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Biomedical subjects

B Chen

Publications and source records attributed to B Chen.

At least 559 records · Page 31Linked to original sources
PubMed1976

Effects of methylation of the beta-galactosidase genome upon in vitro synthesis of beta-galactosidase.

A template DNA from phage lambdah80dlacp5 coding for the in vitro synthesis of beta-galactosidase was used to study the effect of DNA methylation by the alkylating agent, dimethyl sulfate (DMS). Increasing the levels of DMS up to 50 mM concentration in the incubation medium led to an increase of DNA methylation. When incubated for 10 min at 37 degrees C, 3-4% Of nucleotides were methylated. The increase was linear to about 0.6% nucleotide methylation level. A higher yield was obtained at 37 degrees C incubation temperature than at 20 degrees C. Methylation of lambdah80dlacp5 DNA alone without methylation of other factors in the incubation mixture caused inhibition of the synthesis of beta-galactosidase in vitro. Increasing levels of DNA methylation caused greater inhibition of the newly synthesized enzyme activity. Total protein and RNA synthesis was inhibited by the methylated DNA to a much lesser extent than the inhibition of enzyme activity. When the level of nucleotide methylation was 0.74%, only 2% of enzyme activity remained, but total protein and RNA synthetic activities were found to be 72% and 44%, respectively.

Coliphages↗
PubMedDate not supplied

Replication-deficient vaccinia virus gene therpay vector: evaluation of exogenous gene expression mediated by PUV-inactivated virus in glioma cells.

BACKGROUND: Mild psoralen and UV (PUV) treatments inactivate viral DNA replication, but the virus retains its ability to infect cells. Thus, PUV treatment of vaccinia virus (VV) vectors may increase the safety of gene delivery and extend the duration of gene expression. Although the first studies on PUV-inactivated VV (PUV-VV) for the delivery of suicide or cytokine genes to cancer cells were promising, the efficiency and kinetics of exogenous gene expression have not been fully evaluated. Furthermore, these studies should be extended to other gene therapy strategies, e.g. tumor suppressor genes. METHODS: We constructed VV recombinants carrying the luciferase (luc) gene, or the tumor suppressor p53 gene, to analyze exogenous gene expression after PUV treatment. Apoptosis induction and antitumor effects were examined in glioma cell culture and in an animal model, respectively. RESULTS: PUV-VV induced efficient PE/L-driven expression of luc and p53 exogenous genes in infected cells. A surprising prolonged p53 protein production was measured in glioma cells infected with PUV-VV expressing p53 (VV-TK-53) on Days 5-7 post-infection, reaching a maximal level of 9 microg/ml. VV-TK-53 induced apoptosis in 88% and 77.6% of infected C6 and 9L glioma cells, respectively. In contrast, 80% of cells infected with the PUV-inactivated control virus remained viable. Finally, ex vivo infection of C6 glioma cells with PUV-inactivated VV-TK-53 significantly reduced subsequent tumor growth in nude mice. CONCLUSIONS: Replication-deficient PUV-VV is safe and very efficient in prolonged foreign gene expression. Therefore PUV-VVs are recommended as vectors for applications in cancer gene therapy and recombinant vaccine development.

Animals↗
PubMedDate not supplied

Significantly higher frequency of the MspI 2.2 kb allele of the Duchenne muscular dystrophy intragenic probe P-20 in the Chinese population.

The P-20 intragenic marker was used to test for restriction fragment length polymorphisms in unrelated Chinese patients with Duchenne or Becker muscular dystrophy or X-linked mental retardation. In addition to polymorphism at the 6.0/3.5 kb MspI allelic site, we found an independent and high frequency of polymorphism at the 2.2/1.8 kb site. This differs from results found with other populations.

Alleles↗
PubMedDate not supplied

The effect of ethanol drinking on opioid analgesia and receptors in mice.

Recent studies suggest substantial interactions between opioids and ethanol (EtOH). Both in vivo and in vitro experiments indicate that EtOH can regulate opioid systems and that opioids can modify EtOH consumption. In the present studies, we examined if EtOH consumption altered opioid receptors and the potency of opioid analgesics. Mice were given unlimited access to 6-7% EtOH alone for 7 days or were allowed to drink increasing concentrations (3-6%) of EtOH over 13-14 days. Controls had access to water. The EtOH groups drank significantly less volume than controls, although there were no significant differences in body weight or baseline nociception. The analgesic (tail flick) potency of SC morphine was decreased by approximately 1.6-2.0-fold in EtOH-treated mice. A single acute dose of EtOH (1 g/kg) that produced blood alcohol levels in excess of that for 7 day exposure to EtOH, did not change morphine's analgesic ED50, suggesting that chronic exposure to EtOH was necessary for the reduction in potency. The change in morphine potency was not due to pharmacokinetic differences because EtOH consumption did not modify the concentration of morphine in brain and spinal cord. The analgesic potency of a delta-opioid receptor agonist (ICV DSLET) was also decreased by approximately 2-fold. Saturation binding studies indicated no changes in the density or affinity of brain and spinal cord delta-opioid ([3H]DPDPE, [3H]DSLET, [3H]DeltorphinII) and mu-opioid ([3H]DAMGO) receptors. Similarly, there was no significant effect of EtOH on delta-opioid receptor mRNA in either brain or spinal cord preparations. Taken together, these data suggest that EtOH consumption decreases the analgesic potency of opioids in mice through a mechanism that is unrelated to pharmacokinetics or opioid receptor changes in brain and cord.

Alcohol Drinking↗
PubMedDate not supplied

Scorpion BmK venom induces nociceptive response of rats by plantar injection.

In this study, the nociceptive responses induced by subcutaneous injection of scorpion BmK venom into the plantar surface of rat hind paw were quantified. The suitable dose of the venom is between 0.01 and 0.05 mg. The venom injection could induce local edema and tonic-pain responses, which had lasted more than 1 h. The pain scores reached peak intensity within 20 min, and then decreased slowly. Morphine, as a classic analgesic drug, could inhibit the spontaneous nociceptive response induced by the venom. With Morphine pretreatment, the mean pain scores for the entire 60 min were significantly decreased (P<.05). BmK I, a main lethal component, was also found to induce nociceptive responses with dose-dependent means. The nociceptive responses induced by BmK I could be partially inhibited by morphine.

Analgesics, Opioid↗
PubMedDate not supplied

Biological activity of two juvenoids and two ecdysteroids against three stored product insects.

The insecticidal activity of juvenile hormone agonists methoprene and pyriproxyfen, and the ecdysone agonists RH-5849 and tebufenozide was evaluated against susceptible and actellic-resistant strains of Tribolium castaneum and susceptible strains of Rhyzopertha dominica and Sitophilus oryzae. Concentrations ranging from 0.1 to 20 ppm of the analogues were mixed in the food medium to which the tested insects were exposed. The results showed that all these compounds could affect the development of the tested species to differing extents but had no effect on the mortality of parental adults. The two JH analogues did not prolong the life span of R. dominica and S. oryzae, but very greatly extended that of T. castaneum. The extension led to the production of giant larvae and failure to pupate. Actellic-resistant strain of T. castaneum showed some cross-resistance to methoprene and pyriproxyfen, but not to RH-5849 and tebufenozide. Pyriproxyfen was the most effective compound among the four IGRs; a concentration of 0.1 ppm could completely inhibit the F(1) adult occurrence of both S- and R-strains of T. castaneum and its LC(90)s for controlling R. dominica and S. oryzae were 0.1 and 1.2 ppm, respectively. Methoprene was highly effective against R. dominica, but less active on S. oryzae. RH-5849 could achieve almost complete control of F(1) adults of T. castaneum and R. dominica at 10 ppm, but was less potent on S. oryzae. Tebufenozide appeared to be much less active on these three species compared with the other three compounds. The percentage reductions of F(1) adults for S- and R-strains of T. castaneum at a concentration of 20 ppm were 80 and 99%, respectively.

Animals↗
PubMedDate not supplied

Morphological diversity in terminals of W-type retinal ganglion cells at projection sites in cat brain.

The morphological features of retinal terminals in cat brain were examined at sites where projections of W-type ganglion cells predominate. These included the parvicellular C laminae of the dorsal lateral geniculate nucleus, the ventral lateral geniculate nucleus, stratum griseum superficiale of the superior colliculus, and the suprachiasmatic nucleus. Positive identification of retinal terminals was achieved following anterograde transport of intravitreally injected native or wheat germ agglutinin-conjugated horseradish peroxidase. In contrast to the classic features of retinal terminals as defined from sites where X- and Y-type ganglion cells predominate, i.e. round synaptic vesicles, large profiles, and pale mitochondria, substantial numbers of terminals in W-cell rich areas were found to contain dark mitochondria. Synaptic vesicles, although consistently round, were typically smaller in terminals with dark mitochondria than in those with pale mitochondria. These findings indicate a diversity among terminals of W-cells and suggest that such terminals cannot be distinguished on the basis of limited morphological criteria.

Animals↗
PubMedDate not supplied

Identification of the heme adduct and an active site peptide modified during mechanism-based inactivation of rat liver cytochrome P450 2B1 by secobarbital.

The olefinic barbiturate secobarbital (SB) is a sedative hypnotic known to be a relatively selective mechanism-based inactivator of rat liver cytochrome P450 2B1. Previous studies have demonstrated that such inactivation results in prosthetic heme destruction and irreversible drug-induced protein modification, events most likely triggered by P450 2B1-dependent oxidative activation of the olefinic pi-bond. However, the precise structure of the SB-modified heme and/or the protein site targeted for attack remained to be elucidated. We have now isolated the SB-heme adduct from P450 2B1 inactivated by [14C]SB in a functionally reconstituted system and structurally characterized it by electronic absorption spectroscopy and tandem collision-induced dissociation (CID), matrix-assisted laser desorption ionization on time of flight (MALDI-TOF), and liquid secondary ion mass spectrometry in the positive mode (+ LSIMS) as the N-(5-(2-hydroxypropyl)-5-(1-methylbutyl)barbituric acid)protoporphyrin IX adduct. The [14C]SB-modified 2B1 protein has also been isolated from similar inactivation systems and subjected to lysyl endopeptidase C (Lys-C) digestion and HPLC-peptide mapping. A [14C]SB-modified 2B1 peptide was thus isolated, purified, electrotransferred onto a poly-(vinylidene) membrane, and identified by micro Edman degradation of its first N-terminal 17 residues (S277NH(H)TEFH(H)ENLMISLL293) as the Lys-C peptide domain comprised of amino acids 277-323. This peptide thus includes the peptide domain corresponding to the distal helix I of P450 101, a region highly conserved through evolution, and which is known not only to flank the heme moiety but also to intimately contact the substrates. This finding thus suggests that SB-induced protein modification of P450 2B1 also occurs at the active site and, together with heme N-alkylation, contributes to the SB-induced mechanism-based inactivation of P450 2B1.

Amino Acid Sequence↗
PubMedDate not supplied

Solid phase synthesis of combinatorial libraries using anhydrides as templates.

A simple and general approach to the synthesis of chemical libraries based on a universal anhydride template allows the preparation of large number of compounds. Various cyclic/acyclic amines, primary/secondary amines, differentially protected bifunctional amines were used as nucleophiles to react with anhydrides. The free carboxylic acid generated was then coupled with solid-bound amines. The facile and rapid generation of compounds through this multi-component assembly can be accomplished in a combinatorial parallel synthesis.

Acylation↗
PubMedDate not supplied

Continuous passive motion after total knee arthroplasty: a prospective study.

OBJECTIVE: Continuous passive motion (CPM) has been shown to increase significantly the amount of knee flexion for patients with total knee arthroplasty in the acute care hospital. Whether there is any additional benefit to using CPM for these patients who are transferred to a rehabilitation hospital is not known. There have been no prospective, randomized, controlled studies in this area. DESIGN: Fifty-one such patients on an inpatient rehabilitation service were randomly assigned to two groups. Group 1 (n = 23) received CPM for 5 consecutive hours per day plus physical therapy, whereas group 2 (n = 28) received only physical therapy. Knee flexion was measured by a blinded physical therapist on admission, on the third and seventh days of hospitalization, and at the time of discharge. RESULTS: The results indicated no significant difference in passive range of motion between group 1 and group 2. Patients in group 1 achieved an average increase in passive range of motion of 16 degrees, whereas those in group 2 achieved an average of 19 degrees (P = 0.33). CONCLUSION: Although power analysis indicated the need for differences in results for 32 patients per group to achieve significance, the difference between the two groups suggested neither statistical nor clinical significance. We concluded that the use of CPM in the rehabilitation hospital is likely of no added benefit to patients admitted after single total-knee replacement.

Aged↗