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Biomedical subjects

D Kong

Publications and source records attributed to D Kong.

At least 55 records · Page 3Linked to original sources
PubMed1996

Single-stranded DNA binding protein and DNA helicase of bacteriophage T7 mediate homologous DNA strand exchange.

Two proteins encoded by bacteriophage T7, the gene 2.5 single-stranded DNA binding protein and the gene 4 helicase, mediate homologous DNA strand exchange. Gene 2.5 protein stimulates homologous base pairing of two DNA molecules containing complementary single-stranded regions. The formation of a joint molecule consisting of circular, single-stranded M13 DNA, annealed to homologous linear, duplex DNA having 3'- or 5'-single-stranded termini of approximately 100 nucleotides requires stoichiometric amounts of gene 2.5 protein. In the presence of gene 4 helicase, strand transfer proceeds at a rate of > 120 nucleotides/s in a polar 5' to 3' direction with respect to the invading strand, resulting in the production of circular duplex M13 DNA. Strand transfer is coupled to the hydrolysis of a nucleoside 5'-triphosphate. The reaction is dependent on specific interactions between gene 2.5 protein and gene 4 protein.

Bacteriophage T7↗
PubMed1996

Shanghai trial of nifedipine in the elderly (STONE).

OBJECTIVE: To assess the effectiveness of nifedipine treatment in elderly hypertensives. METHODS: A single-blind trial was conducted under the direction of the Shanghai Institute of Hypertension in 1632 subjects aged 60-79 years alternatively allocated to either nifedipine or placebo after a 4-week placebo run-in period between 1987 and 1990 with mean follow-up of 30 months. Clinical events and risk modification were analysed in collaboration with the University of Montreal. Seventy-four patients with severe hypertension were reallocated to active nifedipine treatment after placebo run-in. RESULTS: Cox's proportional hazards model accounting for covariates demonstrated a highly significant decrease in the probability of events: 'original treatment assignment' analysis indicated that 77 events occurred in the placebo and 32 in the nifedipine group. Similar significances were achieved with 'actual treatment' or 'changes excluded' (excluding reallocated subjects) analyses. A significant reduction in relative risk was observed for strokes and severe arrhythmia with an overall decrease from 1.0 to 0.41 (95% confidence interval 0.27-0.61). CONCLUSION: Nifedipine treatment diminished the number of severe clinical outcomes in elderly hypertensives significantly.

Aged↗
PubMed1995

Adenylate cyclase, cyclic AMP and extracellular-signal-regulated kinase-2 in airway smooth muscle: modulation by protein kinase C and growth serum.

Bradykinin and phorbol 12-myristate 13-acetate stimulate adenylate cyclase activity in serum-depleted cultured airway smooth muscle via a protein kinase C (PKC)-dependent pathway. The probable target is the type II adenylate cyclase, which can integrate coincident signals from both PKC and Gs. Therefore, activation of Gs (by cholera-toxin pre-treatment) amplified the bradykinin-stimulated cyclic AMP signal and concurrently attenuated the partial activation of extracellular-signal-regulated kinase-2 (ERK-2) by bradykinin. We have previously demonstrated that, in order to induce full activation of ERK-2 with bradykinin, it is necessary to obliterate PKC-stimulated cyclic AMP formation. We concluded that the cyclic AMP signal limits the magnitude of ERK-2 activation [Pyne, Moughal, Stevens, Tolan and Pyne (1994) Biochem. J. 304, 611-616]. The present study indicates that the bradykinin-stimulated ERK-2 pathway is entirely cyclic AMP-sensitive, and suggests that coincident signal detection by adenylate cyclase may be an important physiological route for the modulation of early mitogenic signalling. Furthermore, the direct inhibition of adenylate cyclase activity enables bradykinin to induce DNA synthesis, indicating that the PKC-dependent activation of adenylate cyclase limits entry of cells into the cell cycle. These studies suggest that the mitogenicity of an agonist may be governed, in part, by its ability to stimulate an inhibitory cyclic AMP signal pathway in the cell. The activation of adenylate cyclase by PKC appears to be downstream of phospholipase D. However, in cells that were maintained in growth serum (i.e. were not growth-arrested), bradykinin was unable to elicit a PKC-stimulated cyclic AMP response. The lesion in the signal-response coupling was not at the level of either the receptor or phospholipase D, which remain functionally operative and suggests modification occurs at either PKC or adenylate cyclase itself. These studies are discussed with respect to the cell signal regulation of mitogenesis in airway smooth muscle.

Adenylyl Cyclase Inhibitors↗
PubMed1995

Whole-virus vaccine development by continuous culture on a complementing host.

We have evaluated an adaptive strategy for generating whole-virus vaccines using a bacteriophage model. Wildtype phage T7 was cultivated in a two-stage continuous stirred-tank reactor (CSTR) utilizing a recombinant E. coli host that constitutively expressed T7 RNA polymerase, an essential enzyme of the early viral metabolism. Over the course of 180 generations a diversity of phage variants emerged, outgrew the wildtype, and were subsequently eclipsed by yet fitter variants, based on host-ranges, restriction patterns, and one-step growth responses of isolated clones. The fittest variant, which required complementation by the recombinant host in order to grow, deleted at least 12 percent of its genome and replicated twice as fast as the wildtype. Moreover, this variant was immunogenically indistinguishable from the wildtype, based on cross-reactivities of antisera raised against both. These results suggest the feasibility of the proposed strategy for the development of safe whole-virus vaccines.

Bacteriophage T7↗
PubMed1995

Secretion of human interleukin 2 by recombinant Mycobacterium bovis BCG.

The human interleukin 2 (huIL-2) gene was introduced into Mycobacterium bovis BCG by using the integrative vector pMV306. To express and secrete huIL-2 from BCG, two different plasmids, CI and CII, were made. In CI, the huIL-2-encoding region was under the control of the alpha-antigen promoter of BCG; in CII, the expression of huIL-2 was regulated by the heat shock protein 60 promoter. A signal peptide sequence isolated from the naturally secreted alpha-antigen of BCG was inserted between the promoter and huIL-2-encoding region to facilitate secretion. Both huIL-2 expression plasmids were integrated into the BCG genome when introduced into the BCG Pasteur strain by electroporation. Approximately 150 U of huIL-2 was secreted into the medium of a BCG-CII culture, while the BCG-CI cells secreted approximately one-sixth of that amount. When the IL-2-expressing BCG strain BCG-CII was injected intravenously into BALB/c mice, the number of BCG cells in the spleens of these mice was significantly less than the number in the control mice. The decreased number of IL-2-expressing BCG cells is likely due to the augmentation of the host immune response by the secreted huIL-2, although the exact mechanism is not known.

Animals↗
PubMed1995

The measurement and application of TSH-IRMA levels among different age groups in areas with iodine deficiency disorders.

Using immunoradiometric TSH assay (TSH-IRMA) to measure whole blood TSH levels spotted onto filter paper, we compared TSH levels among different age groups (neonates, < 1 yr infants, schoolchildren aged 7 to 14 yrs, adults, pregnant women and reproductive-age women) in Guiyang, where iodine deficiency has long been a problem. The results showed: 1) The percentage of subjects with TSH levels equal to or greater than 5 mIU/L in the neonate group was 38.9% while the percentages in the other groups were 0-3.3% (P < 0.01); 2) The TSH levels of the neonates were inversely related to the urinary iodine values of their mothers (pregnant women). (r = -0.5, P < 0.01); 3) 97.6% of the inhabitants in Guiyang ingested salt with less than 20 mg/kg iodine. The results indicate that iodine deficiency remains a problem in Guiyang. Neonates are the only ideal population for monitoring iodine deficiency.

Adolescent↗
PubMed1994

Purification, crystallization and preliminary X-ray analysis of murine interleukin-5.

Wild-type and mutant forms of murine interleukin-5 (mIL-5) have been expressed in the baculovirus expression system, purified, and used in crystallization trials. Attempts to obtain diffraction quality crystals of wild-type protein were unsuccessful. The substitution of glutamine for Asn75 preserved biological activity, while removing one of two predicted N-linked glycosylation sites, and the resulting protein was crystallized from polyethylene glycol 8000 at pH 7.8 in two crystal forms. The orthorhombic crystals, which belong to space group P2(1)2(1)2 with cell dimensions a = 55.9 A, b = 83.0 A and c = 52.3 A, diffract to beyond 2.5 A resolution. The second crystal form belongs to a trigonal space group, either P3(1)21 or P3(2)21, with cell dimensions a = b = 62.1 A, c = 129.9 A, and diffracts to about 3.8 A resolution. Each crystal form probably contains one mIL-5 dimer per asymmetric unit.

Animals↗
PubMed1994

Genes of stress in experimental hypertension.

1. A significant portion of blood pressure variance is modified by the environment. 2. The present report summarizes evidence that: (i) the environmental response is genetically determined; (ii) various stressors can evoke a differential response in hypertensive animals and constitute its intermediate phenotypes; (iii) the response to heat stress can be assigned to a single 'thermosensitivity' locus; (iv) candidate genes of susceptibility to environmental stresses are member(s) of the heat stress gene (HSP) gene families; (v) a restriction fragment length polymorphism of hsp70 and a single base mutation in the 3'-untranslated region of hsp27 are associated with hypertension in recombinant inbred strains. 3. In conclusion, HSP gene variants may be causative in susceptibility to hypertension.

Animals↗
PubMed1994

Deletion between direct repeats in T7 DNA stimulated by double-strand breaks.

An in vitro system based on extracts of Escherichia coli infected with bacteriophage T7 was used to study genetic deletions between directly repeated sequences. The frequency of deletion was highest under conditions in which the DNA was actively replicating. Deletion frequency increased markedly with the length of the direct repeat both in vitro and in vivo. When a T7 gene was interrupted by 93 bp of nonsense sequence flanked by 20-bp direct repeats, the region between the repeats was deleted in about 1 out of every 1,600 genomes during each round of replication. Very similar values were found for deletion frequency in vivo and in vitro. The deletion frequency was essentially unaffected by a recA mutation in the host. When a double-strand break was placed between the repeats, repair of this strand break was often accompanied by the deletion of the DNA between the direct repeats, suggesting that break rejoining could contribute to deletion during in vitro DNA replication.

Bacteriophage T7↗
PubMed1994

[Intracellular recordings and electrophysiological properties of neurons of pancreatic ganglia in vitro].

The work was carried out to investigate electrophysiological properties of neurons of cat intrapancreatic ganglia in vitro by means of intracellular recordings. The mean value of resting membrane potential was -58.5 +/- 8.7 mV (chi +/- s chi, n = 35) with a range from -45 to 72 mV. The mean values of membrane input resistance (Rm), time constant (tau) and capacity (Cm) were 68.6 +/- 5.1M omega, 3.4 +/- 0.2 ms and 50.8 +/- 3.9pF (n = 28), respectively. When depolarizing electrotonic potentials induced by intracellular injection of depolarizing current pulses (0.05-0.5nA, 5ms) reached the threshold potential level, all of the neurons could fire action potentials. The threshold, amplitude, overshoot and duration of spikes were 19.2 +/- 0.5mV, 81.0 +/- 1.7mV, 22.6 +/- 0.9mV and 2.9 +/- 0.1ms (n = 35), respectively. The spike was followed by a prolonged after spike hyperpolarization with amplitude of 20.5 +/- 0.8mV (n = 35). In most of neurons, fast excitatory postsynaptic potentials (f-EPSP) or orthodromic action potentials were recorded during stimulation of nerve trunks attached to the intrapancreatic ganglia. The mean values of amplitude, duration of the f-EPSP and conduction velocity of the nerves were 8.9 +/- 0.7mV, 25.8 +/- 1.9ms and 0.48 +/- 0.04m/s (n = 24), respectively. The ongoing synaptic activity was observed in all of cells. Moreover, f-Epsp, was induced by acetylcholine mediated through nicotinic receptors.

Action Potentials↗
PubMed1993

Deletion between directly repeated DNA sequences measured in extracts of bacteriophage T7-infected Escherichia coli.

An in vitro system based upon extracts of bacteriophage T7 infected Escherichia coli was used to study genetic deletions and to examine the importance of DNA replication in the deletion process. When T7 genomes with gene 1.3 inactivated by a 43-bp insert of random sequence DNA bracketed by 11-bp direct repeats were replicated in vitro the inserts were deleted with a frequency of about 10(-5) deletions per genome replication. Under conditions where deletion could take place only by recombination between direct repeats on distinct DNA molecules deletion frequency was at least an order of magnitude lower. These data demonstrate the utility of the in vitro system as a means for examining deletion mechanisms and underscore the importance of DNA replication in deletions.

Bacteriophage T7↗
PubMed1993

Evidence of mobilization of hobo transposons in a P-element mutagenesis screen.

In Drosophila melanogaster, mutations in Su(var) and En(var) loci either suppress or enhance position effect variegation, respectively. Towards the cloning of these genes we have induced, recovered, and characterized a series of Su(var) mutations located on the third chromosome. These mutations were recovered from a P-element mutagenesis scheme in which the third chromosome P[ry+ delta 2-3](99B) element would provide transposase in trans to mobilize a single nonautonomous P transposon, P[pUCHSNeo](9C), located on the X chromosome. Although this cross scheme induces mobilization of the X-chromosome P transposons and its subsequent reinsertion onto the autosomes, none of the Su(var) mutations recovered could be associated with P-transposon insertions. Further investigation of these mutations by in situ hybridization showed that another mobile element that can be mobilized in hybrids, hobo, was associated with these mutations. We conclude that the mobilization and reinsertion of the hobo elements, through an inadvertent hybrid HE dysgenic cross, is most likely responsible for the Su(var) mutations recovered from this P-element mutagenesis. The association of hobo element with this Su(var) locus will facilitate its cloning and offers the possibility of hobo transposon tagging other Su(var) loci.

Animals↗
PubMed1993

Transvaginal sonography in the diagnosis and management of cervical incompetence.

Cervical incompetence remains a controversial area in obstetric management. The development of transvaginal sonography has resulted in a clearer image of the internal cervical os, a development which may indirectly have beneficial effects on outcome. However, the rapid development of cervical dilatation in some cases necessitates frequent ultrasound monitoring.

Adult↗
PubMed1992

Restriction fragment length polymorphism of hsp70 gene, localized in the RT1 complex, is associated with hypertension in spontaneously hypertensive rats.

Previous studies from our laboratory have demonstrated that the intermediate phenotype of thermosensitivity is present in hypertensive mice and rats. Increased expression of hsp70 caused by increased transcription rate was demonstrated in vivo, in organs, and in cultured cells from spontaneously hypertensive rats and hypertensive mice. In this study, a polymorphism of this gene was revealed with BamHI enzyme by using a human hsp70 probe. A 4.4-kb fragment was visualized in normotensive rats (Brown-Norway BN.lx and Sprague-Dawley), and a 3.0-kb fragment was found in spontaneously hypertensive rats (SHR) of three different origins and in Wistar and Buffalo rats. Both fragments were present in the Wistar-Kyoto rat strain. The present study mapped the polymorphism of hsp70 into the RT1 complex in BN.1K and SHR.1N congenic strains. The hsp70 restriction fragment length polymorphism is associated with a blood pressure difference of 15 mm Hg in recombinant inbred strains. These results justify the search for a mechanism by which hsp70 could influence blood pressure.

Animals↗
PubMed1992

[Preliminary study of changes in plasma prostaglandin E1 in rats with experimental "schizophrenia"].

We referred to Borison's method of making the model schizophrenia. Rats were injected d-amphetamine intraperitoneally once a day so that the schizophrenia models were established within four weeks. After five weeks, rats with schizophrenia were treated with chlorpromazine. Contents of plasma prostaglandin E1(PGE1) in rats were determined before and after treatment respectively. This study demonstrated that contents of plasma PGE1 in rats receiving amphetamine for four weeks were higher than those in control animals (P < 0.05). At this time, male rats' stereotypy scores were 3.5 and female stereotypy scores 5. But contents of plasma PGE1 in rats with schizophrenia approached those in control animals with giving amphetamine once a day continuously. Contents of plasma PGE1 decreased in rats with schizophrenia receiving chlorpromazine.

Alprostadil↗
PubMed1991

The effect of the length of direct repeats and the presence of palindromes on deletion between directly repeated DNA sequences in bacteriophage T7.

The frequency of genetic deletion between directly repeated DNA sequences in bacteriophage T7 was measured as a function of the length of the direct repeat. The non-essential ligase gene (gene 1.3) of bacteriophage T7 was interrupted with pieces of synthetic DNA bracketed by direct repeats of various lengths. Deletion of these 76 bp long inserts was too low to be measured when the direct repeats were less than 6 bp long. However, the frequency of deletion of inserts with longer direct repeats increased exponentially as the length of the repeats increased from 8 to 20 bp. When inverted repeats (palindromes) were designed in the midst of the insert there was essentially no increase in deletion frequency between 10 bp direct repeats. But, the same palindromic sequences increased the deletion frequency between 5 bp direct repeats by at least two orders of magnitude. Thus, in this system homology at the endpoints is a more important determinant of deletion frequency than is the presence of palindromes between the direct repeats.

Base Sequence↗
PubMed1988

Effects of nimodipine on the production of thromboxane A2 following total global cerebral ischemia.

To clarify the contribution of vasoconstrictor prostaglandins to the hypoperfusion state typically following total global cerebral ischemia, 14 mongrel dogs were subjected to 11 minutes of global cerebral ischemia. They were then randomly assigned to receive either no treatment or an intravenous bolus of the calcium channel blocker nimodipine, 10 micrograms/kg, 15 minutes after ischemia followed by a continuous infusion of nimodipine, 1.0 micrograms/kg/min. Thromboxane (Tx) A2 production, as measured by cerebral venous levels of TxB2 (the stable metabolite of TxA2) increased similarly in the two groups. In contrast to previous studies, mean postischemic cerebral blood flow did not increase sufficiently in the nimodipine-treated group to achieve statistical significance. These data suggest that the improved neurological outcome associated with nimodipine treatment following global cerebral ischemia does not relate to reduced levels of the prostaglandin precursor arachidonate.

Animals↗