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Biomedical subjects

D A Knight

Publications and source records attributed to D A Knight.

52 records · Page 3Linked to original sources
PubMed1994

Cytomegalovirus inhibits major histocompatibility class II expression on infected endothelial cells.

Persistent human cytomegalovirus (HCMV) infections are responsible for significant morbidity and mortality in immunocompromised individuals. One mechanism by which HCMV may develop persistence after primary infection is through inhibition of host cell human leukocyte antigen (HLA) class II expression with resultant escape from normal antiviral immune surveillance. Immunofluorescence flow cytometry of human endothelial cell (EC) cultures infected with HCMV AD169 and an EC propagated strain, VHL/E, showed a marked reduction in interferon-gamma (IFN-gamma)-induced surface expression of HLA-DR. This inhibition did not occur when EC were treated with ultraviolet-inactivated virus and IFN-gamma. HCMV, as determined by dual-labeling immunohistochemistry, inhibited induction of surface and cytoplasmic class II antigens specifically in infected cells. HCMV infection also inhibited IFN-gamma and tumor necrosis factor-alpha up-regulation of HLA class I expression. Northern blot analysis of infected, IFN-gamma-treated human umbilical vein endothelial cells revealed an absence of class II mRNA. Persistence of HCMV may result in part from its ability to inhibit HLA class II induction in infected cells.

Blotting, Northern↗
PubMed1993

In vitro induction of endothelial HLA class II antigen expression by cytomegalovirus-activated CD4+ T cells.

CMV has been associated with allograft rejection and transplantation-associated arteriosclerosis. CMV infects endothelium, the interface between allograft tissue and the host immune system. Although endothelial HLA class II expression is a hallmark of vascular rejection, CMV does not directly induce these antigens on infected endothelial cells (EC) and, indeed, renders them refractory to HLA DR induction by IFN-gamma. Our earlier studies have demonstrated, however, that CMV-infected EC are capable of eliciting vigorous activation responses by allogeneic, CMV-seropositive donor-derived CD4+ T cells. We now show that T cells thus activated can induce HLA DR expression on noninfected EC. Human umbilical vein endothelial cells (HUVEC) were inoculated at low titer with CMV strain VHL/E, cocultured with allogeneic, CMV-seropositive or CMV-seronegative donor-derived CD4+ T cells, then dual immunohistochemically stained for CMV antigen and HLA DR, or assayed for HLA DR expression by fluorescence flow cytometry. Results demonstrated that HLA DR was induced in 20-70% of HUVEC in monolayers containing less than 0.5% CMV-infected EC after coculture with CMV-seropositive donor-derived T cells. No such induction was observed in experiments employing T cells isolated from CMV-seronegative individuals. Expression of this antigen was strictly limited to noninfected cells. Rare HLA DR induction was observed in virus-free cocultures. To determine whether endothelial HLA DR was induced by a soluble factor(s) elaborated by activated T cells, noninfected HUVEC monolayers were treated for 72 hr with cell-free supernatant from CMV-infected or noninfected HUVEC/T cell cocultures and assayed as above. Again, HLA DR expression was induced by supernatant from CMV-positive cocultures (only when cocultured T cells were isolated from CMV-seropositive donors), but not by supernatant from CMV-negative cocultures or from T cells cultured alone. The soluble factor was identified as IFN-gamma by the complete attenuation of HLA DR induction by anti-IFN-gamma mAb. These findings suggest that allograft endothelium harboring low level CMV may be capable of eliciting host CD4+ T cell activation, and that consequent release of IFN-gamma is capable of inducing endothelial HLA class II expression, as observed in vascular rejection and transplantation-associated arteriosclerosis. Results of these studies thus provide insight into mechanisms that may help elucidate the association between CMV and rejection-related immune events in the allograft.

Antibodies, Monoclonal↗
PubMed1992

Use of a human enteral feeding preparation for treatment of hyperlipemia and nutritional support during healing of an esophageal laceration in a miniature horse.

Nasogastric infusion of a human enteral feeding preparation was effective in reversing hyperlipemia in an anorectic miniature horse with an esophageal laceration. The nutrient preparation was delivered every 4 hours by gravity flow through a 12 F enteral feeding tube. Within 48 hours of initiating enteral nutrition, the hyperlipemia had resolved. Signs of intolerance to the preparation were not observed, and further weight loss was prevented. The use of a human enteral formula was a convenient and successful alternative for the treatment of hyperlipemia in the horse.

Animals↗
PubMed1992

Histamine tachyphylaxis in human airway smooth muscle. The role of H2-receptors and the bronchial epithelium.

The role of airway epithelium and H2-receptors in the development of histamine tachyphylaxis was studied using human isolated bronchial smooth muscle strips obtained from 18 patients undergoing thoracotomy. In epithelium-intact strips, a 38% reduction in the maximal contractile response (Emax) (p less than 0.002) and a 2.14-fold increase in the EC50 (p less than 0.02; n = 18) was observed after three separate histamine cumulative concentration effect curves (CCEC). In contrast, significant differences were not seen for either Emax (p greater than 0.4; n = 10) or EC50 (p greater than 0.26; n = 10) in epithelium-denuded strips. In separate experiments, both intact and denuded muscle strips were treated with the H2-receptor antagonist ranitidine (60 microM), either 30 min prior to the first or 30 min prior to the second histamine CCEC. In epithelium-intact strips, pretreatment with ranitidine caused a 1.8-fold increase in Emax in the initial CCEC (p less than 0.02), and both ranitidine schedules prevented tachyphylaxis (n = 8). In epithelium-denuded preparations, ranitidine did not enhance the responsiveness to histamine beyond that seen in untreated epithelium-denuded strips (n = 6). These data suggest that histamine-induced tachyphylaxis occurs in human airway smooth muscle and is mediated, at least in part, via H2-receptors resident on airway epithelium. In vivo, this may function as a protective mechanism, but damage to the epithelium and loss of H2-receptors may be significant in the development of histamine bronchial hyperreactivity as seen in asthma.

Bronchi↗
PubMed1992

Effect of protein source in liquid formula diets on food intake, physiologic values, and growth of equine neonates.

The effects of 2 liquid formula diets differing in protein source were evaluated in orphan foals. The response of 7 foals fed a diet containing casein as the protein source, and 6 foals fed a diet containing a combination of whey and casein, was compared with the response in a reference group of 8 mare-raised foals. Orphaned foals were fed 150 kcal/kg of body weight/d, divided into 6 equal feedings of 25 kcal/kg. Formula intake was comparable among the experimental groups, and foals fed the liquid formula diet grew as well as mare-raised foals. There was no difference among groups in mean daily body weight gain, wither height, heart girth, body temperature, pulse, respiration rate, capillary refill time, or skin tenting. Insulin and blood glucose concentrations increased in both groups of foals fed formula diets, returning to prefeeding values within 4 hours. Differences among groups were found for serum alkaline phosphatase, alanine transaminase, cholesterol, creatinine, and glucose values; all other serum chemical values were comparable among groups. Plasma amino acid determinations revealed that arginine and ornithine were significantly lower in foals in both experimental groups than in reference foals, suggesting that arginine may have been the limiting amino acid in these diets. Diarrhea developed in foals in all treatment groups, but in most cases was self-limiting. These results suggest that the protein source of liquid formula diets may be less important in foals than in infants.

Analysis of Variance↗
PubMed1991

A preliminary study of the tolerance of healthy foals to a low residue enteral feeding solution.

After a three day acclimatization period, six healthy, young (aged 4 to 20 days) orphan foals of mixed breeding were fed 100 per cent of their caloric needs (estimated at 523 kjoules/kg bodyweight [bwt] or 125 kcal/kg bwt/day) as a low residue isotonic feeding solution (LRF) for seven days. The solution provided 4.18 kjoules (1 kcal/ml) and was fortified with minerals and protein to meet estimated foal requirements. The solution was fed through an indwelling 12 French feeding tube. Five of the six foals completed the study; the loss of the sixth foal apparently was unrelated to the feeding protocol. The foals tolerated LRF well. Signs of intolerance were noted in two foals and were limited to flatulence, mild bloat and very mild abdominal pain associated with a decreased interval between two feedings during the first 48 h on 100 per cent LRF. Complete recovery without therapy occurred within 6 h and feedings were resumed. Growth in height and weight were comparable to published data for healthy foals raised with their dams. Feeding tubes were easily maintained with no apparent dysphagia, regurgitation or discomfort to foals. This low residue, calorically dense, isotonic feeding solution may be useful for enteral feeding of selected foals aged at least seven days.

Animal Feed↗
PubMed1991

The interaction of acetylcholine and histamine on human bronchial smooth muscle contraction.

The interaction of histamine (Hist) and acetylcholine (ACh) on human isolated bronchial smooth muscle (HIBSM) contraction, and the influence of the epithelium, was assessed using HIBSM obtained from 15 patients undergoing thoracotomy. Cumulative concentration effect curves for ACh and Hist, together with combinations of equipotent concentrations of both agonists, were generated using both epithelium-intact and epithelium-denuded HIBSM. In epithelium-denuded HIBSM both ACh (p less than 0.05) and Hist (p less than 0.005) produced a significantly enhanced maximal response and a 2.1 fold increase in the potency of ACh (p less than 0.02, n = 13). When ACh and Hist were added simultaneously, in equipotent concentrations, to epithelium-intact HIBSM, a significantly less (p less than 0.0005, n = 13) than additive response occurred with only 60% of the predicted maximum response being observed. However, following epithelium removal, an additive interaction between the two agonists (n = 8) occurred. Using HIBSM from five of the original 15 patients, similar experiments were performed to determine the influence of the muscarinic receptor antagonist atropine (0.1 microM) and the H1 receptor antagonist mepyramine (10 microM). Both resulted in a significantly less than additive interaction (40-50% of predicted tensions). Similar experiments were also performed in the presence of the cyclo-oxygenase inhibitor indomethacin (5 microM) and these failed to reverse the inhibition observed in HIBSM contraction (n = 5). The inhibitory interaction between ACh and Hist appears to be epithelium dependent and is not mediated via the release of prostanoids. Thus, there appears to be a complex interaction between contractile agonists and the epithelium, which is not just a simple summation of the activation of individual receptors on HIBSM.

Acetylcholine↗
PubMed1990

The effect of epithelium removal on human bronchial smooth muscle responsiveness to acetylcholine and histamine.

The respiratory epithelium produces a variety of inflammatory mediators which may be influencing the bronchial hyperreactivity observed in patients with asthma. Animal studies have demonstrated that removal of the epithelium from tracheal and bronchial smooth muscle causes enhanced responses to cholinergic agonists and histamine (Hist). In this study the effect of epithelium removal on human bronchial smooth muscle response to acetylcholine (ACh) and Hist was assessed. Bronchial smooth muscle was obtained fresh from the operating theatre from 12 patients undergoing thoracotomy. Cumulative concentration effect curves (CCEC) for Hist and ACh were generated for epithelium intact and epithelium denuded muscle strips. All CCEC's were performed in duplicate and all denuded strips were obtained from the same airway immediately adjacent to the intact strip. The mean (+/- SEM) maximum response for Hist for the intact strip was 8.6 +/- 1.1 (grams/gram wet weight) and 12.0 +/- 1.4 (grams/gram wet weight) for the denuded strip (p less than 0.05). For ACh the values were 9.3 +/- 1.3 (g/g wet weight and 14.3 +/- 1.8 (g/g wet weight), respectively (p less than 0.05). The pD2 (-log EC50) for ACh was increased two-fold following epithelium removal (p less than 0.05). For Hist there was a similar increase in pD2 but this did not reach statistical significance. Thus removal of the epithelium from human isolated bronchial smooth muscle appears to modulate responsiveness to ACh and Hist. This enhanced responsiveness consequent to epithelium loss may prove important with respect to the development of worsening asthma.

Acetylcholine↗
PubMed1990

Biokinetics of 237Pu citrate and nitrate in the rat: implications for Pu studies in man.

Plutonium-237 decays mainly by electron capture with a half-life of 45 d. Alpha particles are emitted in only 5 x 10(-3)% of its disintegrations. This nuclide can now be produced with relatively small amounts of alpha-emitting contaminants so that, in principle, 237Pu can be used for studies of Pu biokinetics in man. However, because of its high specific activity, there was some doubt that its metabolism would be the same as that of the alpha- and beta-emitting isotopes of Pu normally encountered in the nuclear industry. In this study, the biokinetics of nearly "pure," high specific activity 237Pu are compared with those of lower specific activity, "impure" 237Pu containing significant amounts of alpha-emitting Pu, following administration to rats by intravenous injection as the citrate. Both the distribution and excretion of the "pure" and "impure" 237Pu used in the two studies were similar and also in good agreement with the results of previously reported studies using 239Pu and 241Pu citrate, thus validating the use of 237Pu for studies of Pu metabolism in man. Data on the biokinetics of 237Pu nitrate are also included.

Animals↗
PubMed1990

The effects of copper supplementation on the prevalence of cartilage lesions in foals.

The potential role of dietary copper in the development of cartilage defects in foals was investigated. Twenty-one mares were fed rations containing 13 ppm copper (CuC, control) or 32 ppm copper (CuS, supplemented) during the last three to six months of gestation and first three months of lactation. Their foals were fed pelleted concentrate containing 15 or 55 ppm Cu and were destroyed at 90 (5 CuC and 5 CuS foals) or 180 (6 CuC and 5 CuS foals) days. Focal cartilage lesions were found at multiple sites on necropsy. In foals killed at 90 days, there were over twice (9 versus 4) as many lesions of osteochondrosis and more than four times (9 versus 2) as many articular lesions of osteophyte formation or thinning in CuC foals compared with CuS foals. These differences were due predominantly to a higher number of lesions in one CuC foal. Two 90-day CuC foals had osteochondrosis of articular-epiphyseal (A-E) complex, one with thickenings and separation from subchondral bone and one with subchondral fibrosis. One 90-day CuS foal had a cartilage thickening of the A-E complex in the tibiotarsal joint with separation from subchondral bone. In foals killed at 180 days, there were seven times more articular lesions (21 versus 3) of osteophyte formation or thinning, nearly twice as many lesions of osteochondrosis (13 versus 8) [corrected] in the physis and over five times as many involving the A-E complex (11 versus 2) in six CuC foals compared with five CuS foals.(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Feed↗
PubMed1990

The effect of dietary selenium on humoral immunocompetence of ponies.

Fifteen Shetland ponies were used in a 7-wk trial to study the effect of supplemental Se on humoral antibody production. Four 3-yr-old, five 2-yr-old and six yearling ponies were depleted of Se before being assigned randomly to either a low Se (.02 ppm) or higher Se (.22 ppm) diet. Each pony was challenged antigenically with 2 ml of sheep packed red blood cells upon receiving its respective diet and again 2 wk post-treatment. Blood samples were drawn weekly and assayed for glutathione peroxidase activity, Se and immunoglobulin concentration and antibody titers. Compared with those ponies receiving the low Se concentrate, ponies receiving the Se-supplemented diet had higher (P less than .01) glutathione peroxidase activities and blood Se concentrations during the later weeks of the experiment. An enhanced primary response was observed in Se-supplemented ponies as evidenced by increased hemagglutination titers. Higher IgG concentrations (P less than .01) also were observed in the Se-supplemented group. Dietary Se concentration of .02 ppm was inadequate for optimum immune function in the equine.

Animals↗
PubMed1988

Beta-adrenoceptor desensitization in guinea-pig isolated trachea.

Exposure to (-)-isoprenaline (25 microM, 1 h) caused a stereoselective, time and concentration-related decrease in smooth muscle beta 2-adrenoceptor function in guinea-pig trachea. Furthermore, tracheal relaxant responsiveness to the beta-adrenoceptor agonists (+/-)-fenoterol and (-)-noradrenaline was reduced, while that to theophylline and nitroprusside was unaffected. Responsiveness to forskolin was marginally but significantly reduced. Indomethacin, a cyclooxygenase inhibitor and mepacrine, an inhibitor of phospholipid turnover, had no significant effect on the extent of isoprenaline-induced desensitization. Conversely, cortisol (25 microM) significantly reduced desensitization and enhanced the rate of spontaneous recovery of responsiveness to isoprenaline. Desensitization was not accompanied by a reduction in the density of beta-adrenoceptors in the trachea, as assessed by binding and light microscopic autoradiography using [125I]iodocyanopindolol [( 125I]CYP). Thus, desensitization was probably caused primarily by beta-adrenoceptor/adenyl cyclase uncoupling. This model may be useful in investigations of the effect of glucocorticoids on the beta-adrenoceptor dysfunction recognized in severe asthma.

Animals↗
PubMed1985

The effect of artificial rearing on the growth of foals.

Fourteen Quarter Horse foals were used to evaluate the effects of artificial rearing on growth. Seven foals were removed from their dams at 3 d of age and fed a reconstituted 26% crude protein (CP) milk replacer free choice for 1 mo, at which time ad libitum solid feeding began. Controls were weaned from their dams at 2 mo of age and fed a 21% CP concentrate ad libitum until the end of the trial. Variables measured during the 26-wk trial were live body weight, height at the withers and length of body from point of shoulder to point of hip. No significant differences were found between the two groups, except during wk 8 where 2-mo weaned foals were slightly heavier (P less than .10). Average daily gains for artificially reared and 2-mo weaned foals were .95 and .98 kg, respectively.

Animal Feed↗