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Biomedical subjects

G Zhang

Publications and source records attributed to G Zhang.

At least 505 records · Page 28Linked to original sources

Abnormalities in hepatic lipase in chronic renal failure: role of excess parathyroid hormone.

Post-heparin hepatic lipase activity is reduced in chronic renal failure (CRF). This could be due to reduced synthesis, decreased activity, and/or impaired secretion of the enzyme. Further, the factor(s) responsible for such derangements are not elucidated. We examined hepatic lipase metabolism in normal, 6-wk-old CRF rats, CRF-PTX (parathyroidectomized) rats, and CRF and normal rats treated with verapamil (CRF-V, normal-V) using liver homogenate, hepatic cell culture for 8 h, and in vitro liver perfusion. The Vmax of hepatic lipase in liver homogenate was significantly (P < 0.01) reduced and the Km was significantly (P < 0.01) increased in CRF rats, but the values were normal in CRF-PTX, CRF-V, and normal-V rats. Culture of hepatic cells for 8 h was associated with an increase in hepatic lipase activity but the increment in CRF rats was significantly (P < 0.01) lower than that of normal, CRF-PTX, CRF-V, and normal-V rats. Both parathyroid hormone (PTH)-(1-84) and 1-34 inhibited the production of hepatic lipase in cultured cells from normal, CRF-PTX, CRF-V, and normal-V rats. The expression of the mRNA of the hepatic lipase was significantly reduced in CRF animals with the ratio between it and that of house keeping gene G3DPH being 15 +/-3% compared to 40 +/- 1.3% in normal, 44+/-2.9% CRF-PTX, 44 +/- 5.4% in CRF-V, and 39 +/- 3.9% in normal-V rats. Infusion of heparin to the in vitro hepatic perfusion system increased the activity of hepatic lipase in the effluent in all groups of rat except in CRF animals. Infusion of PTH-(1-34) in dose of 10(-6) M into the liver perfusion system inhibited the increase in post-heparin hepatic lipase activity. The data show that in CRF (a) the mRNA of hepatic lipase is downregulated, and hepatic lipase production, activity and release are impaired, (b) that this is due to the state of secondary hyperparathyroidism of CRF since both acute and chronic excess of PTH were associated with these abnormalities, (c) and that prevention of excess PTH by PTX of CRF rats or blocking the effect of PTH by treatment with verapamil corrected the derangement in hepatic lipase metabolism.

Animals↗

v-rel and c-rel induce a membrane protein, p75, on avian hematopoietic cells.

v-rel-induced neoplasias are thought to be caused by the aberrant expression of cellular genes. Using a monoclonal antibody we detected a 75 kDa-protein (p75) on v-rel-transformed chicken hematopoietic cells. This protein is normally expressed only on a certain type of non-T, non-B cells in the chicken caecal tonsils. Overexpression of v-rel in two B-cell lines and in splenic cells induced the expression of p75. In contrast, overexpression of c-rel in these cells resulted in variable expression of p75. These data indicated that p75 is ectopically expressed on v-rel-transformed hematopoietic cells and that it responds differently to v-rel and c-rel induction.

Animals↗

Quantitative distribution of muscle fiber types in the scup Stenotomus chrysops.

Because the mass-specific power generated by myotomal muscle during swimming varies along the length of the fish, a realistic assessment of total power generation by the musculature requires integrating the product of mass-specific power and muscle mass at each position over the length of the fish. As a first step toward this goal, we examined the distribution of red, pink, and white muscle along the length of Stenotomus chrysops (scup) using histochemical and image analysis techniques. The largest cross-sectional area of red fibers occurs at 60% of total fish length and declines both anteriorly and posteriorly. By contrast, white fibers have the largest cross-sectional area in the anterior and decline dramatically moving posteriorly. The proportion of the fishes' cross-section occupied by red fibers increases from 1.37% to 8.42% moving posteriorly along the length of the fish. In contrast, the proportion of cross-sectional area occupied by pink fibers is constant (1.19%), while the proportional cross-sectional area of white fibers falls from 82.5% to 66.3%. The red, pink, and white fibers comprise 2.09, 0.73, and 51.1%, respectively, of total fish weight. We also compared the distribution of muscle in 10 degrees C- and 20 degrees C- acclimated animals. The value for red fiber volume, though slightly higher (13%) in cold-acclimated fish, is not statistically different. No difference was found in pink or white fibers. Finally, the finding that most of the red muscle is in the posterior half of the fish further supports the notion that most power for steady swimming at moderate speeds comes from posterior rather than anterior musculature.

Animals↗

Construction of expressing plasmids of recombinant FN polypeptides with bifunctional-domain and the characterization of the products expressed in E. coli.

Two expressing plasmids have been constructed and used to express two bifunctional-domain recombinant polypeptides of human fibronectin (FN) in E. coli. One was CH50 (Pro1239-Ser1515 of FN linked with Ala1690-Thr1960 of FN through Met) and the other was CH56 (Pro1239-Thr1960 of FN). Both of two polypeptides were capable of binding heparin and were purified by heparin-agarose affinity chromatography. The purified products were capable of binding cells. The production of CH50 and CH56 polypeptides provided a fundamental basis for further study of the anti-metastatic function of recombinant fibronectin polypeptides.

Escherichia coli↗

Long-term compressive property durability of carbon fibre-reinforced polyetheretherketone composite in physiological saline.

In total hip arthroplasty, concerns such as corrosion and stress shielding associated with stiff metallic femoral components have led to the development of low stiffness advanced fibre-reinforced polymer (FRP) composite femoral components. Carbon fibre-reinforced polyetheretherketone (CF/PEEK) composite material is now one of the primary material systems being considered for composite hip stem development. As a hip stem, a composite material must be able to support a complex state of stress in the in vivo environment without failure. Considering the loading conditions of a hip stem (superimposed compression and bending), and the fact that FRP composites typically possess lower compressive than tensile strength, the compressive behaviour of FRP composites becomes very important for femoral component design. This paper presents an investigation of the long-term durability of 0 degree and 90 degrees compressive strengths of CF/PEEK composite following physiological saline saturation. 0 degree and 90 degrees compressive moduli and Poisson ratio (v12) properties are also reported. Samples were tested following conditioning in physiological saline at 37, 65 and 95 degrees C for time periods from 0 to 5000 h. Dry samples were tested as controls. Results show no significant loss in compressive property values of the saline-saturated or the dry control samples as a function of conditioning time or temperature.

Benzophenones↗

Localization of HIV-1 RNA in mammalian nuclei.

The Rev protein of human immunodeficiency virus type 1 (HIV-1) facilitates the nuclear export of unspliced and partially spliced viral RNAs. In the absence of Rev, these intron-containing HIV-1 RNAs are retained in the nucleus. The basis for nuclear retention is unclear and is an important aspect of Rev regulation. Here we use in situ hybridization and digital imaging microscopy to examine the intranuclear distributions of intron-containing HIV RNAs and to determine their spatial relationships to intranuclear structures. HeLa cells were transfected with an HIV-1 expression vector, and viral transcripts were localized using oligonucleotide probes specific for the unspliced or spliced forms of a particular viral RNA. In the absence of Rev, the unspliced viral RNAs were predominantly nuclear and had two distinct distributions. First, a population of viral transcripts was distributed as approximately 10-20 intranuclear punctate signals. Actinomycin D chase experiments indicate that these signals represent nascent transcripts. A second, stable population of viral transcripts was dispersed throughout the nucleoplasm excluding nucleoli. Rev promoted the export of this stable population of viral RNAs to the cytoplasm in a time-dependent fashion. Significantly, the distributions of neither the nascent nor the stable populations of viral RNAs coincided with intranuclear speckles in which splicing factors are enriched. Using splice-junction-specific probes, splicing of human beta-globin pre-mRNA occurred cotranscriptionally, whereas splicing of HIV-1 pre-mRNA did not. Taken together, our results indicate that the nucleolus and intranuclear speckles are not involved in Rev regulation, and provide further evidence that efficient splicing signals are critical for cotranscriptional splicing.

Biological Transport↗

A transcription map of the DiGeorge and velo-cardio-facial syndrome minimal critical region on 22q11.

The majority of patients with DiGeorge syndrome (DGS) and velo-cardio-facial syndrome (VCFS) have a microdeletion of 22q11. Using translocation breakpoints and fluorescence in situ hybridization analysis (FISH), the minimal DiGeorge critical region (MDGCR) has been narrowed to 250 kb in the vicinity of D22S75 (N25). The construction of a detailed transcription map covering the MDGCR is an essential first step toward the identification of genes important to the etiology of DGS/VCFS, two complex disorders. We have identified a minimum of 11 transcription units encoded in the MDGCR using a combination of methods including cDNA selection, RT-PCR, RACE and genomic sequencing. This approach is somewhat unique and may serve as a model for gene identification. Of the 11 transcripts, one is the previously reported DGCR2/IDD/LAN gene, and three revealed a high level of similarity to mammalian genes: a Mus musculus serine/threonine kinase, a rat tricarboxylate transport protein and a bovine clathrin heavy chain. The remaining transcripts do not demonstrate any significant homology to genes of known function. The identification of these transcription units in the MDGCR will facilitate their further characterization and help elucidate their role in the etiology of DGS/VCFS.

Abnormalities, Multiple↗

Analysis of impact of rigid projectiles on compound targets.

We developed a model to simulate a person, with and without protective armor, being subjected to an impact of a high speed, rigid projectile. The complex nature of the head and chest are simulated in our layered, spaced, various thickness targets made of different materials. A rigid projectile-target interaction model is described. Target resistance force was obtained from a generalized Poncelet equation. Good agreement between observed and computed penetration phenomena was obtained.

Computer Simulation↗

Cell death induced by cytopathic bovine viral diarrhoea virus is mediated by apoptosis.

Cells infected with two closely related isolates of bovine viral diarrhoea virus (BVDV), one cytopathic (CP) and one non-cytopathic (NCP), were examined for signs of apoptosis. The results from labelling DNA using terminal transferase and biotinylated dUTP and by observing oligonucleosomal-sized DNA fragmentation indicated that the CP strain of BVDV induced apoptosis in cell culture but the NCP strain did not. Induction of apoptosis correlated with infected cells undergoing apoptosis rather than interactions between infected and uninfected cells and the induction of apoptosis by CP BVDV was a dominant trait in cells co-infected with both types of virus.

Animals↗

Induction of FGF-7 after kidney damage: a possible paracrine mechanism for tubule repair.

A member of the fibroblast growth factor (FGF) family, keratinocyte growth factor (FGF-7 has unique specificity for epithelial cells. We investigated the role of FGF-7 in repair of proximal tubular damage caused by S-(1,1,2,2-tetrafluoroethyl)-L-cysteine (TFEC). In situ hybridization localized FGF-7 to interstitial cells in the medulla and outer stripe of the outer medulla. Interstitial FGF-7 expression increased throughout the kidney 1 day after TFEC treatment. FGFR2 IIIb mRNA was high in the papilla and medulla and also increased after TFEC administration. By in situ hybridization, FGFR2 IIIb was localized to the tubular epithelium, particularly in collecting ducts. Proliferation of collecting duct epithelial cells increased in adult kidney after damage to the proximal tubule. FGFR2 IIIb, but not FGF-7, mRNA was also expressed by rat proximal tubule epithelial (RPTE) cells in vitro, and FGF-7 increased DNA synthesis in RPTE. Thus FGFR2 IIIb and FGF-7 expression is segregated between epithelial and interstitial cells forming a paracrine growth factor loop. These results raise the possibility that a novel paracrine growth loop is activated by chemical damage and regulates epithelial cell growth during tubular repair.

Aging↗

Gene conversion in the CYP11B2 gene encoding P450c11AS is associated with, but does not cause, the syndrome of corticosterone methyloxidase II deficiency.

Cytochrome P450c11AS (aldosterone synthase) has 11 beta-hydroxylase, 18-hydroxylase, and 18-oxidase activities and is expressed solely in the adrenal zona glomerulosa. Corticosterone methyloxidase II (CMOII) deficiency denotes a rare disorder of adrenal steroidogenesis in which only the 18-oxidase activity of P450c11AS is disrupted, while the 11 beta-hydroxylase and 18-hydroxylase activities persist. Such patients have elevated serum concentrations of corticosterone and 18-hydroxycorticosterone and very low or unmeasurable concentrations of aldosterone, often resulting in a clinical salt-losing crisis in infancy. One pair of point mutations, Arg181-->Trp and Val386-->Ala, has been previously characterized to cause this disorder in an inbred Iranian Jewish population. We have sought mutations causing CMOII deficiency in outbred populations. In three of four unrelated P450c11AS alleles from two unrelated patients with CMOII deficiency, we found a gene conversion event in which exons 3 and 4 of the CYP11B2 gene encoding P450c11AS were changed to the sequence of the nearby CYP11B1 gene, which encodes the related enzyme P450c11 beta. This conversion resulted in a mutant P450c11AS protein carrying three changes: Asp141-->Glu, Lys151-->Asn, and Ile246-->Thr. We built seven vectors expressing P450c11AS carrying each mutation singly, each of the three possible pairs of mutations, and the triple mutation as found in the proband. The activities of both the normal P450c11AS and the various mutants in transfected nonsteroidogenic COS-1 cells were very low, but their activities in steroidogenic MA-10 and JEG-3 cells were 10- to 20-fold higher. In these systems all of the mutants retained normal 18-oxidase activity, indicating that the detected gene conversion event is associated with but does not cause CMOII deficiency. None of the four CYP11B2 alleles in these two patients bore other identifiable mutations. These patients might have mutations in the promoters or other noncoding regions, or mutations in genes other than CYP11B2 may cause the syndrome of CMOII deficiency.

Base Sequence↗

Genetic variation in P450c11AS in Chilean patients with low renin hypertension.

Low renin hypertension (LRH), which accounts for 10-20% of patients with idiopathic "essential" hypertension, bears hormonal similarities to mineralocorticoid-induced hypertension, but elevated mineralocorticoid concentrations have not been found. Some patients with LRH have normal, rather than suppressed, plasma aldosterone concentrations, so that the ratio of aldosterone concentration to PRA (Aldo/PRA) is high, suggesting inappropriately increased aldosterone biosynthesis. We characterized the CYP11B2 gene that encodes the aldosterone synthase, P450c11AS, in hypertensive and control populations in a single clinic in Santiago, Chile. We directly sequenced the entire CYP11B2 gene in 12 patients with LRH, 2 high renin hypertensive controls, and 2 normotensive controls. All sequences were identical, except that 8 of 24 LRH alleles encoded arginine rather than lysine at position 173. The Arg173 and Lys173 variants were expressed in transfected MA-10 cells, and their ability to convert deoxycorticosterone to aldosterone was measured; the apparent Michaelis constant (Km) for Lys173 was 2.73 mumol/L; the Km for Arg173 was 2.53 mumol/L. The apparent maximal velocity (Vmax) for Lys173 was 6.5 x 10(-3) micrograms/mL.24 h; the Vmax for Arg173 was 7.8 x 10(-3) micrograms/mL.24 h. The first order rate constant, Vmax/Km was 2.38 for Lys173 and 3.08 for Arg173. As these values were not significantly different, we sought to determine whether Arg173 is a polymorphism linked to LRH. We examined position 173 in 52 unselected patients with idiopathic hypertension and 55 normotensive controls by PCR amplification of CYP11B2 exons 3-5 followed by digestion with Bsu361, which digests the Arg173 sequence, but not the Lys173 sequence. More of the hypertensive alleles (39 of 104, 37.5%) than normotensive alleles (25 of 110, 22.5%) carried Arg173 (chi 2 = 5.57; P < 0.02). Most of the Arg173 alleles (31 of 72, 43.1%) were from hypertensive patients with Aldo/PRA below 30, whereas only 5 of 24 (20.8%) Arg173 alleles were found in patients with Aldo/PRA greater than 30 (chi 2 = 3.79; P = 0.05) Thus, the ARg173 variant of CYP11B2 may be linked to LRH in Chilean patients.

Adult↗

The human genome contains only two CYP11B (P450c11) genes.

The biosynthesis of glucocorticoids and mineralocorticoids requires isozymes of P450c11. Two human isozymes are known: P450c11 beta, encoded by the CYP11B1 gene, has 11 beta-hydroxylase activity; P450c11AS, encoded by the CYP11B2 gene, has 11 beta-hydroxylase, 18-hydroxylase, and aldosterone synthase activities. Recent data show that the rat genome has four CYP11B genes, three of which are functional, and one of which has novel behaviors. As the number of human CYP11B genes was unknown and as the existence of novel P450c11 isozymes might have implications in the study of hypertension, we sought to determine if the human genome, like the rat genome, contained more than two CYP11B genes. Southern blotting of human genomic DNA digested with StuI suggested the existence of at least four human CYP11B genes. Similar analysis of cosmid clones suggested multiple CYP11B genes. However, cloning and sequencing of the multiple hybridizing fragments showed that there are only two CYP11B genes in the human genome, and that the "extra" bands seen were due to spurious hybridization. The absence of additional CYP11B genes in the human genome analogous to those in the rat narrows the search for genes that contribute to low renin hypertension.

Animals↗

Contraction dynamics and power production of pink muscle of the scup (Stenotomus chrysops).

Although the contribution of red muscle to sustained swimming in fish has been studied in detail in recent years, the role of pink myotomal muscle has not received attention. Pink myotomal muscle in the scup (Stenotomus chrysops) lies just medial to red muscle, has the same longitudinal fibre orientation and is recruited along with the red muscle during steady sustainable swimming. However, pink muscle has significantly faster rates of relaxation, and the maximum velocity of shortening of pink muscle (7.26 +/- 0.18 muscle lengths s-1, N = 9, at 20 degrees C, and 4.46 +/- 0.15 muscle lengths s-1, N = 6, at 10 degrees C; mean +/- S.E.M.) is significantly faster than that of red muscle. These properties facilitate higher mass-specific maximum oscillatory power production relative to that of red muscle at frequencies similar to the tailbeat frequency at maximum sustained swimming speeds in scup. Additionally, pink muscle is found in anatomical positions in which red muscle is produces very little power during swimming: the anterior region of the fish, which undergoes the lowest strain during swimming. Pink muscle produces more oscillatory power than red muscle under low-strain conditions (+/- 2-3%) and this may allow pink muscle to supplement the relatively low power generated by red muscle in the anterior regions of swimming scup.

Animals↗

Observations on the vascular structure of malignant tumours of the mandible using specimen arteriography.

OBJECTIVES: To investigate the vascular structure of different malignant tumours of the mandible. METHODS: Specimen arteriography was performed on seven cases of squamous cell carcinoma of gingiva and four sarcomas. RESULTS: Malignant tumours could be divided into two groups: (1) the hypervascular sarcomas in in which the arteries were dilated, distributed evenly and arranged in a bush-like pattern; and (2) hypovascular squamous cell carcinoma with scanty arteries distributed unevenly without dilatation; in some areas, the shape of arteries was irregular and arranged in a network pattern. CONCLUSIONS: The variation in vascular structure of malignant tumours of the mandible may be related to the outcome of regional chemotherapy.

Adolescent↗

Transfection maximizer increases the efficiency of calcium phosphate transfections with mammalian cells.

A variety of methods have been described for introducing DNA into cultured mammalian cells. Among these, calcium phosphate (CaPO4)-mediated transfection has gained wide acceptance due to the procedure's low cost, simplicity and suitability for a wide range of cell types. However, this method is limited by relatively low transfection efficiencies and poor reproducibility. To improve the utility of CaPO4 transfections, we have identified a transfection enhancer (CalPhos Maximizer) that dramatically increases the efficiency of CaPO4 transfections with mammalian cells. The CalPhos Maximizer is a cellular structural protein that improved CaPO4 transfections with all mammalian cell lines tested and yielded greater than 70% transfection efficiency in BHK-21 cells. In the presence of the CalPhos Maximizer, transfection times were decreased to as little as 2 h with no loss in performance. Because of the high transfection efficiency provided by the CalPhos Maximizer transfection system, gene expression can be assayed following overnight incubations, rather than the typical 48-72-h intervals required by conventional transfection methods. Moreover, the Maximizer reagent is compatible with, and improves the performance of, commercially available as well as home-made CaPO4 transfection systems.

Animals↗

Endothelial prostanoids involved in the relaxation produced by acetylcholine in the human pulmonary artery.

In ring preparations of human pulmonary artery contracted with noradrenaline (NA), the application of acetylcholine (ACh) enhanced the tension, and withdrawal produced a large relaxation which was sustained for about 10 min and required over 20 min for recovery; the latter relaxation appeared only in the endothelium-intact preparation. Indomethacin increased the amplitude of NA contractions, changed the ACh-induced contraction to relaxation, and inhibited the ACh-induced sustained relaxation. Nitroarginine increased the amplitude of NA and ACh-induced contractions, with no significant change in the ACh-induced sustained relaxation. These effects of indomethacin and nitroarginine were observed only in the endothelium-intact preparations. In NA-contracted preparations, exogenously applied prostaglandin I2 (PGI2) produced relaxation. Thus, in human pulmonary arteries, NA and ACh activities release vasodilator prostanoids and nitroarginine-sensitive EDRF from the endothelium, and initiate direct contractile actions to smooth muscle. The prostanoid-induced relaxation is sustained for a long time after the withdrawal of ACh stimulation and can be mimicked by exogenously-applied PGI2.

Acetylcholine↗

Mechanical properties of single pellets containing acrylic polymers.

Three aqueous-based acrylic latex dispersions, Eudragit L 30 D, NE 30 D, and RS 30 D, were incorporated as granulating binders into a powder blend of microcrystalline cellulose and anhydrous lactose by wet massing. Spheronized pellets were prepared by extrusion-spheronization and the mechanical properties of single pellets, including the tensile strength at break and the Young's modulus were determined from the stress-strain profiles using a Chatillon TCD-200 tension/compression digital test gauge. The influence of particle size and plasticizer on the mechanical properties of pellets containing Eudragit RS 30 D was investigated. All bead formulations deformed by brittle fracture under a diametral compression force. The mechanical strength was found to be influenced by the adhesive strength between the polymers and the powder particles instead of the cohesive strength of each polymer. The Young's modulus and the tensile strength were also significantly influenced by the type and concentration of polymer, the presence of plasticizer, and the particle size of the beads. The results were related to the properties of the polymers and the fracture mechanisms of the beads. Furthermore, the polymer type and the incorporation of plasticizer influenced the susceptibility of the moistened extruded granules to the shearing forces during the spheronization process, which influenced the surface morphological properties of the pellets.

Acrylates↗