Biomedical subjects

W J Miller

Publications and source records attributed to W J Miller.

At least 127 records · Page 7Linked to original sources

PubMed1986

Production and immunological analysis of recombinant hepatitis B vaccine.

The synthesis of the hepatitis B surface antigen (HBsAG) in cells of Saccharomyces cerevisiae and its subsequent isolation, purification and analysis is described. The final, purified HBsAg particle exhibits close structural and biochemical similarities to particles derived from the plasma of chronically infected humans. Particles of yeast and human origin have been found, by chimpanzee efficacy studies and by various in vitro analyses, to be immunologically equivalent. The antigenic expression of a determinant-specific epitopes, as measured by antibody binding to synthetic peptides, has also been shown to be equivalent.

Animals↗

PubMed1986

G6PD-deficiency infectious haemolysis: a complement dependent innocent bystander phenomenon.

Dramatic haemolysis may accompany viral hepatitis and pneumococcal pneumonia in G6PD-deficient patients. Since red blood cells (RBCs) are richly endowed with receptors for activated complement, particularly C3b, we hypothesized that bulky, complement-activating immune complexes (IC) consisting of microbes and antibody might attract granulocytes (PMNs), facilitating oxidative 'innocent bystander' RBC damage. Indeed, opsonization with only two type-2 pneumococcus (PN3)/anti-PN3/C3b complexes per RBC caused agglutination of RBC, a phenomenon termed immune adherence. Addition of as few as one PMN per 20 opsonized RBCs caused the glutathione (GSH) levels of co-incubated G6PD-deficient RBCs to fall by 30% (from 3.5 to 1.8 +/- 0.8 mumoles GSH/g Hb) compared to identically incubated, but nonopsonized, G6PD-deficient RBCs. GSH levels remained normal (5.2 +/- 0.4 mumoles/g Hb) in PMN-exposed opsonized normal RBCs. GSH depletion in G6PD-deficient RBC was directly related to disease severity--falling a mean 33% in RBCs from two Black G6PD A- subjects but 59% in two Caucasian G6PD deficient RBCs. Prevention of C3b generation (with 10 mM EDTA) during opsonization abrogated both immune adherence and PMN-mediated GSH decline in oxidant-sensitive cells. Similarly, removal of C3b receptors by brief trypsin incubation of RBCs eliminated immune adherence and GSH decline. Thus, both phenomena are dependent on IC complement activation and subsequent binding of the bacterial IC to the RBC complement receptors. Although clearance of IC by RBCs may be beneficial in protecting other tissues from inflammatory damage, G6PD-deficient RBCs are vulnerable to oxidants generated by juxtaposed phagocytes--cells attracted to, and stimulated by, the immune complex/C3b combination. It is suggested that this 'Good Samaritan' activity of RBCs may lead to haemolysis during periods of exuberant antibody response to microbes.

Anemia, Hemolytic↗

PubMed1986

Further search for interaction antigens on erythrocytes in selected species hybrids of birds.

Replicated blood typing reagents were made to detect an interaction antigen (hybrid substance) on the red cells of several species hybrids of birds. Reagents against pigeon X ringneck dove hybrids are compared with other reagents made in an effort to find a similar interaction in the chicken X guinea fowl hybrid and in the ringneck X mourning dove hybrid. Mallard X muscovy hybrids also were tested for their reported red cell interaction product. Both rabbits and parental types were injected with the hybrids' red cells from the chicken X guinea fowl and mallard X muscovy crosses, but no interaction product was disclosed. The single reagent against the ringneck X mourning dove hybrid also failed to detect an interaction product.

Animals↗

PubMed1986

Further characterization of the in vitro tumoricidal activity of staphylococcal protein A.

Treatment of plasma or serum from leukemic patients with solid phase staphylococcal Protein A induced leukemic blast cell lysis in vitro, but this effect was relatively independent of the amount of immunoglobulin G (IgG) removed. Samples with approximately equal cytotoxic activity contained markedly different IgG levels, while samples with similar IgG levels had a wide range of tumoricidal activity. Assays of plasma samples collected during a perfusion of one plasma volume through a Protein A-Sepharose column indicated that the duration of the procedure had a greater effect on cytotoxic activity than did the amount of IgG removed. Neither added leukemic nor normal IgG significantly improved blast cell viability in treated serum. Cytotoxic activity was not dialyzable and concentrated in the Mr less than 100,000 fraction of samples separated by filtration. Treated cytotoxic serum samples did not have important Clq binding activity. These results suggest that the in vitro tumoricidal activity of solid phase Protein A is probably due to a toxic substance added to serum during immunoadsorption rather than to its immunoadsorptive capacity.

Antigen-Antibody Complex↗

PubMed1985

A quantitative Western Blot method for protein measurement.

A radioimmunologic assay method for the quantitation of small amounts of protein in recombinant vaccines at the level of 20-150 ng is evaluated which uses the techniques of SDS-PAGE and electrophoretic protein transfer ("Western Blot'). Known amounts of the protein being determined are included on the same gel as the unknown. After protein blotting, the nitrocellulose membrane is treated with antibody specific for the protein being determined and subsequently with [125I] Protein A. An autoradiogram is produced which corresponds directly to the nitrocellulose blot. It can, therefore, serve as a template to locate the labeled protein which is excised from the blot and measured in a gamma counter. The technique is found especially useful for evaluating cell lysates of recombinant bacteria and yeast for the percentage of the recombinant protein in the total protein mixture.

Autoradiography↗

PubMed1985

Influence of dietary carbohydrate on skeletal muscle glucose uptake.

The effect of altering the percent of dietary carbohydrate on the rate of skeletal muscle glucose uptake was studied using the perfused rat hindlimb preparation. The rats received either a high carbohydrate (HC; 65%), mixed (M; 35%) or low carbohydrate (LC; 10%) isocaloric diet for 7 days. With 0.1 mU/ml of insulin in the perfusate, the muscle of rats on the HC diet had a 33% increase in the rate of glucose uptake and the muscle of rats fed the LC diet a 23% decrease in the rate of glucose uptake when compared to the muscle of rats fed the M diet (3.34 mumol/g/30 min). With 10.0 mU/ml of insulin in the perfusate, ie maximal insulin stimulation, the rate of glucose uptake showed a similar dietary effect as that obtained with 0.1 mU/ml insulin. Compared to the M diet (8.67 mumol/g/30 min), the rate of glucose uptake increased 26% in muscle of rats from the HC group and decreased by 20% in muscle of rats from the LC group. Diet had no effect on the rate of muscle glucose uptake in the absence of insulin. Under both maximal and submaximal insulin stimulation, glycogen accumulation was greatest in muscle from HC fed rats and least in muscle from LC fed rats. During perfusion muscle intracellular free glucose and glucose-6-phosphate accumulation for the three dietary groups was negligible. The groups did not differ significantly in their muscle hexokinase or beta-hydroxyl acyl CoA dehydrogenase activities.(ABSTRACT TRUNCATED AT 250 WORDS)

3-Hydroxyacyl CoA Dehydrogenases↗

PubMed1985

Effects of elevated and exercise-reduced muscle glycogen levels on insulin sensitivity.

The effects of an exercise-induced muscle glycogen reduction and an elevated muscle glycogen concentration on glucose tolerance and the insulin response to an oral glucose tolerance test (GTT) were examined. GTTs were administered to seven male subjects after 3 days on a mixed diet (C), after exhaustive exercise and 1 day on a high-fat protein diet (L-FP), after exhaustive exercise and 1 day on a mixed diet (L-M), and after exhaustive exercise and 3 days on a high-carbohydrate diet (H-CHO). The L-M treatment resulted in a significant reduction in muscle glycogen (C, 79.6 +/- 4.2 mmol/kg wet wt vs. L-M, 53.9 +/- 1.2 mmol/kg wet wt) and a 31.7% reduction in the insulin-glucose (IG) index, a measure of insulin sensitivity in vivo. Muscle glycogen was also significantly reduced by the L-FP treatment (49.1 +/- 2.4 mmol/kg wet wt), but there was no change in the IG index. Preventing a decrease in the IG index during the L-FP treatment may have been a result of elevated free fatty acids (67%) and ketones (552%) prior to the GTT. Muscle glycogen was significantly increased by the H-CHO treatment (124.8 +/- 11.1 mmol/kg wet wt); however, the IG index was not different from that of the C treatment. The results suggest that an exercise-induced reduction in muscle glycogen can improve insulin sensitivity in vivo but that this effect is diet dependent.

Adult↗

PubMed1985

The effect of low to normal dietary phosphorus levels on zinc metabolism and tissue distribution in calves.

Sixteen 10-wk-old, phosphorus (P)-depleted Holstein bull calves were fed for 6 wk a control diet containing .08% P or P-supplemented diets containing .14, .20 or .32% P with supplemental P from two sources (CDP and Dynafos). The diets contained .45, .56, .66 and .87% Ca. After 5 wk of the experiment, the calves were dosed orally with 65Zn, and daily total fecal collections were initiated. At the end of the experimental period, the calves were killed and tissue samples were taken for total Zn and 65Zn analyses. Growth, feed intake and feed efficiency improved with increasing dietary P levels. Level of dietary P and Ca had little or no effect (P greater than .05) on total Zn content of rib, tibia, liver, heart, kidney, muscle or blood. Likewise, 65Zn absorption and content in most tissues were not affected (P greater than .05). The results do not preclude the possibility of some minor effects of P levels on Zn metabolism. However, it is apparent that when adequate Zn is fed, any effects are likely to be of little or no practical importance.

Animal Feed↗

PubMed1985

Effects of dietary aflatoxin and zinc on enzymes and other blood constituents in dairy calves.

Young male Holstein calves were fed diets containing 40 or 640 ppm zinc with 0 or 5 ppm aflatoxin for 3 wk. The aflatoxin mixture contained 80.5% B1 and the calves consumed 143 mg of B1 over 3 wk. Plasma glutamic-oxaloacetic transaminase and alkaline phosphatase concentrations were increased substantially, and lactic dehydrogenase was reduced in aflatoxin-fed calves. Supplemental zinc partially counteracted the effect of aflatoxin on these enzymes. Hemoglobin, packed cell volume, and total solids in blood plasma were increased in aflatoxin-fed calves, but high dietary zinc had no effect on these blood constituents. Glucose in plasma was reduced in calves receiving aflatoxin. High dietary zinc was only partially effective in protecting against the reduced glucose effect for about 1 wk. Total protein, albumin, globulin, ratio of albumin/globulin in blood plasma, and liver lipid were not affected by aflatoxin. Several enzymes and blood constituents are affected by aflatoxin in calves. The protection of zinc against aflatoxicosis appears to be no more than a partial effect.

Aflatoxins↗

PubMed1985

The effects of different levels of dietary lead on zinc metabolism in dairy calves.

The effects of feeding diets containing 500 or 1500 ppm added lead as lead sulfate on zinc and zinc-65 metabolism in Holstein bull calves were investigated. Zinc absorption was slightly (not significantly) reduced in the calves fed lead. Fecal zinc excretion was increased by the lead diets by day 24 of the experiment. Dietary lead had no significant effect on zinc in blood. Except for the tibia, muscle, and brain, stable zinc decreased in all tissues of calves fed the 1500 ppm lead diets, and differences were significant in pancreas, heart, and testicle. A significant decrease was noted in pancreatic zinc in pancreas of calves fed 500 ppm lead. Tissue zinc-65 concentrations were decreased significantly by lead in the tibia and muscle. Intestinal tissue zinc was not affected materially by lead. Dietary lead had very little effect on cellular distribution of zinc in the liver and kidney. In the mucosal cells of the small intestine, lead increased zinc-65 in the cytosol while decreasing it in the crude nuclear fraction. This effect occurred in a linear fashion in all three sections of the small intestine as dietary lead increased.

Animals↗

PubMed1985

Influence of high dietary iron as ferrous carbonate and ferrous sulfate on iron metabolism in young calves.

Twelve intact male Holstein calves averaging 90 kg and 12 wk of age were fed one of three dietary treatments for 28 d. The diets were A) control, B) control plus 1000 ppm iron as ferrous carbonate, and C) control plus 1000 ppm iron as ferrous sulfate monohydrate. Calves were dosed orally on d 15 of the treatment period with 1 mCi of iron-59. Neither source of added iron had a significant effect on weight gains, feed consumption, hemoglobin, packed cell volume, serum total iron, serum total iron-binding capacity, unbound iron-binding capacity, serum copper, tissue copper, fecal dry matter, or a consistent effect on fecal pH. The ferrous carbonate had no significant effect on stable zinc or stable iron in any tissue studied. Calves fed ferrous sulfate had higher average stable iron in most tissues and significantly more in the small intestine. Tissue zinc was lower in spleen and pancreas of ferrous sulfate-fed calves. Both sources of added iron sharply reduced iron-59 in serum, whole blood, and body tissues. The reduction was substantially greater in calves fed the ferrous sulfate iron. Iron in ferrous sulfate had a higher biological availability than that in the ferrous carbonate; however, bioavailability of the ferrous carbonate iron appeared to be substantial and considerably more than that noted in previous studies in which a different source of ferrous carbonate was used. The maximum safe level of dietary iron is materially influenced by the source of iron with a higher tolerance indicated for ferrous carbonated than ferrous sulfate monohydrate.

Animal Feed↗

PubMed1985

Effect of calcium and fat intake on calcium metabolism in calves.

Ten male Holstein calves were fed diets with or without 5% added animal fat in combination with low and high dietary calcium (.15 or .98%) for 4 wk. After 3 wk, the animals were orally dosed with calcium-45. One week later they were killed and tissue samples taken. Except for lower calcium-45 in bile, added dietary fat had no marked influence on calcium metabolism. Net absorption of calcium-45 (not excreted in feces) ranged from 82% for calves fed low calcium to 53% for those given high calcium. Calcium-45 in bone was substantially higher in calves fed .15% calcium. Tailbone biopsies revealed rapid uptake of calcium-45 with approximately as much incorporated during the 1st d as in the following 6 d. Calcium-45 in blood peaked 24 h following dosing. Calves fed .15% calcium had higher calcium-45 in blood and bile than those receiving .98% calcium. Calcium-45 values in soft tissue were low and did not differ materially among treatments. The decreases in radioactive calcium absorption and bone deposition with higher dietary calcium indicated that variable absorption was a major factor in calcium homeostasis. Added fat did not materially effect calcium metabolism with either low or high dietary calcium.

Animals↗

PubMed1985

Effects of plasma treatment with purified protein A and Staphylococcus aureus Cowan I on spontaneous animal neoplasms.

Eleven dogs with spontaneous neoplasms were intensively treated with an immunoadsorption system consisting of a continuous flow centrifuge, Protein A-Sepharose columns, and a semi-automatic elution system. Despite consistent and substantial lowering of immunoglobulin G levels, tumor regression was noted in only one of 11 dogs. In contrast, infusion of small volumes of plasma after incubation with heat and formalin-treated Staphylococcus aureus Cowan I resulted in a tumoricidal response in five of six animals. These results suggest that tumor necrosis is probably not induced by Protein A-mediated removal of humoral "blocking" factors.

Animals↗

PubMed1984

Clinical evaluation in healthy adults of a hepatitis B vaccine made by recombinant DNA.

A vaccine formulated from hepatitis B surface antigen (HBsAg) produced by a recombinant strain of the yeast Saccharomyces cerevisiae was administered to two groups of human volunteers composed of 37 healthy, low-risk adults. Each subject received a 10-micrograms dose of HBsAg at 0, 1, and 6 months. By one month, 27% to 40% of the vaccinees had antibody to HBsAg, and by three months 80% to 100% were antibody positive. Large boosts in titer followed the third dose at six months. The antibody formed is predominantly specific for the a determinant of HBsAg. There have been no serious reactions attributable to the vaccine. The most frequent complaint has been transient soreness at the injection site. As far as we know, this is the first reported use in man of a vaccine prepared by recombinant DNA technology.

Adult↗

PubMed1984

Lead toxicity and metabolism from lead sulfate fed to Holstein calves.

Sixteen Holstein intact male calves averaging 85 kg and 74 days of age were assigned randomly to four dietary lead treatments according to body weight. They were fed for ad libitum consumption a control diet containing no added lead or the control diet supplemented with 500, 1500, or 4500 ppm lead as lead sulfate. One calf fed 1500 ppm lead and all four calves fed 4500 ppm lead died within 6 to 10 days after initiation of treatments. Death was sudden with few or no clinical signs prior to death. Those clinical signs that did appear included muscular tremors, gnashing of teeth, bellowing, and convulsions. Four control, four 500 ppm lead, and two 1500 ppm lead-fed calves survived the 7-wk experimental period. Feed consumption, body weight changes, glutamic oxaloacetic transminase and alkaline phosphatase activity in blood plasma, and hemoglobin were not affected significantly by lead treatments. Packed cell volume in calves fed 500 and 1500 ppm added lead was reduced. Lead concentrations in blood, kidney, liver, bone, brain, and muscle were elevated in lead supplemented calves above those of controls. The highest concentrations of lead were in kidney and liver.

Animals↗

PubMed1984

Effects of high but nontoxic dietary manganese and iron on their metabolism by calves.

Sixteen male Holstein calves were fed one of four diets for 18 days in an experiment consisting of 0 and 1000 ppm supplemental manganese and 0 and 1000 ppm added iron as manganese carbonate and ferrous carbonate. The control diet contained 55 ppm manganese and 220 ppm iron. All calves were dosed orally 48 h prior to sacrifice with 500 muCi of manganese-54. Small intestinal iron was less in calves fed a high manganese diet, a possible interaction of these two elements at the absorption site. Feeding a high manganese diet tended to decrease iron (total) concentrations in liver and pancreas. When the high manganese diet was supplemented with additional iron, antagonistic effects of manganese on iron were eliminated. Neither iron nor manganese concentrations in tissues were affected by an increase of dietary iron. Manganese-54 content of tissue was reduced by the high manganese diet but was not affected by dietary iron. Total manganese and iron in feces fairly closely reflected dietary intake of each element with no evidence of interaction. Calves fed the high iron diet excreted less manganese-54 in their feces over 2 days. Total iron in blood serum was not affected significantly by the dietary treatments.

Animals↗

PubMed1984

Viral enhancement and interference induced in cell culture by hepatitis A virus: application to quantitative assays for hepatitis A virus.

Hepatitis A virus (HAV) growing in human diploid lung fibroblast (MRC5) monolayers can either interfere with or enhance the cytopathic effect of Newcastle Disease virus (NDV) challenge. Enhancement of NDV occurred if HAV-infected monolayers were challenged with a low multiplicity of infection of NDV and incubated at 35 degrees C. Interference occurred if HAV-infected monolayers were given a high NDV multiplicity of infection and incubated at 32 degrees C. These phenomena were applied to assays for quantifying HAV and may be useful in providing new insights into viral interference and enhancement.

Antigens, Viral↗

PubMed1984

Vaccine against human hepatitis B virus prepared from antigen derived from human hepatoma cells in culture.

Vaccine against human hepatitis B was prepared using antigen derived from hepatitis B carrier hepatoma cells grown in the interstices of a Diaflo hollow filter unit. Hepatitis B surface antigen (HBsAg) produced by these cells was purified by immune affinity chromatography, digestion with DNase and pepsin, and Sephadex G-150 separation. The Formalin-treated antigen was formulated in 20-micrograms dose on alum adjuvant with thimerosal added as a preservative. This cell culture vaccine was as potent as human plasma-derived vaccine as measured in a mouse potency assay. The vaccine proved safe in tests in chimpanzees and in human subjects who were in late stages of cancer of the central nervous system and who were receiving therapy for their condition. None of five subjects who received the vaccine developed untoward clinical reactions. Two of the subjects who received all three doses of vaccine developed antibody against HBsAg. Three persons, two given only the primary doses and one who was given all three doses but was lost to follow-up, demonstrated no response. The slow and relatively low antibody responses to the vaccine were similar to those in other immunosuppressed persons who were given vaccine of human plasma origin.

Animals↗