Search PubMed⌕ Search

Biomedical subjects

G Zhu

Publications and source records attributed to G Zhu.

At least 181 records · Page 10Linked to original sources
PubMed1997

Mutations in the small subunit of ribulose-1,5-bisphosphate carboxylase/ oxygenase increase the formation of the misfire product xylulose-1,5-bisphosphate.

The small subunit (S) increases the catalytic efficiency of ribulose-1,5-bisphosphate carboxylase/oxygenase (EC 4.1.1.39) by stabilizing the active sites generated by four large subunit (L) dimers. This stabilization appears to be due to an influence of S on the reaction intermediate 2,3-enediol, which is formed after the abstraction of a proton from the substrate ribulose-1,5-bisphosphate. We tested the functional significance of residues that are conserved among most species in the carboxy-terminal part of S and analyzed their influence on the kinetic parameters of Synechococcus holoenzymes. The replacements in S (F92S, Q99G, and P108L) resulted in catalytic activities ranging from 95 to 43% of wild type. The specificity factors for the three mutant enzymes were little affected (90-96% of wild type), but Km(CO2) values increased 0.5- to 2-fold. Mutant enzymes with replacements Q99G and P108L showed increased mis-protonation, relative to carboxylation, of the 2,3-enediol intermediate, forming 2 to 3 times more xylulose-1,5-bisphosphate per ribulose-1,5-bisphosphate utilized than wild-type or F92S enzymes. The results suggest that specific alterations of the L/S interfaces and of the hydrophobic core of S are transmitted to the active site by long-range interactions. S interactions with L may restrict the flexibility of active-site residues in L.

Amino Acid Sequence↗
PubMed1997

Simultaneous 31P MRS of the soleus and gastrocnemius in Sherpas during graded calf muscle exercise.

The observation that the amount of lactate formed during hypobaric hypoxia decreases with the severity of hypoxia has become known as the "lactate paradox." We used noninvasive 31P magnetic resonance spectroscopy (MRS) to further probe this problem and explore the nature of muscle metabolism during rest-exercise-recovery transitions in Sherpas indigenous to the high Himalayas of Nepal. MRS data were obtained using a whole body 1-m bore, 1.5-T Phillips Gyroscan spectrometer. Muscle-specific localization of MRS data acquisition was achieved by means of a modified image-selected in vivo spectroscopy sequence (ISIS). The spectra acquired from the medial and lateral gastrocnemius muscle, rich in fast-twitch fibers, were well constrained by selective excitation and by the boundary of the leg. The spectra from a third region contained signals predominantly from the soleus, a muscle formed mainly of slow-twitch fibers. We quantified relative concentration changes in phosphocreatine (PCr), Pi, and ATP during a series of calf muscle work bouts; free ADP concentrations were calculated on the assumption that the creatine phosphokinase reaction was always essentially at equilibrium. Hydrogen ion concentrations were calculated from the chemical shift of Pi, which represents the equilibrium between mono- and diprotonated phosphate. Plantar flexion was quantified using a calf muscle ergometer designed for operation within a 1-m whole body magnet. We found that the concentration of ATP was rigorously regulated and thus did not change despite large changes in ATP turnover rates required through exercise. The relative concentrations of PCr and Pi were linear functions of the percent maximum work rate of the lateral and medial gastrocnemius, but on transition to exercise the fractional concentration changes in these metabolites were much less than the fractional change in muscle ATP turnover rates. The relationship between muscle ATP turnover rate and free ADP concentration was complex; again, a kinetic order of 1 was not observed. In contrast to the gastrocnemius, the soleus muscle sustained much smaller changes in the concentrations of these crucial metabolites during rest-work-recovery transitions. Unlike the situation in most other muscles rich in fast-twitch fibers characterized by lactate-associated acidosis during muscle work, the intracellular pH in gastrocnemius of Sherpas was stable through these protocols, which is consistent with the low lactate production (i.e., with the lactate paradox) observed in indigenous highlanders.

Adenosine Diphosphate↗
PubMed1997

Computer program for calculating the melting temperature of degenerate oligonucleotides used in PCR or hybridization.

Degenerate primers or probes have been widely used in molecular cloning, but the calculation of their melting temperatures could not simply be done using thermodynamic parameters because of degeneracy and the lack of a computer program. We present here a simple computer program named dPrimer for the calculation of melting temperature of degenerate oligonucleotides based on the nearest-neighbor model. The program was written in C+2 computer language and implemented in Macintosh with a Symantec C+2 compiler. The degenerate sequencing data were read into a graph data structure. All possible oligonucleotide sequences were then determined by a depth-first search algorithm. Their melting temperature (Tm) values were individually calculated, and output was given as Tm range, mean and standard deviation. These data could help one in the selection of PCR annealing and hybridization temperatures as well as in the design of degenerate oligonucleotides with a desired range of Tm.

Algorithms↗
PubMed1997

[Changes in t-PA, PAI-1 gene expression of coronary arteries with preconditioning in acute myocardial infarction rat].

The changes of t-PA, PAI-1 gene expression and t-PA activity were investigated in coronary arteries of Acute Myocardial Infarction (AMI) rat by nucleic acid molecular hybridization and spectrophotometric assay. The effect of preconditioning on t-PA and PAI-1 was observed in this model. The results indicated that: (1) Tissue t-PA activity in ischemia was significantly decreased than that in control (P < 0.01). t-PA activity was obviously increased in preconditioning group (P < 0.01 compared with ischemia group). (2) t-PA and PAI-1 mRNA expression in ischemia were higher than that in control, especially PAI-1 4.6 times higher than that in control), while PAI-1 expression lower in preconditioning rat. (3) In situ hybridization showed t-PA and PAI-1 were primarily expressed by endothelial cells in vessels of rat heart. These data suggested that abnormal expression of PAI-1 and t-PA was involved in AMI. This fibrinolytic depress was associated with high PAI-1 expression. Preconditioning may be effective to protect myocardium through decreasing PAI-1 mRNA expression.

Animals↗
PubMed1997

[Analysis of chromosome karyotype of Lycium chinense Mill].

The chromosome karyotypes of wild Lycium chinese and its cultivated breeds for vegetable have been studied. The Chromosome 12 of both has a satellite. The former, with 24 chromosomes and 1B karyotype, is a diploid and its karyotype formula is 2n = 2x = 24 = 18m + 6sm. The chromosome number of the latter is 48. As a tetrploid, it has 2B karyotype and a karyotype formula 2n = 4x = 48 = 36m + 12sm.

Chromosomes↗
PubMed1997

[Study on the clinical and biological characteristics of 20 cases of chronic lymphoid leukemias].

OBJECTIVE: To further define the chronic lymphoid leukemia (CLL). METHODS: Tweenty cases of CLL classified after the FAB proposals (1989) were retrospectively analyzed. RESULTS: Based on the clinical features, cell/tissue morphology, immunophenotype, IgH and TCR gamma gene rearrangement serum or urine immunoglobulin levels, 16 cases of B-lymphoid leukemia (12 of CLL, 1 of PLL, 2 of leukemia/lymphoma, and 1 of WM) as well as 4 cases of T-lymphoid leukemia (2 of leukemia/lymphoma, 1 of T-LGLL, and 1 of NK-LGLL) were identified. The diagnostic criteria,cell morphology,biological characteristics and prognosis for CLL were discussed. CONCLUSION: T-leukemia/lymphoma should be incorporated in the classification of CLL.

Chronic Disease↗
PubMed1997

[New advances in liquid chromatography-chemiluminescence detection].

A review on the new development of chemiluminescence (CL) detection for liquid chromatography (LC) is presented. It covers CL detection systems (including luminol, peroxyoxalates, and other chemiluminescence reactions) combined with LC, the design of CL detectors, analytical methods for many kinds of inorganic, organic and biologic samples, applications and the trends of LC-CL in environmental, biomedical, life science and pharmaceutical analysis.

Chromatography, Liquid↗
PubMed1996

Beta1,4-N-acetylgalactosaminyltransferase (GM2 synthase) is released from Golgi membranes as a neuraminidase-sensitive, disulfide-bonded dimer by a cathepsin D-like protease.

Many Golgi membrane-bound glycosyltransferases are released from cells in a soluble form. To characterize this release process, we stably transfected Chinese hamster ovary cells with three myc epitope-tagged forms of cloned beta1, 4-N-acetylgalactosaminyltransferase (GalNAcT); two of these forms resided in the Golgi, while the third was retained in the ER. GalNAcT was released into the culture medium from cells transfected with the Golgi forms but not with the ER form of the enzyme. The medium from cells transfected with the Golgi forms contained disulfide-bonded dimers of GalNAcT, which carried neuraminidase sensitive, complex N-linked carbohydrate chains. This soluble species represented the major degradation product of cellular GalNAcT, which turned over with a half-time of about 1.7 h. The soluble species consisted of a mixture of truncated GalNAcT molecules, the major form of which was produced by cleavage near the boundary between the transmembrane and lumenal domains between Leu-23 and Tyr-24. This cleavage site fits the sequence pattern for sites cleaved by cathepsin D (van Noort, J.M., and van der Drift, A. C.M. (1989) J. Biol. Chem. 264, 14159-14164). These findings suggest that GalNAcT is converted from a membrane-bound to a soluble form as a result of cleavage by a cathepsin D-like protease in a compartment late in the Golgi secretory pathway.

Animals↗
PubMed1996

Nuclear magnetic resonance solution structure of the growth factor receptor-bound protein 2 Src homology 2 domain.

A family of NMR solution structures of the growth factor receptor-bound protein 2 (Grb2) SH2 domain has been determined by heteronuclear multidimensional NMR. Proton, nitrogen, and carbon chemical shift assignments have been made for the SH2 domain of Grb2. Assignments were made from a combination of homonuclear two-dimensional and 15N- and 13C-edited three-dimensional spectra at pH 6.2 and 298 K. Structure-induced proton and carbon secondary shifts were calculated and used to facilitate the spectral assignment process. NOE, scalar coupling, secondary chemical shift, and amide proton exchange data were used to characterize the secondary structural elements and hydrogen-bonding network in the Grb2 SH2 domain. The three-dimensional structure of the Grb2 SH2 domain was calculated using 1112 restraints obtained from NOE, coupling constant, and amide proton exchange data. The rmsd for the 24 calculated structures to the mean structure of the Grb2 SH2 domain was 0.75 A for backbone and 1.28 A for all heavy atoms. The three-dimensional fold of the Grb2 SH2 domain is similar to that observed for other SH2 domains and consists of two alpha-helical segments and eight beta-strands, six strands that make up two contiguous antiparallel beta-sheets, and two strands that form two short parallel beta-sheets. The structure of the phosphotyrosine binding pocket of Grb2 is similar to that observed for other SH2 domains. The hydrophobic binding pocket of Grb2 is similar to that observed for Src with the exception that tryptophan 121 of Grb2 occupies part of the pY+3 binding pocket. Structural implications for the Grb2 SH2 domain selectivity at the pY+2 and pY+3 sites are discussed.

Amino Acid Sequence↗
PubMed1996

Directed mutagenesis of chloroplast ribulose-1,5-bisphosphate carboxylase/oxygenase. Loop 6 substitutions complement for structural stability but decrease catalytic efficiency.

The structure of active-site loop 6 plays a role in determining the CO2/O2 specificity of chloroplast ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco, EC 4.1.1.39). Rubisco from the green alga Chlamydomonas reinhardtii differs from higher plant Rubisco within the loop 6 region, and the C. reinhardtii enzyme has a CO2/O2 specificity 25% lower than that of higher plant enzymes. To examine whether differences in sequence may account for differences in catalytic efficiency, we focused on a conserved pair of residues that are in van der Waals contact at the base of loop 6. C. reinhardtii Rubisco contains Leu-326 and Met-349, whereas higher plant enzymes contain Ile-326 and Leu-349. By employing in vitro mutagenesis and chloroplast transformation, L326I and M349L substitutions were created within the Rubisco large subunit of C. reinhardtii. M349L had little effect, but L326I destabilized the holoenzyme in vivo and in vitro. When present together, the M349L substitution partially alleviated the instability resulting from the L326I substitution, but caused a 21% decrease in CO2/O2 specificity and a 74% decrease in the Vmax of carboxylation. Interactions between loop 6 and other structural regions are likely to be responsible for both holoenzyme stability and catalytic efficiency in higher plant Rubisco enzymes.

Amino Acid Sequence↗
PubMed1996

Cloned beta 1,4N-acetylgalactosaminyltransferase: subcellular localization and formation of disulfide bonded species.

Cloned human beta 1,4N-acetylgalactosaminyltransferase (GalNAcT) catalyzes the synthesis of the glycosphingolipids GM2, GD2, and gangliotriosylceramide. To determine the subcellular location of this enzyme and whether it exists in intermolecular disulfide bonded species, we stably transfected Chinese hamster ovary (CHO) cells with three myc epitope-tagged forms of the GalNAcT gene: the native enzyme; the lumenal domain of GalNAcT fused to the cytoplasmic and transmembrane domains of N-acetylglucosaminyltransferase I (GNT); and the transmembrane and lumenal domains of GalNAcT fused to the cytoplasmic domain of the Iip33 form of human invariant chain in order to retain the enzyme in the endoplasmic reticulum (ER). Immunoelectron microscopic analysis with anti-myc revealed that GalNAcT/myc was present throughout the Golgi stack, the GNT/GalNAcT/myc form was restricted primarily to the medial Golgi cisternae, and the Iip33/GalNAcT/myc form was restricted to the ER. Cells transfected with each of the three constructs contained high levels of GM2 synthase activity in vitro, but only the GalNAcT/myc form and the GNT/GalNAcT/myc forms were able to synthesize the GM2 product in vivo. The enzyme produced by all three constructs was present in the transfected cells in a disulfide bonded form having a molecular size consistent with that of a homodimer or higher aggregate.

Animals↗
PubMed1996

The effect of vincristine-polyanion complexes in STEALTH liposomes on pharmacokinetics, toxicity and anti tumor activity.

Poly(ethylene glycol) (PEG)-derivatized liposome vehicles improve antitumor effectiveness of entrapped anthracyclines and vinca alkaloids. However, the plasma clearance of entrapped vincristine is substantially faster than the lipid phase or other entrapped aqueous markers, suggesting leakage out of the liposome during transit in the blood compartment. We tested the effect of altering the drug's in vivo leakage rate on pharmacokinetics, toxicity, and antitumor activity of entrapped drug in rodent models. Suramin, heparin, and dextran sulfate were tested for their ability to produce a precipitable complex in vitro. PEG-derivatized liposomes were prepared with the complexing agent inside, and vincristine was driven inside using an ammonium gradient. The resulting preparations were found to have plasma distribution half-lives significantly longer than the formulation without a complex-forming agent. There was no increase in acute lethality, and in the case of the suramin-vincristine complex, the acute lethality was significantly reduced at the highest does level. Anti-tumor activity against the mouse mammary carcinoma MC2 was tested in a multiple-dose study. Free vincristine did not affect the tumor growth rate significantly, but at the same dose level all PEG-coated liposome formulations inhibited tumor growth markedly. The suramin containing formulation was as effective as the formulation lacking polyanion, but the heparin and dextran sulfate containing formulations were less effective. Thus, compounds which form insoluble complexes with vincristine alter in vivo plasma distribution phase pharmacokinetics without increasing acute lethality, but without a corresponding increase in anti-tumor activity.

Animals↗
PubMed1996

Carcinoma of the minor salivary glands: results of surgery and combined therapy.

PURPOSE: This article reports the results of treatment of 7 histologic types of minor salivary gland carcinoma. PATIENTS AND METHODS: Of 256 cases, local excision was used in 194, extended excision in 62, excision accompanied by neck dissection in 98, and surgery with adjunctive radiation therapy or chemotherapy in 101. Forty-five of the 98 neck dissection cases had cervical node metastases. RESULTS: Survival rates were analyzed for 233 patients. The 3-year survival rate was (167 of 215) (77.67%), the 5-year survival rate was 134 of 187 (71.76%), and the 10-year survival rate was 79 of 113 (69.91%). CONCLUSIONS: These data confirm that surgery is still the mode of choice in the treatment of minor salivary gland carcinoma, but a combination with radiation therapy or chemotherapy is advisable in some patients.

Adenocarcinoma↗
PubMed1996

Appearance of immune cells and expression of MHC II DQ molecule by fibroblasts in alkali-burned corneas.

Corneal alkali burns are characterized by persistent inflammatory response and recurrent epithelial erosions. We examine whether immune cell types, i.e., T-cells and B-cells, play a role in this devastating process. Rabbit alkali-burned corneas that healed for 1-49 days were subjected to immunostaining with monoclonal antibodies (mAb) L11/135 (anti-T-cells), and 2C4 (anti-MHC II DQ). Serum was collected weekly and subjected to Western blot immunostaining to detect antibodies against denatured corneal proteins. Our observations demonstrated that all injured corneas reepithelialized within 3 days but then developed recurrent erosions. Immunohistochemical studies revealed that PMN, monocytes, and B-cells labeled by 2C4 mAb and T-cells labeled by L11/135 mAb appeared in the periphery to the cornea at 1 day after alkali burn. Many of these myeloid and lymphoid cells invaded the central stroma after 2 weeks of injuries when the alkali-burned corneas were heavily vascularized. In addition, some fibroblastic cells also expressed the MHC II DQ molecules in the alkali-burned corneas that had healed for > 2 weeks. Plasma cells appeared in granulation tissue of injured corneas that had healed for > 3 weeks. Western blot analysis demonstrated a production of heterogeneous antibodies in a majority of the rabbits (11 of 14) to various denatured corneal proteins (between 80 kDa and 25 kDa) at 5 weeks of alkali burn. Inflammatory cell types, i.e., PMN, macrophages could be found underneath the detached epithelium. These observations are consistent with the notion that the myeloid and lymphoid cells may participate in and complicate the healing of corneal alkali burns.

Alkalies↗
PubMed1996

In vivo adenovirus-mediated gene transfer into normal and cystic rat kidneys.

Gene transfer into the mammalian kidney has proved difficult because of the structural complexity of the organ and its low mitotic index. This article describes the use of intra-arterially injected adenovirus to study gene transfer into the rat kidney in vivo. By pre-chilling the kidney, and incubating the virus with the kidney in the cold for extended periods of time, we were able to successfully transfer a beta-galactosidase (beta-gal) reporter gene into the vasculature without ischemic injury to the kidney. Transfer occurred largely in the cortex when cold was used alone, whereas with the use of cold and vasodilators, transfer was accomplished into the outer medulla in both the inner and outer stripes. In the Han:SPRD rat model of autosomal dominant polycystic kidney disease (ADPKD), gene transfer occurred into the vasculature, some epithelial cysts and interstitial cells. This is the first description of substantial in vivo gene transfer into both normal and cystic kidneys. The methodology could find application in the creation of new models of renal disease, for in vivo therapeutic intervention or for genetic modification of an allograft at the time of harvest.

Adenoviridae↗
PubMed1996

[Inhibitory effects of antisense oligonucleotides delivered locally via hydrogel catheter on myointimal hyperplasia of rat carotid artery after nitrogen drying].

OBJECTIVE: To determine if antisense c-myb delivered locally via the hydrogel catheter inhibits the development of myointimal hyperplasia in rat carotid artery after nitrogen desiccation of the endothelium. METHODS: Forty rats were randomized into four groups (ten in each). After endothelial injury, different oligonucleotides were administered locally through the hydrogel catheter. Animals were killed at 14 days following gas drying. The lumen areas, ratios of intimal to medial areas and intimal to medial thickness were measured with the aid of computer. The q test was used for statistical analysis. RESULTS: The lumen area of antisense c-myb group (0.29 +/- 0.01 mm2) was larger than that of sense and mismatch groups (0.17 +/- 0.05 mm2, q = 4.1729; 0.18 +/- 0.05 mm2, q = 3.7489, P < 0.05), and less than that of control group (0.40 +/- 0.14, q = 3.7782, P < 0.05). The ratios of intimal to medial areas and intimal to medial thickness for antisense c-myb group (0.45 +/- 0.24, 0.57 +/- 0.24) were significantly decreased compared with sense and mismatch groups (1.26 +/- 0.25, q = 9.8913; 1.16 +/- 0.38, q = 8.4252; 2.15 +/- 0.85, q = 8.6850; 1.85 +/- 0.79, q = 6.8389, P < 0.01). CONCLUSIONS: Myointimal hyperplasia of rat carotid artery after nitrogen drying can be markedly suppressed by intravascular local delivery of synthetic antisense c-myb via hydrogel catheter. The technique is a promising to prevent artery from obstructive stenosis.

Animals↗