Biomedical subjects

K Yang

Publications and source records attributed to K Yang.

At least 37 records · Page 2Linked to original sources

PubMed2002

The expression of 11 beta-hydroxysteroid dehydrogenase type 2 is induced during trophoblast differentiation: effects of hypoxia.

The intracellular enzyme 11 beta-hydroxysteroid dehydrogenase type 2 (11 beta-HSD2) catalyzes the unidirectional conversion of bioactive glucocorticoids to their inert metabolites. In the human placenta, 11 beta-HSD2 is highly expressed in syncytiotrophoblasts, although cytotrophoblasts also express this enzyme at lower levels. Given that cytotrophoblasts will differentiate into syncytiotrophoblasts in vivo and in vitro, the present study was designed to examine the hypothesis that the expression of 11 beta-HSD2 is induced during in vitro trophoblast differentiation. When Percoll-purified human cytotrophoblast cells were cultured under standard (20% oxygen) conditions, they aggregated and fused to form syncytiotrophoblasts. Within the first 24 h during differentiation, levels of 11 beta-HSD2 protein and activity were increased by 2- to 3-fold, but they did not increase further thereafter. However, when the cells were exposed to hypoxic (1% oxygen) conditions, both the induction of 11 beta-HSD2 and trophoblast differentiation were prevented. Taken together, these results demonstrate for the first time that the expression of 11 beta-HSD2 is induced early during trophoblast differentiation, and hypoxia prevents this induction, indicating that placental 11 beta-HSD2 expression is subjected to regulation by the local oxygen environment. If placental villi respond to hypoxia in a similar fashion in vivo, the present findings would suggest that hypoxia might be a factor contributing to the previously reported decreases in placental 11 beta-HSD2 in pregnancies complicated by intrauterine growth restriction and preeclampsia.

11-beta-Hydroxysteroid Dehydrogenase Type 2↗

PubMed2001

Complementary signaling pathways regulate the unfolded protein response and are required for C. elegans development.

The unfolded protein response (UPR) is a transcriptional and translational intracellular signaling pathway activated by the accumulation of unfolded proteins in the lumen of the endoplasmic reticulum (ER). We have used C. elegans as a genetic model system to dissect UPR signaling in a multicellular organism. C. elegans requires ire-1-mediated splicing of xbp-1 mRNA for UPR gene transcription and survival upon ER stress. In addition, ire-1/xbp-1 acts with pek-1, a protein kinase that mediates translation attenuation, in complementary pathways that are essential for worm development and survival. We propose that UPR transcriptional activation by ire-1 as well as translational attenuation by pek-1 maintain ER homeostasis. The results demonstrate that the UPR and ER homeostasis are essential for metazoan development.

Animals↗

PubMed2001

Maximization of acetic acid production in partial acidogenesis of swine wastewater.

Swine wastewater was biologically treated to produce short-chain volatile organic acids (VOAs) in laboratory-scale continuously stirred tank reactors. The maximum production rates of acetic and butyric acids associated with simultaneous changes in pH and hydraulic retention time (HRT) were investigated, in which the degree of acidification of swine wastewater to the short-chain VOAs was <25% of influent chemical oxygen demand (COD) concentration. A constant inoculum system was used to minimize the experimental error due to the use of inconsistent inoculum. The inoculum system was operated with synthetic wastewater at 6000 mg soluble chemical oxygen demand per liter (pH 6.0) and 35 degrees C at 0.5 day hydraulic retention time. Response surface methodology was applied successfully to determine the optimum physiological condition for which the maximum rate of acetic acid production occurred, which was pH 5.90 and 0.88 day hydraulic retention time at 35 degrees C. The partial acidification process to manage swine waste should be operated in the optimum condition for acetic acid production because the optimum operating condition for butyric acid production approached the washout point.

Acetic Acid↗

PubMed2001

A repressor-response regulator gene pair controlling jadomycin B production in Streptomyces venezuelae ISP5230.

A second regulatory gene (jadR(1)) is located immediately upstream of the putative repressor gene (jadR(2)) in the jad cluster for biosynthesis of the antibiotic jadomycin B in Streptomyces venezuelae ISP5230. It encodes a 234-amino acid polypeptide with a sequence resembling those of response regulator proteins in two-component control systems. Features in the conserved C-terminal domain of JadR(1) place the protein in the OmpR-PhoB subfamily of response regulators. In mutants where jadR(1) was deleted or disrupted, jadomycin B was not produced, implying that the gene has an essential role in biosynthesis of the antibiotic. Cloning jadR(1) from S. venezuelae in pJV73A, and introducing additional copies of the gene into the wild-type parent by plasmid transformation gave unstable strains with pJV73A integrated into the chromosome. The transformants initially showed increased production of jadomycin B but gave lower titers as excess copies of jadR(1) were lost; mature cultures stabilized with a wild-type level of antibiotic production. The mutant from which jadR(1) had been deleted could not be transformed with pJV73A. Altering the composition of jadR genes in the chromosome by integration of vectors carrying intact and disrupted copies of jadR(1) and jadR(2) provided evidence that the two genes form a regulatory pair different in function from previously reported two-component systems controlling antibiotic biosynthesis in streptomycetes.

Alleles↗

PubMed2001

The distinct spectra of tumor-associated Apc mutations in mismatch repair-deficient Apc1638N mice define the roles of MSH3 and MSH6 in DNA repair and intestinal tumorigenesis.

In mammalian cells, mismatch recognition has been attributed to two partially redundant heterodimeric protein complexes of MutS homologues, MSH2-MSH3 and MSH2-MSH6. We have conducted a comparative analysis of Msh3 and Msh6 deficiency in mouse intestinal tumorigenesis by generating Apc1638N mice deficient in Msh3, Msh6 or both. We have found that Apc1638N mice defective in Msh6 show reduced survival and a 6-7-fold increase in intestinal tumor multiplicity. In contrast, Msh3-deficient Apc1638N mice showed no difference in survival and intestinal tumor multiplicity as compared with Apc1638N mice. However, when Msh3 deficiency is combined with Msh6 deficiency (Msh3(-/-)Msh6(-/-)Apc1638N), the survival rate of the mice was further reduced compared to Msh6(-/-)Apc(1638N) mice because of a high multiplicity of intestinal tumors at a younger age. Almost 90% of the intestinal tumors from both Msh6(-/-)Apc1638N and Msh3(-/-)Msh6(-/-)Apc1638N mice contained truncation mutations in the wild-type Apc allele. Apc mutations in Msh6(-/-)Apc1638N mice consisted predominantly of base substitutions (93%) creating stop codons, consistent with a major role for Msh6 in the repair of base-base mismatches. However, in Msh3(-/-)Msh6(-/-)Apc1638N tumors, we observed a mixture of base substitutions (46%) and frameshifts (54%), indicating that in Msh6(-/-)Apc1638N mice frameshift mutations in the Apc gene were suppressed by Msh3. Interestingly, all except one of the Apc mutations detected in mismatch repair-deficient intestinal tumors were located upstream of the third 20-amino acid beta-catenin binding repeat and before all of the Ser-Ala-Met-Pro repeats, suggesting that there is selection for loss of multiple domains involved in beta-catenin regulation. Our analysis therefore has revealed distinct mutational spectra and clarified the roles of Msh3 and Msh6 in DNA repair and intestinal tumorigenesis.

Animals↗

PubMed2001

Splenic and peritoneal B-1 cells differ in terms of transcriptional and proliferative features that separate peritoneal B-1 from splenic B-2 cells.

B-1 cells constitute a distinct B cell subset with characteristic phenotypic and functional features. B-1 cells are highly represented among peritoneal lymphocytes; substantial numbers of B-1 cells are also located within splenic tissue. Here a number of differences in transcription factor and gene expression were identified that separate peritoneal B-1 and splenic B-2 cells, and then splenic B-1 cells obtained from immunoglobulin transgenic mice were tested for these parameters. Splenic B-1 cells resembled splenic B-2 cells rather than peritoneal B-1 cells in terms of nuclear expression of DNA-binding STAT3, CREB, and PU.1, with respect to transcriptional activation of IL-10, and in the failure to enter cell cycle in response to PMA. Splenic B-1 cells (B-1S) appear to constitute a unique population of B-1 cells, which, while sharing with peritoneal B-1 cells (B-1P) certain phenotypic features, differ from them in transcription factor and gene expression and in signaling for cell cycle progression.

Animals↗

PubMed2001

Strain selection of Metarrhizium anisopliae by image analysis of colony morphology for consistency of steroid biotransformation.

Quantitative description of colony morphological images of the fungus Metarrhizium anisopliae was done by means of computer image analysis based on fractal analysis. Compared with simple fractal analysis, the differences in the morphological features of colonies were expressed more accurately by multifractal analysis. The use of multifractal analysis markedly improved the reliability of colony image analysis, which could be extended to morphological characterization for colony recognition and classification. First, the multifractal spectrum was calculated from each morphological image and 14 features were defined on the spectrum to describe the spectral line profile. Second, three features, alpha-right, alpha-width, and f-start, were selected from the 14 features in two-dimensional feature vector spaces and a feature augmented vector (alpha-right, alpha-width, and f-start, 1)(T) was generated and used for classifier design. Third, a statistical least mean square error (LMSE) algorithm was applied to design a piecewise-linear classifier, which consists of two linear classifiers, d(1)(x) and d(2)(x). Representative colony samples were used as training and test sample sets, which were previously classified into a high-activity class and another class by a training person. After passing the training phase, the piecewise-linear classifier could be used for automated classification of unknown colony samples. A correct recognition rate of 96% was achieved in this manner. Compared with conventional strain selection methods, this new method has the advantage of less subjectivity, reduced labor and time, and improved ability to deal with a large number of colonies.

Algorithms↗

PubMed2001

VHL gene alterations in renal cell carcinoma patients: novel hotspot or founder mutations and linkage disequilibrium.

Mutations in the von Hippel-Lindau (VHL) gene are frequently detected in human sporadic renal cell carcinoma (RCC). We analysed 102 Swedish RCCs for VHL mutations by PCR-SSCP and sequencing. In 47 patients (46.1%), 70 different mutations were found, and most of them represented novel variations of the VHL gene. Mutations in the VHL gene were found in 54% of clear cell renal cell carcinomas (CCRCC) and in 18% of chromophilic cancers but in no chromophobe cancers or oncocytomas (P=0.016). Three novel hotspot or founder mutations were detected in our study: four CCRCCs carried a missense mutation (glutamic acid to lysine) at codon 160 which is critical in the stabilization of the H1 helix of the alpha domain and the alpha-beta domain interface in the VHL protein. Five CCRCCs and one chromophilic RCC harbored a 15-nucleotide in-frame deletion (codons 41-45) at a duplex tandem repeat sequence site. Moreover, this deletion was in linkage disequilibrium with a C-->T transition in the promoter region. The frequency of linkage was 17 times more common than chance. Five patients with this linked mutation resided in the same hospital district and at least three of them showed the two sequence variants in the tumor-adjacent tissue. In 5/6 patients the wild-type allele was lost in the tumor samples, suggesting a causal role for the mutations in RCC. These linked mutations might be novel polymorphisms maintained in a relative isolated population. Multiple mutations in VHL were found in 17 tumors out of 47 tumors with the VHL mutation. A higher multiple mutation detected rate (33%) was observed in grade 3 CCRCCs than those in grade 1 (22%) and grade 2 (9%) (P=0.04). This is evidence on the association between VHL mutation and extent of nuclear atypia.

Age of Onset↗

PubMed2001

Dipolar excitons, spontaneous phase coherence, and superfluid-insulator transition in bilayer quantum Hall systems at nu = 1.

The spontaneous interlayer phase coherent (111) state of a bilayer quantum Hall system at filling factor nu = 1 may be viewed as a condensate of interlayer particle-hole pairs or excitons. We show that when the layers are biased in such a way that these excitons are very dilute, they may be viewed as pointlike bosons. We calculate the exciton dispersion relation and show that the exciton-exciton interaction is dominated by the dipole moment they carry. In addition to the phase coherent state, we also find a Wigner crystal/glass phase in the presence/absence of disorder which is an insulating state for the excitons. The position of the phase boundary is estimated and the transition between these two phases is discussed.

Journal Article↗

PubMed2001

Distribution and depression of the GABA(B) receptor in the spinal dorsal horn of adult rat.

gamma-Aminobutyric acid (GABA) is a principal inhibitory neurotransmitter in vertebrate nervous system. The metabotropic receptor for GABA, GABA(B) receptor, is characterized as a G protein-coupled receptor subtype. In the present study, GABA(B) receptor-like immunoreactivity (GABA(B)R-LI) in the rat spinal cord and dorsal root ganglion (DRG), as well as GABA(B) receptor-mediated depression in the spinal dorsal horn were examined by using immunohistochemistry and whole-cell voltage-clamp recording technique, respectively. Under light microscope, GABA(B)R-LI was densely found in laminae I and II of the dorsal horn. DRG cells of various diameters also showed GABA(B)R-LI. Electron microscopy further revealed that GABA(B)R-LI was also localized in terminals of myelinated, unmyelinated fibers as well as the somatodendritic sites of dorsal horn neurons. Bath application of a GABA(B) receptor agonist, baclofen (10 microM, 30 s), induced a slow outward (inhibitory) current in dorsal horn neurons. This slow current was depressed when the postsynaptic G protein-coupled receptor was inhibited, indicating the postsynaptic action of baclofen. Under the condition of postsynaptic GABA(B) receptor being inhibited, baclofen (10 microM, 60 s) depressed large (Abeta) and fine (C, Adelta) afferent fiber-evoked monosynaptic excitatory postsynaptic currents, indicating presynaptic inhibition of GABA(B) receptor on elicited neurotransmitter release. Taken together, the results suggest that baclofen-sensitive GABA(B) receptor is expressed pre- and postsynaptically on primary afferent fibers and neurons in the spinal dorsal horn; activation of GABA(B) receptor in the dorsal horn postsynaptically hyperpolarizes dorsal horn neurons and presynaptically inhibits primary afferents.

Age Factors↗

PubMed2001

Nocistatin, a peptide reversing acute and chronic morphine tolerance.

It has been reported that intracerebroventricular (i.c.v.) injection of nociception/orphanin FQ (OFQ) can antagonize morphine analgesia, whereas i.c.v. OFQ antibody can reverse morphine tolerance. Nocistatin (NST) is a recently characterized neuropeptide possessing an antagonizing effect on OFQ. Here we examine whether i.c.v. NST would result in a reversal of morphine tolerance. The results showed that: (1) i.c.v. NST at doses of 0.005, 0.05, 0.5, 5 or 50 ng per rat produced a bell-shaped dose-dependent reversal of chronic morphine tolerance, with maximum response at 0.5 ng. (2) Acute morphine tolerance could also be reversed, albeit partially, by i.c.v. NST at 0.5 ng. (3) The reversal of acute and chronic morphine tolerance by NST was completely abolished when NST (0.5 ng) was co-injected with (8 microg) OFQ. Since OFQ and NST are derived from the same preprohormone, the profile of its splicing in the CNS may play an important role in determining the effectiveness of morphine analgesia.

Analgesics, Opioid↗

PubMed2001

Calcium inhibits human placental 11beta-hydroxysteroid dehydrogenase type 2 activity.

The effect of Ca2+ on the conversion of cortisol to its inert metabolite cortisone, the reaction catalyzed by the microsomal enzyme 11beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2), was investigated in human placental microsomes. Placental microsomal 11beta-HSD2 activity, as determined by the rate of conversion of cortisol to cortisone, was inhibited up to 50% by increasing free Ca2+ concentrations from 22 to 268 nM. The Ca2+-induced inhibition was reversible since chelation of endogenous Ca2+ with EGTA increased 11beta-HSD2 activity up to 200%. Ca2+ decreased the maximal velocity (Vmax) of the 11beta-HSD2 catalyzed conversion of cortisol to cortisone without altering the Km of 11beta-HSD2 for cortisol, indicating that Ca2+ modulates the catalytic efficiency rather than the substrate binding of 11beta-HSD2. Moreover, the Ca2+-induced inhibition does not appear to involve altered cofactor (NAD+) binding since the inhibition of microsomal 11beta-HSD2 activity by a sub-maximal concentration of free Ca2+ was not overcome by increasing the concentration of NAD+. These findings in the microsomes were then extended to an intact cell system, JEG-3 cells, an established model for human placental trophoblasts. In these cells, an increase in cytosolic free Ca2+ concentration ([Ca2+]i) elicited by a known physiological stimulus, PGF(2alpha), was accompanied by a 40% decrease in the level of 11beta-HSD2 activity. Furthermore, the PGF(2alpha)-induced inhibition of 11beta-HSD2 activity was abrogated when increases in [Ca2+]i were blocked with the intracellular Ca2+ chelator, BAPTA. Collectively, these results demonstrate for the first time that Ca2+ inhibits human placental 11beta-HSD2 activity by a post-translational mechanism not involving substrate or cofactor binding.

11-beta-Hydroxysteroid Dehydrogenase Type 2↗

PubMed2001

Baclofen inhibition of dorsal root-evoked inhibitory postsynaptic currents in substantia gelatinosa neurons of rat spinal cord slice.

We investigated the action of baclofen, an agonist for GABA(B) receptor, on the dorsal root-evoked inhibitory postsynaptic currents (IPSCs) in substantia gelatinosa (SG) neurons in the adult rat spinal cord slice. Most of the dorsal root-evoked IPSCs in SG were mediated by Adelta fibers. Baclofen (10 microM) reduced the amplitude of Adelta fiber-evoked glycine receptor-mediated IPSCs to 20.9+/-3.0% (n=13), and GABA(A) receptor-mediated IPSCs to 18.6+/-3.4% (n=12) of the controls, respectively. The results further suggest modulatory role of GABA(B) receptor in spinal dorsal horn.

Animals↗

PubMed2001

A convenient method for the efficient removal of ruthenium byproducts generated during olefin metathesis reactions.

[reaction in text] An efficient method for removing ruthenium byproducts generated during olefin metathesis reactions with Grubbs catalysts is described. Treatment of the crude reaction products with triphenylphosphine oxide or dimethyl sulfoxide, followed by filtration through silica gel, was found to be a practical and effective method to remove colored ruthenium byproducts.

Alkenes↗

PubMed2001

Cloning and characterization of spliced fusion transcript variants of synovial sarcoma: SYT/SSX4, SYT/SSX4v, and SYT/SSX2v. Possible regulatory role of the fusion gene product in wild type SYT expression.

The synovial sarcoma translocation t(X;18)(p11.2; q11.2) results in the fusion of the SYT gene on chromosome 18 to exon 5 of either SSX1 or SSX2 genes on chromosome X. We recently reported that the SSX4 gene is also involved in such a translocation. In the present investigation we cloned and sequenced the full-length cDNA of SYT/SSX1, SYT/SSX2 and SYT/SSX4 from synovial sarcoma tissues. We isolated a novel fusion transcript type variant involving the fusion of SYT with exon 6 of the SSX4 gene (SYT/SSX4v). The SYT/SSX4 and SYT/SSX2 open reading frame also differed from previously reported SYT/SSX sequences by an in-frame addition of 93bp exon located in the junction between exon 7 and 8 of the SYT. This exon is identical to that reported for the murine SYT but has not been previously found in the human transcript. Two SYT transcripts, with and without the 93 bp exon, were co-expressed in mouse NIH3T3 cells, human malignant cells and human testis tissue, but not in human normal fibroblasts. Stable transfection of an SYT/SSX4 expression vector into human and murine cell lines correlated with a down-regulation of SYT transcripts. This was also observed in a synovial sarcoma tumor expressing SYT/SSX4. This suggests that the SYT/SSX fusion gene may regulate SYT expression from the normal allele and as such alter the normal function of SYT.

Amino Acid Sequence↗

PubMed2001

Immune responses and protection obtained with rotavirus VP6 DNA vaccines given by intramuscular injection.

Intramuscular (i.m.) injection of murine VP6 DNA vaccines raised high titers of rotavirus-specific serum IgG and IgA antibodies in BALB/c mice. A Th1-like antibody response was generated based on the ratio of serum IgG2a to IgG1 antibodies. Rotavirus-specific serum IgA but not fecal IgA was detected in mice prior to rotavirus challenge. Partial protection against rotavirus challenge was achieved as measured by reduction of rotavirus antigen shedding in feces. A similar level of protection was found with a bovine rotavirus VP6 DNA vaccine against a murine rotavirus challenge, suggesting that heterologous protection can be obtained by immunizing with VP6 DNA vaccines. We did not directly test for cytotoxic T lymphocyte (CTL) activity, but in vivo depletion of CD8+ T cells in mice immunized with a murine VP6 DNA vaccine did not significantly change the duration of virus shedding or the pattern of protection obtained. This finding suggested that CD8+ CTL activity was not essential for the partial protection we obtained by i.m. immunization of mice with VP6 DNA vaccines.

Animals↗

PubMed2001

The metabolic evidence of synergistic interaction between DAMGO and DPDPE on undifferentiated SH-SY5Y cells.

Recent studies have demonstrated the analgesic synergy between mu- and delta-opioid receptor, but evidence obtained at the cellular level is scanty. This work was designed to find the evidence of synergy between the actions of D-Ala2-Mephe4-glyol5 enkephalin (DAMGO) and D-Phe2, D-Phe5 enkephalin (DPDPE) on undifferentiated SH-SY5Y cells. Microphysiometer was used to detect the functional activity of cells by measuring the real-time extracellular acidification rate (ECAR). The results demonstrate the unequivocal synergy between DAMGO and DPDPE at least within certain ratios. In addition, combined administration of the two drugs in the synergistic ratios attenuates receptor desensitization. These data indicate that DAMGO and DPDPE have a synergistic effect at cellular level.

Analgesics, Opioid↗

PubMed2001

The metabolic evidence of synergistic effect between ohmefentanyl and [D-Pen2, D-Pen5] enkephalin on differentiated SH-SY5Y cells in humans.

Interactions between selective opioid agonists acting at mu- and delta-opioid receptors were evaluated by co-administering a low-effective dose of the selective mu-opioid receptor agonist ohmefentanyl (OMF) with sequentially increasing doses of the selective delta-opioid receptor agonist [D-Pen(2), D-Pen(5)] enkephalin (DPDPE). Microphysiometer was used to measure the extracellular acidification rate (ECAR) of living cells in real-time, which reflected the functional activity after agonist-receptor binding. The synergy (i.e. a more than additive effect) was observed with combinations of these two opioid agonists on differentiated SH-SY5Y cells functionally expressing both mu- and delta-opioid receptors. The demonstration of the synergy suggests that the agonists of the subtypes of opioid receptors can interact at cellular level.

Acids↗