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Biomedical subjects

Z Dai

Publications and source records attributed to Z Dai.

At least 91 records · Page 5Linked to original sources
PubMed1995

Induction of nopaline synthase promoter activity by H2O2 has no direct correlation with salicylic acid.

Transgenic tobacco (Nicotiana tabacum L.) plants carrying a fusion between the nopaline synthase (nos) promoter and chloramphenicol acetyltransferase (CAT) reporter gene (caf) were tested for their response to treatment with H2O2. The nos promoter-driven CAT activity increased significantly by addition of H2O2, reaching the maximum level at 15 mM. Kinetic analysis for CAT activity showed that induction by H2O2 was similar to that of methyl jasmonate (MJ), but was much slower than induction by salicylic acid (SA). Time-course experiments for mRNA level also revealed that the response to H2O2 treatment was similar to that of MJ. The nos promoter displayed a rapid and transient induction of mRNA with SA treatment, with the maximum levels occurring at 3 h, whereas the levels induced by H2O2 or MJ treatment increased continuously during the 11-h experimental period. The antioxidants N-acetyl-L-cysteine and catechol did not alter the SA effect. The responses of the nos promoter to H2O2, MJ, and wounding were significantly reduced by deletions of the CAAT box region and the sequence between -112 and -101. However, these deletions did not significantly alter the SA response. This suggests that H2O2 may have a different mechanism from that of SA for inducing nos promotor activity.

Acetylcysteine↗
PubMed1995

The atxA gene product activates transcription of the anthrax toxin genes and is essential for virulence.

Bacillus anthracis plasmid pXO1 carries the structural genes for the three anthrax toxin proteins, cya (edema factor), lef (lethal factor), and pag (protective antigen). Expression of the toxin genes by B. anthracis is enhanced during growth under elevated levels of CO2. This CO2 effect is observed only in the presence of another pXO1 gene, atxA, which encodes a transactivator of anthrax toxin synthesis. Here we show that transcription of atxA does not appear to differ in cells grown in 5% CO2 compared with cells grown in air. Using a new efficient method for gene replacement in B. anthracis, we constructed an atxA-null mutant in which the atxA-coding sequence on pXO1 is replaced with an omega km-2 cassette. Transcription of all three toxin genes is decreased in the absence of atxA. The pag gene possesses two apparent transcription start sites, P1 and P2; only transcripts with 5' ends mapping to P1 are decreased in the atxA-null mutant. Deletion analysis of the pag promoter region indicates that the 111 bp region upstream of the P1 site is sufficient for atxA-mediated activation of this transcript. The cya and lef genes each have one apparent start site for transcription. Transcripts with 5' ends mapping to these sites are not detected in the atxA-null mutant. The atxA-null mutant is avirulent in mice. Moreover, the antibody response to all three toxin proteins is decreased significantly in atxA-null mutant-infected mice. These data suggest that the atxA gene product also regulates toxin gene expression during infection.

Animals↗
PubMed1995

Applicability of prostaglandin E1 indirect portal vein angiography in patients with portal hypertension.

The imaging quality of the portal vein was obviously improved with prostaglandin E1 (PGE1) indirect portal vein digital subtraction angiography (DSA) in 23 cases. The time-density curve showed that the occurrence rate of opposite hepatic blood flow of splenic vein (SV) was the highest (17.4%). The total visualization rate of the left gastric vein (LGV) was 78.3%, and the visualization rate of the short gastric vein (SGV) was 36.4%. 38.9% of the LGV and all the SGV were visualized with indirect portal vein DSA through SA. Indirect portal vein angiography through superior mesenteric artery and that through splenic artery were of equal importance. In portal hypertension patients with hemorrhage of the digestive tract, when LGV and SGV could not be visualized in PGE1 indirect portal vein DSA, the possibility of non-varices vein bleeding should be considered. When opposite hepatic blood flow with obvious dilation appeared in LGV and SGV, devascularization of the pericardial blood vessels would be justifiable.

Adult↗
PubMed1995

Presynaptic differentiation induced in cultured neurons by local application of basic fibroblast growth factor.

Recent studies have suggested a role for molecules residing at the muscle surface in signaling presynaptic development at the neuromuscular junction (NMJ). Since heparan sulfate-proteoglycan is a major component of the extracellular matrix of skeletal muscle, factors that are bound to this proteoglycan, such as basic fibroblast growth factor (bFGF), are in a strategic position for neuronal signaling. To test this idea, we applied bFGF to cultured Xenopus spinal cord neurons and monitored the change in intracellular Ca2+ level with fura-2 ratio imaging. In one-third of the neurons, local application of bFGF elicited a 30-140% increase in intracellular Ca2+ level. Ca(2+)-free medium or suramin abolished this change. Latex beads coated with bFGF induced clustering of synaptic vesicles at the bead-neurite contacts as evidenced by anti-synaptotagmin antibody labeling and electron microscopy. This response was also blocked by Ca(2+)-free medium and by suramin. Uncoated beads or beads coated with PDGF were ineffective. This induction was also inhibited by a tyrosine kinase inhibitor, tyrphostin RG-50864, suggesting the role of receptor tyrosine kinase activation in this process. In addition, bFGF-coated beads also induced the localization of depolarization-dependent Ca2+ influx to the bead-neurite contacts. In contrast, depolarization caused a distributed Ca2+ elevation in untreated neurites. These results suggest that local presentation of bFGF can mimic the muscle target in signaling the development of both a cytoplasmic and a membranous specialization for excitation-secretion coupling observed at the NMJ.

Animals↗
PubMed1995

The role of heparin-binding growth-associated molecule (HB-GAM) in the postsynaptic induction in cultured muscle cells.

The heparan sulfate proteoglycan (HSPGs) is a components of the extracellular matrix of skeletal muscle that is concentrated at the neuromuscular junction (NMJ). Recent studies have suggested that HSPG, together with its bound peptide growth factors, plays important roles in autocrine or paracrine types of regulation of cell growth and differentiation. Heparin-binding growth-associated molecule (HB-GAM; also known as pleiotrophin, or p18) is a newly discovered HSPG-bound factor that is expressed at high levels in the developing CNS and PNS. In this study, we examined the role of this factor in NMJ development by examining its relationship to the formation of ACh receptor (AChR) clusters. Using an antibody against recombinant rat brain HB-GAM, we found that this protein is present prominently on the surface of cultured Xenopus myotomal muscle cells by immunocytochemistry. It is associated with HSPGs as evidenced by the fact that heparin and heparinase treatment greatly diminished the antibody labeling. HB-GAM is concentrated at preexisting AChR hot spots as well as at those induced by polystyrene beads. In addition, this molecule is also concentrated at AChR clusters induced by spinal cord neurons in nerve-muscle cocultures. To assess its function in synaptic induction, we applied recombinant HB-GAM-coated beads to cultured muscle cells to effect its focal presentation. Over 70% of these beads induced the formation of AChR clusters as shown by fluorescent alpha-bungarotoxin labeling. Furthermore, bath application of HB-GAM inhibited the nerve-induced formation of AChR clusters. Thus, HB-GAM is an endogenous muscle-derived factor that may be a component of the molecular mechanism in postsynaptic induction.

Animals↗
PubMed1995

Effect of perindopril and metoprolol on left ventricular hypertrophy and performance in essential hypertension.

The effects of perindopril and metoprolol on left ventricular hypertrophy (LVH) and function were studied in 47 essential hypertensive patients with LVH. Previous antihypertensive drugs were discontinued for at least 2 weeks, after which patients were randomly divided into 2 groups. 25 subjects were treated with perindopril 4 to 8 mg once daily in the morning (Group A) and 22 subjects with metoprolol 25 to 62.5 mg twice daily (Group B). The subjects were evaluated before and after 4 and 8 weeks of treatment by use of echocardiography. Before treatment LV mass indexes (LVMI) of two groups were respectively 143.2 +/- 21.3 g/m2 and 140.6 +/- 23.7 g/m2 (P > 0.05). In Group A, reduction of LVMI occurred after 4 weeks of treatment, and more pronounced after 8 weeks (from 143.2 +/- 21.3 g/m2 to 126.6 +/- 15.3 g/m2, P < 0.001), whereas reduction of LVMI occurred only after 8 weeks in Group B (from 140.6 +/- 23.7 g/m2 to 133.4 +/- 13.2 g/m2, P < 0.001). In addition, there was a significant (P < 0.05) difference in LVMI between the two groups after 8 weeks. LV systolic function remained unchanged, whereas E/A increased significantly (P < 0.001) in two groups after 8 weeks. In conclusion, antihypertensive treatment with perindopril and metoprolol induced a significant regression of LVH associated with improvement in LV diastolic performance. Perindopril, compared with metoprolol, was more effective in reversing LVH.

Adult↗
PubMed1994

Brain growth retardation due to the expression of human insulin like growth factor binding protein-1 in transgenic mice: an in vivo model for the analysis of igf function in the brain.

Three lines of transgenic (Tg) mice carrying a fusion gene linking the mouse metallothionein-I promoter to a cDNA encoding human insulin-like growth factor binding protein-1 (hIGFBP-1) were found to express the transgene in brain. As judged by comparing Tg brain weights to those of non-transgenic littermates, adult hemizygotic Tg mice of each line exhibited brain growth retardation (16.2%, 14.4% and 8.1% reductions in weight, respectively in each line). In two lines, total brain DNA and protein content were decreased. Further analysis indicated that the brain growth retardation was manifested in the second week of postnatal life. Given that the insulin-like growth factors (IGFs) stimulate cell proliferation and/or survival in neural cultures and that hIGFBP-1, when present in a molar excess, inhibits IGF interactions with their cell surface receptors, the brain growth retardation in hIGFBP-1 Tg mice likely results from hIGFBP-1 inhibition of IGF-stimulated growth-promoting actions. These hIGFBP-1 Tg mice should prove useful in defining IGF actions during postnatal brain maturation.

Aging↗
PubMed1994

Regulation of the Bacillus anthracis protective antigen gene: CO2 and a trans-acting element activate transcription from one of two promoters.

The pag gene of Bacillus anthracis, located on plasmid pXO1 (185 kb), encodes protective antigen, a component of the anthrax lethal and edema toxins. Synthesis of protective antigen is enhanced during growth of the organism with elevated levels of CO2. The CO2 effect is at the level of transcription, and pXO1-encoded regulatory factors have been implicated in control of pag expression. We used a Tn917-LTV3 insertion mutant of B. anthracis in which the wild-type pag gene on pXO1 was replaced with a pag-lacZ transcriptional fusion to monitor pag promoter activity. Expression of the pag-lacZ fusion is induced five- to eightfold during growth in 5% CO2 compared with growth in air. Growth in 20% CO2 increases transcription up to 19-fold. By monitoring pag-lacZ expression in atmospheres with different O2 and CO2 concentrations, we demonstrated definitively that the CO2 effect is specific and not simply a result of increased anaerobiosis. The results of 5' end mapping of pag transcripts indicate multiple sites of transcript initiation. We have determined two major apparent start sites, designated P1 and P2, located at positions -58 and -26 relative to the translation initiation codon, respectively. Analysis of total RNA from late-log-phase cells shows comparable initiation from P1 and P2 in wild-type strains grown in aerobic conditions. However, initiation from P1 is increased approximately 10-fold in cultures grown with an elevated level (5%) of CO2. We have identified a locus on pXO1, more than 13 kb upstream from the pag gene, which enhances pag transcription. When added in trans, this locus increases the level of transcripts with 5' ends mapping to P1 but has no effect on the level of transcripts with 5' ends mapping to P2. The CO2 effect on P1 is observed only in the presence of the activator locus.

Amino Acid Sequence↗
PubMed1994

Human insulin-like growth factor-binding protein-1 (hIGFBP-1) in transgenic mice: characterization and insights into the regulation of IGFBP-1 expression.

Three hemizygous transgenic (Tg) mouse lines were generated with a fusion gene composed of the mouse metallothionein promoter (mMT-I) and a full-length human insulin-like growth factor binding protein-1 (hIGFBP-1) complementary DNA that was truncated in its 3'-untranslated region. Despite high serum hIGFBP-1 levels (120-2570 micrograms/liter) before puberty in two of these lines, no significant alterations were observed in somatic growth, nor were marked alterations noted in fasting or random serum glucose or in the response of young adult Tg mice to ip glucose. The transgene was expressed in a number of tissues from each line, but liver was a significant site of transgene expression in only one line. Unexpectedly, liver hIGFBP-1 messenger RNA (mRNA) expression in this line was regulated in fashion similar to the native liver IGFBP-1 mRNA: 1) its abundance waned with advancing postnatal age and became minimal in early adult life, despite continuous zinc supplementation to stimulate its transcription; and 2) fasting increased its abundance 3- to 4.3-fold. The decline in transgene expression with aging was not due to a deletion, rearrangement, or a change in the methylation of liver transgene DNA. Transcriptional mechanisms also were not likely to account for the observed regulation of the transgene mRNA, because liver expression of the mMT-I gene, which shares identical genomic 5'-regulatory elements with the transgene, was not similarly altered by aging or fasting. Because cycloheximide (CHX) treatment of cultured rat H4IIE cells has been shown to prolong IGFBP-1 mRNA half-life while decreasing its transcription, Tg mice were treated with CHX to test the possibility that instability of the liver transgene mRNA influenced its abundance. After CHX and under conditions of chronic zinc supplementation, liver transgene mRNA abundance increased in parallel with that of the native IGFBP-1 mRNA. Although CHX is known to activate mMT-I transcription by mechanisms involving the 5'-regulatory regions contained in the transgene, CHX-induced transcription only in part accounted for the increase in liver transgene mRNA, because CHX induced an earlier and greater increase in liver transgene mRNA than in mMT-I mRNA. Taken together, these data indicate that both transgene and native IGFBP-1 liver mRNA are regulated by factors that alter mRNA stability. The finding that native liver IGFBP-1 mRNA abundance is influenced by transgene expression further supports the concept that both mRNAs share some common mechanisms of regulation.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1994

[Study on evolutionary genetics of Drosophila auraria species complex--cladistic analysis and phenetic analysis].

In the present paper, seventeen geographic strains of D. auraria species complex and five geographic strains of other four species, which belong to the montium species subgroup were biochemically examined for the construction of phylogenetic tree based on phenetic and cladistic analysis. Eighteen isoenzymes were analyzed by isoelectric focusing polyacrylamide gel electrophoresis in the above twenty two geographic strains. The phenogram of the D. auraria species complex is supported by UPGMA program of phenetic analysis. It was found that: 1. D. auraria, D. triauraria and D. quadraria grouped as an type are more closely related to one another, and D. biauraria, D. subauraria are more closely related to each other, while the former three species are distantly related to the latter two species. 2. The genetic identity between two geographic strains of same species is higher than the genetic identity between different species, except the geographic strain T-KM of D. triauraria. 3. The geographic strain T-KM, one of the D. triauraria from the southeast isolated islands Kametoku [symbol: see text] of Japan is far more distantly related to the other geographic strains of it. We suggest that the new species or subspecies are diverged from D. triauraria will be due to the geographic speciation mechanisms. 4. The genetic differentiation among eight geographic strains of D. triauraria are much larger than those among three geographic strains of D. auraria, which may be related to their different inhabitation environment as well as their flying ability. On the other hand, a cladistic analysis of the D. auraria species complex are made in the present paper.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

[Auto-analysis of corneal endothelium with photoelectrical scans and computer].

A quantitative analysis of corneal endothelial cells was made with photoelectrical scans and computer in 67 eyes. No difference was found as regards to the mean cell area and density between the endothelial analysis system and routine square counting. On the other hand, we analyzed corneal endothelial cells between 30 eyes wearing hard contact lens and 30 eyes of age- and sex-matched nonwearing control. The parameters, including cell density individual cell area, area, mean cell area, standard deviation, coefficient of variation of cell size (CV), maximum cell, minimum cell, the ratio of the maximum to minimum cell, and percentage of the hexagoal cells, were automatically displayed. The cell density and mean cell area also showed no significant difference (P > 0.05) between the wearers and nonwearing control, but the CV and the frequency of the hexagonal cells showed significant difference (P < 0.01). The ratio of maximum to minimum cell, the polymegethism and pleomorphism were obviously increased in the wearers. It is highly suggested that computer analysis system is of great value in evaluating corneal endothelial cells.

Adolescent↗
PubMed1993

BCR-ABL-induced oncogenesis is mediated by direct interaction with the SH2 domain of the GRB-2 adaptor protein.

BCR-ABL is a chimeric oncoprotein that exhibits deregulated tyrosine kinase activity and is implicated in the pathogenesis of Philadelphia chromosome (Ph1)-positive human leukemias. Sequences within the first exon of BCR are required to activate the transforming potential of BCR-ABL. The SH2/SH3 domain-containing GRB-2 protein links tyrosine kinases to Ras signaling. We demonstrate that BCR-ABL exists in a complex with GRB-2 in vivo. Binding of GRB-2 to BCR-ABL is mediated by the direct interaction of the GRB-2 SH2 domain with a phosphorylated tyrosine, Y177, within the BCR first exon. The BCR-ABL-GRB-2 interaction is required for activation of the Ras signaling pathway. Mutation of Y177 to phenylalanine (Y177F) abolishes GRB-2 binding and abrogates BCR-ABL-induced Ras activation. The BCR-ABL (Y177F) mutant is unable to transform primary bone marrow cultures and is impaired in its ability to transform Rat1 fibroblasts. These findings implicate activation of Ras function as an important component in BCR-ABL-mediated transformation and demonstrate that GRB-2 not only functions in normal development and mitogenesis but also plays a role in oncogenesis.

Adaptor Proteins, Signal Transducing↗
PubMed1993

Molecular mimicry in Lyme disease: monoclonal antibody H9724 to B. burgdorferi flagellin specifically detects chaperonin-HSP60.

A monoclonal antibody (H9724), specific for the 41-kDa flagellar protein of the Lyme disease pathogen Borrelia burgdorferi, cross-reacts with human axons and detects one major protein in human neuroblastoma cell extracts. The homologous cross-reacting protein has now been isolated from calf adrenal and identified as chaperonin-HSP60 by N-terminal sequencing.

Amino Acid Sequence↗
PubMed1993

Covalent attachment of peptides to membranes for dot-blot analysis of glycosylation sites and epitopes.

Noncovalent binding of proteins to membranes is often employed for dot-blot analysis with various visualization techniques. These techniques are usually not applicable to peptide dot-blot analysis due to peptide wash-off during the staining procedure. As exemplified with a synthetic peptide and peptides produced by proteolysis of a protein, it is possible to achieve efficient covalent attachment to Immobilon-AV membranes. The utility of this membrane has been demonstrated with immunostaining and carbohydrate staining procedures.

Amino Acid Sequence↗
PubMed1993

Elevation in presynaptic Ca2+ level accompanying initial nerve-muscle contact in tissue culture.

Nerve-muscle cocultures were used to study the relationship between neuronal Ca2+ level and the earliest nerve-muscle interaction. Xenopus spinal cord neurons were loaded with Ca2+ indicators and monitored by digital video microscopy as a myoball was manipulated into contact with it. Transmitter release was measured from the myoball by whole-cell recording. We observed a 1.5- to 6-fold increase in Ca2+ level in the neurite upon contact with a myoball. Fifty percent of the pairs showing Ca2+ elevation were also positive for neurotransmission. This Ca2+ rise was suppressed by lifting away the myoball, by Ca(2+)-free solution, or by suramin, a polyanionic compound that interferes with cell surface receptors. This suppression was accompanied by a reduction in transmitter release. The specificity of the nerve-muscle contact-induced Ca2+ rise was shown by its absence upon neuron-neuron contact. Naturally formed nerve-muscle contacts also showed an elevation in presynaptic Ca2+ level. Thus, this elevation appears to be a physiological step in the early stage of synaptogenesis and is likely mediated by muscle-derived molecules.

Animals↗