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Biomedical subjects

R L Wu

Publications and source records attributed to R L Wu.

At least 37 records · Page 2Linked to original sources
PubMed1992

Two pharmacologically and kinetically distinct transient potassium currents in cultured embryonic mouse hippocampal neurons.

Transient potassium currents in mammalian central neurons influence both the repolarization of single action potentials and the timing of repetitive action potential generation. How these currents are integrated into neuronal function will depend on their specific properties: channel availability at the resting potential, activation threshold, inactivation rate, and current density. We here report on the voltage-gated transient potassium currents in embryonic mouse hippocampal neurons dissected at embryonic days 15-16 and grown in dissociated cell culture for up to 3 d. Two transient potassium currents, A-current and D-current, were isolated based on steady state inactivation and sensitivity to 4-aminopyridine (4-AP) and dendrotoxin (DTx). A-current had an activation threshold of approximately -50 mV and was half-inactivated at approximately -81 mV. A-current relaxations at voltages between -40 and +40 mV could be fit by single exponential functions with time constants of 20-25 msec; these time constants showed little sensitivity to voltage. In contrast, D-current had an activation threshold of between -40 and -30 mV and was half-inactivated at approximately -22 mV. D-current inactivation was voltage dependent; time constants of fitted exponential functions ranged from approximately 7 sec at -40 mV to 200 msec at +40 mV. A slower component of inactivation was also evident. D-current was preferentially blocked by 4-AP (100 microM) and DTx (1 microM). Operationally, A- and D-currents could be cleanly separated based on conditioning pulse potential and 4-AP sensitivity. Total transient potassium current amplitude increased during the time that neurons were in culture (recordings were made between 2 hr after dissociation and 3 d in culture). When normalized for cell capacitance (an index of membrane area), A-current density (pA/pF) decreased and D-current density increased, even during a period between days 1 and 3 when total transient current density remained constant. This observation suggests that A- and D-currents may be reciprocally modulated. Since blockade of D-current (with 100 microM 4-AP) increased both action potential duration and repetitive firing in response to constant current stimulation, long-term modulation of the A-current:D-current ratio may affect the excitability of hippocampal neurons.

Animals↗
PubMed1991

Age-related changes in the synaptic plasticity of rat superior cervical ganglia.

A few seconds of tetanic preganglionic stimulation of rat sympathetic ganglia results in potentiation of cholinergic synaptic transmission lasting several hours. However, ganglia from aged (28-32 months) rats did not develop as much potentiation as did ganglia from young (3-6 months) rats. This potentiation appears to decay exponentially but in two phases. The early component decays in 15 minutes, a time course consistent with the phenomenon of post-tetanic potentiation (PTP). The later component decays over several hours and is a form of long-term potentiation (LTP). A double exponential decay model was employed to quantitatively resolve the decay of potentiation and allow quantitative comparison of the decay parameters in both aged and young rats. It is clear that the magnitude and duration of PTP was the same in both age groups. However both the magnitude and decay time-constant for LTP were 30-50% smaller in the aged group of rats. Several agents which mimic cyclic AMP or stimulate cyclic AMP production in the ganglion enhance nicotinic transmission for several hours. However, these agents, 8-bromo cAMP, forskolin, isoproterenol, and secretin were equally effective in potentiating transmission in ganglia from both young and aged rats. Previous studies demonstrated that stimulus induced LTP and cyclic AMP induced potentiation was the result of enhanced release of acetylcholine (ACh) and not from increased postsynaptic responsiveness. Presumably these agents act through the same pathway and by the same mechanism that generates LTP following preganglionic tetany. These observations demonstrate that there is a selective age-dependent decline in the capacity for sympathetic ganglia to generate long-term changes in synaptic efficacy.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗
PubMed1991

Capillary gas chromatographic method for mandelic and phenylglyoxylic acids in urine.

In support of an occupational investigation of styrene exposure, a capillary gas chromatographic method was developed for the quantitation of the styrene metabolites mandelic and phenylglyoxylic acids. The method was based on that of Guillemin and Bauer, in which phenylglyoxylic acid was converted to mandelic acid by reduction before instrumental analysis. The earlier method had to be modified for use with capillary columns; the resulting method was sensitive, selective and reproducible. The detection limit was approximately 0.001 mg/ml urine. Approximately less than 5% relative precision was achieved in the range of 0.05-2 mg/ml urine. Mandelic acid was resolved from other components of urine and from by-products of derivatization.

Air Pollutants, Occupational↗
PubMed1988

Preparation and properties of a biomaterial: HEMA grafted SBS by gamma-ray irradiation.

Solvent-cast films of styrene-butadiene-styrene triblock copolymer (SBS) were synthesized and a poly(2-hydroxyethyl methacrylate) (PHEMA) layer was grafted onto these films using Co 60 gamma-ray irradiation. The IR absorption spectrum of the graft copolymer was taken and its surface morphology studied by a scanning electron microscope. The degree of grafting and the mechanical properties of SBS, SBS-g-HEMA, and PHEMA were measured. In order to study its biocompatibility, contact angle and blood clotting time experiments were performed. The degree of grafting was related to the soaking time, film thickness and total irradiation dosage. However, it levelled off at 7% when the total dosage reached 1 Mrad. It was found that the mechanical properties of SBS-g-HEMA were the same as those of SBS and were superior to those of PHEMA. The blood compatibility of SBS-g-HEMA as measured by the Lee-White clotting test was better than that of SBS and polystyrene and was quite similar to that of PHEMA.

Acrylates↗
PubMed1988

[Morphology, diagnosis and differential diagnosis of mesothelioma].

Forty cases of mesothelioma biopsied and autopsied from 1961 to 1985 were studied and analysed as to their morphology, diagnosis and differential diagnosis. Mesothelioma had complex histologic types. The tumor cells varied in size and shape and were characterized morphologically by the coexistence of biphasic differentiation, adenoid and microvilli formation. The pathologic diagnosis is made according to the feature of biphasic differentiation, or coexistence of epithelioid and spindle tumor cells. It should be emphasized that the tissue sections are taken from multiple areas. Usually, the correct diagnosis is made in correlation to the clinical manifestations. For difficult cases of the diffuse mesothelioma, electron microscopy is needed to differentiate from metastatic adenocarcinoma and spindle cell sarcoma etc.

Adult↗
PubMed1988

Biological function of modified nucleotides in tRNA molecules--synthesis and biological activity of the analogues of yeast alanyl tRNA with I34 replaced by A34 or G34.

Analogues of yeast alanyl tRNA with I34 replaced by A34 or G34 were synthesized. Synthetic analogues of yeast alanyl tRNT occupy the same position as the natural yeast alanyl tRNA on polyacrylamide gel electrophoresis, and their purity is about 95% after electrophoresis on a 10% or 20% polyacrylamide gel. The two terminal and nearest neighbour nucleotides of the analogues are all correct. The accepting activity of the synthetic analogues is similar to that of the reconstituted natural yeast alanyl tRNA. The incorporation activity of alanine into proteins of the synthetic analogues is about 30% of that of the natural of reconstituted natural yeast alanyl tRNA when I34 is replaced by A, and is 90% when I34 is replaced by G. The reason of the variation in biological function of the analogues of yeast alanyl tRNA after I34 replaced by A or G was discussed.

RNA, Fungal↗
PubMed1987

Effect of the anticodon loop size of yeast alanyl tRNA on its biological activity.

Three analogs of yeast alanyl tRNA with anticodon loops of different sizes, tRNA75 (no G35 and 5'-terminal phosphate), tRNA77 (one more C between G35 and C36, no 5'-terminal phosphate), and ptRNA79 (with Cm1I psi between G35 and C36), were synthesized. In comparison with the reconstituted natural yeast tRNA, the charging activities of the three analogs were 90% (tRNA75), 94.7% (tRNA77), and 104% (ptRNA79). These results supported the conclusion (Yang De-ping and Wang De-bao (T. P. Wang) (1983) Acta Biochim. Biophys. Sin. 15, 83-90) that the anticodon loop of yeast alanyl tRNA was not involved in the interaction between alanyl-tRNA synthetase from rat liver and yeast alanyl tRNA. In contrast, in the rabbit reticulocyte lysate system, the incorporation of alanine in the charged analogs was 0% (tRNA75 and ptRNA79) and 100% (tRNA77). There were significant differences between the incorporation activities of analogs and those of the reconstituted molecule. The reason for these differences is discussed.

Anticodon↗
PubMed1987

[Significance of ultrastructure of peripheral nerve tumors in pathological and differential diagnoses].

Biopsy material fixed with formalin in 10 cases of peripheral nerve tumors and 5 soft tissue tumors were studied with special reference to the significance of the ultrastructure of peripheral nerve tumors in histopathological and differential diagnosis. The results demonstrated that electron microscopic examination of formalin-fixed tissue may still reveal their ultrastructural characteristics. This is of certain practical value in pathological and differential diagnosis in some difficult peripheral nerve tumors.

Diagnosis, Differential↗
PubMed1984

alpha-Tocopherol in the developing rat retina: a high pressure liquid chromatographic analysis.

High pressure liquid chromatography was used to measure alpha-tocopherol in the retinas of rats reared in a cyclic light or dark environment. These measurements were performed on extracts of whole retinas during the developmental period, 18-60 days, and on isolated ROS from adult animals. Similar alpha-tocopherol determinations were performed on retinas and isolated ROS following exposure of rats to intense visible light for 24 hr periods. The results show that alpha-tocopherol is chromatographically separated from the vitamin A derivatives found in the retina and is pure, as judged by mass spectrometry. In the retinas of cyclic light and dark reared rats, alpha-tocopherol accumulates in an age dependent fashion, so that at 60 days the level is nearly double that of animals at 18-20 days of age (P less than 0.001). Because the age dependent accumulation of rhodopsin is greater in dark reared rats, the average molar ratio of rhodopsin to alpha- tocopherol in the retina of dark reared animals is 25% higher than in cyclic light rats. Following exposure of rats to intense visible light for 24 hr periods, alpha-tocopherol concentrations in the retina were unchanged from the levels in control animals. In adult animals the concentration of alpha-tocopherol in ROS is 2.5-3.5 times higher than in whole retina. ROS from adult cyclic light reared rats also contain an average of 43% more alpha-tocopherol per mg protein than ROS from dark maintained animals (P less than 0.005).(ABSTRACT TRUNCATED AT 250 WORDS)

Age Factors↗
PubMed1983

Total synthesis of yeast alanine transfer ribonucleic acid.

By a combination of chemical and enzymatic methods, small oligonucleotides with lengths varying from 2 to 8 nucleotides were synthesized from mononucleotides. The small oligonucleotides were then ligated with T4 RNA ligase into six large oligonucleotides (9 to 19 nucleotides long) which were further ligated to form two half molecules with 35 and 41 nucleotides respectively. Finally, the two synthetic half molecules were annealed and ligated to obtain the whole molecule of yeast alanine tRNA (tRNAAlay). Prior to this, two semi-syntheses were performed, i.e. ligation of the synthetic 5'-half molecule with the natural 3'-half molecule and that of the natural 5'-half molecule with the synthetic 3'-half molecule. Both the semi-synthetic tRNAAlay and the synthetic tRNAAlay occupy the same position as the natural tRNAAlay after electrophoresis on a 20% polyacrylamide gel. They have the same chemical composition (containing 9 modified nucleotides of 7 different species) and structure as the natural tRNAAlay and are biologically active, i.e. accepting and transferring alanine into proteins in a cell-free protein synthesizing system, the accepting activity of the synthetic product is 52-66% of that of the natural tRNAAlay and 91-106% of that of the reconstituted product of the two natural half molecules. The incorporation activity of alanine into proteins of the synthetic 3H-alanine tRNAAlay is 63%, corresponding to 91% of that of the natural tRNAAlay and 115% of that of the reconstituted product of the two natural half molecules. To the best of our knowledge, this is the first time that a natural RNA with biological activity is synthesized.

Base Sequence↗
PubMed1983

Synthesis of the 3'-half molecule of yeast alanine tRNA.

This paper deals with the synthesis of the 3'-half molecule of yeast alanine transfer RNA (tRNAAlay) by ligation with T4 RNA ligase of three component oligonucleotide fragments corresponding to nucleotides 36-45(I), 46-57(II) and 58-76(III) in succession extending from the 3'-end to the 5'-end. First, in a ratio of acceptor to donor at 1.5 to 1, we adopted a method of three successive reactions, namely, the 5'-phosphorylation of the nonadecamer (III), ligation with the dodecamer (II) and the 5'-phosphorylation of the ligation product formed; with one isolation step and obtained the 5'-phosphorylated 31mer(46-76) (IV) in an overall yield of 70%. Then the 31mer(IV) as a donor was ligated with 3 times of decamer (I) to form the 41mer(36-76) (V), the 3'-half molecule of tRNAAlay. The yield was 67%. After 5'-phosphorylation, (V) was ligated with the natural 5'-half molecule to form the semi-synthetic tRNAAlay, which was biologically active, i.e. accepting and transferring (3H)-alanine into proteins.

Base Sequence↗
PubMed1983

Synthesis of the 5'-half molecule of yeast alanine tRNA.

In this paper, we report the synthesis of the 5'-half molecule of yeast alanine tRNA (tRNAAlay) by ligating three oligonucleotide fragments corresponding to the nucleotide sequences 1-13, 14-22 and 23-35 respectively under the catalysis of T4 RNA ligase (Fig. 1). Because of the high purity of the oligonucleotide fragments and the excellent quality of T4RNA ligase and polynucleotide kinase we prepared, the isolation steps were simplified and the overall yields were much higher. The ligating yield of the docosamer (IV) was 75%, that of the pentatriacontamer (V), 90%, and the isolated yield of the final product was 21% calculated on the basis of the tridecamer (III) used in the first reaction. Under the action of T4 RNA ligase the synthetic 5'-half molecule was joined with the natural 3'-half molecule forming a semi-synthetic tRNAAlay, which possessed the biological activities of both accepting (3H)-alanine and incorporating it into proteins. The correctness of the structure of the synthetic 5'-half molecule was verified by both chemical analysis and biological activity assay.

Base Sequence↗