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Biomedical subjects

S M Wu

Publications and source records attributed to S M Wu.

At least 163 records · Page 9Linked to original sources
PubMed1986

Analysis of six distinct integrated hepatitis B virus sequences cloned from the cellular DNA of a human hepatocellular carcinoma.

Six distinct hepatitis B virus (HBV) integrations and the flanking cellular sequences were cloned from a hepatoma DNA preparation. None of the cloned fragments retains the entire HBV sequences but the surface antigen (HBsAg) gene and the HBV enhancer are retained in three of the six clones. The other three clones carry only short and possibly highly rearranged HBV genomic sequences and seem to contain some GC-rich clusters. Members of the repetitive Alu family are also found in the vicinity of five of the six integration regions which may have contributed to genome instability. In these six clones, the preferred integration sites are shown to lie within the single-strand region of the HBV genome. None of the clones carries in the flanking cellular sequences any of the 17 oncogenes tested, although the possibility still exists that an oncogene may be found on the side of the genome which has not been cloned. This work thus paves the way for detailed sequence analysis of virus-host junctions, for transfection studies of the HBV integration events, and for a search of genes in the flanking cellular sequences which may have been activated by the retained HBV enhancer using the clones described.

Base Sequence↗
PubMed1986

Effects of gamma-aminobutyric acid on cones and bipolar cells of the tiger salamander retina.

The gamma-aminobutyric acid (GABA) system in the tiger salamander retina was studied using autoradiographic and electrophysiological techniques. A high-affinity uptake mechanism for GABA has been localized in about 60% of the horizontal cells and about 30% of the amacrine cells. Effects of exogeneously applied GABA on the membrane conductance of cones and hyperpolarizing bipolar cells (HBC) were examined using the two electrode current-clamp technique in the living retinal slices. In both cell types, 1 mM of GABA caused a conductance increase. In perfused eyecups, 2 mM of GABA selectivity abolished the surround response of the HBC and left the center response unchanged. These results are consistent with the notion that a population of horizontal cells and a population of amacrine cells in the salamander retina may use GABA as their neurotransmitter.

Ambystoma↗
PubMed1986

Restriction fragment length polymorphism of the human beta-globin gene cluster: analysis of a beta-thalassemic family in Taiwan.

We have analysed seven polymorphic restriction sites of the human beta-globin gene cluster of six members of a Chinese family with a beta +-thalassemic sibling. The seven polymorphic sites analysed are the HincII site at the 5'-end of the epsilon-globin gene, the HindIII sites in the two gamma-globin genes, two HincII sites within and at the 3'-end of the psi beta 1 pseudogene, the AvaII site in the beta-globin gene and the BamHI site located at the 3' side of the beta-globin gene. The beta thal chromosome has been identified to have a haplotype of +----++ with respect to these seven polymorphic sites. This is also the most predominant haplotype associated with beta +-thalassemia in Mediterranean and Chinese populations (Chen et al., 1984; Orkin et al., 1982). Of the seven sites analysed in this family, four will be useful in prenatal diagnosis of beta-thalassemia in subsequent pregnancies in the family.

DNA Restriction Enzymes↗
PubMed1985

The effect of light on the spread of signals through the rod network of the salamander retina.

Adjacent rods in the amphibian retina are electrically coupled to each other by gap junctions. By injecting current pulses into one rod and recording the voltage change produced in nearby rods, we have studied the extent to which signals spread between rods in the presence and absence of illumination. Light has little effect on the steady potentials produced in nearby rods by the injection of a hyperpolarizing current, but does affect the propagation of transient signals through the rod network. The responses to injection of depolarizing current are increased by light. These effects of light were mimicked by hyperpolarizing the rod network (non-uniformly) by injecting continuous current (on top of which current pulses were superimposed to monitor signal spread). This suggests that the effects of light are due solely to the rod hyperpolarization produced by light. The effects of light are not completely predicted from computer simulations based on a previous characterization of the properties of isolated rods; these experiments thus reveal an inadequacy in the description of the rod membrane currents in that model. Light-induced hyperpolarization of cones has no effect on signal spread between rods. The functional significance of these results is discussed.

Ambystoma↗
PubMed1985

Characteristics of hepatitis B surface antigen produced in yeast.

We have constructed an expression plasmid for regulated expression of the hepatitis B surface antigen gene in yeast using promoter of the yeast Pho5 gene. In the yeast transformants, the monomeric HBsAg (22K dalton) was estimated to constitute approximately 3% of the total proteins. On extraction, the HBsAg was found to have a buoyant density of 1.18 g/ml and an Sw.20 value of 54. Electron microscopy revealed particles of heterogeneous size ranging from 18-28 nm. When the yeast HBsAg was used to immunize guinea pigs, the anti-HBsAg antibodies produced could react with human serum HBsAg.

Animals↗
PubMed1985

Chloroplast biogenesis. Molecular structure of chlorophyll b (E489 F666).

Nuclear magnetic resonance analysis established the presence of two vinyl groups/macrocycle of chlorophyll b (E489 F666). The latter is the only chlorophyll b that accumulates in a corn mutant (ON 8147). On the other hand the presence of only one vinyl group/macrocycle of authentic monovinyl chlorophyll b (E475 F660) was confirmed.

Chlorophyll↗
PubMed1985

State of hepatitis B virus DNA in leucocytes of hepatitis B patients.

Hepatitis B virus (HBV) DNA in leucocytes from 50 hepatitis patients with various patterns of HBV serological markers and serum HBV DNA and 13 normal controls were examined by Southern blot hybridization with 32P-labeled 3.2 Kb HBV DNA. A free form of HBV DNA was observed in leucocytes of 8 patients, 7 of whom were positive for serum HBeAg, and in 6 patients an integrated form of HBV DNA was identified. HBV DNA was not identified in leucocytes from 13 normal controls. The free form of HBV DNA in leucocytes existed as a heterogeneous smear from 2.0 to 3.2 Kb, similar to the pattern in liver and hepatocellular carcinoma cells but different from serum HBV DNA in which the 3.2 Kb band was absent. The banding pattern of the integrated form of HBV DNA in leucocytes varied among different patients. During preparation of white blood cells and purification of HBV DNA probes, it was important to remove plasma contamination and traces of pBR322, respectively. The presence of extrachromosomal DNA sequences partially homologous to pBR322 could cause false results. The presence of a free and integrated form of HBV DNA in leucocytes is important for explaining the biology of HBV, the harbouring and replication sites of extrahepatic origin, the mechanism of recurrent infection, and the rationale of the treatment of hepatitis B.

Carrier State↗
PubMed1985

Synaptic transmission from rods to bipolar cells in the tiger salamander retina.

Synaptic transmission between rods and depolarizing bipolar cells (DBC) was studied by using simultaneous recording techniques in the living retinal slice preparation. Current injection into the rod elicited a sign-inverting, sustained voltage change in the DBC. Voltage "tails" after the termination of a bright flash were observed in dark-adapted rods and DBC but not in cones. These simultaneously recorded voltage tails were used to isolate the rod input from the cone input and to study the input-output relation of the rod-DBC synapse. Within the voltage range between 0 and -10 mV from the rod dark potential (-39 +/- 1.2 mV), the input-output relation of the rod-DBC was approximately linear, with an estimated gain of about 3.7.

Animals↗
PubMed1985

Measurement of individual aflatoxin exposure among people having different risk to primary hepatocellular carcinoma.

The establishment of the carcinogenic role of aflatoxins has been impeded by the lack of suitable tests to measure individual exposure for long-term studies. It was shown that the use of immuno-concentration followed by high pressure liquid chromatography (HPLC) or immunoassay can regularly detect aflatoxins down to pg/ml in fluids including urine. Aflatoxin M1 (AFM1) in free form was identified as the major metabolite in urine suitable for use as an approximate dosimetric indicator of recent exposure to aflatoxin B1 (AFB1). A panel of monoclonal antibodies of IgG class were developed in the murine system against AFB1 and/or AFM1. Their affinity constants are at the level of 10(8)-10(9) L/mol, and they are suitable for use in either effective immuno-concentration or in high sensitivity immunoassays. It was shown in a high risk area of liver cancer (Qidong of China) that 10% or more of the local inhabitants had a urinary output of AFM1 one or two orders of magnitude higher than that of Beijing people, especially during the wet seasons. The ingestion of AFB1 among these local people was estimated to exceed 1 mg/year. The major source came from contaminated corn and rice, but local alcoholic beverages also contributed. The increased excretion of AFM1 was more pronounced in patients with chronic active hepatitis. This observation of very low AFM1 content in urine of a significant percentage of local people implies their food storage practices may offer a feasible method for such prevention. The value and possible problems in conducting long-term studies on primary prevention are discussed.

Aflatoxins↗
PubMed1985

Microbiological examination of foods sold via vending machines.

A total of 153 food specimens obtained from vending machines in Taipei city were subjects for the microbiological examination. Based on the standards for total microbial counts and total coliform counts established by the Department of Public Health, Republic of China, the unsatisfactory percentage of such counts for foods sold from vending machines were: 14% for carbonated beverages, 71% for non-carbonated beverages, 73% for ice cubes, and 100% for soft ice cream. Among the non-carbonic acid beverages examined, the unsatisfactory percentage for iced coffee, cocoa, chocolate milk, and fruit milk reached 90%. Escherichia coli and fecal streptococci each were found once. None of the pathogens causing food-poisoning were found during the study. As for the sanitation control of vending machines examined, 66 to 74% percent were unsatisfactory.

Bacteria↗
PubMed1984

Activation of chemical carcinogens by cultured human fetal liver, esophagus and stomach.

Cultured fetal human stomach, esophagus and liver activated benzo[a]pyrene (BP), aflatoxin B1 (AFB) and certain N-nitrosamines into metabolites that bound to cellular DNA. When the 3 organs were compared the highest level of activity was observed in the stomach. The interindividual variation was 10-fold and the amount of carcinogen-DNA adducts did not correlate with the sex or age of the fetus. The reaction products between BP or AFB and cellular DNA were investigated in liver explants. The carcinogen-DNA adduct patterns were identical to those observed in adult human tissues; BPDEI-Gua being the major adduct formed by BP and 2,3-dihydro-2-(7'-guanyl)-3-hydroxy-AFB by AFB. The results indicate that fetal organs can metabolize those oncogenic compounds at an early stage of the development, and that the metabolic pathways and DNA adducts are quite similar to those in experimental animals in which the compounds are carcinogenic.

Aflatoxin B1↗
PubMed1984

A quantitative analysis of interactions between photoreceptors in the salamander (Ambystoma) retina.

A quantitative description of the electrical properties of the photoreceptor layer in the salamander retina was obtained from earlier data on the characteristics of isolated rods and cones and on rod-rod coupling, and from new data on rod-cone and cone-cone coupling and on the rod photocurrent. Injecting -1 nA current into a rod elicits hyperpolarizations of about 20 mV in an adjacent rod and 4 mV in an adjacent cone. Responses of more distant receptors are smaller. Injecting -1 nA into a cone elicits hyperpolarizations of about 4 mV in an adjacent rod and 0.4 mV in a nearby cone. Depolarizing current evokes smaller responses. Assuming, in agreement with anatomical evidence, that each rod is electrically coupled to four rods and to four cones around it, and that there is no direct electrical coupling between cones, we found these results could be predicted from the properties of isolated rods and cones if adjacent rods are coupled by a resistance of 300 M omega and adjacent rods and cones are coupled by a resistance of 5000 M omega. The small cone-cone coupling seen is due to coupling via intervening rods. The two halves of double cones are not electrically coupled. The spectral sensitivity of both halves is a maximum around 620 nm wave-length. The rod photocurrent has been characterized by voltage-clamping rods isolated from the retina. In agreement with Bader, MacLeish & Schwartz (1979) we found the time course of the photocurrent to be approximately independent of voltage between -35 and -85 mV. The voltage responses of rods, single cones and double cones isolated from the retina obey the principle of univariance. Responses of receptors in the retina do not obey univariance. The main deviations from univariance observed can be explained if adjacent rods and cones are coupled by a resistance of 5000 M omega. Our data demonstrate that rod-cone coupling is relatively weak. We simplified our description of the photoreceptor network, by omitting cones, to investigate the spatiotemporal processing that the rod network is capable of. Computer simulations predict, as is found experimentally, that the rod voltage response to a large spot of bright light should show a much more pronounced initial transient hyperpolarization than the response to a small spot of light of the same intensity. This difference is produced by the combination of electrical coupling of the rods with the existence of a voltage-gated current, IA, in the rod membrane.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗
PubMed1983

Chlorophyll a biosynthetic routes and chlorophyll a chemical heterogeneity in plants.

A six-branched chlorophyll a biosynthetic pathway instead of a four-branched pathway has been proposed as being responsible for the formation of chlorophyll a in green plants. The several biosynthetic routes that make up the pathway have been described as leading to the formation of ten chemically different groups of chlorophyll a species. The latter differ from one another by one or more of the following modifications: (a) by having a vinyl or ethyl group at position 4 of the macrocycle, (b) by the nature of the long-chain fatty alcohols at position 7 of the macrocycle, and (c) by having a 6-membered lactone ring instead of a 5-membered cyclopentanone ring. The chemical structure of several of the metabolic intermediates of that pathway and of some of the chlorophyll a species have now been determined by primary chemical derivatization methods coupled to spectrofluorometric, nuclear magnetic resonance and mass spectral analyses. The formation of highly organized photosynthetic membranes in which some of the chlorophyll alpha molecules are specifically oriented is ascribed to the multiplicity of chlorophyll biosynthetic routes which result in the formation of multiple chlorophyll alpha chemical species. Proper orientation of chlorophyll in the photosynthetic membranes is visualized as being controlled by peripheral group modifications that either modulate the polarity of the Chl chromophore or control the magnitude of the net positive charge on the central Mg atom. Finally it is proposed that in addition to the proper orientation of chlorophyll a, chemical heterogeneity of the chlorophyll chromophores in the photosynthetic reaction centers is mandatory for efficient charge separation, and proper vectorial electron transfer.

Chlorophyll↗
PubMed1983

A sign-reversing pathway from rods to double and single cones in the retina of the tiger salamander.

Signal transmission between rods and cones was studied by passing current into a rod and recording the voltage response in a nearby double or single cone and vice versa. Two types of rod-cone interaction were found. Between immediately adjacent rods and cones, passage of current into either receptor elicited in the other receptor a sustained voltage response of the same sign as the injected current. These signals were still seen in the presence of Co2+, and are probably mediated by the electrical synapses which have been seen anatomically between adjacent rods and cones. In addition to this short-range sign-preserving interaction, passing current into a rod elicited a transient sign-inverted signal in cones up to at least 80 micron from the injected rod. No such response was seen in rods for current injection into cones. This signal was greatly reduced by Co2+ ions. Hyperpolarization of the cone to about -65 mV, with about 0.1 nA current, reversed this signal, which is presumed to be mediated by a chemical synaptic input to cones. Light flashes suppressed the sign-inverted signal for a period which was longer for brighter flashes. The time of reappearance of the signal was correlated with the return of the rod and horizontal cell potentials to their dark levels. This suppression could also be produced by an annulus of light which produced no light response in the receptors at the centre of the annulus, but which did polarize horizontal cells under the centre of the annulus. The wave form of the sign-inverted signal was similar to that produced in horizontal cells by current injection into rods, but of opposite sign. If an electrode was left in a cone for some time, the normal hyperpolarizing light response diminished, leaving a depolarizing response produced, presumably, by feed-back from horizontal cells. This signal was reversed when the cone was hyperpolarized with about 0.1 nA current. These data suggest that the sign-inverted response is mediated by feed-back from horizontal cells and, assuming that depolarization increases the rate of release of horizontal cell synaptic transmitter, then the feed-back transmitter opens channels in the cone membrane whose currents have a reversal potential around -65 mV.

Action Potentials↗