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Biomedical subjects

C Wu

Publications and source records attributed to C Wu.

At least 541 records · Page 30Linked to original sources
PubMed1988

Analysis of the assembly of laminin and the laminin-entactin complex with laminin chain specific monoclonal and polyclonal antibodies.

Antibodies specific for the A, B1, and B2 chains of laminin have been obtained and characterized. Lam V, a rat X mouse monoclonal antibody, was obtained by immunizing Lewis rats with the extracellular matrix derived from the mouse endodermal line M1536-B3. The antibody was shown to recognize a conformation-sensitive epitope present on the A chain of laminin. The antibody exhibited high avidity for native laminin and uncomplexed newly synthesized laminin A chains. cDNA clones in the vector lambda-gt11 containing sequences for the B1 and B2 chains of laminin were shown to synthesize beta-galactosidase fusion proteins in the host cells induced with IPTG. The fusion protein F3 contained amino acid residues 822-1765 of the B1 chain of mouse laminin, and the fusion protein E4 contained 219 amino acids at the carboxyl terminus of the B2 chain of rat laminin. These two fusion proteins were used to obtain rabbit polyclonal antibodies which were characterized for their specificity and ability to immunoprecipitate laminin and the B chains of laminin. The chain-specific antibodies were used to analyze the assembly and processing of laminin in the mouse endodermal cell line M1536-B3. The results indicated that the covalent assembly of the A and B chains of laminin was initiated as early as 3 min after labeling cells. At this time point uncomplexed A chain of laminin could be observed even though there was an excess of B1 and B2 chains. As early as 4 min after labeling monomeric, dimeric, and oligomeric forms of the B chains of laminin were observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1987

Purification and properties of Drosophila heat shock activator protein.

Drosophila heat shock activator protein, a rare transacting factor which is induced upon heat shock to bind specifically to the heat shock regulatory sequence in vivo, has been purified from shocked cells to more than 95 percent homogeneity by sequence-specific duplex oligonucleotide affinity chromatography. The purified protein has a relative molecular mass of 110 kilodaltons, binds to the regulatory sequence with great affinity and specificity, and strongly stimulates transcription of the Drosophila hsp70 gene. Studies with this regulatory protein should lead to an understanding of the biochemical pathway underlying the heat shock phenomenon.

Animals↗
PubMed1987

Isolation of RNA for dot hybridization by heparin-DNase I treatment of whole cell lysate.

We have developed a new procedure for the rapid preparation of undegraded total RNA from cultured cells for specific quantitation by dot blotting analysis. Pelleted cells are resuspended in hypotonic solution containing a ribonuclease inhibitor and heparin and disrupted by freeze-thaw. Heparin is employed as an agent for nuclear lysis, dissociation of chromosomal protein, and release of mRNA from rough endoplasmic reticulum. We eliminate chromosomal DNA by digestion with DNase I and denature the RNA in the lysate with formaldehyde. After centrifugation to remove debris, the supernatant is used directly for dot blotting. All manipulations are performed in the same microfuge tube and recovery of RNA is quantitative. The procedure is especially useful for processing large numbers of samples. We illustrate its versatility by analysis of specific RNAs in Drosophila, rat, and human cell lines. In reconstruction experiments, less than 80 molecules per cell of a small RNA (beta-globin) can be detected under highly stringent hybridization conditions, using only moderately labeled double-stranded plasmid DNA probes and short film exposures.

Animals↗
PubMed1987

Venous hypercarbia in canine hemorrhagic shock.

The venous-arterial PCO2 gradient may increase in certain low-flow states, such as CPR and canine endotoxemia. To determine whether venous hypercarbia also occurs in hemorrhagic shock, we studied 12 anesthetized, mechanically ventilated dogs. We performed laparotomies on the animals, inserting catheters into their renal, superior mesenteric, and external iliac veins. Flow in the corresponding arteries were determined using electromagnetic flow probes. The dogs were randomized into a control group (n = 6), and a hemorrhagic shock group (n = 6) which was bled to a mean arterial pressure of 45 to 50 mm Hg and maintained at this pressure for the 6-h study. The results demonstrated a significant (p less than .05) increase in lactate and venous-arterial PCO2 gradient systemically and in all three regional beds. A significant decrease of venous blood pH accompanied these changes which are consistent with our previous findings in low-flow, canine endotoxemia. We conclude that venous hypercarbia is a nonspecific phenomenon, common to low-flow states. The increased CO2 represents both an increased CO2 production and a decreased removal, secondary to low-flow.

Animals↗
PubMed1987

Comparison of motor cortex-induced flexor muscle activity inhibition by hard pressure on various parts of the body and by light pinch of abdomen of animals with gastro-duodenal ulcers.

The flexsor muscle electromyogram (EMG) of the upper extremities in response to the motor cortex stimulation was inhibited by hard hand pressure on base of the ear lobe and lumbar perivertebral region and by electrical stimulation of these regions. Similar inhibition was produced by electrical stimulation around the brachium conjuctivum and locus coeruleus (BC-LC) and the reticulogigantocellular nucleus (NRGC). Inhibition of the flexor muscle EMG due to hard pressure on the body parts was abolished by electrical lesioning of the BC-LC and NRGC. The light pinch with hand on the restricted abdominal region did not inhibit the flexor muscle EMG induced by the motor cortex stimulation in normal condition, while such stimulation inhibited the flexor muscle EMG in the ulcer suffering animals after treatment with cysteamine. This inhibition was not influenced by destruction of the NRGC. From these results, it was concluded that inhibition of the motor cortex induced-flexor muscle activities caused by light pinch stimulation of the restricted abdominal region, as the model of the voluntary finger flexion inhibition in O-ring test, was produced by spinal reflex inhibition.

Animals↗
PubMed1987

The levamisole receptor, a cholinergic receptor of the nematode Caenorhabditis elegans.

We describe a glass fiber filter binding assay for the levamisole receptor, a putative acetylcholine receptor of the nematode Caenorhabditis elegans, and we show that receptor detected in vitro binds both levamisole derivatives and cholinergic agonists with the pharmacological specificity expected of the physiologically functional nematode receptor. The receptor is detected by the binding of tritiated meta-aminolevamisole ([3H]MAL, 27 Ci/mmol). In extracts of the wild-type nematode, there is a saturable, high affinity binding activity for [3H]MAL (Kd approximately 5-10 nM). Well fed wild-type worms contain as much as 3 fmol of high affinity binding activity per mg of extract protein (0.14 pmol/g of wet weight of worms) and dauer larvae, a special juvenile stage, contain as much as 15 fmol of activity per mg of protein. Specific binding activity per mg of protein is highest in larval stages and decreases severalfold in the adult worm. The rates of formation and dissociation of the [3H]MAL-receptor complex are relatively slow (dissociation half-life, 17 min), in agreement with physiological studies of levamisole on Ascaris muscle strips. Levamisole derivatives and cholinergic agonists have the same relative potencies in inhibiting [3H]MAL binding as they do in causing nematode muscle contraction. Vertebrate cholinergic antagonists do not inhibit [3H]MAL binding, but several antagonists (mecamylamine, alpha-bungarotoxin, and cobra venom) potentiate the binding of [3H]MAL and can be used to demonstrate more clearly the presence of a second, lower affinity binding activity whose ligand-binding affinity is also potentiated by these agents. Both high and low affinity wild-type binding components are missing in the extremely levamisole-resistant mutant unc-74(x19).

Age Factors↗
PubMed1986

Conversion of leukotrienes A4 to C4 in cell-free systems.

A procedure for assaying leukotriene C4 synthase activity in cell-free extracts has been presented. Leukotriene A4 methyl ester was as active a substrate as leukotriene A4 (Na salt) for the synthesis. The methyl ester is the substrate of choice, because (1) it is more stable than the sodium salt, (2) it is not a substrate of epoxide hydrolase for leukotriene B4 synthesis, and (3) it gives a lower blank than an equimolar concentration of leukotriene A4. The enzyme activity in rat liver, guinea pig and human lungs, and human nasal polyp was chiefly membrane-bound, although the cytosol contained some activity.

Animals↗
PubMed1986

Evidence for a pool of non-recycling transferrin receptors in peripheral sheep reticulocytes.

Sheep reticulocytes from phlebotomized animals have a total transferrin binding potential that may exceed by an order of magnitude the surface binding capacity. Steady state uptake of transferrin at 37 degrees C is generally less than 50% of the total transferrin binding capacity. During long-term incubation of the reticulocytes, all transferrin binding ability is lost, the ability to internalize being lost most rapidly. The loss in ability to bind transferrin during long-term incubation is independent of the number of surface transferrin binding sites, since removal of surface receptors with pronase does not affect the rate of loss of the internal pool of receptors during long-term incubation. Moreover, after removing surface receptors with pronase, only a fraction of the original number of receptors is restored to the surface, despite the presence of a large pool of internal receptors. These data suggest that only a fraction of the internal pool of receptors is capable of recycling to the cell surface in sheep reticulocytes.

Animals↗
PubMed1986

Generation of drug metabolite antigenicity in the intestinal mucosa.

Both nitroreductase and transglutaminase activities have been assayed in the 10,000 X g supernatant fluids of rat intestine homogenates after Triton X-100 treatment. Incubation of 14C-nitrofurantoin in the intestine extract yielded protein-bound 14C-labeled products. Injection into rabbits of the conjugated protein similarly prepared with unlabeled nitrofurantoin elicited formation of antibodies against nitrofurantoin. These results suggest that intestinal metabolism and conjugation to protein of orally administered drugs may serve as a probable mechanism of drug allergy, and this may be accomplished by enzymatic coupling of relatively stable drug metabolites to protein carriers.

Animals↗