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Biomedical subjects

G Liu

Publications and source records attributed to G Liu.

At least 577 records · Page 32Linked to original sources
PubMed1994

Regulation of myosin regulatory light chain phosphorylation via cyclic GMP during chemotaxis of Dictyostelium.

Previous studies on the chemotactic movement of Dictyostelium have indicated a role for cyclic GMP in regulating the association of myosin II with the cytoskeleton. In this study we have examined the part played by phosphorylation of the 18 kDa myosin regulatory light chain in this event. Using streamer F mutant NP368 (which is deficient in the structural gene for cyclic GMP-specific phosphodiesterase) we find that, for the regulatory light chain kinase, the major peak of phosphorylation is delayed compared to the parental control strain XP55, occurring at 80 seconds rather than about 30 seconds in XP55. In two independently derived mutants that are unable to increase their cellular concentration of cyclic GMP (above basal levels) in response to a chemotactic stimulus of cyclic AMP (KI-10 and SA219), no increase in the phosphorylation of the light chain occurred, or movement of myosin II to the cytoskeleton. We also find a smaller peak of light chain phosphorylation that occurs within 10 seconds of cyclic AMP stimulation of the amoebae, and which is absent in the cyclic GMP-unresponsive strains. We conclude that cyclic GMP is involved in regulating light chain phosphorylation in this system. The possible significance of these findings is discussed and a model that relates these findings to published data on cytoskeletal myosin changes during chemotaxis is presented.

3',5'-Cyclic-GMP Phosphodiesterases↗
PubMed1994

Long-term comparisons of eight-week body weight for a randombred line of chickens reared at two locations.

For 36 generations, 8-wk body weight was measured in Athens-Canadian Randombred males and females reared at the Southern Regional Poultry Genetics Laboratory (SRPGL) and at Virginia Polytechnic Institute and State University. Regressions of this trait on generations ranged from -.7 g for females reared in Virginia to -2.6 g for males reared at the SRPGL. Chickens were consistently heavier at the SRPGL, where a broiler diet was fed, than at Virginia, where the diet was considerably lower in protein and energy. Highly significant (P < or = .01) location by generation interactions reflected fluctuations at one location that were independent of those at the other location. Correlations between body weight at the two locations increased when chicks were vaccinated for Marek's disease.

Animals↗
PubMed1994

Responses to long-term divergent selection for eight-week body weight in chickens.

Thirty-six generations of divergent selection for body weight at 8 wk of age resulted in approximately an eightfold difference between the high (HWS) and low (LWS) lines for this trait. In both White Plymouth Rock lines, selection intensities declined over generations, with responses about five times greater during the first 18 than the last 18 generations of selection in Line HWS and about two times greater in Line LWS. Also, responses to selection were more irregular in the later than in earlier generations. Sexual dimorphism of body weight (male:female) was greater for Line LWS than HWS. A randombred control line (AC) maintained concurrently with the selected lines did not exhibit environmental trends across generations. Relaxed lines, established at different generations of selection, were used to evaluate effectiveness of selection and monitor environmental changes over short periods.

Animals↗
PubMed1994

Comparison studies of passive and active shunting during cross-clamping of the descending thoracic aorta.

The effects of passive (no pump) and active (using centrifugal pump) shunting on hemodynamics and metabolisms during cross-clamping of the descending thoracic aorta were compared in mongrel dogs. In Group I (control), the aorta was cross-clamped for 2 hours without shunting; in Group II (passive), a temporary shunt was placed between the left common carotid and left femoral arteries during cross-clamping; and in Group III (active), left heart by-pass, matching flow to that in group II, was performed. Hemodynamic and metabolic parameters were recorded prior to cross-clamping and following every 30 min for 4 hours. All animals in Group I suffered from hemodynamic instability, metabolic abnormal and neurologic injury and died within 12 hours. Hemodynamic changes were unstable in Group III than II, and three dogs in Group III, but none in Group II, suffered from neurologic injury. Metabolic changes between Group II and III were not significant. We conclude that passive shunting is superior to active shunting for aortic by-pass.

Animals↗
PubMed1994

[Gas chromatographic analysis of C1-C6 fatty acids in rat fecal samples].

A gas chromatography procedure was performed to determine C1-C6 fatty acids in rat feces. Acidated fecal homogenate was extracted with ethyl ether-petroleum ether (1 + 1). After adding tetramethylammonium hydroxide to the extract, the quaternary ammonium salts of the fatty acids could react with ethyl bromide in the presence of N,N-dimethyl acetamide producing corresponding esters. The derived products were separated by the column packed with ethylene glycol succinate polyester and were determined with Flame ionization detector (FID). The extractive efficiencies of C1-C6 fatty acids from the fecal samples were 88.5%-100%, and the recoveries ranged from 82.4% to 110.8% with the coefficients of variation of 3.2%-10.9%. The detection limits were at 1ppm. The optimum conditions of esterification are discussed. The method is safe and rapid, and has been applied to the analysis of C1-C6 fatty acids in rat feces.

Animals↗
PubMed1994

[A new species of Thermoplasma].

An extremely thermoacidophilic strain, ES-23, was isolated from a self-heating coal spoil pile, Sichuan, China. The new isolate grows between 50 and 75 degrees C with optimum 70 degrees C and pH 1.0-5.0 with optimum 2.0. Strain ES-23 is aerobic and facultative autotrophic depending both heterotrophically on organic compounds and autotrophically on elemental sulfur as energy source and CO2 as carbon source. G-, spherical or oval with 0.9-1.5 microns in diameter. The autotrophic growth showed peri pilus-like structure. Cells Lack a rigid cell wall and surrounded only by a single triple layed membrane containing ether lipids. GC content of DNA is 38 mol%(Tm). It has been determined as a new species of Thermoplasma genus and named Thermoplasma thiooxidans sp. nov.

Terminology as Topic↗
PubMed1994

Lipoprotein lipase and hepatic lipase: the role of asparagine-linked glycosylation in the expression of a functional enzyme.

Lipoprotein lipase (LPL) and hepatic lipase (HL) share two conserved asparagine-linked glycosylation sites, located at the amino- and carboxy-terminal domains of the protein. Human HL contains two additional sites, preceding each conserved site by 36 and 35 amino acids, respectively. The utilization of these sites for glycan-binding and the role of each glycan chain for the catalytic function of human LPL, rat HL, and human HL was investigated. To accomplish this aim, potential Asn glycosylation sites were changed to Gln by site-directed mutagenesis and the resulting constructs were expressed in a mammalian (COS) cell system. We demonstrate the following. 1) All potential glycosylation sites in human LPL, rat HL, and human HL are utilized. 2) Lack of glycosylation at the two non-conserved sites in human HL has no effect on enzyme expression. 3) Glycosylation at the conserved Asn sites in the N-terminal domain of LPL and HL is required for the synthesis of a fully active and secreted lipase. While this is an absolute requirement for LPL, a portion (approximately 25%) of HL molecules lacking glycosylation at this essential site still becomes active and secreted. However, the simultaneous elimination of both glycosylation sites at the N-terminal domain of human HL results in the virtual abolishment of enzymatic activity and secretion. 4) Glycosylation at the conserved sites in the C-terminal domain is not essential for the expression of active lipases. 5) Eliminating all glycosylation sites in LPL and HL results in the synthesis of inactive enzymes that are retained intracellularly; however, a small portion (2%) of unglycosylated rat HL was active and secreted. We conclude that glycosylation overall plays an important role in the formation of functional LPL and HL.

Animals↗
PubMed1994

Effectiveness of assistance circulations for distal circulatory support during cross-clamping of the descending thoracic aorta.

In order to find the ideal distal circulatory support during cross-clamping of the descending thoracic aorta, the author compared the effects of simple shunting (SS) and centrifugal pump (CP) on hemodynamics and metabolisms in mongrel dogs. In group I (control), the aorta was cross-clamped for two hours without SS or CP; in group II (SS), a temporary shunt was placed between the left common carotid and left femoral arteries during cross-clamping; in group III (CP), with left heart bypass, the flow was maintained about the same as that in group II; in group IV (CP), proximal pressure was maintained unchanged. Hemodynamic and metabolic parameters were recorded prior to cross-clamping and every 30 min for four hours during and after cross-clamping. All animals in group I suffered from hemodynamic instability, metabolic abnormalities and neurologic injury and died within 12 hours. Hemodynamic changes were more unstable in group III than in groups II and IV. Three dogs in group III and also in group IV but none in group II, suffered from neurologic injury. Metabolic changes in groups II, III and IV were not significant. The author conclude that hemodynamic and metabolic abnormalities can be minimized through the efficient use of a shunt in cross-clamping of the descending thoracic aorta and postoperative complications such as paraplegia, renal failure and hepatic dysfunction can be prevented. The centrifugal pump as a distal circulatory support device is able to maintain stable hemodynamics and normal distal organic metabolisms if aortic pressure proximal to the clamp is maintained essentially unchanged through regulation of pump flow. However, it is unable to prevent paraplegia during cross-clamping of the descending thoracic aorta for two hours. Shortening the time of cross-clamping of the descending thoracic aorta is necessary to prevent paraplegia.

Animals↗
PubMed1994

Direct evidence for the existence and functional role of hyperreactive sulfhydryls on the ryanodine receptor-triadin complex selectively labeled by the coumarin maleimide 7-diethylamino-3-(4'-maleimidylphenyl)-4-methylcoumarin.

The fluorogenic sulfhydryl probe 7-diethylamino-3-(4'-maleimidylphenyl)-4-methylcoumarin (CPM) (1-50 nM) is used to characterize the functional role and location of highly reactive thiol groups on the ryanodine-sensitive Ca2+ release channel complex [i.e., ryanodine receptors (RyRs)] of skeletal and cardiac junctional sarcoplasmic reticulum (SR). The kinetics of forming fluorescent CPM adducts with junctional but not longitudinal SR membrane proteins (0.02-1 pmol of CPM/microgram of SR protein) are found to be markedly dependent on the presence of physiological and pharmacological modulators of the RyR Ca2+ channel. RyR agonists, micromolar Ca2+, and nanomolar ryanodine promote a slow SR thiol-CPM reaction, with an apparent rate constant k of 0.0021 +/- 0.0002 sec-1, and > 89% of the fluorescence is associated with the 110-kDa Ca2+ pump, which constitutes 68% of the protein in the SR preparations. However, in the presence of Ca2+ channel antagonists (millimolar Mg2+, millimolar Ca2+, or micromolar ryanodine), CPM rapidly forms adducts with a single class of highly reactive (hyperreactive) SR thiols (k = 0.025 +/- 0.002 sec-1). Nonreducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis of CPM-labeled SR protein and Western blot analyses with antiryanodine or antitriadin antibodies reveal that the hyperreactive thiols labeled by CPM under conditions favoring channel closure are localized principally to the RyR protomer and triadin, which constitute < 6% of the protein in the SR preparation. Immunoprecipitation experiments with antiryanodine and antitriadin monoclonal antibodies confirm the location of CPM-labeled thiol groups on RyR and triadin, respectively. The results indicate that the RyR and triadin contain a small number of highly reactive cysteine residues that selectively conjugate with CPM only when channel closure is favored. It is shown that either 1) the redox state (sulfhydryl/disulfide status) or 2) the accessibility of the hyperreactive thiols on the RyR and triadin is determined by the conformational state of the channel. Covalent modification of hyperreactive thiols with nanomolar CPM inhibits both Ca(2+)-induced Ca2+ release and the gating activity of single channels reconstituted in bilayers, revealing the essential functional importance of hyperreactive thiols on channel-associated proteins. 1,4-Naphthoquinone (0.4-40 pmol/micrograms of protein) selectively oxidizes hyperreactive thiols on RyR and triadin and releases Ca2+ from SR vesicles, without inhibiting Ca(2+)-ATPase activity. The results provide direct evidence of the existence and functional role of hyperreactive cysteine residues on the RyR and triadin in regulating the gating of ryanodine-sensitive intracellular Ca2+ channels and strongly suggest that these important Ca2+ regulatory channels may be an important target for oxidative cell damage mediated by quinones.

Animals↗
PubMed1993

Purification and characterization of a macromolecular-translocation inhibitor III of activated glucocorticoid-receptor-complex binding to nuclei from rat liver.

Macromolecular-translocation inhibitors (MTI) of binding of the activated glucocorticoid-receptor complex (GRC) to nuclei from rat liver are separated into at least three components (MTI-I-III) by DEAE-cellulose column chromatography [Okamoto, K., Isohashi, F., Horiuchi, M. & Sakamoto, Y. (1982) Biochem. Biophys. Res. Commun. 108, 1655-1660]. In this study, we have purified MTI-III from the livers of adrenalectomized rats to apparent homogeneity, as determined by SDS/PAGE. The purification procedure consisted of DEAE-cellulose chromatography, acid treatment and sequential chromatographies using gel filtration, S-Sepharose and Mono S columns. The purified protein had a molecular mass of approximately 69 kDa, as estimated by SDS/PAGE, and the molecular mass of the inhibitor was approximately 68 kDa, as estimated by gel filtration. Thus, MTI-III exerts its inhibitory activity as a monomer. The sedimentation coefficient of MTI-III was approximately 3.7 S. Purified MTI-III was fairly stable at 4 degrees C but at higher temperatures, especially above 25 degrees C, it was rapidly inactivated. Under low-salt conditions, MTI-III was associated with activated GRC (4.2 S) and the resulting complex was detected on sucrose density gradients as a larger species (6.8 S). Initial treatment of nuclei or DNA-cellulose with MTI-III did not alter their abilities to bind activated GRC. These results indicate that MTI-III acts through an interaction with GRC.

Animals↗
PubMed1993

Prevention of metastasis by inhibition of the urokinase receptor.

The plasminogen activator urokinase (u-PA) mediates proteolysis by a variety of human tumor cells. Competitive displacement of u-PA from cellular binding sites results in decreased proteolysis in vitro, suggesting that the cell surface is the preferred site for u-PA-mediated protein degradation. We studied the effect of u-PA receptor blockade on the metastatic capacity of human PC3 prostate carcinoma cells, using transfectants which expressed chloramphenicol acetyl-transferase (CAT). Eight weeks after subcutaneous inoculation of these cells into nude mice, CAT activity was detected in regional lymph nodes, femurs, lungs, and brain, thereby mimicking the organ tropism observed for naturally occurring metastases of prostate cancer. In a second transfection, CAT-expressing PC3 cells received cDNA encoding a mutant u-PA (Ser356-->Ala) which lacks enzymatic activity but which retains full receptor binding affinity. Three mutant u-PA expressors, each with < 5% of wild-type cell-associated u-PA activity, were compared in vivo with independently derived controls. Primary tumor growth was similar in each group of animals and all tumors expressed comparable CAT activity. In contrast, metastasis (as assessed by CAT activity) was markedly inhibited when cell surface u-PA activity was blocked. Levels of CAT activity were reduced by a factor of > 300 in regional lymph nodes, 40-100 in brain tissue, and 10-20 in lung tissue. Metastatic capacity was inhibited similarly when animals were given intermittent intraperitoneal injections of a u-PA/IgG fusion protein capable of displacing u-PA activity from the tumor cell surface. Our results indicate that cell surface u-PA activity is essential to the metastatic process. In addition, the assay system employed in these experiments may be generally useful in testing other therapeutic modalities to limit the spread of primary tumors.

Adenocarcinoma↗
PubMed1993

Assembly of lipoprotein lipase in perfused guinea-pig hearts.

It has been suggested that lipoprotein lipase (LPL) can be assembled into its catalytically active dimeric form only after its oligosaccharide chains have been processed in the Golgi. To study this in a complete organ, LPL was metabolically labelled with [35S]methionine in perfused guinea-pigs hearts. After 10 min pulse-labelling, LPL protein was eluted as two peaks from heparin-agarose: peak 1 at about 0.65 M NaCl, peak 2 at about 0.95 M NaCl. Catalytic activity was associated only with peak 2. Model studies with bovine LPL showed that active dimeric LPL is eluted in peak 2, but after treatments that dissociate the enzyme into inactive monomers it is eluted in peak 1. Pulse-labelled LPL in both peak 1 and peak 2 was fully sensitive to treatment with endoglycosidase (Endo) H. With chase, peak 1 disappeared and peak 2 acquired resistance to Endo H. These findings suggest that core glycosylated LPL is assembled into dimers already in the endoplasmic reticulum and that processing of the oligosaccharide chains occurs after dimerization.

Animals↗
PubMed1993

Expression, targeting, and assembly of functional Na,K-ATPase polypeptides in baculovirus-infected insect cells.

Deciphering the roles of the individual subunits of the heteromeric Na,K-ATPase in the structure, function, and assembly of this enzyme has been complicated because most expression systems have endogenous levels of Na,K-ATPase activity. This problem has become even more significant in light of the recent identification of multiple isoforms for both the alpha and beta subunits. The baculovirus expression system, which uses the baculovirus Autographica californica to infect insect cells, affords two distinct advantages for the expression of the Na,K-ATPase; some insect cells have little or no levels of Na,K-ATPase, and baculovirus-infected cells produce extremely high levels of foreign protein. We have made two separate recombinant baculoviruses containing the rodent alpha 1 or beta 1 cDNAs and used them to infect the insect cell line Sf-9. The infected Sf-9 cells produce Na,K-ATPase subunit protein on the order of 5-10 micrograms of protein/ml of cultured cells. The rodent alpha 1 polypeptide produced in the Sf-9 cells is indistinguishable electrophoretically and antigenically from the native subunit. The expressed beta 1 subunit is also antigenically identical but has a higher electrophoretic mobility due to differential glycosylation by the infected Sf-9 cell. In contrast to other systems, when expressed alone, each individual Na,K-ATPase subunit is targeted to the infected Sf-9 plasma membrane. In contrast, when infected with a virus that induces the heavy chain of murine IgG, the infected Sf-9 cell retains the polypeptide in the endoplasmic reticulum. However, when both IgG light and heavy chains are expressed, the polypeptides are properly processed and secreted. When the Na,K-ATPase alpha 1 and beta 1 polypeptides are simultaneously expressed, they form detergent-resistant complexes that are functional. Ouabain-sensitive ATPase activity on the order of 5 mumol Pi/mg/h in infected Sf-9 membranes was dependent on the expression of both the alpha 1 and beta 1 subunits. Sodium-dependent phosphorylated intermediates were detected that were potassium- and ouabain-sensitive. No increase in ouabain-sensitive activity or phosphorylated intermediates was detected when either subunit was expressed alone. The alpha 1 beta 1-coinfected cells were also able to transport ions, as detected in 86Rb uptake experiments. Thus, the recombinant Na,K-ATPase expressed in insect cells is biologically active and is suitable for structural and functional analysis.

Animals↗
PubMed1993

Fast echo-shifted gradient-recalled MRI: combining a short repetition time with variable T2* weighting.

The principles of a fast T2*-sensitized MR imaging method (Moonen et al., Magn. Reson. Med. 26, 184 (1992)) are extended to further increase T2* sensitivity. It is shown that the period of T2*-weighting can be lengthened by n TR-periods by appropriate gradient schemes without RF refocusing resulting in progressively delayed gradient-recalled echoes. This extension of the echo-shifting concept thus introduces large flexibility in the choice of T2*-weighting without changing total imaging time. The coherence pathway formalism is used to evaluate and describe the selection of the desired echo and the attenuation of unwanted coherences. The new techniques are demonstrated for tracking a bolus of susceptibility contrast agent in cat brain. Relative blood-volume maps are derived with expected contrast between white and gray matter.

Animals↗
PubMed1993

A functional MRI technique combining principles of echo-shifting with a train of observations (PRESTO).

We present a fast MRI technique sensitized to microscopic susceptibility effects. The method combines elements of echo-shifted gradient-recalled MR imaging (TE > TR) with the acquisition of multiple k-space lines within a single TR-period. The sequence results in a much reduced imaging time as compared with conventional gradient-echo MRI methods. The feasibility of the method is demonstrated for susceptibility bolus tracking in the cat brain using an imaging time of 153 ms. The relative cerebral blood volume maps created with this method are comparable with those obtained with conventional methods.

Animals↗
PubMed1993

The vascular pathology of human hepatic allografts.

Studies of hepatic transplant vasculopathy (TV) have described arterial lesions in selected cases but have not sought to establish the relative prevalence of various arterial and venous lesions. We examined 20 unselected liver allografts and 12 controls to establish the spectrum and distribution of transplant-related arterial and venous lesions. Sections of 1,175 arteries and 936 portal and hepatic veins were reviewed and several histologic parameters were tabulated. Fibrotic (18 cases: 184 arteries and 17 veins) and cellular (10 cases: 353 arteries and 33 veins) lesions were seen. Three allografts had evidence of severe TV with abundant foam cell, myointimal, and mixed fibrocellular lesions in the intima of arteries. These three grafts also had foam cells in portal veins. Allografts with TV had more stenotic arteries (P < .05) and more intimal cellular proliferation (P < .05) compared with non-TV allografts and with controls. Foam cells often demonstrated immunoperoxidase staining for actin, lysozyme, or UCHL-1. Two of the TV cases had marked paucity of bile ducts and required retransplantation.

Adult↗
PubMed1993

Nursing diagnoses for psychiatric patients in China.

This report describes the development of a preliminary Chinese psychiatric classification system with nine categories and 211 separate diagnoses, and the corresponding diagnostic interview that enables psychiatric nurses in China to systematically assess the presence or absence of the various diagnoses. Using this interview with 30 representative inpatients, the authors identified 152 of the 211 (72%) diagnoses. On average, each patient was given 26 different nursing diagnoses. The interrater reliability of the four nurses who independently coded the diagnostic interview for these 30 patients was excellent: in over 77% of the identified diagnoses the generalized kappa was greater than 0.75. These findings show that further work on this classification of nursing diagnoses is warranted.

Adolescent↗