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Biomedical subjects

B A Watkins

Publications and source records attributed to B A Watkins.

At least 55 records · Page 3Linked to original sources
PubMed1996

Dietary (n-3) and (n-6) polyunsaturates and acetylsalicylic acid alter ex vivo PGE2 biosynthesis, tissue IGF-I levels, and bone morphometry in chicks.

This study examined the effects of dietary (n-6) and (n-3) polyunsaturated fatty acids (PUFA) and acetylsalicylic acid (ASA) on bone ash content, morphometry, fatty acid composition, ex vivo PGE2 biosynthesis, tissue IGF-I concentration, and serum alkaline phosphatase (ALPase) activity in chicks. Newly hatched chicks were fed a semipurified diet containing soybean oil (S) or menhaden oil / safflower oil (M) at 90 g/kg. At 4 days of age, chicks were divided into four equal treatment groups receiving 0 mg [symbol: see text] or 500 mg [symbol: see text] of ASA/kg of diet: S[symbol: see text]ASA, M[symbol: see text]ASA, S[symbol: see text]ASA, and M[symbol: see text]ASA. Lipid and ASA treatments did not affect bone length, bone ash, or bone mineral content in chicks. Chicks fed M had increased fractional labeled trabecular surface and tissue level bone formation rates, independent of ASA treatment, compared with those given S. A significant fat x ASA interaction effect was found for trabecular bone volume, thickness, separation, and number. Chicks fed S had higher 20:4(n-6) but lower 20:5(n-3) concentrations in liver and bone compared with those given M. Ex vivo PGE2 biosynthesis was higher in liver homogenates and bone organ cultures of chicks fed S compared with the values for those given M at 17 days. ASA treatment decreased ex vivo PGE2 production in liver homogenates and bone organ cultures of chicks, independent of the dietary lipids. Chicks fed ASA had a lower concentration of IGF-I in tibiotarsal bone compared with those not given ASA at 19 days. Serum ALPase activity was higher in chicks given M compared with those fed S, but the values were reversed with ASA feeding. This study demonstrated that both dietary fat and ASA modulated bone PGE2 biosynthesis, and that (n-3) PUFA and fat x ASA interactions altered bone morphometry.

Alkaline Phosphatase↗
PubMed1996

Susceptibility to hepatic oxidative stress in rabbits fed different animal and plant fats.

OBJECTIVE: This study was designed to determine the effect of diets enriched with plant and animal fats on oxidative stress and glutathione metabolism in rabbit liver tissues. This study was conducted to investigate whether the type of dietary fat will impact fatty acid composition and oxidant/antioxidant status in tissues. METHODS: Rabbits were fed diets containing 2 g corn oil/100 g diet (low fat diet, LF) and LF supplemented with 16 g/100 g diet of either corn oil (CO), CO with added cholesterol (CO + C), milk fat (MF), chicken fat (CF), beef tallow (BT), or lard (L) for 30 days. After the feeding period, livers were analyzed for total fatty acid composition, thiobarbituric acid reactive substances (TBARS), conjugated dienes, and reduced glutathione (GSH), as well as for activities of glutathione peroxidase (GP) and glutathione reductase (GR). Moreover, to fully determine the oxidative stability and free radical trapping capacity, TBARS levels were measured after additional exposure of liver homogenates to 10 mM 2,2(1)-azo-bis-amidinopropane- hydrochloride (AAPH) for up to 21 hours. RESULTS: CO and CF, but not saturated fats such as MF, increased liver conjugated diene and TBARS levels and decreased liver GSH levels and GP activity. In tissues additionally exposed to AAPH, the maximum oxidation, measured as TBARS, was reached between 6 and 7 hours of treatment, independent of dietary fat. In addition, there was a marked effect of AAPH on the maximum rate of TBARS formation with the following descending order: CO > CF > CO + C > L > MF > BT > LF. This high susceptibility to oxidative stress in liver tissues of rabbits fed the CO diet may be explained in part by the significant elevation in linoleic acid (18:2n-6). DISCUSSION: There appears to be an inverse correlation between dietary fat-mediated oxidative stress and antioxidant enzyme activities. The present data suggest that high levels of dietary unsaturated fat should be avoided if oxidative stress is a critical issue in nutrition-related diseases. In addition, these data support our hypothesis that diets rich in MF provide a lipid environment with low susceptibility to oxidative stress.

Amidines↗
PubMed1996

Dietary polyunsaturated fatty acids modulate responses of pigs to Mycoplasma hyopneumoniae infection.

Polyunsaturated fatty acids (PUFA) are immunomodulators, but few studies have examined how these dietary components influence infectious respiratory disease. Groups of nine pigs were fed casein and corn starch-based diets containing 10.5 g/100 g corn oil (CO), linseed oil (LO), menhaden oil (MO), linseed + corn oil (LC, 1:1) and menhaden + corn oil (MC, 1:1). As a methodological control, one group of pigs (n = 15) was fed a commercial ration (control diet; C). Pigs inoculated intratracheally with Mycoplasma hyopneumoniae after 4 wk of consuming the diets were killed 3 wk later. Gross lung lesions in MO-fed pigs were less (P < 0.05) than those in LC- and MC-fed pigs. Pigs fed MO had less peribronchial inflammation (P < 0.05) than all other groups. Gross lung lesions correlated negatively with basal in vitro alveolar macrophage tumor necrosis factor (TNF) production in pigs fed diets that contained negligible levels of (n-3) PUFA (C and CO). Basal macrophage TNF production did not correlate with lung lesion scores for diets containing more (n-3) PUFA than C or CO (LO, MO, LC and MC). For pigs fed the LO, MO, LC and MC diets, mean gross lung lesions increased as the mean ratio of (n-3):(n-6) PUFA in alveolar macrophage lipids decreased. Serum levels of alpha1 acid glycoprotein (AGP) were less (P < 0.05) in pigs fed MO, and there was a rise in mean lung lesions scores for each PUFA-fed group as mean AGP levels increased. These results indicate that dietary PUFA can affect disease pathogenesis and that the (n-3):(n-6) PUFA ratio may modulate the host response.

Animals↗
PubMed1996

Resistance of human immunodeficiency virus type 1 to neutralization by natural antisera occurs through single amino acid substitutions that cause changes in antibody binding at multiple sites.

The ability of human immunodeficiency virus type 1 (HIV-1) to replicate in the presence of strong immune responses to the virus may be due to its high mutation rate, which provides envelope gene variability for selection of neutralization-resistant variants. Understanding neutralization escape mechanisms is therefore important for the design of HIV-1 vaccines and our understanding of the disease process. In this report, we analyze mutations at amino acid positions 281 and 582 in the HIV-1 envelope, where substitutions confer resistance to broadly reactive neutralizing antisera from seropositive individuals. Neither of these mutations lies within an antibody-binding site, and therefore the mechanism of immune escape in both cases is by alteration of the shape of the envelope proteins. The conformation of the CD4-binding site is shown to be critical with regard to presentation of other discontinuous epitopes. From our analysis of the neutralization of these variants, we conclude that escape from polyclonal sera occurs through alterations at several different epitopes, generally resulting from single amino acid substitutions which influence envelope conformation. Experiments on a double mutant showed that the combination of both mutations is not additive, suggesting that these variants utilized alternate pathways to elicit similar alterations of the HIV-1 envelope structure.

Animals↗
PubMed1996

Linoleate impairs collagen synthesis in primary cultures of avian chondrocytes.

The effects of supplemental fatty acids, vitamin E (VIT E), and iron-induced oxidative stress on collagen synthesis, cellular injury, and lipid peroxidation were evaluated in primary cultures of avian epiphyseal chondrocytes. The treatments included oleic and linoleic acids (O or 50 microM) complexed with BSA and dl-alpha-tocopheryl acetate (VIT E at 0 or 100 microM). After 14 days of preculture, the chondrocytes were enriched with fatty acids for 8 days then cultured with VIT E for 2 days. The chondrocytes were then treated with ferrous sulfate (O or 20 microM) for 24 hr to induce oxidative stress. Collagen synthesis was the lowest and the activity of lactate dehydrogenase (LDH) was the highest in chondrocyte cultures treated with 50 microM linoleic acid and 0 VIT E. In contrast, VIT E supplemented at 100 microM partially restored collagen synthesis in the chondrocytes enriched with linoleic acid and lowered LDH activity in the media. The iron oxidative inducer significantly increased the values of thiobarbituric acid-reactive substances (TBARS) in the culture medium. The data showed that linoleic acid impaired chondrocyte cell function and caused cellular injury but that VIT E reversed these effects. Results from a previous study demonstrated that VIT E stimulated bone formation in chicks fed unsaturated fat, and the present findings in cultures of epiphyseal chondrocytes suggest that VIT E is important for chondrocyte function in the presence of polyunsaturated fatty acids. VIT E appears to be beneficial for growth cartilage biology and in optimizing bone growth.

Animals↗
PubMed1995

Vitamin E stimulates trabecular bone formation and alters epiphyseal cartilage morphometry.

The effects of dietary vitamin E (VIT E) and lipids on tissue peroxidation and fatty acid composition, epiphyseal growth plate cartilage development, and trabecular bone formation were evaluated in chicks. A 2 x 2 factorial design was followed using two levels (30 and 90 IU/kg of diet) of dl-alpha-tocopheryl acetate and two different dietary lipids. The basal semipurified diet contained one of the following lipid treatments: anhydrous butter oil (40 g/kg) + soybean oil (60 g/kg), [BSO], or soybean oil (100 g/kg), [SBO]. After 14 days of feeding, the level of alpha tocopheryl in plasma was higher and thiobarbituric acid reactive substances (TBARS) were less in plasma and liver of chicks supplemented with 90 IU of VIT E compared with those given 30 IU of VIT E. Body weights and tibiotarsal bone lengths were not affected by the dietary treatments. Saturated fatty acids (14:0, 15:0, 16:0, 17:0, and 18:0) were increased in the tibiotarsal bone of chicks fed the BSO diet. In contrast, total polyunsaturated fatty acids and the ratio unsaturated fatty acids/saturated fatty acids were higher is plasma of chicks fed SBO compared with the values from chicks fed BSO. The thickness of the entire growth plate cartilage and the lower hypertrophic chondrocyte zone was significantly greater in chicks fed 90 IU/kg of VIT E. Kinetic parameters on bone histomorphometry indicated that mineral apposition rate was higher in chicks fed 90 IU of VIT E. The interaction effect between the VIT E and BSO treatments led to the highest trabecular bone formation rate among the groups. These data suggest that VIT E protects against cellular lipid peroxidation in cartilage to sustain normal bone growth and modeling.

Animals↗
PubMed1995

Electron spin resonance studies of fatty acid-induced alterations in membrane fluidity in cultured endothelial cells.

Endothelial cell dysfunction has been implicated in the development of atherosclerosis. Of vital importance to the maintenance of endothelial cell integrity is the preservation of membrane functional and structural properties, such as membrane fluidity. The aim of this study was to develop a model for studying the relationship between endothelial cell integrity and membrane fluidity alterations in a well-defined cell culture setting. Alterations in membrane fluidity were assessed using electron spin resonance after labeling endothelial cells with the lipid-specific spin labels, CAT-16 and 12-nitroxide stearic acid. Endothelial cells were exposed to various 18-carbon fatty acids, i.e. stearic (18:0), oleic (18:1), linoleic (18:2), or linolenic (18:3), in addition to lipolyzed HDL (L-HDL) and benzyl alcohol. Membrane phospholipid fatty acid composition of endothelial cells supplemented with these fatty acids was analyzed using gas chromatography. All fatty acids, except 18:0, decreased membrane fluidity. A relationship between membrane fluidity and fatty acid compositional alterations in cellular phospholipids was observed. In particular, the arachidonic acid content decreased following exposure to 18:1, 18:2, or 18:3. Exposure of endothelial cells to L-HDL, lipoprotein particles which contain high levels of 18:1 and 18:2, also decreased membrane fluidity. The stabilization of cytoskeletal actin filaments by phalloidin partially prevented 18:2-induced increases in albumin transfer, thus implicating a cytoskeletal involvement in the 18:2-induced membrane fluidity changes involved in endothelial cell dysfunction. The present study shows that the exposure of endothelial cells to various lipids causes membrane fluidity alterations which may contribute to endothelial cell dysfunction and atherosclerosis.

Animals↗
PubMed1995

Essential fatty acid metabolism in boys with attention-deficit hyperactivity disorder.

Attention-deficit hyperactivity disorder (ADHD) is the term used to describe children who are inattentive, impulsive, and hyperactive. The cause is unknown and is thought to be multifactorial. Based on the work of others, we hypothesized that some children with ADHD have altered fatty acid metabolism. The present study found that 53 subjects with ADHD had significantly lower concentrations of key fatty acids in the plasma polar lipids (20:4n-6, 20:5n-3, and 22:6n-3) and in red blood cell total lipids (20:4n-6 and 22:4n-6) than did the 43 control subjects. Also, a subgroup of 21 subjects with ADHD exhibiting many symptoms of essential fatty acid (EFA) deficiency had significantly lower plasma concentrations of 20:4n-6 and 22:6n-3 than did 32 subjects with ADHD with few EFA-deficiency symptoms. The data are discussed with respect to cause, but the precise reason for lower fatty acid concentrations in some children with ADHD is not clear.

Attention Deficit Disorder with Hyperactivity↗
PubMed1995

V-region and class specific RT-PCR amplification of human immunoglobulin heavy and light chain genes from B-cell lines.

We have designed and tested primers that amplify complete human kappa and lambda light chain genes, and human Fd fragments from gamma, mu and alpha heavy chain genes. These primers were tested for efficiency and specificity on monoclonal sources of human immunoglobulin RNA, obtained from human B-cell lines of known immunoglobulin gene expression. Analysis of the sequences derived from these B-cells confirms the specificity of the PCR primers and the extent of somatic mutation seen in different B-cell malignancies supports existing concepts for differing aetiologies in the tumours concerned.

Amino Acid Sequence↗
PubMed1995

Role of dietary lipids on fetal tissue fatty acid composition and fetal survival in swine at 42 days of gestation.

Eighty-six crossbred (Duroc x Yorkshire) gilts were used in two trials to assess the effect of supplemental dietary fats during early gestation on fetal survival, fetal development, and fatty acid (FA) composition in gilt plasma and fetal head and body at d 37 to 45 after breeding. Three diets contained 4% (wt/wt) added fat either as coconut, soybean (SO), or menhaden oils (MO). A fourth diet containing corn starch approximately isoenergetic to the quantity of the oils was used as a control. Across both trials, fetal survival percentage did not differ (P > .10) according to treatment, but in Trial 2 fetal survival was higher (P < .06) for gilts fed the MO diet than for gilts fed the control diet. The FA composition of plasma of gilts at slaughter and the conceptus tissues were similar; both were influenced by the FA composition of the diets. The ratio of n-3:n-6 FA was higher in conceptus tissue than in maternal plasma and the ratio was higher (P < .05) for the MO diet than for the other diets. Subsequently, a total of 46 multiparous females were used in three trials to further evaluate the effects of MO on percentage of fetal survival relative to the corn starch control diet; the results demonstrated no differences (P > .10) in percentage of fetal survival due to the addition of MO. The high percentage of fetal survival observed in gilts on all the treatments and the unequal ovulation rate in sows on different oil diets precluded definitive conclusions regarding the effects of supplemental oil on fetal survival.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Evaluating effects of fish meal on milk fat yield of dairy cows.

Experiment 1 measured the effect of different amounts of dietary fish meal on milk yield and composition. Milk fat percentage and yield were decreased by increased fish meal intake, but this change was not associated with changes in ruminal fermentation patterns. Plasma long-chain n-3 polyunsaturated fatty acids were increased as intake of fish meal increased. Ruminal disappearance of DM, CP, and lipid in fish meal was measured in situ. Seventy percent of lipid disappeared by 8 h. Intraruminal administration of fish oil did not alter ruminal fermentation and only slightly changed fatty acid profiles in duodenal digesta, plasma, or milk. Duodenal infusion increased plasma n-3 fatty acids but did not affect composition of fatty acids in milk. Experiment 2 compared effects of dietary fish meal and fish oil on milk production and composition. Fish meal increased n-3 fatty acids in plasma compared with those of the fish oil treatment. No changes were found in milk yield or composition because of experimental treatments. Cows fed fish meal or fish oil differed significantly in plasma fatty acid profiles but did not differ in ruminal VFA concentrations or milk fat yield.

Animal Feed↗
PubMed1994

Dietary lipids modify the fatty acid composition of cartilage, isolated chondrocytes and matrix vesicles.

The effects of dietary lipids on the fatty acid composition of hyaline cartilage, epiphyseal chondrocytes (EC) and matrix vesicles (MV) were evaluated in chicks. A basal semipurified diet was fed to chicks containing one of the following lipid sources at 70 g/kg: soybean oil, butter+corn oil, margarine+corn oil or menhaden oil+corn oil (MEC). Articular and epiphyseal growth cartilage were isolated from the proximal tibiotarsus; EC and MV were subsequently released by trypsin (EC 3.4.21.4) and collagenase (EC 3.4.24.3) digestion followed by ultracentrifugation. The fatty acid composition of polar lipids in chick epiphyseal cartilage at three and six weeks, as well as articular cartilage, EC and MV at eight weeks of age revealed the presence of high levels of saturated and monounsaturated fatty acids (up to 85.5%) but low levels of n-6 polyunsaturated fatty acids (PUFA) (2.6-10.2%). Mead acid (20:3n-9, > 3%) was also present in cartilage, EC and MV lipids, and was unaffected by the dietary lipid treatments. Total n-3 PUFA concentrations were the highest in cartilage, EC and MV of chicks consuming MEC. Feeding MEC lowered the levels of 20:4n-6 in cartilage, but increased 20:5n-3 levels. The data are consistent with those reported previously which showed that cartilage tissues are low in n-6 PUFA and that they contain 20:3n-9. We furthermore demonstrated that the PUFA composition of cartilage can be modified by dietary lipids.

Age Factors↗
PubMed1994

Dietary and ruminally derived trans-18:1 fatty acids alter bovine milk lipids.

The objectives of this study were to determine whether dietary fish oil enhances production of trans-18:1 isomers in the bovine rumen and whether flow of ruminally derived and/or dietary trans-18:1 to the small intestine is correlated with milk fat production. Four lactating Holstein cows with cannulas in the rumen and proximal duodenum were fed diets with 0% supplemental fat (control diet), 3% hydrogenated tallow fatty acids (HTFA diet), 1.5% menhaden oil plus 1.5% stearic acid (MO+SA diet), or 1.5% soybean oil plus 1.5% partially hydrogenated soybean oil (SBO+HSBO diet) in a 4 x 4 Latin square design. Intakes of trans-18:1 were 0, 12, 0 and 69 g/d when cows were fed the control, HTFA, MO+SA and SBO+HSBO diets, respectively. However, the extent of 18:1 + 18:2(n-6) + 18: 3(n-3) biohydrogenation in the rumen was reduced by consumption of the MO+SA and SBO+HSBO diets. As a result, flow of trans-18:1 to the duodenum was 163 g/d for MO+SA-fed cows and 152 g/d for SBO+HSBO-fed cows compared with 38 g/d for cows fed the HTFA and control diets. Incomplete biohydrogenation accounted for all of the trans-18:1 flow when the MO+SA diet was fed. Compared with results when the HTFA and control diets were fed, milk fat percentage was lower and concentration of trans-18:1 in milk fat was higher when the MO+SA and SBO+HSBO diets were fed. Across all treatments, milk fat percentage decreased linearly with the amount of trans-18:1 flowing to the duodenum and the concentration of trans-18:1 in milk fat. Results indicate that bovine milk fat percentage is depressed by trans-18:1, whether derived from the diet or from incomplete biohydrogenation of unsaturated fatty acids.

Animals↗
PubMed1993

Functional properties of retinal Müller cells following transplantation to the anterior eye chamber.

Two types of glial cells occur in the retina, Müller cells and astrocytes. These cells share several structural features such as extending endfeet onto blood vessels of the retina. Retinal vessels express a tight blood-retinal barrier which is comparable to the blood-brain barrier (BBB) of the CNS. While astrocytes have been implicated in the induction of the BBB, the role of Müller cells in the blood-retinal barrier is unknown. To determine if Müller cells are capable of influencing vascular permeability, we have prepared Müller cells that are free of astrocytes and transplanted them to a peripheral target, the anterior eye chamber. Müller cells were identified 2 weeks to 3 months after injection and were predominantly localized within the connective tissue of the ciliary body. The Müller cells occurred as dense clusters of cells closely associated with ciliary blood vessels. The ciliary vessels adjacent to Müller cells were freely permeable to circulating horseradish peroxidase (HRP), suggesting that Müller cells did not induce tight barrier properties from these leaky peripheral vessels. In contrast, cortical astrocytes injected into the anterior eye chamber preferentially formed a monolayer on the anterior surface of the iris, a region known to contain blood vessels that are impermeable to circulating tracers (e.g., Raviola, Exp Eye Res [Suppl] 25:27, 1977). Müller cells were rarely associated with the iris and the few cells that were present were located deep within the iris stroma rather than on the surface. The behaviour of guinea pig Müller cells transplanted to the anterior eye chamber contrasts sharply with that of cortical astrocytes in terms of: 1) the ocular compartment to which Müller cells migrate; 2) the tissue invasiveness of the cells; and 3) the degree of permeability of blood vessels adjacent to transplanted cells. The results of this study emphasize the functional distinctness of the two types of retinal glia and suggest that Müller cells from guinea pig retina may not be active in modifying the permeability properties of peripheral blood vessels, a function that has been suggested for astrocytes.

Animals↗
PubMed1993

Selective disruption of endothelial barrier function in culture by pure fatty acids and fatty acids derived from animal and plant fats.

Endothelial cell integrity has been suggested to play a role in the development of atherosclerosis. The effects of fatty acids on endothelial barrier function were tested by measuring albumin transport across endothelial monolayers cultured on polycarbonate filters. Compared with control cultures, a 24-h exposure to 90 mumol/L lauric (12:0) and linoleic acid (18:2) but not to butyric (4:0), hexanoic (6:0), octanoic (8:0), decanoic (10:0), myristic (14:0), palmitic (16:0) or stearic acid (18:0) caused an increase in albumin transfer across endothelial monolayers. Selective enrichment of a "physiological" serum fatty acid mixture (FA-Mix; 90 mumol/L) with 90 mumol/L of 12:0 or 18:2 significantly increased albumin transfer, whereas enrichment with 90 mumol/L of 4:0, 16:0 or 18:0 significantly decreased albumin transfer relative to 180 mumol/L FA-Mix. Only 12:0- or 18:2-treated cultures showed increased Ca(++)-ATPase activity and the presence of lipid droplets. Fatty acids (60 mumol/L) extracted from butter fat and beef tallow had no effect on albumin transfer, whereas fatty acids extracted from chicken fat and corn oil consistently disrupted endothelial barrier function. This fat-induced disruption of endothelial barrier function seems to be related to the amount of 18:2 present in each fat source. These data indicate that unsaturated fats cause cellular perturbations that result in a decrease in endothelial barrier function in this model system, and that high dietary levels of unsaturated fats may be detrimental to cell integrity.

Albumins↗
PubMed1993

Local modulation of skeletal growth and bone modeling in poultry.

Skeletal growth and bone modeling in poultry are regulated by complex interactions between the animal's genetic potential and a host of systemic and localized factors (growth factors and cytokines) influencing bone biology. The objective of these interactions is to orchestrate the achievement of bone architecture that balances functionally appropriate morphology with the skeleton's involvement in mineral homeostasis. Within this context, bone modeling in the growing animal represents an adaptive process that is distinct from bone remodeling, which is the term used to describe the resorption and formation of mineralized tissue that maintains skeletal mass and morphology in the adult. As many of the skeletal lesions that afflict poultry are the consequence of abnormalities in bone modeling, not bone remodeling, an appreciation of the differences between these two contrasting processes is a prerequisite for understanding the pathogenesis of skeletal lesions in poultry.

Animals↗