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Biomedical subjects

X Gu

Publications and source records attributed to X Gu.

At least 235 records · Page 13Linked to original sources
PubMed1995

Regulation of sialyltransferase activities by phosphorylation and dephosphorylation.

The composition of tissue gangliosides is thought to result mainly from the active regulation and selective expression of specific enzymes responsible for their metabolism. In the last few years, we have purified several rat brain sialyltransferases to homogeneity; the availability of these highly purified enzymes enabled us to investigate their regulation and expression at the molecular level. Thus, we studied the regulation of sialyltransferase activities, in particular, CMP-NeuAc:GM1 and CMP-NeuAc:LacCer sialyltransferases by a phosphorylation/dephosphorylation mechanism. Protein kinase C was added to a standard enzyme assay mixture containing [gamma-32P]ATP, and the activity of the enzyme was measured after various incubation times. We found that treatment of several sialyltransferases by protein kinase C decreased their activities in a time-dependent manner. Analyses of 32P-labeled amino acids revealed that the major phosphorylation site of CMP-NeuAc:GM1 alpha 2-->3 sialyltransferase (ST-IV) was serine and that for CMP-NeuAc:LacCer alpha 2-->3 sialyltransferase (ST-I) was primarily threonine. Partial recovery of the enzyme activity could be achieved by treatment of the phosphorylated sialyltransferases with rat brain protein phosphatase. We conclude that the activities of sialyltransferases can be modulated by protein kinase C and protein phosphatase and this may represent a potential regulatory mechanism for ganglioside biosynthesis.

Adenosine Triphosphate↗
PubMed1995

Maximum likelihood estimation of the heterogeneity of substitution rate among nucleotide sites.

This paper presents a maximum likelihood approach to estimating the variation of substitution rate among nucleotide sites. We assume that the rate varies among sites according to an invariant+gamma distribution, which has two parameters: the gamma parameter alpha and the proportion of invariable sites theta. Theoretical treatments on three, four, and five sequences have been conducted, and computer program have been developed. It is shown that rho = (1 + theta alpha)/(1 + alpha) is a good measure for the rate heterogeneity among sites. Extensive simulations show that (1) if the proportion of invariable sites is negligible, i.e., theta = 0, the gamma parameter alpha can be satisfactorily estimated, even with three sequences; (2) if the proportion of invariable sites is not negligible, the heterogeneity rho can still be suitably estimated with four or more sequences; and (3) the distances estimated by the proposed method are almost unbiased and are robust against violation of the assumption of the invariant + gamma distribution.

Computer Simulation↗
PubMed1995

Signalling pathways in cardiac failure.

UNLABELLED: 1. Cardiac failure in humans and in animal models is associated with a marked desensitization of the catecholamine signalling pathway. 2. Beta 1- and beta 2- and possibly beta 3-adrenoceptors (beta-AR) are found in the hearts of humans and common laboratory animals such as rats and guinea-pigs. In rats and guinea-pigs chronic stimulation of cardiac beta-AR leads to a rapid loss of beta 2-AR whereas heart failure in humans is associated with a loss of beta 1-AR or beta 1-AR and beta 2-AR. 3. Desensitization is also associated with phosphorylation of beta-AR by beta-AR kinase (beta-ARK) and uncoupling of receptors from the signalling pathway. Beta-ARK but not beta-arrestin activity and mRNA are markedly increased in heart failure. 4. Chronic beta-AR stimulation and heart failure are associated with increases in Gi alpha but little if any change in Gs alpha. 5. The roles of beta gamma subunits of G-proteins, adenylate cyclase subtypes and cAMP dependent protein kinase A in heart failure are unclear at present. ABBREVIATIONS: beta-ARK - beta-adrenoceptor kinase AR - adrenoceptor G-protein - GTP binding protein

Animals↗
PubMed1995

Effect of pH on potassium and proton conductance in renal proximal tubule.

The effect of intracellular and extracellular pH on potassium conductance (GK) was examined in isolated amphibian (Rana pipiens) proximal tubule cells under whole cell voltage clamp conditions. Internal perfusion of the patch pipette was used to precisely control intracellular pH. In the region of normal resting potential (-51 +/- 3 mV), raising cell pH from 6.5 to 8.0 did not significantly increase GK (1.1 +/- 0.3 vs. 1.3 +/- 0.3 nS; P > 0.08, n = 8). Similar elevations in external (bath) pH had even less of an effect on GK. In contrast, when cells were voltage clamped to 30 mV more negative than the resting potential, raising internal pH from 6.5 to 8.0 did increase GK from 1.05 +/- 0.3 to 1.8 +/- 0.5 nS (P < 0.04; n = 8). These results suggest that modest changes in pH have little effect on GK, except at large negative potentials. In the process of examining the pH dependence of GK, a slowly activating, voltage-dependent conductance of 7.5 +/- 1 nS (n = 20; for 20 microns cells) was observed during cell depolarization. Although the instantaneous current-voltage relation of this conductance was linear, its marked voltage dependence produced an apparent steady-state rectification, with Gm = 0.5 +/- 0.2 nS and Gout = 9.0 +/- 1 nS (n = 11). Outward current was reversibly blocked by 3 mM Cu, Cd, or Co. In the absence of Na, K, and Ca (and only trace amounts of Cl), rapid changes in bath pH from 6.5 to 8.0 shifted the steady-state reversal potential (Erev) by -37 +/- 4 mV (n = 9) and the instantaneous Erev by -56 +/- 4 mV (n = 9). These shifts in Erev were consistent with a hydrogen ion conductance (GH), similar to what has been reported for snail neuron, neutrophils, alveolar epithelial cells, and phagocytes. Since the magnitude of this GH would be insignificant at resting cell pH and membrane potential, its role in renal proximal tubule under normal conditions is somewhat obscure. Nonetheless, in pathological situations, GH could function to prevent acid overload during any process that depolarizes the cell, such as low temperature or metabolic inhibition.

Animals↗
PubMed1995

Insulin-like growth factor-I promotes cell proliferation in the absence of modulation of collagen phenotypes and utilizes IRS-1, not PLC-gamma 1, in corneal endothelial cells.

Corneal endothelial cells are differentiated cells and are thus incapable of physiologic regeneration. In a search for a growth factor that would promote optimal proliferation of corneal endothelial cells in the absence of other modulating activities, the effect of insulin-like growth factor-I (IGF-I) on rabbit corneal endothelial cells was studied. In addition, cellular effector molecules responsible for the signal pathway for IGF-I were studied. IGF-I at 50 ng/ml stimulated corneal endothelial cell proliferation after at least 8 h of treatment. IGF-I did not change cell shape of corneal endothelial cells: the cells treated with IGF-I at 50 ng/ml maintained polygonal morphology regardless of the duration of exposure. IGF-I did not alter collagen phenotypes either qualitatively or quantitatively: the treated cells continued to synthesize types IV and VIII collagen, as did the control cells. The steady-state levels of alpha 2(I) collagen RNA and alpha 2(IV) RNA were not altered by IGF-I treatment. Immunohistochemical analysis showed that IGF-I is present in corneal endothelium in vivo, while the underlying Descemet's membrane demonstrated no staining. Corneal endothelial cells also produce IGF binding protein-2 (IGFBP-2), which appears to bind IGF-I that has been introduced exogenously in the medium. Further investigation as to how the signals of IGF-I were transmitted for the biological activities demonstrated that the expression of insulin receptor substrate-1 (IRS-1) is up-regulated by IGF-I treatment, while PLC-gamma 1 expression is not altered by this growth factor.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

[Causes of postoperative recurrent varicose veins of lower extremities].

We analysed venograms of recurrent varicose veins in 180 cases (206 lower limbs). Deep vein valvular insufficiency resulted in varicose veins in 52.9% cases, and post-thrombotic syndrome in 18.9%. The causes of postoperative recurrent varicose veins varied: blindly performed single superficial veins operation, low ligation of the long saphenous trunk, incomplete stripping of varicose veins, missing of the short varicose saphena or incomplete ligation of the perforating veins and failure of deep vein valvular repairs. Venography is of value in making correct diagnosis and choosing appropriate operations.

Adult↗
PubMed1995

Chemotropism in nerve regeneration studied in tissue culture.

The neurotropic effect of the distal stump of transected sciatic nerve on regenerating axons emerging from the proximal stump was investigated in rats by the coculture of excised neonatal dorsal root ganglia and segments of degenerated and nondegenerated sciatic nerves. In all cultures, neurites from the ganglion extended directly towards the degenerated distal stump and not towards the undegenerated nerve segment. Analysis of the supernatants of nerve homogenates by native polyacrylamide gel electrophoresis demonstrated that 250 and 18 kDa proteins were upregulated in the degenerated distal stumps and that 89, 67 and 65 kDa proteins were expressed by these stumps but not by normal nerve. Placement of the individual protein strips, cut from the gels, in dissociated cultures of dorsal root ganglion cells showed that the 89 and 18 kDa strips possessed strong neurotropic activity. Neurons survived selectively on these strips and outgrowing neurites elongated on the strips, parallel to the direction of the band. When explanted dorsal root ganglia were cultured along with the strips, outgrowing neurites extended selectively towards those containing the 89 and 18 kDa bands. It is concluded that these bands contain chemotropic substances, the nature of which requires further investigation.

Animals↗
PubMed1995

Encircling suture method in the treatment of femoral venous valvular incompetency.

Many patients with chronic venous insufficiency syndrome caused by the "relative valvular incompetency" with venous dilatation. Eighty-six per cent of the cases can be cured by reducing the femoral venous diameter by simply using the encircling suture method just distal to the first femoral valvular sinus. Our preliminary results suggest that such a method of surgical treatment is simple and efficient.

Adult↗
PubMed1994

A panel of radiation hybrids for human chromosome 8.

We have developed a panel of radiation hybrids containing fragments of chromosome 8 as the only human material. The human chromosome content of each cell line was determined relative to an ordered map of sequence tagged sites (STSs) specific to chromosome 8. Between one and four fragments of chromosome 8 were identified in each cell line, with an average of 25% of the STSs retained in each line. Subclones of one radiation hybrid were examined to determine whether all cells within a line are homogeneous with respect to chromosome 8 sequence content. There was considerable variability between subclones, with retention rates for individual STSs ranging from 5 to 100% in different clones. Furthermore, a gradient of retention of sequences along the length of one large chromosome fragment was found, suggesting that sequence loss involved deletions from one end of the fragment at early stages in the establishment of the cell line. We have also made use of the radiation hybrids to develop novel sequence tagged sites for the pericentromeric region of chromosome 8.

Adenine Phosphoribosyltransferase↗
PubMed1994

In vitro methylation of Escherichia coli 16S rRNA by tRNA (m5U54)-methyltransferase.

16S rRNA, isolated from Escherichia coli or synthesized in vitro, is methylated by tRNA (m5U54)-methyltransferase (RUMT) and S-adenosyl-L-methionine to give ribothymidine (m5U). By methylation studies of 16S rRNA fragments, nearest-neighbor analysis, and nuclease protection experiments, the site of methylation was identified as U788. We have previously shown that the substrate consensus sequence for the T-arm of tRNA consists of a 2-5 base-pair stem and a 7-base loop, with certain constraints on base substitutions within the loop, and in the first two bases which close the loop [Gu, X., & Santi, D. V. (1991) Biochemistry 30, 2999-3002]. U788 of 16S rRNA is within a 9-base loop of a predicted stem-loop structure of 16S rRNA. If Ado substitution is allowed at the third and seventh positions of the loop and the first and ninth bases of the loop form an A-C base pair, the resulting stem-loop falls within the RUMT consensus sequence of the T-arm of tRNA. Individual mutants of the tRNA T-arm at these positions confirm that the substitutions are allowable, and expand the previous consensus sequence. Further, prevention of 7-base loop formation by requiring C-C base-pair formation at the loop closure abolishes substrate activity. RUMT forms a complex with Syn 16S rRNA which can be isolated on nitrocellulose filters or by SDS-PAGE electrophoresis. The enzyme also catalyzes exchange of tritium of [3H]Ura-16S rRNA for protons of water. By analogy with studies with tRNA [Gu, X., & Santi, D. V. (1991) Biochemistry 31, 10295-10302], the mechanism of methylation is proposed to involve formation of a covalent, albeit reversible, Michael adduct with the target U788 of 16S rRNA.

Base Sequence↗
PubMed1994

Lung cancer among tin miners in southeast China: silica exposure, silicosis, and cigarette smoking.

The Dachang Tin Mine is an industrial facility with high rates of lung cancer compared to the resident population in Guangxi province in southern China. Historically, exposure to silica dust was very high in the 1950s, falling in recent years to levels in keeping with international standards. Radon levels in the mine are low. We report the findings of a case-control study nested into a cohort study on miners working in Dachang. Cases of lung cancer among miners incident from 1973-1989 were obtained from local comprehensive medical records covering workers employed at the mine. These were matched approximately 3 to 1 with miners randomly chosen from the district surrounding the mine within the same birth decade. Matched odds ratios of 2.42 (95% confidence limit [CL] 1.3, 4.4) for underground employment, 3.52 (95% CL 1.7, 7.5) for smoking, and 2.04 (95%) CL 1.2, 3.7) for silicosis as determined on chest film were noted. Multifactor analysis of unconditional logistic regression showed that among the risk factors for excess mortality from lung cancer only the years spent drilling underground and the cumulative smoking index (product of daily cigarette consumption and number of years smoking) were independent contributors to risk and there was no interaction observed. The presence of silicosis did not contribute to predicting risk independently of the years spent underground.

Case-Control Studies↗
PubMed1994

Patterns of biosynthesis and accumulation of hydrocarbons and contact sex pheromone in the female German cockroach, Blattella germanica.

De novo synthesis of contact female sex pheromone and hydrocarbons in Blattella germanica was examined using short in vivo incubations. Accumulation of pheromone on the epicuticular surface and the internal pheromone titer were related to age-specific changes in hydrocarbon synthesis and accumulation in normal and allatectomized females. The incorporation of radiolabel from [1-14C]propionate into the cuticular methyl ketone pheromone fraction was positively related to corpora allata activity during two gonotrophic cycles. During peak pheromone production the total internal lipid fraction contained greater titers of pheromone than the cuticular surface, and it too exhibited a cycle internally, preceding the rise in external pheromone. This suggests that synthesis and accumulation of pheromone internally are followed by transport of pheromone to the epicuticular surface where it accumulates. Radiolabel was incorporated efficiently into both cuticular and internal hydrocarbons after the imaginal molt and until the peak of pheromone synthesis, but it declined to lower levels before ovulation and throughout pregnancy. The internal hydrocarbon titer decreased 58% after oviposition, suggesting deposition in the egg case. It remained relatively unchanged during pregnancy and increased again during the second gonotrophic cycle. In allatectomized females, hydrocarbon synthesis was reduced relative to control females until oviposition in the latter. However, subsequent rates of hydrocarbon synthesis in allatectomized females (without oothecae) exceeded the rates in sham-operated females (with oothecae). In the absence of ovarian uptake of hydrocarbons, the internal titer increased without the decline found in control females at oviposition. As internal hydrocarbons increased, so did cuticular hydrocarbons and both internal and cuticular methyl ketone pheromones. These patterns corresponded well with feeding patterns in sham-operated and allatectomized females, suggesting that pheromone production is normally regulated by stage-specific feeding-induced hydrocarbon synthesis (precursor accumulation internally) and juvenile hormone-induced conversion of hydrocarbon to pheromone. They also suggest that both the cuticle and the ovaries might be target sites for hydrocarbon and possibly methyl ketone deposition.

Aging↗
PubMed1994

A model for the correlation of mutation rate with GC content and the origin of GC-rich isochores.

Based on the biochemical kinetics of DNA replication and mutagenesis, including misincorporation and correction, a model has been developed for studying the relationships among the mutation rate (u), the G+C content of the sequence (f), and the G+C proportion in the nucleotide precursor pool (N). Also a measure for the next-nucleotide effect, called the maximum capacity of the next-nucleotide effect (MC), has been proposed. Under the normal physiological conditions of mammalian germ cells, our results indicate: (1) the equilibrium G+C content in a sequence is approximately equal to the G+C proportion in the nucleotide precursor pool, i.e., f approximately N, which is independent of the next-nucleotide effect; (2) an inverted-V-shaped distribution of mutation rates with respect to G+C contents is predicted, when the next-nucleotide effect is week, i.e., MC approximately 1; (3) the distribution becomes flatter (i.e., inverted-U-shaped) as MC increases, but the peak at 50% GC is still observed when MC < 2; and (4) the peak disappears when MC > 2.8, that is, when the next-nucleotide effect becomes strong. Our results suggest that changes in the relative concentrations of nucleotide precursors can cause variations among genes both in mutation rate and in G+C content and that compositional isochores (DNA segments with a homogeneous G+C content) can arise in a genome due to differences in replication times of DNA segments.

Base Composition↗
PubMed1994

Tityustoxin-K alpha, from scorpion venom, blocks voltage-gated, non-inactivating potassium current in cultured central neurons.

Whole-cell voltage-clamp was used to examine the effects of tityustoxin-K alpha (TsTX-K alpha), from the venom of the scorpion Tityus serrulatis, on voltage-gated K+ currents in cultured hippocampal neurons and cerebellar granule cells. Slowly activating, noninactivating outward currents (IK) were generated by depolarizing steps from a holding potential of -60 mV to potentials positive to -40 mV. TsTX-K alpha produced a dose-dependent block of the sustained outward current. The fraction of total current blocked ranged from 10 to 60% over a concentration range of 2.5-120 nM in both cerebellar and hippocampal neurons. A hyperpolarizing prepulse to -100 mV was used to generate a rapidly inactivating current with properties like those of IA. When IA was isolated pharmacologically (with 5-10 mM TEA to block IK) or by subtracting IK from total outward current, TsTX-K alpha had no effect on the IA in either cell type. TsTX-K alpha also had no apparent effect on the leak conductance or on the inward rectifier current in these cells. The data indicate that TsTX-K alpha in cultured mammalian neurons is a potent and selective blocker of a voltage-gated, non-inactivating K+ current with properties like those of a delayed rectifier.

Animals↗
PubMed1994

Enzymatic mechanism of tRNA (m5U54)methyltransferase.

tRNA (m5U54)methyltransferase (RUMT) catalyzes the methylation of uridine 54 of transfer RNA by S-adenosyl-L-methionine. In this report, we present the enzymatic mechanism of RUMT, including the stereochemical course of the methylation reaction, and discuss RUMT-tRNA recognition. As part of its enzymatic mechanism, we postulate that RUMT catalyzes the disruption of RNA-RNA contacts. We also show that many nucleotide substitutions can be made in the T-loop of tRNA without affecting RUMT binding, indicating that the recognition of the T-loop by RUMT is not stringent.

Amino Acid Sequence↗
PubMed1994

Action potentials, calcium transients and the control of differentiation of excitable cells.

Calcium influx via action potentials in differentiating nerve and muscle is regulated principally by the expression of potassium currents. Transient elevations of intracellular calcium in spontaneously active cells are necessary for normal neuronal development. The mechanisms that connect calcium elevations to long term developmental change are likely to be utilized in the mature nervous system.

Action Potentials↗
PubMed1994

Further studies of a maximum-likelihood yes-no procedure.

Absolute thresholds were estimated for pure tones of 500, 1000, and 2000 Hz using both a maximum-likelihood yes-no procedure and a traditional three-down one-up forced-choice adaptive procedure. Threshold estimates obtained with the two procedures were highly correlated. In the maximum-likelihood yes-no procedure, the false-alarm probability was poorly estimated. A simple change in that procedure produced less-biased estimates of the false-alarm rate. In a second experiment, the influence of the false-alarm rate on the threshold estimates was investigated. The listeners' absolute thresholds were estimated when they adopted either a liberal or conservative criterion for detecting the signals. Data were collected using both a constant-stimulus yes-no procedure and a maximum-likelihood yes-no procedure. Threshold estimates are 5 to 6 dB higher for the low than for the high false-alarm rates. Psychometric functions based on a stimulus-power model provided the best fit to the data obtained with the constant-stimulus method. This stimulus-power model was used to produce the assumed psychometric function in the maximum-likelihood yes-no procedure. Computer simulations were conducted to determine how threshold estimates were affected by the listener's inattention or by mismatches between the slope parameter of the listener's psychometric function and that assumed in the maximum-likelihood procedure.

Attention↗