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Biomedical subjects

D W Wright

Publications and source records attributed to D W Wright.

At least 19 recordsLinked to original sources

Monoclonal antibodies that distinguish avian type I and type III collagens: isolation, characterization and immunolocalization in various tissues.

Monoclonal antibodies were prepared that were specific for chicken type I and type III collagens. The specificity of these antibodies was determined by ELISA, inhibition ELISA, and immunoblot assays. The results showed that the monoclonal antibodies were specific for their respective antigens without significant cross reactivity to other types of collagen. An analysis of the location of the epitopes by rotary shadowing that a monoclonal antibody for type I collagen (called DD4) recognized type I procollagen close to the large globular domain at the carboxyl terminus of the molecule. A monoclonal antibody for type III collagen (called 3B2) recognized both the intact type III molecule and also the TCA fragment of type III collagen after mammalian collagenase digestion. The epitope was located approximately one-fifth of the distance from the amino-terminus of the intact molecule. The monoclonal antibodies were used for immunolocalization of type I and type III collagens in cryosections of heart, aorta, kidney, liver, thymus, skin, gizzard and myotendinous junction. In heart, aorta, kidney, liver, thymus and skin, type I and III collagens were colocalized in the connective tissue of each organ. In contrast, gizzard and myotendinous junction showed distinctly different staining patterns for the distribution of type I and type III collagen. The two monoclonal antibodies reported here are potentially useful reagents to study fibril formation involving type I and type III collagens.

Animals

Effect of including relationships in the estimation of genetic parameters of beef calves.

Variances and covariances for birth weight, gain from birth to weaning (ADG), and 205-d weight were obtained from a sire-dam model and a sire-maternal grandsire model for a herd of Angus and a herd of Hereford cattle. Estimates of direct additive genetic variance (sigma 2A), maternal additive genetic variance (sigma 2M), covariance between direct and maternal additive genetic effects (sigma AM), permanent environmental variance (sigma 2PE), and residual variance (sigma 2e) were obtained both with and without the inverse of the numerator relationship matrix (A-1) included. Estimates of heritability for direct genetic effects (h2A), maternal genetic effects (h2M), and the correlation between direct and maternal effects (rAM) for birth weight were .37, .18, and -.01 in Angus and .53, .23, and -.19 in Herefords, respectively, for the analyses without A-1. For the analyses with A-1, estimates of h2A, h2M, and rAM were .42, .22, and -.12 for Angus and .58, .22, and -.13 for Herefords, respectively. Estimates of h2A, h2M, and rAM for ADG were .43, .15, and -.44 in Angus and .52, .38, and -.03 in Herefords, respectively, without A-1. With A-1, estimates of h2A, h2M, and rAM were .57, .15, and -.32 for Angus and .58, .39, and -.05 for Herefords, respectively. Estimates of h2A, h2M, and rAM for 205-d weight were .49, .15, and -.46 for Angus and .58, .43, and -.06 for Herefords, respectively, without A-1. With A-1, estimates of h2A, h2M, and rAM were .63, .16, and -.36 for Angus and .66, .43, and -.08 for Herefords, respectively. Estimates of h2A were higher with A-1 than without A-1, but estimates of h2M were similar. Using variances and covariances obtained from analyses including A-1 generally gave higher estimates of direct breeding values than using variances and covariances obtained from analyses not including A-1. Both Pearson product-moment and Spearman rank correlations were high (.99) between estimates of breeding values from the two analyses, although some changes in rank did occur.

Analysis of Variance

Structural and functional comparison of type IX collagen-proteoglycan from chicken cartilage and vitreous humor.

Type IX collagen-proteoglycan is a major component of hyaline cartilages where it is located on the surface of the collagen fibrils so that a collagenous domain of the molecule (called COL3) and a non-collagenous domain (called NC4) project at periodic distances away from the surface of the fibril. Type IX collagen-proteoglycan is also present on the surface of the collagen fibrils of the adult chicken vitreous but, unlike cartilage, lacks the NC4 domain and possesses a very long chondroitin sulfate chain which provides an extensive coat to the fibril. A monoclonal antibody (called 4D6) is described which will distinguish cartilage from vitreous type IX collagen. To form the epitope for 4D6 two peptides called C2 and C5 derived, respectively, from the alpha 1(IX) and alpha 3(IX) chains are required. Further analysis shows that specificity for 4D6 resides only in the C2 peptide from cartilage and not in C5. These results are entirely consistent with recent evidence that there are two promoters for transcription of the alpha 1(IX) chain which will result in an alpha 1(IX) chain in which the NC4 domain is either present or absent and that expression of these two promoters has tissue specificity (Nishimura, I., Muragaki, Y., and Olsen, B. R. (1989) J. Biol. Chem. 264, 20033-20041). In addition, the function of type IX collagen in cartilage and vitreous may differ with the long chondroitin sulfate chains of vitreous type IX collagen being responsible for the gel-like matrix of this tissue.

Animals

Rubidium (86Rb+) influx into dorsal root ganglia and sciatic nerve endoneurium of control and streptozotocin-diabetic rats: comparison with enzymatic Na,K-ATPase activity.

Streptozotocin (STZ)-induced diabetes in the rat causes a significant reduction in ouabain-sensitive Na,K-ATPase pumping activity measured by 86Rb+ influx, in sciatic endoneurium (by 54%) and dorsal root ganglia (by 22%). For endoneurium, the change is similar to that of ouabain-sensitive enzymatic Na,K-ATPase activity (42%), but in dorsal root ganglia, the decrease in enzymatic Na,K-ATPase activity was much greater. 86Rb+ efflux from dorsal root ganglia showed no difference between diabetic and control animals, confirming that the abnormal 86Rb+ influx reflects Na,K-ATPase function and not abnormal membrane permeability. The significance of these findings to pathogenetic mechanisms in diabetic neuropathy is discussed.

Animals

Variance and covariance estimates for weaning weight of Senepol cattle.

Variance and covariance components were estimated for weaning weight from Senepol field data for use in the reduced animal model for a maternally influenced trait. The 4,634 weaning records were used to evaluate 113 sires and 1,406 dams on the island of St. Croix. Estimates of direct additive genetic variance (sigma 2A), maternal additive genetic variance (sigma 2M), covariance between direct and maternal additive genetic effects (sigma AM), permanent maternal environmental variance (sigma 2PE), and residual variance (sigma 2 epsilon) were calculated by equating variances estimated from a sire-dam model and a sire-maternal grandsire model, with and without the inverse of the numerator relationship matrix (A-1), to their expectations. Estimates were sigma 2A, 139.05 and 138.14 kg2; sigma 2M, 307.04 and 288.90 kg2; sigma AM, -117.57 and -103.76 kg2; sigma 2PE, -258.35 and -243.40 kg2; and sigma 2 epsilon, 588.18 and 577.72 kg2 with and without A-1, respectively. Heritability estimates for direct additive (h2A) were .211 and .210 with and without A-1, respectively. Heritability estimates for maternal additive (h2M) were .47 and .44 with and without A-1, respectively. Correlations between direct and maternal (IAM) effects were -.57 and -.52 with and without A-1, respectively.

Analysis of Variance

Behavioral effects of cocaine and its interaction with d-amphetamine and morphine in rats.

Drugs of abuse are commonly co-abused, and frequently these combinations produce effects which cannot be predicted by studying the effects of the individual drugs. To investigate the behavioral interactions which occur following combinations of cocaine plus amphetamine or cocaine plus morphine, rats were trained to respond under a differential reinforcement of low rates (DRL) schedule (10-14 sec). Cocaine (0.1-10 mg/kg) and d-amphetamine (0.1-3 mg/kg) decreased the percentage of reinforced responses (efficiency) at doses which had no effect on overall rate of responding. Following moderate doses of either drug, the interresponse time (IRT) distribution showed an increase in the percentage of shorter (less than 10 sec) IRT's. Morphine (0.1-10 mg/kg) also decreased efficiency, but the decrease which occurred was only observed at doses which also decreased overall response rates. As might be expected, the IRT distribution for morphine showed a dose-related increase in the percentage of long IRT's (greater than 14 sec). When doses of morphine which had no significant effect when administered alone (1 or 3 mg/kg) were combined with cocaine, the cocaine dose-response curve for efficiency was shifted down and to the left and response rates were increased. Analysis of the IRT distribution showed that the combination of an ineffective dose of cocaine, 1 mg/kg, plus 3 mg/kg morphine produced a shift in the IRT distribution to the left (an increase in the percentage of short IRT's). When cocaine was combined with 0.3 mg/kg d-amphetamine, a dose which had no effect when given alone, no significant interactions were observed on efficiency or overall rate of responding.(ABSTRACT TRUNCATED AT 250 WORDS)

Amphetamines

Disruption of performance under a titrating matching-to-sample schedule of reinforcement by drugs of abuse.

Five adult, male White Carneaux pigeons were trained to respond under a titrating delayed matching-to-sample schedule of reinforcement. Under this schedule, the length of the delay was changed as a function of the pigeons' performances in a manner that resulted in approximately 80% accuracy in matching performance. During the first five trials of each session, the delay value was fixed at either 3 sec or 30 sec (starting delay). On the 6th and all subsequent trials, the length of the delay value was increased by a fixed amount (the titration step), did not change or was decreased by an amount equal to the titration step. Initially, the starting delay and the titration step were set at 3 sec. These values were then systematically changed to a starting delay of 30 and a titration step of 3, and finally, a starting delay of 3 and a titration step of 1. Using this procedure, the effects of pentobarbital (0.3-10 mg/kg), cocaine (0.01-10 mg/kg), d-amphetamine (0.003-3 mg/kg) and phencyclidine (0.03-3 mg/kg) were determined under the different titration parameters. Pentobarbital was shown to decrease matching performance at doses (3, 5.6 and 10 mg/kg) that did not decrease response rates. Phencyclidine produced a similar effect at a dose of 1 mg/kg under two of the three phases of the experiment. No such specific effects were observed with cocaine or d-amphetamine. Thus, of the four drugs studied, pentobarbital showed the most pronounced effect on matching performance. Whether this represents merely a loss of stimulus control or a specific effect on memory is yet to be determined. In addition, the results of these experiments clearly show that when the delay value is allowed to change as a function of the pigeons' performance, pigeons are able to perform at or above 80% accuracy at longer delay values than those generally reported using fixed-delay values.

Animals

Vitreous humor of chicken contains two fibrillar systems: an analysis of their structure.

An analysis of the structure of chicken vitreous humor after brief homogenization of the tissue was performed. Electron micrographs prepared after rotary shadowing with platinum showed the presence of two distinct fibrils. The collagen fibril was coated by glycosaminoglycan which could be removed by chondroitinase ABC digestion. In addition, individual molecules of tenascin were observed wrapped around some of the collagen fibrils. A second beaded fibril was present and several fine filaments were observed to extend from each bead. The beaded fibril is formed by the overlap of these filaments, and beaded fibrils were observed in either a "closed" or an "open" form dependent on whether all of the filaments are brought together to form the overlap. A schematic diagram is presented for the structure of the beaded fibril. The potential relationship of the beaded fibril to the zonular fibrils and the elastin microfibrils is briefly discussed.

Animals

Amino acid uptake by dorsal root ganglia from streptozotocin-diabetic rats.

Dorsal root ganglia from streptozotocin-diabetic rats and age- and weight-matched control animals were incubated with 4-amino 3H-butyric acid. A significant reduction in uptake was observed in the diabetic animals. The incorporation of 3H-leucine into protein was also significantly reduced but the results did not allow a conclusion as to whether this was a secondary or an independent effect. These findings are discussed in relation both to the abnormalities known to develop in diabetic rats and to the causation of human diabetic neuropathy.

Animals

Insulin resistance in older rats.

Insulin-stimulated glucose utilization was estimated in vivo in 1.5-, 4-, and 12-mo-old rats with an insulin suppression test wherein the height of the steady-state plasma glucose ( SSPG ) concentration, at similar steady-state plasma insulin levels, provides a direct reflection of the efficiency of insulin-stimulated glucose disposal. In parallel studies, the effect of age on in vitro insulin-stimulated glucose uptake was assessed in perfused hindlimb preparations. In addition, changes in the activity of enzymes that regulate muscle glycolysis, glycogenesis, and glycogenolysis were determined in isolated soleus muscle. The results indicated that rats got heavier as they became older, and changes in weight were associated with parallel increases in mean (+/- SE) SSPG concentrations as rats grew from 1.5 (56 +/- 3 mg/dl) to 4 (172 +/- 6 mg/dl) to 12 mo of age (194 +/- 8 mg/dl). The age-related decline in in vivo insulin action was associated with a reduction in insulin action on muscle, and maximal insulin-stimulated glucose uptake by perfused hindlimbs of 12-mo-old rats was approximately 50% of the value seen with perfused hindlimbs from 1.5-mo-old rats. Soleus muscle enzyme activity also varied with age, with significant increases in glycogen synthase and decreases in glycogen phosphorylase documented. Furthermore, muscle glycogen phosphorylase activity, which fell during an insulin infusion in 1.5-mo-old rats, did not change when 12-mo-old rats were infused at comparable insulin levels. Finally, glycogen content was significantly increased (P less than 0.01) in soleus muscle from 12-mo-old rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging

Sucrose-induced insulin resistance in the rat: modulation by exercise and diet.

Isocaloric substitution of sucrose for starch results in hyperinsulinemia and deterioration of glucose tolerance, suggesting a loss of insulin sensitivity. In this study we have quantitated the insulin resistance which develops with sucrose feeding, and evaluated the ability of dietary fiber, or an increase in skeletal muscle activity, to inhibit, or even prevent, the detrimental effect of sucrose feeding on in vivo insulin action. Thus, 6-wk-old rats were fed one of the following regimens for three weeks: a 64% cornstarch diet (C), a 32% cornstarch + 32% sucrose diet (S), the (S) diet containing added wheat bran fiber (S/F), and the (S) diet given to rats running spontaneously in exercise wheel cages (S/ET). Insulin sensitivity was evaluated by comparing steady-state plasma glucose (SSPG) concentrations at constant plasma insulin levels approximately 70 microU/ml attained during the continuous infusion of epinephrine (0.08 micrograms/kg/min), propranolol (1.7 micrograms/kg/min), glucose (8 mg/kg/min), and insulin (2.5 mU/kg/min) to each experimental group. The results show that rats fed the S diet had a significant increase (p less than 0.01) in mean (+/- SEM) SSPG concentration compared with rats fed the C diet (255 +/- 14 versus 165 +/- 3 mg/dl). SSPG concentrations, although lower (p less than 0.05) in rats fed S/F (205 +/- 8 mg/dl), were still higher (p less than 0.05) than the C levels (165 +/- 3 mg/dl). However, S/ET completely inhibited the increase in SSPG concentration seen in rats fed S and the values were actually lower (p less than 0.05) than in rats fed C (100 +/- 10 versus 165 +/- 3 mg/dl). In conclusion 1) sucrose feeding results in a loss of insulin sensitivity in normal rats; 2) addition of fiber attenuates, but does not completely prevent, the loss of insulin sensitivity associated with feeding sucrose; 3) exercise training prevents the loss of insulin sensitivity seen in sucrose-fed rats, and actually improves glucose uptake beyond that seen in the control group. These results document the profound effect of environmental factors on in vivo insulin action.

Animals