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Biomedical subjects

B Yu

Publications and source records attributed to B Yu.

At least 73 records · Page 4Linked to original sources
PubMed1999

Transport of fluid by lens epithelium.

We report for the first time that cultured lens epithelial cell layers and rabbit lenses in vitro transport fluid. Layers of the alphaTN4 mouse cell line and bovine cell cultures were grown to confluence on permeable membrane inserts. Fluid movement across cultured layers and excised rabbit lenses was determined by volume clamp (37 degrees C). Cultured layers transported fluid from their basal to their apical sides against a pressure head of 3 cmH2O. Rates were (in microliter. h-1. cm-2) 3.3 +/- 0.3 for alphaTN4 cells (n = 27) and 4.7 +/- 1.0 for bovine layers (n = 6). Quinidine, a blocker of K+ channels, and p-chloromercuribenzenesulfonate and HgCl2, inhibitors of aquaporins, inhibited fluid transport. Rabbit lenses transported fluid from their anterior to their posterior sides against a 2.5-cmH2O pressure head at 10.3 +/- 0.62 microliter. h-1. lens-1 (n = 5) and along the same pressure head at 12.5 +/- 1.1 microliter. h-1. lens-1 (n = 6). We calculate that this flow could wash the lens extracellular space by convection about once every 2 h and therefore might contribute to lens homeostasis and transparency.

4-Chloromercuribenzenesulfonate↗
PubMed1999

Mobilization of GLUT-4 from intracellular vesicles by insulin and K(+) depolarization in cultured H9c2 myotubes.

The insulin-responsive glucose transporter, GLUT-4, moves from an intracellular compartment to the cell surface in response to insulin and/or muscle contraction. Treatment of H9c2 myotubes with insulin significantly increased uptake of 2-deoxyglucose. Depolarization of the myotubes by increasing extracellular [K(+)], which mimics the initial phases of excitation-contraction coupling, also increased 2-deoxyglucose uptake. The K(+)- but not insulin-evoked increase was blocked by dantrolene, an inhibitor of Ca(2+) release from the sarcoplasmic reticulum. In contrast, wortmannin, an inhibitor of phosphatidylinositol 3-kinase, blocked insulin- but not K(+)-stimulated 2-deoxyglucose uptake. Increased glucose uptake in response to insulin or K(+) depolarization was associated with increased GLUT-4 in plasma membranes and depletion of a population of small intracellular GLUT-4-containing vesicles. Similarly, in H9c2 cells transfected with c-myc-tagged GLUT-4, translocation of c-myc GLUT-4 to the cell surface was increased after stimulation with insulin or K(+) depolarization. Taken together, these data demonstrate that insulin and K(+) depolarization increase glucose uptake by recruiting GLUT-4 from intracellular vesicles to the plasma membrane of H9c2 myotubes via distinct signaling mechanisms.

Androstadienes↗
PubMed1999

Different subtypes of GABAB receptors are present at pre- and postsynaptic sites within the rat dorsolateral septal nucleus.

GABAB receptor activation modulates neuronal activity mediated by multiple CNS transmitters and can occur at pre- and postsynaptic sites. In low concentrations, baclofen acts presynaptically to diminish transmitter release via both hetero- and autoreceptors, whereas at increasing concentrations, the same compound alters postsynaptic membrane excitability by inducing a membrane hyperpolarization. We have utilized electrophysiological techniques in vitro to focus on the possibility that pharmacologically different subtypes of GABAB receptors are present on presynaptic sites of glutamatergic terminals when compared with GABAB receptors on postsynaptic sites within the dorsolateral septal nucleus (DLSN). The glutamatergic terminal within the DLSN originates from a pyramidal cell body located within the hippocampus and most likely terminates on a GABAergic neuron from which recordings were made. Whole cell patch voltage-clamp methods were employed to record pharmacologically isolated excitatory postsynaptic currents (EPSCs) from DLSN neurons as an index of glutamatergic transmission. Using a modified internal pipette solution containing QX-314 and in which CsGluconate and GDPbetaS replaced Kgluconate and GTP, respectively, we recorded isolated monosynaptic EPSCs. The GABAA receptor antagonists bicuculline and picrotoxin were included in the external standard superfusion solution. Application of the GABAB receptor agonists, (+/-)-baclofen, CGP44533, and CGP35024 (10 nM to 10 microM) depressed glutamate-mediated EPSCs in a concentration-dependent manner. With the use of this combination of solutions, CGP44533 did not produce postsynaptic membrane property changes. Under these conditions, both (+/-)-baclofen and CGP35024 still induced increases of postsynaptic membrane conductance associated with an outward current. The GABAB receptor antagonist CGP55845A (1 microM) blocked the presynaptic CGP44533-mediated depressant effects of EPSCs, whereas CGP35348 (100 microM) or barium (2 mM) was ineffective. Furthermore, both CGP35348 (100 microM) and CGP55845A (1 microM) were effective in blocking the postsynaptic conductance changes associated with baclofen and CGP35024, whereas barium was ineffective. Our results demonstrate a distinct pharmacology for GABAB agonists acting at putative subtypes of GABAB receptors located on presynaptic sites of a glutamatergic terminal versus GABAB receptors on postsynaptic sites of a DLSN neuron. Furthermore, our results also suggest a different pharmacology and/or coupling of a GABAB receptor to different effectors at postsynaptic sites within the DLSN. Thus there may be three or more pharmacologically distinct GABAB receptors or receptor complexes associated with DLSN neurons: at least one pre- and two postsynaptic. If this distinct pharmacology and GABAB receptor distribution also extends to other CNS structures, such differences could provide development of selective drugs to act at these multiple sites.

Animals↗
PubMed1999

Morphological and biochemical analysis of anti-nuclear matrix protein antibodies in human sera.

Autoimmune sera have been used in the diagnosis of autoimmune diseases as well as the analysis of nuclear substructures. In an attempt to study the biological characteristics of the nuclear matrix, we screened human sera using immunofluorescent staining and immunoblot. We detected antibodies against nuclear matrix (NM), a remnant nonchromatin protein compartment after the treatment of detergent, salt and nuclease, in 212 out of 284 tested sera (74.6%) by immunoblot. Peptides with molecular weights of 70 kDa, 50 kDa and 25 kDa were detected in the order of frequency. Clinical informations of 198 out of 212 cases were available and went as follows: 38 cases were autoimmune diseases, such as systemic lupus erythematosus and rheumatoid arthritis; 132 non-autoimmune and non-neoplastic diseases; 16 neoplastic diseases and 12 cases unclassified. The immunofluorescent staining intensity by anti-nuclear matrix protein (NMP) antibodies decreased variably, but fibrillogranular, speckled and nucleolar immunolocalization patterns were retained after in situ fractionation. Ku70 and La protein were detected by anti-NMP antibodies. Immunolocalization by anti-NMP antibodies indicates that the NMPs constitute a variety of characteristic nuclear substructures and may serve as autoantigens in diverse human diseases. In addition, the presence of Ku70 and La protein as NMPs suggests that the NM can be functionally active in association with DNA or RNA.

Antigens, Nuclear↗
PubMed1999

[Study of the relation between mutant gene in bile and hepatic metastasis of large intestinal carcinoma].

OBJECTIVE: To study the value of point mutant genes in the diagnosis of hepatic metastasis of large intestinal carcinoma. METHODS: Techniques of PCR-SSCP and DNA sequencing were used to detect point mutations of p53 and k-ras genes in primary focus, hepatic metastatic focus, and bile. RESULTS: 50 cases of large intestinal cancer were detected. 38 cases (76.0%) had p53 and k-ras gene mutations. p53 gene mutations were detected in 28 cases (56.0%) and k-ras gene mutations in 16 cases (32.0%). The same point mutant genes in bile with primary focus were detected in 10 out of 12 patients with hepatic metastasis (83.3%). CONCLUSIONS: The tumor cells of hepatic metastasis show the same clonal origin with primary focal cells that had same point mutant genes. Detection of the point mutant genes in bile is of value in the diagnosis of hepatic metastasis of large intestinal carcinoma.

Bile↗
PubMed1999

[The effect of sandostatin and 5-fluorouracil on apoptosis in breast cancer].

OBJECTIVE: To investigate the effect of 5-FU plus sandostatin on apoptosis in breast cancer. METHODS: Breast cancer samples from 16 patients were treated with 5-FU (10 microg/ml) and sandostatin (0.1 microg/ml) in vitro using the in situ terminal deoxynucleotidyl transferase assay (TDT) to detect apoptosis. The patients were female (mean age 54.4 years). In the 16 patients, 10 had positive estrogen receptor (ER) and positive progestogen receptor (PR), and the rest had negative ER and PR. RESULTS: 18 hours after addition of 5-FU, sandostatin and both, the apoptotic rate of tumor cells was (21.8 +/- 6.6)%, (14.3 +/- 8.8)%, and (26.7 +/- 6.9)% respectively. When samples were divided into 2 groups according to the sensitivity of ER and PR, the apoptotic rate of 5-FU plus sandostatin was (32.2 +/- 7.9)%, in positive ER and PR group, but (20.5 +/- 5.5)% in negative ER and PR group (P < 0.05). CONCLUSIONS: 5-FU plus sandostatin is more sensitive in the positive ER and PR group than in the negative ER and PR group. 5-FU plus sandostatin would be helpful in the biotherapy of breast cancer patients with positive ER and PR.

Adult↗
PubMed1999

[The correlation of free radicals induced by cadmium toxicess with erythrocyte injury and band3 expression].

OBJECTIVE: To study the correlation of hydrogen peroxide induced by cadmium ion with erythrocyte injury and band3 expression. METHODS: It was application immunoscanning electronmicroscopy and cytochemistry and EDX methods. RESULT: Cadmium ion not only induced a lot of hydrogen peroxide but also caused serious distortion of erythrocyte. CONCLUSION: Cadmium ion-induced free radicals lead to oxygen free radical damage of membrane and crytoskelecton of erythrocyte, affecting the stability and deformability and band3 expression in erythrocyte.

Animals↗
PubMed1999

[Application of cerium cytochemical method in observation of renal injury caused by oxygen free radical with cadmium induction].

OBJECTIVE: To study acute renal injury caused by oxygen free radical with cadmium induction. METHODS: Electron microscopy cytochemical and energy dispersive X-ray (EDX) analysis techniques were used. RESULTS: Cadmium ions could enter the epithelial cells of renal proximal convoluted tubules. Excessive production of oxygen free radical was found in the kidney tissues, which located mainly in the endothelial cells of blood vessels, mitochondria of epithelial cells of renal proximal convoluted tubules, microvilli and basement membrane. Swelling of epithelial cells and mitochondria and drop of epithelial cells could occur in renal proximal convoluted tubules. CONCLUSION: It suggests that cadmium ions can induce the formation of large amount of oxygen free radical and cause renal injury.

Acute Kidney Injury↗
PubMed1998

Unsymmetric nonpeptidic HIV protease inhibitors containing anthranilamide as a P2' ligand.

A series of novel unsymmetrical anthranilamide-containing HIV protease inhibitors was designed. The structure-activity studies revealed a series of potent P2-P3' inhibitors that incorporate an anthranilamide group at the P2' position. A reduction in molecular weight and lipophilicity is achieved by a judicious choice of P2 ligands (i.e., aromatic, heteroaromatic, carbamate, and peptidic). A systematic investigation led to the 5-thiazolyl carbamate analog 8 m, which exhibited a favorable Cmax/EC50 ratio (> 30), plasma half-life (> 8 h), and potent in vitro antiviral activity (EC50 = 0.2 microM).

Amides↗
PubMed1998

Interaction of the xanthine nucleotide binding Goalpha mutant with G protein-coupled receptors.

We constructed a double mutant version of the alpha subunit of Go that was regulated by xanthine nucleotides instead of guanine nucleotides (GoalphaX). We investigated the interaction between GoalphaX and G protein-coupled receptors in vitro. First, we found that the activated m2 muscarinic cholinergic receptor (MAChR) could facilitate the exchange of XTPgammaS for XDP in the GoalphaXbetagamma heterotrimer. Second, the GoalphaXbetagamma complex was able to induce the high affinity ligand-binding state in the N-formyl peptide receptor (NFPR). These experiments demonstrated that GoalphaX was able to interact effectively with G protein-coupled receptors. Third, we found that the empty form of GoalphaX, lacking a bound nucleotide and betagamma, formed a stable complex with the m2 muscarinic cholingeric receptor associated with the plasma membrane. Finally, we investigated the interaction of GoalphaX with receptor in COS-7 cells. The empty form of GoalphaX bound tightly to the receptor and was not activated because XTP was not available intracellularly. We tested the ability of GoalphaX to inhibit the activities of several different G protein-coupled receptors in transfected COS-7 cells and found that GoalphaX specifically inhibited Go-coupled receptors. Thus the modified G proteins may act as dominant-negative mutants to trap and inactivate specific subsets of receptors.

Animals↗
PubMed1998

UBE3A "mutations" in two unrelated and phenotypically different Angelman syndrome patients.

Angelman syndrome (AS) is a rare neurodevelopmental disorder. Recently, several mutations have been found in the E6-AP ubiquitin protein ligase gene (UBE3A) in a group of patients who are nondeleted and do not have uniparental disomy or imprinting defects. Most of the reported mutations cluster within exons 9 or 16 of the UBE3A gene, and nearly all are predicted to give rise to truncated E6-AP ligases. Here, we describe two AS patients with dissimilar phenotypes. At the molecular level, they are both nondeleted, do not display uniparental disomy, and have normal imprint patterns. One has the typical AS phenotype and carries the previously reported 1344delAG de novo mutation involving a functionally significant region of UBE3A. The other expresses an atypical phenotype in that she has less severe ataxia, no inappropriate laughing, or epilepsy, and her EEG was normal at an early age. A 14-bp deletion in the 3' untranslated region of exon 16 (3'UTRdel14) adjacent to the poly(A) signal was identified. Further investigation revealed that the DNA change was a neutral polymorphism. Haplotype analysis indicated that both the AS patient and her normal sibling had inherited the same maternal UBE3A gene and its 5' flanking region. Although the 14-bp change has no functional significance, it assists with counseling to determine future risks of recurrence in this family.

Angelman Syndrome↗
PubMed1998

Elite endurance athletes and the ACE I allele--the role of genes in athletic performance.

Genetic markers that might contribute to the making of an elite athlete have not been identified. Potential candidate genes might be found in the renin-angiotensin pathway, which plays a key role in the regulation of both cardiac and vascular physiology. In this study, DNA polymorphisms derived from the angiotensin converting enzyme (ACE), the angiotensin type 1 receptor (AT1) and the angiotensin type 2 receptor (AT2) were studied in 64 Australian national rowers. Compared with a normal population, the rowers had an excess of the ACE I allele (P<0.02) and the ACE II genotype (P=0.03). The ACE I allele is a genetic marker that might be associated with athletic excellence. It is proposed that the underlying mechanism relates to a healthier cardiovascular system.

Adult↗
PubMed1998

Structure of cubic insulin crystals in glucose solutions.

X-ray structures of cubic insulin crystals in high concentrations of glucose at different pH levels and temperatures have been refined to high resolution. We have identified one glucose-binding site near the N-terminus of the A-chain whose occupancy is pH dependent. The effects of reduced water activity on the ordered protein and solvent structures have been examined. Our analysis showed no notable conformational changes in the ordered protein structures or ordered solvent molecules near the protein surface, but the presence of glucose does have a significant effect on the overall density distribution of the bulk solvent in the solvent-accessible volume. We compared the structure of cubic insulin at room temperature and liquid-nitrogen temperature, under identical solvent conditions, using glucose as a cryoprotectant. In this case, we found that the average temperature factor of the protein is reduced and more water molecules can be identified, but there are no significant changes in the protein conformation.

Animals↗
PubMed1998

Indirect laser photoacoustic detection for trace samples on membranes.

An indirect laser photoacoustic technique is described. The basic principle, the experimental set-up and the influencing factors are discussed in detail. This method was used to quantify phenylamine, arginine and methylene blue on membranes. The linear ranges of the calibration curve are up to two orders of magnitude. With 1 microliter of sample solution, the detection limits are 0.25, 1.52 and 0.14 pmol for phenylamine, arginine and methylene blue, respectively.

Absorption↗
PubMed1998

Effect of different sources of dietary fibre on growth performance, intestinal morphology and caecal carbohydrases of domestic geese.

1. The aim of this study was to examine the effects of dietary fibre sources on growth and on the development of the gastro-intestinal tract in growing geese. 2. Six-week-old female White Roman geese were divided at random into 6 groups with 6 dietary treatments. Diets were isoenergetic and isonitrogenous and contained alfalfa meal, barley bran, rice hulls, cellulose, pectin or lignin as the major dietary source of fibre. 3. Food intake was significantly higher in the barley bran group than in the other groups. Both daily weight gain and food conversion of the pectin and alfalfa meal groups were lower than those of the other groups. 4. In the 9-week-old geese, the relative weight and the length of the small intestine did not significantly differ among treatment groups. However, the relative weight was significantly lightest and the caecal length was significantly shortest (P < 0.05) in the pectin group. 5. Activities of amylase and cellulose hydrolases of the caecal contents were also highest in the pectin group. 6. From SEM micrographs, no morphological damage of the villi was observable in the different intestinal segments of the geese in any of the treatments.

Amylases↗
PubMed1998

Capillary electrophoresis: new technology for DNA diagnostics.

New innovations in the diagnostic laboratory achieve their full potential when they can be automated. Increasingly molecular biology (DNA) techniques are being utilised in traditional pathology disciplines, as well as the more recent ones of cytogenetics and molecular genetics. Molecular biology was first exploited for diagnostic purposes when Southern blotting became established. However the time-consuming nature of the methodology, as well as the skills required, made it difficult for Southern blotting to be used routinely in the service laboratory. Subsequently the invention of PCR facilitated the approach to DNA diagnostics. Today PCR in commercial or home-made kits is used for a range of procedures. The steps required to amplify DNA with PCR can also be fully automated. However the analysis of PCR products, which frequently requires slab gel electrophoresis and toxic chemicals or radioisotopes for visualisation, remains difficult to automate. An alternative way for analysing PCR products is now available through capillary electrophoresis. With this technique, automation can be extended to sample loading, electrophoresis and data analysis. The use of toxic chemicals or radioisotopes can be avoided.

Chromosome Mapping↗