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Biomedical subjects

A Chen

Publications and source records attributed to A Chen.

At least 289 records · Page 16Linked to original sources
PubMed1995

Activation of monocyte effector genes and STAT family transcription factors by inflammatory synovial fluid is independent of interferon gamma.

Activated monocytes play an important role in the pathogenesis of inflammatory arthritis. Blood monocytes which enter the inflamed joint become activated upon adherence to extracellular matrix and exposure to a complex inflammatory environment. We have analyzed the mechanism of monocyte activation by soluble factors present in inflammatory synovial fluid (SF). Greater than 75% of inflammatory SFs tested (a total of 22 fluids to date) increased cell surface expression and dramatically increased mRNA levels of monocyte activation markers Fc gamma RI, Fc gamma RIII, and HLA-DRA. This induction was not triggered by adherence, a known activating stimulus, and several lines of evidence showed that induction was not dependent upon interferon gamma (IFN-gamma). Induction was not prevented by neutralizing anti-IFN-gamma antibodies and IFN-gamma was not detected in the SFs using a sensitive enzyme-linked immunosorbent assay. The SFs also were not able to activate the IFN-gamma-activated transcription factor Stat1, thus providing further support for the absence of IFN-gamma. SFs did activate a related signal transducer and activator of transcription (STAT) family factor, termed Stat-SF, which bound specifically to the IFN-gamma response region (GRR), a well-characterized transcription element in the Fc gamma RI promoter. Based upon DNA-binding specificity and mobilities in gel shift assays, and reactivity with specific antisera, Stat-SF likely contains Stat3, or a closely related STAT family member. Neutralization of interleukin 6, a cytokine present in SFs which is known to activate Stat3, abolished the activation of Stat-SF and inhibited the induction of Fc gamma RI expression by SFs. These results demonstrate the activation of monocytes by inflammatory SF and suggest that monocyte activation at an inflammatory site may occur in the absence of IFN-gamma through the triggering of signal transduction pathways that activate STAT transcription factors.

Arthritis, Rheumatoid↗
PubMed1995

Clinical and molecular characterization of a rare syndrome of acute promyelocytic leukemia associated with translocation (11;17).

Analysis of a variant translocation t(11;17) in a case of acute promyelocytic leukemia (APL) led to discovery of a novel zinc finger gene, PLZF, fused to the retinoic acid receptor-alpha (RAR alpha) gene. We reviewed the clinical and molecular features of five additional patients with t(11;17)-associated APL. The clinical course of three patients was characterized by early death and three experienced disseminated intravascular coagulation. Morphologically all of the patients fell in a unusual morphologic spectrum of APL, with features intermediate between M2 and M3 AML. All six patients had PLZF-RAR alpha gene fusion as detected by reverse transcription/polymerase chain reaction assay, Southern blotting, or pulsed-field gel electrophoresis. Five of the six patients failed to achieve complete remission after initial chemotherapy or differentiation therapy with all-trans retinoic acid (ATRA). A sixth patient responded to initial chemotherapy, but on relapse failed to respond to ATRA. When tested in vitro, cultured cells from three of the patients failed to differentiate in response to ATRA. APL associated with t(11;17) and fusion of the PLZF and RAR alpha genes is a discrete clinico-pathologic syndrome with a distinctly worse prognosis than t(15;17) APL.

Adult↗
PubMed1995

Microdialysis sampling coupled on-line to fast microbore liquid chromatography.

A system which combines intravenous microdialysis sampling on-line with fast microbore liquid chromatographic analysis is described. Using a 14 mm x 1 mm I.D. ODS column, caffeine, theobromine and paraxanthine could be resolved in under 1 min and acetaminophen and its two main metabolites, the glucuronide and sulfate conjugates, could be resolved in 30 s. This fast separation allowed on-line microdialysis sampling to be conducted with a 1-min sampling interval. The system was evaluated by monitoring the pharmacokinetics following intravenous administration of acetaminophen and caffeine.

Acetaminophen↗
PubMed1995

Loss of heterozygosity at chromosome 1q loci in rat mammary tumors.

To better characterize abnormalities affecting rat chromosome 1 during mammary carcinogenesis, tumors were induced by nitrosomethylurea in F1 hybrid rats polymorphic at multiple chromosome 1 loci. By means of restriction fragment length polymorphism and microsatellite length polymorphism analyses, we observed loss of heterozygosity or allelic imbalance affecting various loci on the q arm of chromosome 1 in a high percentage of the 49 tumors analyzed. Fifty percent of the tumors showed loss or imbalance affecting the most distal (1q55) INS1 (rat insulin 1 gene) locus. The MT1PA (metallothionein-1 pseudogene a) locus was observed to be affected in 58% of tumors induced in BUF/NCr x ACI/Vsp rats. Most of the losses appeared to have occurred by mitotic recombination. No parental bias was observed on the affected chromosome 1. Tumors were also screened for mutations in codon 12 of the Ha-ras-1 gene, which is located on 1q. We observed an association between the presence of mutation and allelic imbalance. These studies confirm our previous cytogenetic observations of a high level of nonrandom instability affecting rat chromosome 1 during mammary carcinogenesis. The observed loss of heterozygosity may indicate the existence of a putative tumor suppressor gene within the distal half of the 1q arm. These abnormalities, however, could also be related to the early stages of Ha-ras amplification.

Adenocarcinoma↗
PubMed1995

In vitro effects of interleukins on human mesangial cells: implications for glomerulonephritis.

Intrinsic glomerular cells, especially mesangial cells, are considered to be actively involved in the pathogenesis of glomerulonephritis (GN), but the precise mechanism(s) remains elusive. We have previously demonstrated that nephritogenic IgA immune complex can stimulate human mesangial cells (HMCs) to increase their production of interleukin-1 (IL-1) and interleukin-6 (IL-6). In order to evaluate the roles of cytokines such as IL-1 and/or IL-6 and mesangial cells as mediators of renal injury in GN, we have now examined the changes of HMCs and their secreted products in vitro, after stimulation with various concentrations of IL-1 and IL-6. Cytokine-activated HMCs showed the following changes: (1) increased cell size, with intracytoplasmic vacuoles, dilated endoplasmic reticulum, increased free ribosomes and polysomes, and mitochondrial swelling; (2) increased cell proliferation, reflected in thymidine incorporation and an increased proportion of S and G2/M phase cells by cell cycle analysis; (3) enhancement of IL-6 mRNA expression in HMCs with stimulation of IL-6 alone or IL-1 plus IL-6; and (4) release of large amounts of platelet activating factor (PAF), thromboxane B2 (TxB2), and superoxide anion. Taken together, these results strongly suggest that mesangial cell proliferation and increased production of immune/chemical mediators and superoxide anion can be directly induced by IL-1 plus IL-6. These changes may lead to ongoing renal injury.

Adolescent↗
PubMed1995

Assessment of p53 expression in nasopharyngeal carcinoma.

We analyzed the expression of the p53 protein by immunohistochemical methods from 101 patients with nasopharyngeal carcinoma (NPC): 24 with NPC and dysplastic lesions adjacent to carcinoma and 14 with primary and metastatic specimens. Ninety-six of 101 lesions (95%) had detectable p53 protein in the nuclei of tumor cells, indicating that overexpression of the p53 protein might be closely associated with NPC. Among 24 patients who had NPC and dysplastic lesions adjacent to carcinoma, 19 of the dysplastic lesions (79.2%) and 22 of the carcinomas (91.7%) showed positive staining for the p53 protein. In dysplastic epithelia p53 antigenicity was generally in a basal location. The significant association of p53 expression in NPC and dysplastic lesions adjacent to carcinoma (P < .0001, Fisher's exact probability test) suggests that p53 overexpression seems to occur at an early stage in the development of NPC. p53 expression in NPC does not correlate with histological grading, degree of lymphocytic infiltration between tumor cells, clinical stage, sex, or age (P > .05, chi-squared test). A comparison of p53 expression between primary and metastatic NPC was performed in 14 lesions. Although the p53 protein was consistently expressed in primary and metastatic tumor cells, there was no significant difference in p53 expression in both distinct but related lesions (P > .05, paired t-test). Our results suggest that the association of overexpression of the p53 protein in NPC may not be indicative of a mutant type p53 protein.

Adult↗
PubMed1995

A simple technique for fabrication of transmission block for concomitant treatment of bilateral anterior inguinal lymphatics and pelvic/perineal structures.

The efficacy of transmission block technique with contiguous photon irradiation of inguinal as well as pelvic and perineal structures has been reported in literature. In this paper, a simple and accurate method for the proper fabrication of transmission block has been described and also demonstrated to be effective and useful. The procedures for a precise geometrical alignment and dose verification are also detailed.

Anus Neoplasms↗
PubMed1995

Quantitation of reduced disulfide groups in monoclonal antibodies using 5-iodoacetamidofluorescein: a novel size exclusion-HPLC technique.

The interference of stannous in the quantitation of sulfhydryl in pretreated MAb using the Ellman's method is described. A HPLC method which effectively overcomes the interference by stannous ion in measuring sulfhydryls by the commonly used Ellman's technique has been developed. The method involves reacting the reduced monoclonal antibody with 0.1 M EDTA followed by the sulfhydryl specific reagent 5-iodoacetamidofluorescein and incubating the mixture at 37 degrees C for 1.0-1.5 h. The 5-IAF labeled protein is separated from the unbound free 5-IAF and its secondary degraded products by size exclusion HPLC using a TSK G3000 SWXL column. The method allows quantitation of MAb sulfhydryl present as monomeric MAb, aggregates and fragments and is a major improvement over the spectroscopic techniques.

Antibodies, Monoclonal↗
PubMed1995

Teachers' value orientations in urban and rural school settings.

The purpose of this research was to describe physical educators' value orientations for curricular decision making in urban and rural teaching settings. The revised Value Orientation Inventory (Ennis & Chen, 1993) was used to collect data from 495 physical educators in urban and rural districts. Data were analyzed descriptively using a full-design and a nested MANOVA model. Results indicated that teachers in urban school districts placed a higher priority on self-actualization and social responsibility than did teachers in rural school districts. Conversely, teachers in rural school districts placed a higher priority on disciplinary mastery and learning process than did their colleagues in urban schools. The discussion focused on differences in school contexts in urban and rural schools. Teachers appeared to shape their curriculum to reflect the opportunities and constraints within their school settings.

Adult↗
PubMed1995

Consanguineous nuclear families used to identify a new locus for recessive non-syndromic hearing loss on 14q.

Hearing impairment is inherited most frequently as an autosomal recessive isolated clinical finding (non-syndromic hearing loss, NSHL). Extreme heterogeneity and phenotypic variability in the audiometric profile preclude pooling of affected families and severely hamper gene mapping by conventional linkage analysis. However, in instances of consanguinity, homozygosity mapping can be used to identify disease loci in small nuclear families. This report demonstrates the power of this technique by identifying a locus for recessive NSHL on 14q (DFNB4).

Chromosome Mapping↗
PubMed1995

Induction of proteinuria by adriamycin or bovine serum albumin in the mouse.

To establish models of proteinuria in the mouse, BALB/c mice were injected with puromycin aminonucleoside (PAN, 1.5 or 4.5 mg/10 g body weight), adriamycin (AD, 0.2 mg/10 g body weight) intravenously or bovine serum albumin (BSA, 100 mg/10 g body weight) intraperitoneally. Proteinuria was measured as the ratio of urinary albumin (micrograms/ml) to creatinine (mg/dl) and further characterized by isotyping the immunoglobulin. Although not obtained with PAN (followed for 4 weeks), proteinuria was readily attained in the mouse after treatment with AD or BSA. Most AD-treated mice (5/7) developed an abrupt increase of proteinuria at day 2 after injection, with the ratio of urinary albumin to creatinine in the range 0.28-0.45. The degree of proteinuria increased with time and all mice tested (7/7) showed overt proteinuria at day 4. These mice became anuric at day 5 and died at days 6 and 7. For BSA, 4 h after administration, four of seven mice showed enhanced proteinuria, lasting 8 h with urinary albumin and creatinine in a ratio < 0.05. Isotyping of urine samples collected at the time of heavy proteinuria (ratio of urinary albumin to creatinine: > 0.40 for AD-treated mice, > 0.15 for BSA-treated mice) showed that all of the mice (7/7) with AD-induced proteinuria (ADp) and three of four mice with BSA-induced proteinuria (BSAp) revealed urinary IgG2b and IgA, while only one of seven control mice showed IgG2b alone in urine. The mice were sacrificed at the time they presented with heavy proteinura for pathologic and anionic studies on renal tissues.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

A point mutation in the integrin beta 6 subunit abolishes both alpha v beta 6 binding to fibronectin and receptor localization to focal contacts.

The integrin alpha v beta 6 was initially identified from primary cultures of airway epithelial cells. This integrin is expressed in bronchiolar and alveolar epithelium during development and in settings of injury and/or inflammation and mediates attachment of epithelial cells to fibronectin and tenascin. Like other integrins, this receptor localizes to structures called focal contacts in cells plated on appropriate ligands. In the present study, we produced a mutant beta 6 cDNA (beta 6m) containing a single substitution of Asp140 with Ala and transfected mutant (or wild-type) beta 6 cDNA into the human colon carcinoma cell line SW480. In parallel, we used cDNAs truncated just proximal to the transmembrane domain to generate secreted forms of mutant alpha v beta 6 in Chinese hamster ovary (CHO) cells. The mutant beta 6, like the wild type, formed heterodimers with human alpha v that were expressed on the cell surface of SW480 cells and secreted by CHO cells. Secreted alpha v beta 6 containing this point mutation did not bind to fibronectin-Sepharose. Furthermore, in contrast to wild-type beta 6, the mutant form did not allow SW480 cells to bind to fibronectin in the presence of beta 1-blocking antibody and did not localize to focal contacts. Our results confirm that the Asp140 of beta 6, like the corresponding residues in beta 1 (Asp130) and beta 3 (Asp119), is critical for interactions of alpha v beta 6 with ligand, and also suggest that ligand binding to alpha v beta 6 is necessary for localization of this receptor to focal contacts.

Alanine↗
PubMed1995

Investigation of the role of beta 1 integrins in cell-cell adhesion.

Various beta 1 integrins (VLA-2, VLA-3, VLA-4) have been suggested to bind directly to themselves or to each other, thus mediating cell-cell adhesion. Here we expressed the human alpha 2 and alpha 3 subunits in three different cell lines (human erythroleukemia K562, human rhabdomyosarcoma RD and Chinese hamster ovary CHO cells). Although cell surface alpha 2 beta 1 and alpha 3 beta 1 in the transfectants mediated adhesion to matrix ligands (collagen or laminin 5, respectively), in no case did we observe enhanced cell-cell adhesion. In the presence of a range of different divalent cation concentrations, stimulatory anti-beta 1 antibodies or anti-alpha 3 antibodies, VLA-2 and VLA-3 still did not appear to interact directly, through either heterophilic (i.e. VLA-3/VLA-2) or homophilic (i.e. VLA-3/VLA-3) mechanisms, to mediate cell-cell adhesion. Furthermore, in some but not all alpha 3 transfectants we observed an unexpected decrease in cell-cell adhesion, suggesting a novel anti-adhesive function. This inhibitory effect was not observed for alpha 2 transfection nor when the alpha 3 cytoplasmic tail was exchanged with that of another integrin alpha subunit. Finally, no evidence for VLA-4/VLA-4 mediated cell-cell adhesion was observed using alpha 4-transfected K562 and CHO cells. In conclusion, using many different combinations of cell lines, we found that cell-cell adhesion mediated by direct integrin/integrin interaction is not a widespread phenomenon, and is not observable in standard cell-cell adhesion assays. Furthermore, in some cell combinations, alpha 3 expression may actually cause diminished cell-cell adhesion.

Amino Acid Sequence↗
PubMed1995

The selectins: vascular adhesion molecules.

The selectin family of adhesion molecules mediates the initial attachment of leukocytes to venular endothelial cells before their firm adhesion and diapedesis at sites of tissue injury and inflammation. The selectin family consists of three closely related cell-surface molecules with differential expression by leukocytes (L-selectin), platelets (P-selectin), and vascular endothelium (E- and P-selectin). The selectins have characteristic extracellular regions composed of an amino-terminal lectin domain that binds a carbohydrate ligand, an epidermal growth factor-like domain, and two to nine short repeat units homologous to domains found in complement binding proteins. In contrast to most other adhesion molecules, selectin function is restricted to leukocyte interactions with vascular endothelium. Multiple studies indicate that the selectins mediate neutrophil, monocyte, and lymphocyte rolling along the venular wall. The generation of selectin-deficient mice has confirmed these findings and provided further insight into how the overlapping functions of these receptors regulate inflammatory processes. Selectin-directed therapeutic agents are now proven to be effective in blocking many of the pathological effects resulting from leukocyte entry into sites of inflammation. Future studies are focused on how the selectins interact with the increasing array of other adhesion molecules and inflammatory mediators.

Animals↗
PubMed1995

Coexisting IgA nephropathy and leukocytoclastic cutaneous vasculitis associated with ankylosing spondylitis: a case report.

A patient with ankylosing spondylitis and coexisting IgA nephropathy and leukocytoclastic cutaneous vasculitis is described. Renal biopsy demonstrated mesangial proliferative glomerulonephritis with prominent IgA, C3 and fibrin deposition in the glomeruli. Simultaneously, leukocytoclastic cutaneous vasculitis with prominent IgG, IgA and C3 deposition of dermal vessel wall was also observed in the skin biopsy specimen. Such associations have been previously reported in only four cases. This report once again indicates that antigenic mucosal stimulation may play an important role in the pathogenesis of ankylosing spondylitis.

Adult↗
PubMed1994

Isolation and characterization of idsA: the gene for the short chain isoprenyl diphosphate synthase from Methanobacterium thermoautotrophicum.

The gene that encodes the bifunctional short chain isoprenyl diphosphate synthase (idsA) for synthesis of farnesyl diphosphate and geranylgeranyl diphosphate in Methanobacterium thermoautotrophicum, a strict archaebacterial anaerobe, was isolated from a genomic DNA library by colony-lift hybridization and sequenced. Amino acid sequences were obtained for the N-terminus of the enzyme and for internal peptide fragments generated by proteolysis and treatment with cyanogen bromide. Degenerate primers based on the amino acid sequences were used in PCR to synthesize a 220-bp probe from genomic DNA. The probe was radiolabeled and used to isolate idsA. DNA sequencing revealed a 975-bp open reading frame located within an operon. The encoded 325-amino-acid protein contained five conserved regions found in eubacterial and eukaryotic farnesyl diphosphate and geranylgeranyl diphosphate synthases, including aspartate-rich motifs commonly found in prenyltransferases.

Alkyl and Aryl Transferases↗