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Biomedical subjects

B A Watkins

Publications and source records attributed to B A Watkins.

At least 73 records · Page 4Linked to original sources
PubMed1993

Immune escape by human immunodeficiency virus type 1 from neutralizing antibodies: evidence for multiple pathways.

Sera from many HIV-1-infected individuals contain broadly reactive, specific neutralizing antibodies. Despite their broad reactivity, variant viruses, resistant to neutralization, can be selected in vitro in the presence of such antisera. We have previously shown that neutralization resistance of an escape mutant with an amino acid substitution in the transmembrane protein (A582T) occurs because of alteration of a conformational epitope that is recognized by neutralizing antibodies directed against the CD4 binding site. In this report we demonstrate that immune escape via a single-amino-acid substitution (A281V) within a conserved region of the envelope glycoprotein gp120 confers neutralization resistance against a broadly reactive neutralizing antiserum from a seropositive individual. We show this alteration affects V3 and additional regions unrelated to V3 or the CD4 binding site. Together with previous studies on escape mutants selected in vitro, our findings suggest that immune-selective pressure can arise by multiple pathways.

Amino Acid Sequence↗
PubMed1993

Early feed restriction of broilers: effects on abdominal fat pad, liver, and gizzard weights, fat deposition, and carcass composition.

A total of five experiments were conducted to investigate the effects of early feed restriction on organ weights, fat deposition, and carcass composition in broilers. In Experiments 1 and 2, broiler chicks were reared in litter pens for 49 days, whereas Experiments 3, 4, and 5 were conducted in battery cages for 28 days. Feed restriction in all experiments was accomplished by providing male broiler chicks 40 kcal per bird per day for 7 (Experiments 1 and 2) or 6 (Experiments 3, 4, and 5) days, starting at 4 days of age. Feed restriction of broiler pullets in Experiment 1 was imposed from 4 to 9 days of age. Furthermore, the starter and grower diets in Experiment 2 contained either 1 or 4% added fat and the starter diets in Experiments 3, 4, and 5 contained either 21 or 26% protein. No significant differences were observed for abdominal fat pad and gizzard weights, liver and carcass fat, or carcass protein between early restricted birds and ad libitum controls at 49 days of age in Experiments 1 and 2. Furthermore, with the exception of Experiment 4, no differences in abdominal fat pad weights were observed among the different feeding regimens. Significant increases in abdominal fat pad weights, percentage liver, and percentage carcass fat were noted for females at 49 days of age when compared with male broilers. Carcass fat was significantly higher and carcass protein was significantly lower in broilers fed commercial versus reduced fat diets. Broilers fed a 21% protein diet had significantly heavier abdominal fat pad weights at 28 days of age than birds fed a 26% protein diet.

Adipose Tissue↗
PubMed1993

Evaluation of two novel biochemicals on plasma and egg yolk lipid composition and laying hen performance.

PD132301-2, an inhibitor of acyl-coenzyme A:cholesterol O-acyltransferase (ACAT; EC 2.3.1.26), and 1-stearylboronic acid (SBA), a fatty acid analogue, were orally administered to White Leghorn hens in separate experiments to evaluate their effects on layer performance and plasma and egg yolk lipids. Five 60-wk-old hens each were fed either a corn-soybean meal basal layer ration, or the basal diet supplemented with .0121, .0363, or .1089% PD132301-2. In a second experiment, 12 37-wk-old hens each were fed either a basal layer ration, or the basal diet supplemented with .20 or .40% SBA. The duration of the experiments were 21 and 16 days, respectively. Neither compound significantly affected hen-day production, egg weight, yolk weight, BW gain, feed consumption, feed efficiency, plasma total cholesterol and triglyceride concentrations, or egg yolk cholesterol content. PD132301-2 had no effect on yolk fatty acid profiles, and C22:6n3 was the only fatty acid altered by SBA. Although 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors have been successful in reducing egg cholesterol, ACAT inhibitors and fatty acid analogues apparently hold little promise in this regard. The results of the present work also support the concept that, in order to pharmacologically alter the cholesterol content of eggs, direct inhibition of key enzymes in the cholesterol biosynthetic pathway is necessary.

Administration, Oral↗
PubMed1993

Characterization of lipogenic and lipolytic activity, muscle tissue composition, and DNA and RNA levels of broilers eating ad libitum or severely restricted at an early age.

Four experiments were conducted to characterize the rates of lipogenic and lipolytic activities and the composition and DNA and RNA concentrations in muscle tissue of broilers that were feed restricted at an early age. In all experiments, feed restriction (40 kcal/day per bird) was initiated at 4 days of age with ad libitum consumption of feed resuming at either 11 (Experiments 1 and 2) or 10 (Experiments 3 and 4) days of age. Female chicks in Experiment 1 were feed restricted at the same level from 4 to 9 days of age. Birds consuming feed ad libitum served as controls in all experiments. Broilers restricted for 7 days had significantly lower plasma very low density plus low density lipoproteins (VLDL+LDL) than the ad libitum birds at 49 days of age in Experiments 1 and 2. However, the reverse was observed at 28 days of age, as plasma triglycerides and lipoproteins and lipogenesis were greater in restricted chicks when compared with controls. No significant differences were observed in plasma triglycerides at 49 days of age or lipolysis, DNA and RNA concentrations, muscle protein, and muscle fat at 28 days of age between restricted and ad libitum broilers. Results from these studies indicate that early feed restriction may have induced a metabolic shift in restricted broilers that increased lipogenesis at 28 days of age, followed by a partitioning of excess nutrients to support anabolic activity at market age.

Adipose Tissue↗
PubMed1993

A rapid method for site-specific mutagenesis using larger plasmids as templates.

To facilitate the introduction of specific point mutations in plasmids that are too large to be amplified efficiently by a single PCR, we have developed a method for site-directed mutagenesis by generating partial plasmid fragments, which introduces changes as simply as conventional techniques. Plasmids containing a fragment of the human immunodeficiency virus type-1 (HIV-1) envelope gene were subjected to PCR with four pairs of PCR primers for each desired point mutation. One primer in each of two of these four pairs contained the desired mutation. The four pairs of primers were designed so that four overlapping fragments were amplified from the plasmid template, two of which contained the mutation. These fragments were then reannealed and electroporated directly into Escherichia coli. The desired mutation was typically found in 66% to 83% of the resulting colonies. The technique is almost as simple as previous techniques, shows similar efficiency and is applicable to plasmids that would normally be too large for efficient site-specific mutagenesis. The entire procedure, from PCR amplification to transfection into E. coli, can be completed in one day.

Base Sequence↗
PubMed1992

Disruption of endothelial barrier function by lipolytic remnants of triglyceride-rich lipoproteins.

Remnants, resulting from the lipolysis of triglyceride-rich lipoproteins, injured cultured endothelial cells and resulted in decreased barrier function of the vascular endothelium. Endothelial cells were cultured on micropore filters. Albumin transfer across endothelial cell monolayers was measured after a 24-h exposure to media enriched with control or in vitro-lipolyzed samples of various hypertriglyceridemic (HTG) sera and its isolated lipoprotein (VLDL, LDL and HDL) and serum free protein (d greater than 1.21 g/ml) fractions. Compared with control cultures, neither control HTG serum nor its isolated lipoprotein and serum-free protein fractions had any effect on albumin transfer. In contrast, lipolyzed HTG (L-HTG) serum and all of its isolated lipoprotein fractions (L-VLDL, L-IDL, L-LDL and L-HDL) caused a marked decrease in endothelial barrier function, evidenced by a significant increase in albumin transfer across endothelial monolayers. The L-IDL and L-HDL fractions were more effective in increasing albumin transfer than the L-VLDL and L-LDL fractions. The extent of the L-IDL and L-HDL mediated increases in albumin transfer was concentration dependent. An exposure of 12 h was required for L-HDL to increase albumin transfer. The L-HDL mediated increase in albumin transfer was reversible only after a 12-h exposure at low concentrations. The free protein fraction from L-HTG serum had no significant effect on the barrier function of endothelial cells. The presence of normolipidemic HDL in culture medium prevented disruption of the endothelial barrier induced by L-IDL but not by L-HDL. The decrease in endothelial barrier function induced by lipolyzed samples of HTG serum or lipoproteins appeared to be correlated with the level of free fatty acids contained in lipolytic remnants. Enrichment of LDL, and in particular HDL, with fatty acid significantly increased albumin transfer. Compared with lipolyzed samples, sera/lipoproteins oxidized in vitro by Cu2+ ions had little effect on endothelial barrier function, which did not correlate with their respective thiobarbituric acid-reacting substance (TBARS) values. TBARS remained within normal range after L-HDL incubation with endothelial cells for up to 48 h. At most concentrations tested, exposure to lipolyzed but not oxidized lipoproteins resulted in morphological perturbations of cell monolayers. These data suggest that lipolytic remnants of triglyceride-rich lipoproteins may play an important role in the development of atherosclerosis by decreasing the barrier function of the vascular endothelium. The remnant-induced injury of the arterial wall may permit the entry of cholesterol-rich lipolytic remnants as well as LDL into the arterial wall.

Animals↗
PubMed1992

The effect of triacylglycerol chain length on food intake in domestic fowl.

Triacylglycerols of varying chain lengths were tested for their effects on food intake in fasted Single-Comb White Leghorn cockerels 9-12 weeks of age. Tributyrin, tricaprylin, triolein, or saline were intubated intragastrically or infused intrahepatically. All triacylglycerol administrations were in 10 ml volumes with 10 kcal being infused intrahepatically and 10 or 20 kcal being infused intragastrically. When administered intragastrically, the short-chained triacylglycerol tributyrin decreased food intake at most times tested; the medium chain-length tricaprylin and the long chain-length triolein affected food intake sporadically. When infused into the liver, tributyrin and tricaprylin decreased food intake at most times tested, whereas triolein decreased food intake at 60 and 180 minutes only. Satiating effectiveness was related to triacylglycerol chain length when intubated, which may reflect rate of absorption from the gut; however, hepatic satiety receptors were also differentially sensitive to triacylglycerol chain length indicating that more than rate of assimilation from the gut must be considered.

Animals↗
PubMed1992

Effect of beta, beta'-tetramethyl-substituted hexadecanedioic acid (MEDICA 16) on laying hen performance and egg yolk lipid composition.

1. beta, beta'-tetramethyl-substituted hexadecanedioic acid (MEDICA 16), an inhibitor of hepatic cholesterogenesis and lipogenesis in rats, was orally administered to 24-week-old White Leghorn hens for a period of 16 d. Hens were fed maize-soya-bean meal diets containing 0, 1.5, or 3.0 g MEDICA 16/kg. 2. Although MEDICA 16 did not affect egg weight, yolk weight, egg cholesterol content, or the efficiency of food utilisation, egg production was significantly reduced in birds fed 3.0 g MEDICA 16/kg compared to those fed the other two diets. 3. Total yolk monounsaturated fatty acids were significantly higher in eggs of hens fed both inclusion rates of MEDICA 16 compared to those of the control birds. In contrast, egg yolk total polyunsaturated fatty acid content and the ratio of polyunsaturated to saturated fatty acids were both inversely related to the dietary content of MEDICA 16. 4. These results suggest that MEDICA 16 primarily altered hepatic fatty acid metabolism, but not cholesterol synthesis, in laying hens.

Animals↗
PubMed1992

Chronic low-dose exposure of sodium nitrite in VM-strain mice: central nervous system changes.

1. There is suggestive evidence that nitrite may be a causative factor in cerebral glioma. 2. To test this hypothesis we selected the VM mouse strain, known for its susceptibility to spontaneous glioma formation, and exposed 300 animals to 0.2% sodium nitrite in their drinking water. One hundred of this group were exposed both in utero and throughout their adult lives. The remaining 200 animals received nitrite from the time of weaning. A further 200 mice were used as controls and received distilled water. 3. All animals were maintained until their natural death and were then subjected to autopsy and routine histological examination. 4. There was no excess of nervous system tumours in the experimental groups.

Administration, Oral↗
PubMed1992

Effect of early feed restriction on growth, feed conversion, and mortality in broiler chickens.

Two floor pen and two battery experiments were conducted to determine the effects of early feed restriction on the performance of commercial broilers. Feed restriction was induced in all experiments by providing chicks with 40 kcal of ME per bird per day, commencing at 4 days of age. Male chicks were feed-restricted for 7 (Experiments 1 and 2) or 6 days (Experiments 3 and 4), whereas broiler females were restricted for 5 days (Experiment 1). Ad libitum feeding was resumed after the restriction periods, and continued through the conclusion of the experiments at 49 (Experiments 1 and 2) or 28 (Experiments 3 and 4) days of age. Broilers provided ad libitum access to feed for the entire experimental period served as the controls in each study. Broilers subjected to an early feed restriction had significantly (p less than or equal to .05) lower mean body weights than controls for all ages measured in the four experiments. However, feed conversion ratios for restricted broilers were significantly lower at 28 (Experiments 1 through 4) and 49 (Experiments 1 and 2) days of age than for birds consuming feed ad libitum. Through regression analyses, it was estimated that male broilers in Experiments 1 and 2 would require approximately 2 additional days to obtain body weights similar to those observed in control broilers and would still maintain a lower feed conversion ratio at this older age. In Experiments 1 and 2, weekly body weight gains for restricted broilers were significantly lower than for controls from 0 through 28 days of age. However, restricted broilers (7 days) in Experiment 2 had significantly higher rates of gain from 29 to 49 days of age than unrestricted controls. Total pen body weights for restricted and ad libitum groups were similar at 49 days of age in Experiments 1 and 2, which reflected the significant difference in mortality observed between the two groups.

Age Factors↗
PubMed1992

Influence of dietary n-6 and n-3 polyunsaturates on lipids in chickens divergently selected for body weight.

The effects of dietary lipid sources rich in n-6 (soybean oil) or n-3 (menhaden oil) polyunsaturated fatty acids (PUFA) on blood triacylglycerol levels and tissue fatty acid compositions in female chickens from two genetic lines were examined. Triacylglycerol concentrations in the plasma very low density and low density lipoprotein fractions were elevated in the low-weight (LW) chickens compared with the high-weight (HW) group. Chicken heterophil to lymphocyte ratios and total antibody titers to sheep red blood cells at 84 days were not influenced by genetic line or by the dietary lipid fed. Concentrations of C18:1 fatty acid isomers and total monounsaturates were highest in liver and heart tissues of HW chickens. Feeding menhaden oil enriched the plasma, liver, and heart with n-3 PUFA in both genetic lines. The data indicate a difference in hepatic fatty acid metabolism between the HW and LW lines that is unaffected by dietary n-6 or n-3 PUFA.

Animals↗
PubMed1991

Response of Müller cells to growth factors alters with time in culture.

We have developed an explant culture technique, using the retinae of newborn guinea pigs, that reliably yields cultures of Müller cells showing uniform morphology and phenotype. Since the guinea pig retina is avascular and lacks astrocytes, Müller cells are the only glial cell-type and the only vimentin-positive population present. Virtually all passaged cells (greater than 98%) contain vimentin-positive intermediate filaments and no glial fibrillary acidic protein (GFAP) has been detected using a range of GFAP antibodies known to label astrocytes in the guinea pig optic nerve. Most vimentin-positive cells were also labeled with an antibody to carbonic anhydrase II, an enzyme which in the retina is specific for Müller cells. Proliferating Müller cells were identified within the inner nuclear layer of retinal fragments as early as 2 days in culture using bromodeoxyuridine (BrdU) and vimentin double labeling. Cultured Müller cells change their growth characteristics with successive passaging. The length of the cell cycle increases from 25.4 h for cells at first passage, to 66.7 h for cells at fourth passage. Altered responses to mitogens were also observed with passaging. First-passage cultures responded to basic fibroblast growth factor (bFGF) but not to several other factors tested including interleukin-2 (IL-2). In contrast, older cultures were highly responsive to IL-2 but showed a minimal response to bFGF. The altered responsiveness to mitogens observed in vitro may be relevant to changes in growth control of Müller cells in the developing and mature retina. The guinea pig retina provides an ideal mammalian tissue for generating Müller cell cultures that are free of astrocytes, endothelial cells, and pericytes, the most frequent contaminants of retinal glial cultures. The monolayers obtained show a high degree of homogeneity and are well suited for studies of Müller cell function.

Animals↗
PubMed1991

Hydrogenated oil decreases tissue concentrations of n-6 polyunsaturated fatty acids and may contribute to dyschondroplasia in broilers.

1. In a factorial design of dietary treatments, male Ross broilers were given diets containing soyabean oil, hydrogenated soyabean oil (as a source of trans-fatty acids) or feed fat with either 0 or 300 micrograms of added D-biotin/kg. 2. Growth to 28 d was not influenced by the dietary treatments. 3. Length of tibiotarsal bones was reduced (P less than 0.05) and severity of leg bone cartilage lesions, characteristic of dyschondroplasia, was highest (P less than 0.05) in broilers fed on diets containing hydrogenated soyabean oil. 4. Feeding hydrogenated soyabean oil lowered (P less than 0.05) the concentrations of C20:4n6 and the ratios of C20:4n6/C18:2n6 in liver and growth plate cartilage. 5. Growth plate cartilage from birds affected with dyschondroplasia contained lower proportions of prostaglandin precursor fatty acids compared with normal growth plate. 6. It is speculated that an inhibition of prostaglandin biosynthesis brought about by the presence of trans-fatty acids might contribute to the occurrence of lesions similar to dyschondroplasia.

Animal Feed↗
PubMed1991

Importance of essential fatty acids and their derivatives in poultry.

More than thirty-five years have passed since essential fatty acid requirements were demonstrated for poultry. Subsequent studies were conducted to establish a recommended level of linoleic acid for growing poultry. Although a requirement of 1% linoleic acid is suggested, a critical evaluation of the nutritional and metabolic significance of essential fatty acids is lacking. Perhaps alpha-linolenic acid is also essential, but direct evidence is not yet available. Recent literature indicates that several individual fatty acids, including the polyunsaturates derived from essential fatty acids, are biologically active. Furthermore, the numerous eicosanoids biosynthesized from specific polyunsaturates act as either cellular autocrine or paracrine signaling agents. Formation of eicosanoids is ubiquitous in tissues and organs, and these compounds influence nearly all systems in mammalian species. However, although such compounds have been studied extensively in mammalian systems, the formation of polyunsaturates, the biosynthesis of eicosanoids and the biological effects of these compounds have not been intensely studied in domestic poultry.

Animal Nutritional Physiological Phenomena↗
PubMed1991

Infection of brain microglial cells by human immunodeficiency virus type 1 is CD4 dependent.

In the central nervous system of AIDS patients, human immunodeficiency virus (HIV) infects primarily microglia, a cell type of bone marrow origin. Moreover, microglial cells isolated from adult human brain support the replication of macrophage-adapted strains of HIV type 1 (HIV-1) (B.A. Watkins, H.H. Dorn, W.B. Kelly, R.C. Armstrong, B. Potts, F. Michaels, C.V. Kufta, and M. Dubois-Dalcq, Science 249:549-553, 1990). To determine whether the CD4 receptor, which is expressed in brain, mediates the entry of HIV-1 in microglial cells, we analyzed CD4 transcript expression in cultured microglia using highly sensitive polymerase chain reaction detection of cDNAs synthesized from RNA. With this method, CD4 transcripts could be detected in cultured microglia--as well as in various human brain regions and cultured macrophages used as positive controls--along with transcripts for the LDL and Fc receptors which are characteristic of cells of the macrophage lineage. We then attempted to block viral entry into microglial cells using anti-CD4 antibodies or soluble CD4 (sCD4), which recognize binding sites on CD4 and HIV-1 glycoprotein gp120, respectively. Cultures were pretreated with blocking antibodies (Leu-3a, OKT4A) or virus was preincubated with sCD4 prior to infection with HIV-1 strain AD87(M) or BaL. With either viral strain, these treatments resulted in the prevention of infection or significant and dose-dependent reduction in the number of infected cells and in the levels of reverse transcriptase or p24 antigen released in the medium. Thus, brain-derived microglial cells, which are the primary target of HIV-1 infection in the brain, express the CD4 receptor and this receptor is effectively used for viral entry in vitro.

Acquired Immunodeficiency Syndrome↗
PubMed1990

Specific tropism of HIV-1 for microglial cells in primary human brain cultures.

Human immunodeficiency virus (HIV) frequently causes neurological dysfunction and is abundantly expressed in the central nervous system (CNS) of acquired immunodeficiency syndrome (AIDS) patients with HIV encephalitis or myelopathy. The virus is found mostly in cells of the monocyte-macrophage lineage within the CNS, but the possibility of infection of other glial cells has been raised. Therefore, the effects of different HIV-1 and HIV-2 strains were studied in primary cultures of adult human brain containing microglial cells, the resident CNS macrophages, and astrocytes. These cultures could be productively infected with macrophage-adapted HIV-1 isolates but not with T lymphocyte-adapted HIV-1 isolates or two HIV-2 isolates. As determined with a triple-label procedure, primary astrocytes did not express HIV gag antigens and remained normal throughout the 3-week course of infection. In contrast, virus replicated in neighboring microglial cells, often leading to their cell fusion and death. The death of microglial cells, which normally serve immune functions in the CNS, may be a key factor in the pathogenesis of AIDS encephalitis or myelopathy.

Adult↗
PubMed1990

Effects of feeding aspirin and soybean oil to weanling pigs.

Three trials were conducted with 216 Yorkshire x Duroc x Hampshire crossbred pigs (equal males and females) weaned at 3 to 4 wk of age (avg 7.4 kg initially) to determine the effectiveness of aspirin in an 18% CP corn-soybean meal starter diet (with and without soybean oil in Trial III) for improving postweaning performance and reducing scours. When aspirin levels common to all trials (0, 250 ppm) or common to Trials II and III (0, 125 and 250 ppm) were compared, improvements in ADG (P less than .10) and daily feed intake (P less than .05) were observed with no effect on efficiency of feed utilization. There was a quadratic effect of aspirin (P less than .10) in Trials II and III, the trend suggesting that the response to 125 ppm was as effective as the response to 250 ppm aspirin. The addition of soybean oil in Trial III did not enhance the response. In all trials, scouring was reduced (P less than .05) when pigs were fed 125 or 250 ppm aspirin; the improvement was greatest for 125 ppm aspirin after 5 d on test. Pooled data suggested no effect (P greater than .10) of aspirin on hemoglobin concentrations and blood clotting times; slightly higher (P less than .10) hematocrit values and DM content of colon digesta occurred in pigs fed 125 ppm aspirin vs controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Feed↗