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A Zhang

Publications and source records attributed to A Zhang.

At least 163 records · Page 9Linked to original sources
PubMed1992

Structure of HIV-1 reverse transcriptase/DNA complex at 7 A resolution showing active site locations.

AIDS, caused by human immunodeficiency virus (HIV), is one of the world's most serious health problems, with current protocols being inadequate for either prevention or successful long-term treatment. In retroviruses such as HIV, the enzyme reverse transcriptase copies the single-stranded RNA genome into double-stranded DNA that is then integrated into the chromosomes of infected cells. Reverse transcriptase is the target of the most widely used treatments for AIDS, 3'-azido-3'-deoxythymidine (AZT) and 2',3'-dideoxyinosine (ddI), but resistant strains of HIV-1 arise in patients after a relatively short time. There are several nonnucleoside inhibitors of HIV-1 reverse transcriptase, but resistance to such agents also develops rapidly. We report here the structure at 7 A resolution of a ternary complex of the HIV-1 reverse transcriptase heterodimer, a monoclonal antibody Fab fragment, and a duplex DNA template-primer. The double-stranded DNA binds in a groove on the surface of the enzyme. The electron density near one end of the DNA matches well with the known structure of the HIV-1 reverse transcriptase RNase H domain. At the opposite end of the DNA, a mercurated derivative of UTP has been localized by difference Fourier methods, allowing tentative identification of the polymerase nucleoside triphosphate binding site. We also determined the structure of the reverse transcriptase/Fab complex in the absence of template-primer to compare the bound and free forms of the enzyme. The presence of DNA correlates with movement of protein electron density in the vicinity of the putative template-primer binding groove. These results have important implications for developing improved inhibitors of reverse transcriptase for the treatment of AIDS.

Base Sequence↗
PubMed1992

The neurospora CYT-18 protein suppresses defects in the phage T4 td intron by stabilizing the catalytically active structure of the intron core.

The Neurospora CYT-18 protein, a tyrosyl-tRNA synthetase, which functions in splicing group I introns in mitochondria, promotes splicing of mutants of the distantly related bacteriophage T4 td intron. In an in vivo assay, wild-type CYT-18 protein expressed in E. coli suppressed mutations in the td intron's catalytic core. CYT-18-suppressible mutations were also suppressed by high Mg2+ or spermidine in vitro, suggesting they affect intron structure. Both the N- and C-terminal domains of CYT-18 are required for efficient splicing, but CYT-18 with a large C-terminal truncation retains some activity. Our results indicate that CYT-18 interacts with conserved structural features of group I introns, and they provide direct evidence that a protein promotes splicing by stabilizing the catalytically active structure of the intron RNA.

Base Sequence↗
PubMed1992

Magnesium regulates intracellular free ionized calcium concentration and cell geometry in vascular smooth muscle cells.

Regulatory effects of extracellular magnesium ions ([Mg2+]o) on intracellular free ionized calcium ([Ca2+]i) were studied in cultured vascular smooth muscle cells (VSMCs) from rat aorta by use of the fluorescent indicator fura-2 and digital imaging microscopy. With normal Mg2+ (1.2 mM)-containing incubation media, [Ca2+]i in VSMCs was 93.6 +/- 7.93 nM with a heterogeneous cellular distribution. Lowering [Mg2+]o to 0 mM or 0.3 mM (the lowest physiological range) resulted in 5.8-fold (579.5 +/- 39.99 nM) and 3.5-fold (348.0 +/- 31.52 nM) increments of [Ca2+]i, respectively, without influencing the cellular distribution of [Ca2+]i. Surprisingly, [Mg2+]o withdrawal induced changes of cell geometry in many VSMCs, i.e., the cells rounded up. However, elevation of [Mg2+]o up to 4.8 mM only induced slight decrements of [Ca2+]i (mean = 72.0 +/- 4.55 nM). The large increment of [Ca2+]i induced by [Mg2+]o withdrawal was totally inhibited when [Ca2+]o was removed. The data suggest that: (1) [Mg2+]o regulates the level of [Ca2+]i in rat aortic smooth muscle cells, and (2) [Mg2+] acts as an important regulatory ion by modulating cell shapes in cultured VSMc and their metabolism to control vascular contractile activities.

Animals↗
PubMed1992

Extracellular magnesium regulates intracellular free Mg2+ in vascular smooth muscle cells.

Regulatory effects of extracellular magnesium ions ([Mg2+]o) on intracellular free ionized magnesium ([Mg2+]i) were examined in cultured vascular smooth muscle cells (VSMCs) from rat aorta by digital imaging microscopy using the Mg2+ fluorescent probe, Mag-fura-2. With normal Mg2+ (1.2 mM)-containing incubation media, [Mg2+]i in VSMCs was 0.63 +/- 0.09 mM. The ratio of [Mg2+]i/[Mg2+]o was 0.52 +/- 0.07. Elevation of [Mg2+]o up to 4.8 mM induced consistent increments in [Mg2+]i (to a mean values of 1.63 +/- 0.08 mM) in 5 min and lowered the ratio of [Mg2+]i/[Mg2+]o to 0.34 +/- 0.02. Our data suggest that [Mg2+]o can regulate [Mg2+]i, which may be related to its effects on intracellular Ca2+ ([Ca2+]i) and tone of VSMCs.

Animals↗
PubMed1992

Ethanol decreases cytosolic-free calcium ions in vascular smooth muscle cells as assessed by digital image analysis.

Effects of ethanol on intracellular-free Ca2+ concentration in cultured rat aortic smooth muscle cells were examined by digital imaging fluorescence microscopy using the Ca2+ fluorescence indicator, fura-2. Ethanol induced dose-dependent decrements in cytosolic-free Ca2+ concentration at 45 mM and 90 mM, which was consistent with previously reported observations of relaxation in intact rat aortic tissues. However, ethanol at high pharmacological concentrations (e.g., 450 mM) failed to induce any further inhibition in cytosolic-free Ca2+ concentration. Our results suggest that the vasodilator effects of ethanol, observed on intact blood vessels, may result in part from an interference with the availability of Ca2+ for excitation-contraction coupling in vascular smooth muscle cells.

Animals↗
PubMed1992

[A mid-infrared spectra study of a series of benzocrown ether and macrocyclic polyether diester].

With a Nicolet 20 SX B Fourier Transform infrared spectrometer, the mid-infrared spectra in region of 4000-400 cm-1 were observed for a series of benzocrown ether and macrocyclic polyether diester. A difference in the wavelength of absorption of the ether bond was found between benzocrown ether and macrocyclic polyether diester. The wavelength of absorption of the ether bond of the compounds I-IV moved down about 25 cm-1, as compared with that of the compounds V-IX.

Ethers, Cyclic↗
PubMed1991

Interactions of magnesium and chloride ions on tone and contractility of vascular muscle.

Replacement of extracellular chloride ions ([Cl-]o) by other anions, on contractility and the effects of extracellular magnesium ions ([Mg2+]o) on spontaneous mechanical activity, as well as on agonist-induced responses of rat aorta and portal vein, were studied. Replacement of [Cl-]o with acetate (Ac-) or isethionate (Ise-) ions resulted in an increase and decrease, respectively, of the spontaneous mechanical activity frequency in portal vein; the amplitudes of the spontaneous mechanical activity were attenuated by Ac- and Ise- substitution. Withdrawal of [Mg2+]o in Cl(-)-containing media resulted in elevation of tension development in rat aortas, whereas a similar maneuver in media with Ac- or Ise-, substituted for [Cl-]o, resulted in abrogation of this tension development. Use of disulfonic stilbene anion-channel blockers, DIDS (4,4'-diisothiocyano-2,2'-stilbene disulfonate, 400-600 microM) and SITS (4,4'-acetamido-4'-isothiocyano-2,2'-stilbene disulfonic acid, 400-600 microM), failed to influence either spontaneous mechanical activity or basal tone of rat portal portal vein or aortas. Incubation of DIDS or SITS in Mg(2+)-free media also failed to influence mechanical responses to withdrawal of [Mg2+]o. Use of the Cl- cation transport inhibitor bumetanide (30-80 microM) also failed to alter spontaneous mechanical activity or basal tone in either the presence or absence of [Mg2+]o. Ac- and Ise- substitution attenuated norepinephrine- and K(+)-induced contractile responses in portal vein and aorta, Caffeine-induced contractions of aortas were potentiated by withdrawal of [Mg2+]o in Cl(-)-containing media but inhibited in Ac(-)- or Ise(-)-substituted solutions. In the presence of [Mg2+]o, substitution of foreign anions resulted in alterations in the agonist contractile dose-response curves; EC50s were increased whereas maximum tensions were depressed. Withdrawal of [Mg2+]o amplified these effects. Substitution of Ac- or Ise- for [Cl-]o in the presence or absence of [Mg2+]o depressed contractions induced by Ca acetate in aortas and portal vein. These results suggest that: (1) Cl- plays an important role in regulating spontaneous mechanical activity, basal tone, and contractility in rat aorta and portal vein; and (2) Cl- probably physiologically mediates some of the effects and actions of [Mg2+]o on intracellular release of and influx of Ca2+ in these smooth muscles.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1991

Sexual dimorphism of vascular smooth muscle responsiveness is dependent on anions and estrogen.

Alteration of the extracellular anion environment by replacement of chloride ions (Cl-) with thiocyanate ions (SCN-) in normal Krebs-Ringer bicarbonate solution (NKRB) induced sustained development of basal tension in isolated aortas from male adult Sprague-Dawley and Long-Evans rats, but not from females of these strains. However, aortic smooth muscle isolated from sexually immature male and female Wistar rats underwent more marked tension development under such treatment, exhibiting no gender differences. Such SCN(-)-induced contractile responses are not tachyphylactic, completely dependent on extracellular calcium ([Ca2+]0), and could be aborted (relaxed to basal tone) by readmission of Cl-. Castration and replacement of male sex hormones with estradiol inhibited SCN(-)-induced contractions in isolated aortas from adult Wistar male rats, while castration and testosterone supplementation failed to induce contraction in aortic muscle of adult female Wistar rats exposed to SCN-. We believe these data are compatible with the notion that gender differences in vascular activity may be modulated by actions of estradiol on the metabolism of Cl- and other anions in vascular smooth muscle, which may be linked to transport of Ca2+ across the vascular muscle cell membrane.

Animals↗
PubMed1990

Endothelial cells are required for inhibition of contractile responses induced by reduction in extracellular magnesium and sodium ions in rat aortic smooth muscle [corrected].

The possible importance of facilitation of sodium-calcium (Na(+)-Ca2+) exchange by removal of extracellular magnesium ions ([Mg2+]o) in expression of endothelium-dependent relaxation was investigated in aortic rings isolated from female rats. Simultaneous [Mg2+]o withdrawal (0 mM Mg2+) and reduction in extracellular Na+ (Total [Na+]o = 84 mM), by replacement of NaCl with isosmolar amounts of sucrose in normal Krebs-Ringer bicarbonate (NKRB), induced significant increases of basal tone of denuded rat aortic rings, but not in tissues with intact endothelium. These vascular effects were not affected by indomethacin, phentolamine or atropine in any of the tissues tested. Reintroduction of 1.2 mM Mg2+ or removal of extracellular Ca2+ ([Ca2+]o) from the Mg2+ and Na(+)-deficient incubation media induced complete relaxation of the denuded tissues. Methylene blue (10(-5) M), an inhibitor of endothelium-derived relaxant factor (EDRF), potentiated tension development in intact tissues. These results suggest that: (1) as in vascular muscle, Mg2+ plays an important role in Ca2+ homeostasis in endothelial cells (EC), probably via Na(+)-Ca2+ exchange; and (2) such Mg(+)-regulated internal Na(+)-dependent Ca2+ entry participates in the expression of endothelium-dependent relaxation.

Animals↗
PubMed1989

Sister chromatid exchanges in the human active and inactive X chromosomes.

Four human female fibroblast strains with an i(Xq) or derivative X chromosome as a cytological marker for the inactive X chromosome were used to determine the frequency of sister chromatid exchanges (SCEs) in the active and inactive X chromosomes. No significant difference in SCE frequency between the active and inactive X chromosomes was observed. Therefore, the state of chromatin condensation and the late DNA replication in the facultative heterochromatin of the inactive X chromosome do not appear to influence the SCE frequency.

Cells, Cultured↗
PubMed1988

DA/DAPI-fluorescent heteromorphism of human Y chromosome.

Variation of DA/DAPI intensity in the Yq12 band was observed in five amniotic cell specimens and one blood specimen from the father of one fetus. Three distinct classes of Yq heterochromatin were identified by distamycin A (DA) treatment of the cell cultures and various staining techniques. The heterochromatin in the Yq11.23 sub-band does not undercondense when exposed to DA, and shows pale fluorescence with quinacrine staining, positive C-banding, and bright fluorescence with DA/DAPI technique. This class of heterochromatin was consistently observed in all specimens studied. The other two classes of heterochromatin are in the Yq12 band. Both show undercondensation when exposed to DA, quinacrine-bright fluorescence, and positive C-banding; however, one class of heterochromatin shows DA/DAPI-bright fluorescence and the other shows pale fluorescence. The size and banding intensity of the two classes of heterochromatin in Yq12 are variable. These results provide cytological evidence of heterogeneity within the Y heterochromatin region containing AT-rich DNA.

Chromosome Banding↗
PubMed1988

Size characterization of Mycobacterium bovis BCG (Bacillus Calmette Guérin) vaccine, Tice substrain.

Reconstituted, lyophilized, attenuated Mycobacterium bovis, Bacillus Calmette Guérin (BCG) vaccine, Tice substrain, was characterized using a Coulter Multisizer and a HIAC/Royco counter. The primary organism has an equivalent spherical diameter approximating 1 micron but the BCG cell suspension is heavily aggregated. The cumulative size distribution of the suspension fits a log-probit plot and this information can be used to determine the total number of particles per ampoule. The instrumental count may be related to the viable count. The state of dispersion was unaffected by mild shear (syringe aspiration or ultrasound) and only slightly affected by the addition of cetylpyridinium chloride or sodium tauroglycolate.

BCG Vaccine↗
PubMed1988

The surface charge of cells of Mycobacterium bovis BCG vaccine, Tice substrain.

The zeta potential of cells of an attenuated vaccine Mycobacterium bovis, BCG, Tice substrain, was measured over a pH range of 1.5-11.0 at low electrolyte concentration. There was a marked electro-positive charge at low pH, the zero charge point, pH 4.4, being similar for viable and heat killed vaccine. At pH 6.5 the addition of sodium dodecyl sulphate or sodium tauroglycolate made little difference to the zeta potential. However, progressive addition of cetylpyridinium chloride reduced and finally reversed the charge to a maximum of +60 mV at a concentration of 1.5 x 10(-2) mol dm-3 surfactant. Higher concentrations reduced the charge although it remained positive. The nature of the adsorbing species on the cell surface is discussed since the Tice substrain has both cationic and anionic surface charges whereas the Glaxo strain is reported to only possess electro-negative phosphate surface groups. Some hydrophobic interaction involving lipid within the surface may also be involved.

BCG Vaccine↗
PubMed1987

Chronic lymphocytic leukemia in China.

Chronic lymphocytic leukemia (CLL) is a very common form of leukemia among middle-aged and elderly persons in Western countries. In the Japanese, or in those with Japanese ancestry born and living in the USA, CLL is rather rare. It is also rare in China, but convincing data to support this statement are found primarily in the Chinese literature. We have reviewed previously unpublished data from Peking Union Medical College (PUMC) concerning the frequency of CLL among adult patients with the four common types of leukemia. Of 4,174 such patients (1952-1986), 4.6% had CLL; 22.9% chronic myeloid leukemia (CML); 50.3% acute myeloid leukemia (AML); and 22.3% acute lymphoblastic leukemia (ALL). This low frequency of CLL is none the less higher than that found in a multiinstitutional collection of Chinese patients with leukemia. Various clinical and laboratory aspects of a carefully studied, previously published group of patients with CLL from PUMC were compared to similar published findings in patients from the University of Utah. No differences in the overall nature of the disease were detected between these groups. However, not unexpectedly, the duration of symptoms before the diagnosis was made longer in Chinese than in USA patients and the proportion diagnosed at a time when no CLL related symptoms had been noted was less in Chinese. Neither differences in life span in the two populations nor an excess of CML, AML, and ALL at the expense of CLL were viable hypotheses to explain the rarity of CLL in China. We suggest that the dearth of CLL in Chinese is on a genetic basis, as it is thought to be in the Japanese.

Adult↗