receptors.

OBJECTIVE AND DESIGN: To demonstrate the role of bile acids in immune modulation we examined the ability of select bile acids to inhibit leukocyte migration and chemoattractant receptor function. MATERIALS: To elucidate this mechanism, we employed primary human monocytes, neutrophils and cell lines transfected to express either the high affinity fMLP receptor (FPR) or the low affinity fMLP receptor like 1 (FPRL1). TREATMENT: Cells were treated with chenodeoxycholic acid (CDCA) and related bile acids in a 0-400 micromolar range. METHOD: Cell viability, chemotaxis and calcium flux analysis were preformed. RESULTS: We observed that pathophysiological levels (< or = 150 micromolar) of CDCA competitively inhibited 3H-fMLP binding to human monocytes, FPR and FPRL1 transfected cells. Additionally, CDCA reduced both the chemotactic and calcium flux responses induced by fMLP or "W" peptide. Further, CDCA inhibited anti-FPR antibody binding to monocytes. CONCLUSIONS: CDCA selectively inhibited human leukocyte chemotaxis and calcium flux induced by fMLP, but not other chemoattractants, suggesting a mechanism for inhibition of inflammation and suppression of innate immune response.

Calcium↗
PubMed2000

Microdetermination of proteins using m-carboxychlorophosphonazo as detection probe by enhanced resonance light scattering spectroscopy.

Proteins can be determined using a common spectrofluorometer to detect the intensity of resonance light scattering (RLS). Under acidic conditions, the reaction between m-carboxychlorophosphonazo (CPA-mK) and proteins enhances the weak light scattering of CPA-mK drastically. This enhanced intensity is proportional to the concentration of proteins. The linear ranges for human serum albumin are 0.5-35.0 microg/mL, with detection limits of 0.104 microg/mL. The method yields results comparable to those of the calorimetric method using Coomassie Brilliant Blue G-250 (CBB) with relative standard deviations of 0.72-2.10% (n = 10). There is almost no interference by amino acids and most of the metal ions.

Algorithms↗
PubMed2000

Application of a modified simplex method to the multivariable optimization of a new FIA system for the determination of osmium.

A methodology based on the coupling of experimental design and a modified simplex method is proposed for the optimization of a new flow injection-kinetic system for the spectrophotometric determination of Os (IV) with m-acetylchlorophosphonazo, which has for the first time been used as chromogenic reagent in the quantitative analysis of this element. An orthogonal array design is utilized to design the experimental protocol, in which six variables are varied simultaneously, and obtain the initial simplex using 25 experiments. A modified simplex method is applied to continuously optimize the data of the orthogonal array design; the search for optimum conditions of 6 variables using the modified simplex method required only 25 experiments. The efficiency and simplicity of the coupling of the experimental design and the modified simplex method are attractive for the development of new analytical methods. The method has been applied to the determination of Os (IV) in a refined ore as well as in a secondary alloy and provided satisfactory results.

Journal Article↗
PubMed2000

Spectroelectrochemical investigation of direct electron transfer between resting horseradish peroxidase and its oxidation states promoted by DNA.

Direct electron transfer between resting horseradish peroxidase (HRP) and its oxidation states was observed at a gold mesh electrode in a spectroelectrochemical cell in the presence of DNA. The conversion between HRP and the oxidized species induced electrochemically was found to be reversible and parallel to that initiated chemically. DNA played an important role as electron carrier and promoted the electron transfer between HRP and the electrodes. Voltammetric results and CD spectra indicated an interaction between HRP and DNA. Moreover, the secondary structure of HRP was slightly disturbed upon mixing with DNA. The direct spectroelectrochemistry of HRP at a gold mesh electrode presented new information on its bioelectrochemical characteristics.

Circular Dichroism↗
PubMed2000

Effect of interleukin-3, stem cell factor and granulocyte-macrophage colony-stimulating factor on committed stem cells, long-term culture initiating cells and bone marrow stroma in a one-step long-term bone marrow culture.

Long-term bone marrow culture (LTBMC) supports both differentiation and conservation of hematopoietic progenitor cells. During the culture period, the frequency and proliferative potential of early repopulating stem cells that give rise to progenitors detectable after 5 weeks in LTBMC--so-called long-term culture-initiating cells (LTC-IC)--can be investigated. The adherent stroma cell layer was modified by interleukin-3 (IL-3) + granulocyte-macrophage colony-stimulating factor (GM-CSF)+ stem-cell factor (SCF) with an increased cellularity and higher percentage of differentiated myeloid cells and a reduced percentage of stroma cells. We have studied the effects of stimulative cytokines, such as GM-CSF, SCF, and IL-3, on proliferation of committed colony-forming unit cells (CFU-C), LTC-IC, and stroma cell formation in LTBMC of unseparated bone-marrow cells. IL-3, GM-CSF, and SCF significantly stimulated the cumulative proliferation of nucleated cells and committed CFU-C. Weekly stimulation of LTBMC during the culture period did not exhaust the proliferative capacity of stem cells as seen in maintenance of LTC-IC after 5 weeks in LTBMC. In contrast, no LTC-IC were seen in limiting dilution analyses from LTBMC cultured 5 weeks without cytokine supplementation. Our data indicate that an increased proliferation of committed stem cells can be achieved by the addition of stimulatory growth factors, and maintenance of LTC-IC is possible over a period of 5 weeks. A net expansion of LTC-IC if compared with the starting bone-marrow suspension could not be obtained after a 5-week LTBMC period.

Bone Marrow↗
PubMed2000

Comparative genomic sequence analysis of the Williams syndrome region (LIMK1-RFC2) of human chromosome 7q11.23.

Williams syndrome (WS) is a complex neurodevelopmental disorder arising from a microdeletion at Chr band 7q11.23, which results in a hemizygous condition for a number of genes. Within this region we have completely characterized 200 kb containing the genes LIMK1, WBSCR1, and RFC2. Evidence was also found for WBSCR5 in this region, but not the previously proposed genes WSCR2 and WSCR6. The syntenic region in mouse was also sequenced (115 kb) and characterized, and a comparative sequence analysis with a percent identity plot (PIP) easily allowed us to identify coding exons. This genomic region is GC rich (50.1% human, 49.9% mouse) and contains an unusually high abundance of repetitive elements consisting primarily of Alu (45.4%, one of the highest levels identified to date) in human, and the B family of SINES (30.6% of the total sequence) in mouse. WBSCR1 corresponds to eukaryotic initiation factor 4H, identified in rabbit, and is herein found to be constitutively expressed in both human and mouse, with two RNA and protein products formed (exon 5 is alternatively spliced). The transcription pattern of WBSCR5 was also examined and discussed along with its putative amino acid sequence.

Amino Acid Sequence↗
PubMed2000

On the conservation of function of the Drosophila fat facets deubiquitinating enzyme and Fam, its mouse homolog.

Fat facets is a Drosophila deubiquitinating enzyme required for eye development and early embryogenesis. Genetic evidence suggests that Fat facets deubiquitinates and thereby prevents the proteasomal degradation of specific substrates. The Drosophila Liquid facets protein is implicated as the critical substrate of Fat facets in the eye. A mouse homolog of Fat facets, called Fam, has been identified. The results of biochemical experiments implicate two different proteins, Af-6 and beta-catenin, as substrates for Fam. Here, the functional relationship between Fat facets and Fam is explored. We show that Fam can substitute for Fat facets in all of its essential functions in Drosophila. In addition, we tested the hypothesis that Canoe and Armadillo, the Drosophila homologs of Af-6 and beta-catenin, respectively, are important substrates for Fat facets in the Drosophila eye. We found no genetic evidence to support a role for either Canoe or Armadillo in the essential Fat facets pathways in Drosophila eye development. The significance of these results is discussed in light of the biochemical experiments that suggest that Af-6 and beta-catenin are substrates of Fam.

Animals↗
PubMed2000

Determining the growing season of land vegetation on the basis of plant phenology and satellite data in Northern China.

The objectives of this study are to explore the relationships between plant phenology and satellite-sensor-derived measures of greenness, and to advance a new procedure for determining the growing season of land vegetation at the regional scale. Three phenological stations were selected as sample sites to represent different climatic zones and vegetation types in northern China. The mixed data set consists of occurrence dates of all observed phenophases for 50-70 kinds of trees and shrubs from 1983 to 1988. Using these data, we calculated the cumulative frequency of phenophases in every 5-day period (pentad) throughout each year, and also drew the cumulative frequency distribution curve for all station-years, in order to reveal the typical seasonal characteristics of these plant communities. The growing season was set as the time interval between 5% and 95% of the phenological cumulative frequency. Average lengths of the growing season varied between 188 days in the northern, to 259 days in the southern part of the research region. The beginning and end dates of the surface growing season were then applied each year as time thresholds, to determine the corresponding 10-day peak greenness values from normalized difference vegetation index curves for 8-km2 pixels overlying the phenological stations. Our results show that, at the beginning of the growing season, the largest average greenness value occurs in the southern part, then in the northern, and finally the middle part of the research region. In contrast, at the end of the growing season, the largest average greenness value is measured in the northern part, next in the middle and lastly the southern part of the research region. In future studies, these derived NDVI thresholds can be applied to determine the growing season of similar plant communities at other sites, which lack surface phenological data.

China↗
PubMed2000

Genomic organization of Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus.

The genome organization of the Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus (HearNPV) was investigated and compared to that of other baculoviruses. A detailed physical map was constructed for the restriction enzymes BamHI, BglII, EcoRI, HindIII, KpnI, PstI, SacI and XhoI. The 130.1-kilobase pairs (kb) long viral genome was cloned as restriction fragments into a plasmid library, which covered 97.5% of the viral genome. From this plasmid library about 45 kb of dispersed sequence information was generated. Fifty-three putative open reading frames (ORFs) with homology to ORFs of other baculoviruses were identified and their location on the genome of HearNPV was determined. The arrangement of the ORFs along the HearNPV genome is significantly different from the multiple-nucleocapsid NPVs, AcMNPV, BmNPV and OpMNPV, which have a highly collinear genome, and the granulovirus XcGV. In contrast the genomes of HearNPV and SeMNPV are highly collinear. This close relatedness between an MNPV and an SNPV is supported by the phylogeny of specific genes of these two viruses and suggests that the morphotype (S or M) is only a taxonomic and not a phylogenetic denominator. The data presented here also give further support to the view that the gene distribution along baculovirus genomes can be used as a character to describe baculovirus relatedness independent of gene phylogeny. Five homologous regions (hrs) were located on the genome of HearNPV. This is also the first report of the existence of hrs in SNPVs and this observation suggests that hrs are a common feature of baculovirus genomes.

Animals↗
PubMed2000

Spectroscopic characterization of Co(II)-, Ni(II)-, and Cd(II)-substituted wild-type and non-native retroviral-type zinc finger peptides.

The nucleocapsid protein (NCP) from Mason-Pfizer monkey virus (MPMV) contains two evolutionary invariant Cys-X2-Cys-X4-His-X4-Cys retroviral-type zinc finger structures, where the Cys and His residues provide ligands to a tetrahedrally coordinated Zn(II) ion. The N-terminal zinc finger (F1) of NCP from MPMV contains an immediately contiguous Cys in the -1 position relative to the start of this conserved motif: Cys-Cys-X2-Cys-X4-His-X4-Cys. Metal complexes of 18-amino acid peptides which model the native zinc finger sequence, SER-Cys-X2-Cys-X4-His-X4-Cys (F1-SC), and non-native Cys-SER-X2-Cys-X4-His-X4-Cys (F1-CS) and SER-SER-X2-Cys-X4-His-X4-Cys (F1-SS) sequences have been spectroscopically characterized and compared to the native two-zinc-finger protein fragment, MPMV NCP 21-80. All Co(II)-substituted peptide complexes adopt tetrahedral ligand geometries and have S- -->Co(II) ligand-to-metal charge-transfer (LMCT) transition intensities consistent with three Co(II)-S bonds for F1-SC and F1-CS. The non-native F1-CS peptide binds Co(II) with KCo= 1.5 x 10(6) M(-1), comparable to that of the native complex, and approximately 100-fold tighter than F1-SS. Like the Co(II) derivative, the absorption spectrum of Ni(II)-substituted NCP 21-80 is most consistent with tetrahedral Ni(II) complexes with multiple thiolate donors. In contrast, Ni(II) complexes of F1-SC and F1-CS exhibit a single absorption band in the 400-550 nm region (epsilon approximately 200-300 M(-1) cm (-1), distinct in the two complexes, assignable to a degenerate d-d transition envelope characteristic of non-native square-planar coordination geometry, and an intense LMCT transition in the UV (epsilon255 approximately 14,000 M(-1) cm(-1)). Cd(II) complexes have intense absorption in the UV (lambda(max)=233nm), with absolute intensities consistent with approximately 5000 M(-1) cm(-1) per Cd(II)-S bond. 113Cd NMR spectroscopy of 113Cd MPMV NCP gives delta=649 ppm, consistent with S3N coordination. Co(II) and Cd(II) complexes of non-native F1-CS peptides are more sensitive to oxidation by O2, relative to F1-SC, suggestive of a higher lability in the non-native chelate. The implications of these findings for the evolutionary conservation of this motif are discussed.

Amino Acid Sequence↗
PubMed2000

Branch retinal artery occlusion after thyroid artery interventional embolization.

PURPOSE: To report a case of branch retinal artery occlusion after thyroid artery interventional embolization. METHODS: A 33-year-old man with hyperthyroidism complained of visual loss and scotoma in the left eye after thyroid artery interventional embolization. He underwent a full ophthalmologic examination, including fluorescein angiography. RESULTS: Visual acuity was 20/25, with inferior and superior scotomas present in the left eye. Fluorescein angiography of the left eye revealed delayed filling of a superotemporal branch retinal artery and nonfilling of an inferotemporal branch retinal artery. CONCLUSION: A small, but definite risk of retinal artery occlusion after thyroid artery interventional embolization should be considered.

Adult↗
PubMed2000

Torque-speed relationship of the flagellar rotary motor of Escherichia coli.

The output of a rotary motor is characterized by its torque and speed. We measured the torque-speed relationship of the flagellar rotary motor of Escherichia coli by a new method. Small latex spheres were attached to flagellar stubs on cells fixed to the surface of a glass slide. The angular speeds of the spheres were monitored in a weak optical trap by back-focal-plane interferometry in solutions containing different concentrations of the viscous agent Ficoll. Plots of relative torque (viscosity x speed) versus speed were obtained over a wide dynamic range (up to speeds of approximately 300 Hz) at three different temperatures, 22.7, 17.7, and 15.8 degrees C. Results obtained earlier by electrorotation (, Biophys. J. 65:2201-2216) were confirmed. The motor operates in two dynamic regimes. At 23 degrees C, the torque is approximately constant up to a knee speed of nearly 200 Hz, and then it falls rapidly with speed to a zero-torque speed of approximately 350 Hz. In the low-speed regime, torque is insensitive to changes in temperature. In the high-speed regime, it decreases markedly at lower temperature. These results are consistent with models in which torque is generated by a powerstroke mechanism (, Biophys. J. 76:580-587).

Biomechanical Phenomena↗
PubMed2000

Solvent-isotope and pH effects on flagellar rotation in Escherichia coli.

We studied changes in speed of the flagellar rotary motor of Escherichia coli when tethered cells or cells carrying small latex spheres on flagellar stubs were shifted from H(2)O to D(2)O or subjected to changes in external pH. In the high-torque, low-speed regime, solvent isotope effects were found to be small; in the low-torque, high-speed regime, they were large. The boundaries between these regimes were close to those found earlier in measurements of the torque-speed relationship of the flagellar rotary motor (, Biophys. J. 65:2201-2216;, Biophys. J., 78:1036-1041). This observation provides direct evidence that the decline in torque at high speed is due primarily to limits in rates of proton transfer. However, variations of speed (and torque) with shifts of external pH (from 4.7 to 8.8) were small for both regimes. Therefore, rates of proton transfer are not very dependent on external pH.

Animals↗