Search PubMed⌕ Search

Biomedical subjects

W Yan

Publications and source records attributed to W Yan.

At least 91 records · Page 5Linked to original sources
PubMed1999

Velocardiofacial syndrome in childhood-onset schizophrenia.

OBJECTIVES: Deletion of chromosome 22q11 (velocardiofacial syndrome) is associated with early neurodevelopmental abnormalities and with schizophrenia in adults. The rate of 22q11 deletions was examined in a series of patients with childhood-onset schizophrenia (COS), in whom early premorbid developmental and cognitive impairments are more pronounced than in adult-onset cases. METHOD: Through extensive recruiting and screening, a cohort of 47 patients was enrolled in a comprehensive study of very-early-onset schizophrenia. All were tested with fluorescence in situ hybridization for deletions on chromosome 22q11. RESULTS: Three (6.4%) of 47 patients were found to have a 22q11 deletion. All 3 COS patients with 22q11 deletions had premorbid impairments of language, motor, and social development, although their physical characteristics varied. Brain magnetic resonance imaging revealed increased midbody corpus callosum area and ventricular volume in relation both to healthy controls and to other COS patients. CONCLUSIONS: The rate of 22q11 deletions in COS is higher than in the general population (0.025%, p < .001) and may be higher than reported for adult-onset schizophrenia (2.0%, p = .09). These results suggest that 22q11 deletions may be associated with an earlier age of onset of schizophrenia, possibly mediated by a more salient neurodevelopmental disruption.

Abnormalities, Multiple↗
PubMed1999

The glycine-phenylalanine-rich region determines the specificity of the yeast Hsp40 Sis1.

Hsp40s are ubiquitous, conserved proteins which function with molecular chaperones of the Hsp70 class. Sis1 is an essential Hsp40 of the cytosol of Saccharomyces cerevisiae, thought to be required for initiation of translation. We carried out a genetic analysis to determine the regions of Sis1 required to perform its key function(s). A C-terminal truncation of Sis1, removing 231 amino acids but retaining the N-terminal 121 amino acids encompassing the J domain and the glycine-phenylalanine-rich (G-F) region, was able to rescue the inviability of a Deltasis1 strain. The yeast cytosol contains other Hsp40s, including Ydj1. To determine which regions carried the critical determinants of Sis1 function, we constructed chimeric genes containing portions of SIS1 and YDJ1. A chimera containing the J domain of Sis1 and the G-F region of Ydj1 could not rescue the lethality of the Deltasis1 strain. However, a chimera with the J domain of Ydj1 and the G/F region of Sis1 could rescue the strain's lethality, indicating that the G-F region is a unique region required for the essential function of Sis1. However, a J domain is also required, as mutants expected to cause a disruption of the interaction of the J domain with Hsp70 are inviable. We conclude that the G-F region, previously thought only to be a linker or spacer region between the J domain and C-terminal regions of Hsp40s, is a critical determinant of Sis1 function.

Amino Acid Sequence↗
PubMed1999

Possible novel mechanism for bitter taste mediated through cGMP.

Taste is the least understood among sensory systems, and bitter taste mechanisms pose a special challenge because they are elicited by a large variety of compounds. We studied bitter taste signal transduction with the quench-flow method and monitored the rapid kinetics of the second messenger guanosine 3',5'-cyclic monophosphate (cGMP) production and degradation in mouse taste tissue. In response to the bitter stimulants, caffeine and theophylline but not strychnine or denatonium cGMP levels demonstrated a rapid and transient increase that peaked at 50 ms and gradually declined throughout the following 4.5 s. The theophylline- and caffeine-induced effect was rapid, transient, concentration dependent and gustatory tissue-specific. The effect could be partially suppressed in the presence of the soluble guanylyl cyclase (GC) inhibitor 10 microM ODQ and 30 microM methylene blue but not 50 microM LY 83583 and boosted by nitric oxide donors 25 microM NOR-3 or 100 microM sodium nitroprusside. The proposed mechanism for this novel cGMP-mediated bitter taste signal transduction is cGMP production partially by the soluble GC and caffeine-induced inhibition of one or several phosphodiesterases.

Aminoquinolines↗
PubMed1999

The pattern of inhibin/activin alpha- and betaB-subunit messenger ribonucleic acid expression in rat testis after selective Leydig cell destruction by ethylene dimethane sulfonate.

To further investigate the regulatory mechanisms responsible for the control of testicular inhibin/activin subunit gene expression, inhibin-alpha, -betaA, and -betaB messenger RNA (mRNA) levels were assessed after ethylene dimethane sulfonate (EDS)-induced destruction of Leydig cells (LC) in different animal models: the intact rat, the rat treated with high doses of testosterone, and the unilaterally cryptorchid rat. In intact rats, EDS selectively eliminates the mature adult-type LCs, activating the proliferation and differentiation of preexisting LC precursors into a new population of functionally active LCs. In this model, a single dose of EDS (75 mg/kg BW, ip) induced a significant increase in testicular inhibin-alpha and -betaB mRNA levels 5 days after treatment (5.0- and 5.5-fold increases, respectively), whereas inhibin-betaA mRNA remained undetectable upon Northern hybridization in control and EDS-treated testes. Moreover, in situ hybridization analysis demonstrated that the increased expression of inhibin-alpha and -betaB mRNAs observed 5 days after EDS takes place mainly in Sertoli cells. Along with LC repopulation, the expression level of inhibin-alpha and -betaB messages declined, and inhibin-alpha mRNA returned to control values on day 40 after EDS. This treatment, however, failed to alter the pattern of testicular expression of FSH receptor and androgen-binding protein mRNAs, thus suggesting selectivity for the above effects. In EDS-treated rats supplemented with high doses of testosterone, the preexisting mature LCs are destroyed, but, due to elevated testosterone concentrations, disruption of spermatogenesis is attenuated, and the post-EDS rise in serum gonadotropins is blocked; the latter prevents LC regeneration. In this model, a 5.0-fold increase in inhibin-alpha mRNA levels, similar to that found in intact animals, was detected 5 days after EDS administration, but the rise in inhibin-betaB levels was partially delayed. In addition, the blockade of LC repopulation resulted in permanent elevation of inhibin-alpha and -betaB messages throughout the study period. In unilaterally cryptorchid rats, the abdominal testis shows disrupted spermatogenesis and altered paracrine environment that expedites LC repopulation after EDS treatment. In this model, the abdominal testes showed a significant 2.5-fold increase in inhibin-alpha mRNA levels 5 days after EDS, but no effect was found in those of inhibin-betaB. Further, the faster rate of LC repopulation resulted in precocious decline of inhibin-alpha mRNA levels. Finally, the expression of inhibin/activin subunit mRNAs was monitored during postnatal testicular development, specifically at the time of regression of fetal-type LCs and appearance of those of the adult type. High levels of expression of inhibin-alpha and -betaB mRNAs were detected in neonatal and infantile testes. A sharp decline in both messages took place between days 15-20, i.e. at the time when fetal-type Leydig cells are replaced by adult-type cells. From this time point onward, inhibin-alpha and -betaB mRNA levels remained low, ranging between 15-30% of the maximum. In conclusion, our results suggest that the adult-type LCs differentially modulate the expression of inhibin/activin subunit genes and point to a major inhibitory role in this cell type on expression of the inhibin-alpha gene.

Activins↗
PubMed1999

Retinol-binding protein is produced by rabbit chondrocytes and responds to parathyroid hormone (PTH)/PTH-related peptide-cyclic adenosine monophosphate pathway.

PTH and dibutyryl cAMP [(Bu)2cAMP] induced the expression of a 19-kDa protein in the conditioned media of rabbit growth plate chondrocyte cultures. The 19-kDa protein was identified as plasma retinol-binding protein (RBP) by aminoterminal sequence analysis and immunoblot analysis with an anti-RBP monoclonal antibody. Northern blot analysis showed that PTH, PTH-related peptide (PTHrP), and (Bu)2cAMP increased the RBP messenger RNA (mRNA) level in chondrocyte cultures. Further, both PTH and (Bu)2cAMP markedly induced the expression of RBP mRNA by about 10-fold at 3 h and by about 40-fold at 24 h, indicating a pretranslational regulation. The level of the mRNA expression induced by PTH, PTHrP, and (Bu)2cAMP was as high as that by retinoic acid (RA), known as a potent inducer of RBP in hepatoma cells. RBP mRNA was also detected in cartilage tissues at higher levels than in the other tissues examined except liver. Both RBP and PTH/PTHrP inhibited the dedifferentiative activity of RA on growth plate chondrocytes when added to the culture medium. These results demonstrate that chondrocytes synthesize and secrete RBP in vivo and in vitro and suggest that PTH/PTHrP modulates the effect of RA by means of RBP production in chondrocytes.

Amino Acid Sequence↗
PubMed1999

Function of stem cell factor as a survival factor of spermatogonia and localization of messenger ribonucleic acid in the rat seminiferous epithelium.

To address the possibility that stem cell factor (SCF) is a paracrine regulator of germ cell development in the adult rat testis, stage-specific distribution of SCF messenger RNA (mRNA) was investigated with Northern blot and in situ hybridization analyses. The highest levels of SCF mRNA were found in stages II-VI of the rat seminiferous epithelial cycle, whereas the lowest levels were in stages VII-VIII. Intermediate levels of SCF mRNA were detected in stages IX-XIV-I of the cycle. The expression of the SCF gene was found to be developmentally regulated, and the expression pattern followed the process of Sertoli cell proliferation and differentiation during postnatal life. The effect of mouse recombinant SCF on spermatogonial DNA synthesis was studied using an in vitro tissue culture system for stage-defined seminiferous tubules. A significant increase in DNA synthesis in spermatogonia could be detected when tubule segments from stage XII were cultured in the presence of 100 ng/ml SCF for 48 h (P < 0.05) and 72 h (P < 0.01). This observation was further confirmed with autoradiographic analyses; almost a 100-fold increase in thymidine incorporation in the SCF-treated (100 ng/ml) tubule segments was observed compared with that in untreated samples. The results of the present study suggest that SCF is a Sertoli cell-produced paracrine regulator and acts as a survival factor for spermatogonia in the adult rat seminiferous epithelium in a stage-specific manner.

Animals↗
PubMed1999

Stage-specific regulation of stem cell factor gene expression in the rat seminiferous epithelium.

To assess the regulation of stem factor factor (SCF) gene expression during spermatogenesis, we tested the effects of hormones (FSH, testosterone, and 17beta-estradiol) and some growth factors [transforming growth factor-beta (TGF beta), TGF alpha, tumor necrosis factor-alpha, and activin] on SCF gene expression by using a transillumination-assisted microdisection technique, a seminiferous tubule culture system, and Northern hybridization. Our results showed that FSH (10 ng/ml) increased steady state levels of SCF messenger RNA (mRNA) in a stage-specific and time-dependent manner. 8-Bromo-cAMP could increase the SCF mRNA level in a similar way as FSH, whereas phorbol 12-myristate 13-acetate had no effect. Actinomycin D could abolish the stimulatory effect of FSH, whereas cyclohexamide could not. The half-life of SCF mRNA was apparently prolonged after FSH stimulation (FSH-treated tubules, 15.6 +/- 1.2 h; controls, 8.6 +/- 2.7 h). Nuclear run-on assay revealed 5- and 10-fold increases in the transcription rate after FSH stimulation for 8 and 30 h, respectively. Neither testosterone nor estradiol had significant effects on SCF gene expression in our tissue culture system. Activin, TGF beta, TGF alpha, and tumor necrosis factor-alpha had no effect on SCF gene expression in vitro. In conclusion, SCF gene expression in the rat seminiferous tubule is regulated by FSH through the cAMP/protein kinase A pathway. FSH regulates SCF gene expression at both transcriptional and posttranscriptional levels involving the increase in transcription rate and prolongation of half-life of SCF mRNA, but is independent of de novo protein synthesis.

Animals↗
PubMed1999

Expression of peroxisome proliferator-activated receptor alpha messenger ribonucleic acid and protein in human and rat testis.

Peroxisome proliferator-activated receptor a (PPARalpha), a member of the steroid hormone receptor superfamily, has been linked to lipid homeostasis and tumorigenesis in tissues with high expression of receptor protein. On the other hand, the role of PPARalpha in tissues with a lower expression is not well known. Here we demonstrate the localization of PPARalpha messenger RNA (mRNA) and protein in developing and adult rat testis. Additionally, we demonstrate the expression of PPARalpha protein in adult human testis. Our experiments with Northern analysis, in situ hybridization and immunocytochemistry reveal a complex distribution of PPARalpha in tubular and interstitial cells of both adult and developing rat testis. The overall expression is rather low but may be modified by exogenous or endogenous stimuli. An up-regulation of PPARalpha mRNA could be observed after stimulation with FSH. In the developing rat testis, a clear expression of PPARalpha mRNA was present from the first days after birth. Additionally, PPARalpha mRNA and protein increased toward adulthood. In adult human testis PPARalpha immunoreactivity (IR) was present in interstitial Leydig cells and tubular cells. In the seminiferous epithelium of adult human testis the expression of PPARalpha-IR could be seen in meiotic spermatocytes, spermatids and myoid peritubular cells. The findings of our study suggest that PPARalpha may be involved in the regulation of growth and differentiation of tubular and interstitial cells in rat and human testis.

Adult↗
PubMed1999

Killing effect of interleukin-2 and interferon-alpha activated leukemic bone marrow in remission on K562 leukemic cells.

OBJECTIVE: To evaluate the killing effect of interleukin-2 (IL-2) and interferon-alpha (IFN-alpha) activated bone marrow cells on K562 cells. METHODS: Semi-solid colony and 3H-TdR incorporating method were used. RESULTS: Bone marrow from leukemia patients in remission was activated in vitro with IL-2 for 3 days. The activated bone marrow (ABM) displayed killing effects of 0.31-2.30 logs on K562 cells. This killing effect was further increased to 0.30-3.15 logs when IFN-alpha added with IL-2 to the marrow for activation. IL-2 alone or in combination with IFN-alpha showed no inhibition of CFU-GM and K562 cells. Compared with IL-2 or IFN-alpha alone, the combination of the two cytokines could more effectively maintain the killing effect of ABM on leukemic cells. CONCLUSIONS: IFN-alpha can augment the purging effect of IL-2 ABM and combination of the two cytokines can effectively maintain the cytotoxicity of ABM.

Acute Disease↗
PubMed1999

[An analysis of nutritional and harmful components of vegetables grown in plastic greenhouses].

OBJECTIVE: To study the changes in nutritional and harmful components of vegetables grown in plastic greenhouses. METHODS: In plastic greenhouses, microclimate and air concentrations of carbon monoxide, carbon dioxide, fluoride and respirable particulate were measured, and chlorophyll, total sugar, crude fiber, nitrite, fluoride, arsenic and some mineral elements in vegetables were determined as compared with those grown in the open-air fields. RESULTS: Greenhouse appeared a lower wind speed and darker illumination. Contents of chlorophyll a an b, total chlorophyll, reduced vitamin C, crude fiber in vegetables grown in greenhouse all were lower than those grown in open-air fields. Contents of potassium, calcium, magnesium, iron, zinc, copper and phosphorous were all lower in the vegetables grown in greenhouse than those grown in open-air fields. The contents of chlorophyll reducing Vitamin C. CONCLUSION: Lower wind speed and inadequate illumination in greenhouse affected photosynthesis and uptake of water in vegetables causing changes in their nutritional components. But, no contamination of burning coal was found in vegetables grown in greenhouse.

Air Pollutants↗
PubMed1998

A method to assess invasion and intracellular replication of Trypanosoma cruzi based on differential uracil incorporation.

Screening of candidate trypanocidal compounds or factors affecting invasion of mammalian cells by the infective stages of Trypanosoma cruzi in tissue culture models has primarily involved labor-intensive microscopic counting of the parasites. A very efficient method for quantitating the inhibitory effect of antimicrobial agents or signaling pathways inhibitors on T. cruzi grown in L6E9 myoblasts was devised. This assay takes advantage of the selective incorporation of [3H]uracil into nucleic acids by replicating T. cruzi amastigotes. L6E9 rat myoblasts are submitted to gamma irradiation to inhibit their replication. Uracil uptake by uninfected cells is considerably decreased by this method. Nifurtimox, benznidazole, fexinidazole, MK-436, and megazol are drugs known to have activity against T. cruzi and were used in growth inhibition assays. The results demonstrated that [3H]uracil incorporation in the presence of different concentrations of nifurtimox and benznidazole closely correlated with the number of amastigotes per 100 myoblasts in Giemsa-stained monolayers under the conditions used. This method also has the advantage to differentiate between the effects of the compounds on the invasion and the replication steps of the infection with T. cruzi, as shown by the inhibitory effect of genistein when added in invasion assays.

Animals↗
PubMed1998

[A study of correlation between essential hypertension and HLA-DQA1 alleles].

OBJECTIVE: To analyse the hereditary susceptibility by genotyping of HLA-DQA1 alleles in essential hypertensives. METHODS: The allelic types of HLA-DQA1 were detected by PCR-SSP technic in 52 cases of essential hypertensives and 86 normal individuals as the control. RESULTS: The frequency of HLA-DQA1*0302 allele in hypertensive group was markedly higher than that in normal control group (17.9572 vs 3.5531), but the frequency of HLA-DQA1*0103 allele in normal group was higher than that in hypertensive group. CONCLUSION: HLA-DQA1*0302 may be a correlative gene of essential hypertension, whereas HLA-DQA1*0103 may be a defence gene of hypertension.

Adult↗
PubMed1998

Expression of membrane-bound transferrin-like protein p97 on the cell surface of chondrocytes.

A concanavalin-A-binding protein of 76 kDa was purified from the plasma membrane fraction of rabbit chondrocyte cultures. Amino acid sequencing of the N-terminal region and of tryptic peptides of the protein, in addition to sequencing of its cDNA revealed that this protein is highly similar to the tumour-associated antigen p97. Hence, it was concluded that this protein is the rabbit form of p97. Western blotting, Northern blotting and reverse-transcription PCR analyses indicated that rabbit p97 is expressed at high levels in cartilage and chondrocytes, but is barely detectable in the bone, liver, kidney, small intestine, eye, pancreas, heart, testis, skeletal muscle, spleen and fibroblasts. Immunocytochemical and immunohistochemical analyses demonstrated that p97 is expressed in the plasma membrane of chondrocytes. p97 transcript was detected in all zones of the cartilage but the level was relatively low in the hypertrophic zone. These findings suggest that p97 is involved in maintaining the cell surface characteristics of chondrocytes.

Amino Acid Sequence↗
PubMed1998

Zuotin, a ribosome-associated DnaJ molecular chaperone.

Correct folding of newly synthesized polypeptides is thought to be facilitated by Hsp70 molecular chaperones in conjunction with DnaJ cohort proteins. In Saccharomyces cerevisiae, SSB proteins are ribosome-associated Hsp70s which interact with the newly synthesized nascent polypeptide chain. Here we report that the phenotypes of an S.cerevisiae strain lacking the DnaJ-related protein Zuotin (Zuo1) are very similar to those of a strain lacking Ssb, including sensitivities to low temperatures, certain protein synthesis inhibitors and high osmolarity. Zuo1, which has been shown previously to be a nucleic acid-binding protein, is also a ribosome-associated protein localized predominantly in the cytosol. Analysis of zuo1 deletion and truncation mutants revealed a positive correlation between the ribosome association of Zuo1 and its ability to bind RNA. We propose that Zuo1 binds to ribosomes, in part, by interaction with ribosomal RNA and that Zuo1 functions with Ssb as a chaperone on the ribosome.

Cytosol↗
PubMed1998

Functional characterization of transforming growth factor beta type II receptor mutants in human cancer.

We recently identified missense mutations at amino acid residues 526 and 537 located within the highly conserved subdomain XI of the transforming growth factor beta type II receptor (TbetaR-II) serine-threonine kinase in two human squamous carcinoma cell lines. These cell lines are resistant to transforming growth factor beta-mediated inhibition of growth. Moreover, treatment with transforming growth factor beta fails to increase the levels of type 1 plasminogen activator inhibitor and fibronectin synthesis. To test the effects of the mutations on receptor function, mutant TbetaR-II cDNAs were expressed in TbetaR-II-deficient T47D cells. Cyclin A promoter activity was reduced by 50% in cells expressing wild-type TbetaR-II but increased 2-fold in cells transfected with either of the two mutant receptors. Conversely, plasminogen activator inhibitor type 1 promoter activity was increased 6-fold in cells transfected with wild-type receptor but not with either of the two mutant receptors. Moreover, the activity of both mutant serine-threonine kinases was strongly reduced compared to that of the wild-type receptor. Thus, the amino acid residues at positions 526 and 537 seem to be essential for kinase function and signaling activity of the TbetaR-II.

Carcinoma, Squamous Cell↗
PubMed1998

Role of chondroitin sulfate-hyaluronan interactions in the viscoelastic properties of extracellular matrices and fluids.

The purpose of this study was to investigate the role of chondroitin sulfate-hyaluronan interactions in the viscoelastic properties of tissues and fluids, using capillary and cone-on-plate viscometers. Chondroitin sulfate markedly increased the viscosity of hyaluronan solutions at a wide range of hyaluronan mass (50-1900 kDa) under physiological conditions of pH, temperature, ionic strength and glycosaminoglycan concentration (0.5-40 mg/ml), although the viscosity of the chondroitin sulfate solutions themselves was very low. In the assay using a cone-on-plate viscometer, chondroitin sulfate increased the viscosity of hyaluronan solutions at various shear rates. At low shear rates, the viscosity of a chondroitin sulfate (5 mg/ml)-hyaluronan (0.5 mg/ml) mixture was about 40% of that of an aggrecan (5 mg/ml)-hyaluronan (0.5 mg/ml) mixture, and at 2.8-fold higher concentrations, chondroitin sulfate elicited the same effect on the viscosity of hyaluronan solutions (5 mg/ml) as an aggrecan monomer. In the presence of oscillatory motion, the addition of aggrecan increased the elasticity (storage) modulus G' and the viscosity (loss) modulus G" of hyaluronan solutions and markedly decreased the loss tangent G"/G' at frequencies corresponding to normal joint movements. In contrast, chondroitin sulfate had only a marginal effect on the loss tangent G"/G', although it increased G' and G". These findings demonstrated that chondroitin sulfate, as well as aggrecan, increases the viscosity of hyaluronan solutions, although chondroitin sulfate has less effect on the elasticity of hyaluronan solutions than that of aggrecan, and suggest that chondroitin sulfate may play an important physiological role in determining the viscoelastic properties of extracellular matrices and fluids.

Aggrecans↗