Chemical doping and intergranular magnetic-field effects in bulk thallium-based superconductors.
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Journal Article↗
Biomedical subjects
M Yang
Publications and source records attributed to M Yang.
Structure of human apolipoprotein D: locations of the intermolecular and intramolecular disulfide links.
We have determined the primary structure of human apolipoprotein D (apoD) by aligning peptides derived from digestions by cyanogen bromide, trypsin, and chymotrypsin. Our results confirm the primary structure derived from cDNA [Drayna et al. (1986) J. Biol. Chem. 261, 16535-16539]. ApoD consists of 169 amino acid residues, including 5 cysteines. Tryptic peptide analysis indicated that Cys41 and Cys16 are joined by a disulfide bridge. Using a combination of manual Edman degradations and mass spectrometric analysis on a purified cluster of chymotryptic fragments, we identified an intramolecular disulfide bridge between Cys8 and Cys114 and an intermolecular bridge between Cys116 of apoD and Cys6 of apoA-II. In addition, sites of N-glycosylation were found at Asn45 and Asn78. Because apoD contains two intramolecular disulfide linkages and has a high content of proline to disrupt alpha-helical structures, formation of the amphipathic helical regions that characterize the other soluble apolipoproteins is unlikely. We conclude that apoD binds to lipoprotein surfaces through structures other than alpha-helices, such as disulfide links.
Amino Acid Sequence↗
Origin of multiple-peak photoluminescence spectra of light-emitting porous silicon.
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Journal Article↗
Different mechanisms of regulation of the human stromelysin and collagenase genes. Analysis by a reverse-transcription-coupled-PCR assay.
Tissue-remodeling processes are largely controlled by matrix metalloproteinases that degrade the extracellular components of connective tissues. In this study, gene regulation of two human matrix metalloproteinases, stromelysin and collagenase, was investigated by a reverse-transcription-coupled (RT)-PCR assay. Here, signals from both the heterogenous nuclear RNA (hnRNA) and mRNA are amplified, allowing the regulation of gene expression to be divided between transcriptional and/or post-transcriptional control. In confluent human lung fibroblast cultures, tumor-necrosis factor-alpha and 12-O-tetradecanoyl-phorbol 13-acetate induce stromelysin and collagenase genes transcriptionally. Interleukin-1 beta (IL-1 beta) induces stromelysin gene transcription but has little, if any, effect on the collagenase gene transcription in cells cultured in the presence of 10% serum. By a competitive RT-PCR assay, the IL-1 beta-reated cultures contain an average of 60 molecules of stromelysin mRNA/cell and the untreated cultures about 1.9 molecules/cell. In serum-starved cells, both IL-1 beta and serum induce transcription of the collagenase gene. Also, in serum-starved cells type II collagen can induce collagenase mRNA but not stromelysin mRNA. Inhibition of protein synthesis with cycloheximide induces stromelysin gene transcription but has no effect on the collagenase gene. These data indicate different mechanisms of regulation of the human stromelysin and collagenase genes in cultured cells.
Base Sequence↗
Contrasting actions of cocaine, local anaesthetic and tetrodotoxin on discharge properties of rat aortic baroreceptors.
1. Effects of cocaine, lignocaine, benzocaine and tetrodotoxin (TTX) on the simultaneously measured pressure- and diameter-discharge frequency relations of single fibre baroreceptors were compared in rat in vitro aortic arch-aortic nerve preparations. 2. Between 1 and 10 microM, cocaine produced selective increases in the pressure threshold shifting the pressure-response curve without altering the gain or threshold frequency. At near-blocking concentrations, gain was depressed as well. Cocaine experiments were done in nitroprusside (NP, 1 microM). Neither NP or NP with cocaine altered diameter (P > 0.36). 3. Lignocaine (at > 10 microM) and benzocaine (at > 100 microM) shifted pressure-response curves to higher pressures and generally depressed discharge by increasing pressure threshold and decreasing maximum discharge frequency (P < 0.05). Gain decreased and threshold frequency increased at higher concentrations. Diameter was unaffected by lignocaine or benzocaine (P > 0.14). 4. TTX increased thresholds and discharge frequencies at threshold but did not shift pressure-discharge curve locations. This produced superimposable discharge curves with changes occurring as losses of discharge points in the threshold region. Diameter was unaffected by TTX (P > 0.80). 5. The contrasting patterns of effects between TTX and local anaesthetics suggest that blockade of TTX-sensitive sodium channels alone may not be responsible for the effects of cocaine, lignocaine and benzocaine.
Anesthetics, Local↗
Deletion of QCR6, the gene encoding subunit six of the mitochondrial cytochrome bc1 complex, blocks maturation of cytochrome c1, and causes temperature-sensitive petite growth in Saccharomyces cerevisiae.
It was previously reported that disruption or deletion of QCR6, the nuclear gene encoding subunit 6 of the cytochrome bc1 complex, does not impair growth of yeast on non-fermentable carbon sources (Schoppink, P. J., Hemrika, W., Reyne, J. M., Grivell, L.A., and Berden, J. A. (1988) Eur. J. Biochem. 113, 115-122; Crivellone, M. D., Wu, M. M., and Tzagoloff, A. (1988) J. Biol. Chem. 262, 14323-14333; Schmitt, M. E., and Trumpower, B. L. (1990) J. Biol. Chem. 265, 17005-17011). We have discovered that deletion of QCR6 results in a temperature-sensitive petite phenotype, manifested at 37 degrees C, and that this phenotype can be masked by spontaneously arising suppressor mutations. Mitochondrial membranes from the deletion strain grown at 37 degrees C lack ubiquinol-cytochrome c reductase activity, and optical spectra reveal an extensive decrease in cytochrome b absorption, but little or no decrease in cytochrome c1 absorption. Immunoblots of mitochondrial membrane proteins from the deletion strain indicate that processing of cytochrome c1 from intermediate to mature size is blocked coincident with the loss of subunit 6. This is the first example where mutation of a subunit within the bc1 complex blocks maturation of cytochrome c1.
Cytochromes c1↗
Expression of mouse metallothionein-I gene confers cadmium resistance in transgenic tobacco plants.
Transgenic tobacco plants containing a mouse metallothionein-I (MT-I) gene fused to the cauliflower mosaic virus 35S (CaMV 35S) promoter and nopaline synthase (nos) polyadenylation site were obtained by transforming tobacco leaf discs with an Agrobacterium tumefaciens strain carrying the chimaeric gene. Transformants were directly selected and rooted on medium containing cadmium and kanamycin. A total of 49 individual transgenic tobacco plants were regenerated. Among them 20% showed a very high expression level and their growth was unaffected by up to 200 microM cadmium, whereas the growth of control plants was severely affected leaf chlorosis occurred on medium containing only 10 microM cadmium. The concentration of microM cadmium. The concentration of MT-I in leaves of control and transgenic tobacco was determined with Cd/haemoglobin saturation assay, a polarographic method and western blotting. In addition, seeds from self-fertilized transgenic plants were germinated on medium containing toxic levels of cadmium and scored for tolerance/susceptibility to this heavy metal. The ratio of tolerant to susceptible plants was 3:1 indicating that the metallothionein gene is inherited as a single locus.
Agrobacterium tumefaciens↗
A Gln-281 to Arg substitution in alpha-L-fucosidase is responsible for a common polymorphism detected by isoelectric focusing.
A common polymorphism of human alpha-L-fucosidase consists of three phenotypes (Fu 1, Fu 2, and Fu 2-1) assigned by isoelectric focusing. The phenotypes are determined by two codominant alleles (Fu1 and Fu2). Isozymes with the Fu 2 phenotype have more basic pI than Fu 1, while Fu 2-1 is a mixture of Fu 2 and Fu 1. Recently, a missense mutation (A860-->G) in the alpha-L-fucosidase gene was described that did not affect alpha-L-fucosidase activity. The mutation causes the substitution of Arg (pKaGuan = 12.5) for Gln-281, which has no ionizable side chain. Isoelectric focusing profiles of extracts of COS-1 cells transfected with wild-type or mutant alpha-L-fucosidase cDNAs had phenotypes of Fu 1 and Fu 2, respectively. Next, 20 human lymphoid cell lines were examined for the occurrence of the A860-->G mutation and expression of the Fu 1, Fu 2, and Fu 2-1 phenotypes. Eight lines with Fu 2 were homozygous for the A860-->G mutation; six lines with Fu 1 were homozygous for the normal nucleotide (A860); and six lines with Fu 2-1 were heterozygous. Thus, the A860-->G mutation is the molecular basis for the protein phenotypes and the Fu1 and Fu2 alleles. The normal nucleotide (A860) is responsible for Fu1 and the A860-->G mutation for Fu2.
Animals↗
Alpha-domain of human metallothionein IA can bind to metals in transgenic tobacco plants.
With a view to exploring its use as a metal-binding factor in transgenic plants we prepared the alpha-domain of metallothionein by reconstitution of rabbit apometallothionein and proteolysis of MT-1 and MT-2 with subtilisin. The isolated alpha-domains were characterised by UV and CD spectroscopy Double-Stranded. DNA encoding the alpha-domain (106 bp) of the human MT-IA was constructed from chemically synthesized oligomers by repair synthesis and enzymatic ligation, cloned into pUC19 and sequenced. A expression construct containing the cloned alpha-domain was introduced into tobacco cells on a disarmed Agrobacterium tumefaciens Ti-plasmid. Transformed tobacco cells were selected and regenerated on medium containing cadmium and kanamycin. The growth of roots and shoots of transformants was unaffected by up to 100 microM cadmium, whereas control plants showed severe inhibition of root and shoot growth, and chlorosis of leaves on medium containing only 10 microM cadmium. Southern hybridization confirmed the presence of the transgene in the transformed plant tissues. The concentration of human alpha-domain peptides in transgenic tobacco leaves was determined by the Cd/hemoglobin saturation assay and polarography using the rabbit alpha-domain as standard. The results indicate that the alpha-domain, one of two domains in MT molecules, is not only stable in vitro, but is also expressed efficiently and functions independently in transgenic plant cells.
Agrobacterium tumefaciens↗
Primary structure of apoB-100.
Apolipoprotein B-100 (apoB-100) is the major protein in low-density lipoprotein (LDL) and contains the ligand for binding LDL to its cell surface receptor. Lipoprotein [a] (Lp[a]) is a lipoprotein that consists of LDL and apolipoprotein [a] (apo[a]). The primary structure of apoB-100 has been determined by a combination of recombinant DNA and protein sequencing methods. Using high-performance liquid chromatographic techniques, we have identified sulfhydryl and disulfide groups of apoB-100 from LDL. Sixteen of the 25 cysteine residues in apoB-100 exist in disulfide form. All 14 cysteine residues within the N terminal end of apoB-100 are linked in disulfide bridges. Using the fluorescent sulfhydryl probe, 5-iodoacetoamidofluoresceine, two free sulfhydryls of apoB-100 on LDL were identified at positions 3734 and 4190. Based on its differential susceptibility to trypsin, apoB-100 can be divided into five domains: domain 1 (residues 1-1000), largely trypsin-releasable (TR); domain 2 (residues 1001-1700), alternating TR and trypsin non-releasable (TN); domain 3 (residues 1701-3070), largely TN; domain 4 (residues 3071-4100), mainly TR and mixed; and domain 5 (residues 4101-4536), almost exclusively TN. Based on our data, we propose that the structure of apoB-100 in LDL is probably an elongated form that wraps around the LDL particle, and that Cys3734 of apoB-100 may be the cysteine residue linked to a cysteine of apo[a].
Apolipoprotein B-100↗
Reverse genetics of Drosophila brain structure and function.
A set of molecular genetic technologies are described, which will have far reaching consequences for the study of brain structure, function and development in Drosophila melanogaster. Site selected mutagenesis (a PCR-based screen for P-element insertion events) allows insertion mutants to be isolated for any cloned gene, and is being used in this laboratory to ask questions about the rolls of particular cellular components in learning and memory. Transposants have been isolated in genes encoding a regulatory (RI) and a catalytic (DCO) subunit of cAMP-dependent protein kinase, and in a gene encoding a Gi-like alpha subunit. The alternative use of I factors is described. The PKA RI homozygous mutants display a significant decrement in initial learning ability. Enhancer-trap strategies, for which the GAL-4 P-element system is particularly convenient, allow the identification of genes expressed in the developing fly brain. Strategies for the efficient detection of such events are described.
Animals↗
Complete androgen insensitivity due to a splice-site mutation in the androgen receptor gene and genetic screening with single-stranded conformation polymorphism.
OBJECTIVE: To characterize the genetic defect in a family with complete androgen insensitivity syndrome and to determine whether single-stranded conformation polymorphism (SSCP) can be used to detect subtle mutations in the androgen receptor (AR) gene. DESIGN: Amplification, subcloning where appropriate, and sequencing of the AR gene in members of the affected family and to use SSCP to differentiate rapidly mutant from normal alleles. SETTING: Reproductive endocrinology clinic and laboratory in a university hospital. PATIENTS: A family of which two sisters (46XY) have complete androgen insensitivity syndrome. RESULTS: A novel single base (G --> A) mutation in the exon G-intron 7 junction of the AR gene caused an abnormal donor splice site leading to complete androgen insensitivity in both affected siblings. Their mother was demonstrated to be the heterozygous carrier of this mutation while the other two males in the family carried the normal allele. Single-stranded conformation polymorphism proved useful for defining the normal, mutant, and heterozygous carrier status of each member of this family. CONCLUSIONS: This new mutation of the human AR gene illustrates the importance of exon G in receptor function. Single-stranded conformation polymorphism is a simple and rapid screening technique that can be used to detect unknown subtle mutations in the AR gene.
Adolescent↗
Extreme evolutionary conservation of QM, a novel c-Jun associated transcription factor.
QM is a 214 amino acid polypeptide, encoded by a gene (DXS648) in Xq28, that contains a high percentage of charged amino acids and has been found to bind c-Jun and DNA. Searches of the GenBank database revealed no matches between QM and any other known transcription factors. However, we and others have isolated QM homologs from a diverse array of eukaryotes. Alignment of these sequences indicated a high degree of conservation throughout the first 175 residues of the protein and revealed several interesting features. Most notable is the considerable conservation of charged amino acids within specific regions of the protein. Secondary structure analysis suggests that two of these regions form amphipathic alpha-helices, one basic and one acidic. A third conserved charged domain, comprising the N-terminal 30 amino acids, is both basic and proline rich. The rate of sequence divergence of the various homologs was found to be slow (of the order of 1% change every 22 million years), consistent with a critical role for QM in eukaryotic cells. A role for QM as a novel class of transcription regulatory protein is suggested.
Amino Acid Sequence↗
Dynamic and steady-state behaviors of reverse saturable absorption in metallophthalocyanine.
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Journal Article↗
A comparison of the growth patterns of Chinese infants in Chengdu and Hong Kong: 0-2 years.
Factors affecting geographical differences in the growth of Chinese infants were examined by comparing data obtained from longitudinal growth studies on growth and feeding practices performed in two Chinese populations: Chengdu and Hong Kong infants from birth to 2 years. Their mean growth curves were fitted by the Reed model for the first year and the Count model for the second year. Chengdu mothers were significantly taller than Hong Kong mothers. Chengdu infants grew more rapidly than the Hong Kong infants during the first 6 months; it is likely that this is related to genetic differences in growth potential and earlier introduction of solids in Chengdu infants.
Child, Preschool↗
Intrauterine pressure: comparison of extra vs intra amniotic methods using a transducer tipped catheter.
Fourteen women admitted in early labor, with intact membranes, were studied. In each woman, two catheters were inserted transcervically; one catheter was inserted into the extraamniotic space before rupture of membranes, and the second catheter was inserted, after artificial rupture of membranes into the intraamniotic cavity. The contraction to contraction difference in active pressure (pressure above the baseline) recorded by the two catheters, as well as the cumulative total active pressure generated (calculated by adding the active pressure of all contractions) throughout labor by the two catheters were compared. Of the 606 contractions analysed, 43.2% showed pressure difference of 0-5 mmHg between the intra amniotic and extraamniotic catheter; 21.3% showed 6-10 mmHg difference, 12.2% showed a difference of 11-15 mmHg, and 23.3% showed a difference greater than 15 mmHg. When total active pressure generated by each catheter for individual patients was calculated, the percentage difference varied from 2.3% to 53.7%. Intrauterine pressure measurements in labour using transducer-tipped catheters inserted in the extraamniotic space may not provide comparable information to a similar catheter inserted intraamniotically when membranes are ruptured.
Amnion↗
A nonliner poroelastic model for the trabecular embryonic heart.
A theoretical model is presented for the primitive right ventricle of the stage 21 chick embryo. At this stage of development, the wall of the heart is trabecular with direct intramyocardial blood flow. The model is a pressurized fluid-filled cylinder composed of a porous inner layer of isotropic myocardium and a relatively thin compact outer layer of transversely isotropic myocardium. The analysis is based on nonlinear poroelasticity theory, modified to include residual strain and muscle activation. Correlating theoretical and experimental pressure-volume loops and epicardial strains gives first-approximation constitutive relations for stage 21 embryonic myocardium. The results from the model suggest three primary conclusions: (1) Some muscle fibers likely are aligned in the compact layer, with a fiber angle approximately + 10 deg from the circumferential direction. (2) Blood is drawn into the wall of the ventricle during diastolic filling and isovolumic contraction and is squeezed out of the wall during systolic ejection, giving a primitive intramyocardial circulation before the coronary arteries form. As the heart rate increases, the transmural blood-flow velocity increases, but the volume of blood exchanged with the lumen per beat decreases. (3) Residual strain affects transmural stress distributions, producing nearly uniform stresses in the porous layer, where the peak end-systolic stress occurs. These results improve our understanding of the relation between form and function in the developing heart and provide directions for biological experiments to study cardiac morphogenesis.
Animals↗
Processing of the LH/CG receptor and bound hormone in rat luteal cells after hCG-induced down-regulation as studied by a double immunofluorescence technique in conjunction with confocal laser scanning microscopy.
We developed a double immunofluorescence technique for detection of the rat luteinizing hormone/choriogonadotropin (LH/CG) receptor and bound hCG in the same rat ovarian section and used it in conjunction with confocal laser scanning microscopy (CLSM) to study the fate of the receptor-hormone complex in luteal cells during the hCG-induced down-regulation. Pseudopregnant immature females rats were perfusion-fixed before (0 hr) and 2, 6, 12, 24, or 36 hr after a down-regulating dose of hCG (500 IU IV). The cryosections were stained for the LH/CG receptor and bound hormone by sequential incubations with a polyclonal rabbit antiserum to purified rat LH/CG receptor and a mouse monoclonal antibody (MAb) to hCG, followed by sequential incubation with TRITC- and FITC-conjugated secondary antibodies to rabbit and mouse immunoglobulins, respectively. The results were semiquantitatively analyzed by a pseudo-three-dimensional (3D) plotting of the intensities of the receptor and hormone-specific fluorescence in luteal cells by CLSM. The analysis suggested that the majority of the LH/CG receptors are located on the luteal cells before induction of the down-regulation and that their content seem to vary not only among cells but also on the surface of single cells, thus supporting the previous concept of the functional heterogeneity among the cells and their functional compartmentation. At 2 hr after injection of the down-regulating dose of hCG, the LH/CG receptor-specific and hCG-specific fluorescences clearly co-localized on the luteal cells. Both the LH/CG receptor- and hCG-specific fluorescences disappeared from the luteal cell surfaces in a parallel fashion within 36 hr without a detectable accumulation of either fluorescence deep in the cell interior. These results suggest that the LH/CG receptor and bound hCG do not differ in their manner of in vivo processing in luteal cells. Therefore, the disappearance of the receptor and bound hormone occurs in a parallel fashion and without detectable internalization.
Animals↗