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Biomedical subjects

D B Morton

Publications and source records attributed to D B Morton.

At least 73 records · Page 4Linked to original sources
PubMed1989

DNA fingerprinting of spontaneously hypertensive and Wistar-Kyoto rats: implications for hypertension research.

Probes to hypervariable minisatellite regions of DNA identify multiple loci scattered over the autosomal chromosomes and produce a complex Southern blot pattern of fragments termed a DNA 'fingerprint'. As concern has been raised that different stocks of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY) may not be biologically identical, we have compared the DNA of SHR and WKY from several sources using two such probes which identify different sets of minisatellite sequences. While the DNA fingerprints of SHR from the various sources were identical, variability was observed in those of WKY, indicating genetic heterogeneity between different WKY stocks. In animals from one of the commercial suppliers even inter-rat variability in DNA fingerprints was seen, suggesting genetic heterogeneity within that single colony. These observations indicate that experimental results obtained using WKY from different sources may not be directly comparable and could provide an explanation for some of the conflicting data that exist on the comparative characteristics of SHR and WKY. In separate studies, direct comparisons both of the DNA fingerprints of SHR and WKY and of SHR and stroke-prone spontaneously hypertensive rats (SHRSP) showed multiple differences between the strains. The polymorphisms seen could provide useful linkage markers in locating the chromosomal sites of the genetic loci responsible for raised blood pressure in the SHR and the propensity to strokes in the SHRSP.

Animals↗
PubMed1989

Identification and characterization of a pancreatic intrinsic factor in the dog.

An intrinsic factor has been identified in the canine pancreas, and output and properties of this protein have been compared with those of gastric intrinsic factor in the dog. Mean concentrations of intrinsic factor and peak outputs per minute were approximately 5- to 10-fold higher in pure pancreatic juice after stimulation with secretin and cholecystokinin, respectively, than in pentagastrin-stimulated gastric juice. Purified gastric and pancreatic intrinsic factors had an identical molecular mass of 65 kDa, estimated by gel filtration on Sephacryl S-200, while sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrated single bands corresponding to 53 kDa. Immunoblots showed that rabbit polyclonal antiserum to canine gastric intrinsic factor cross-reacted with canine pancreatic intrinsic factor. Gastric and pancreatic intrinsic factor-cyano[57Co]cobalamin complexes exhibited comparable association constants for ileal receptors in canine brush-border vesicles, while there was minimal binding to jejunal vesicles. These findings demonstrate that the canine pancreas is an important source of an intrinsic factor that closely resembles gastric intrinsic factor in the dog.

Animals↗
PubMed1989

Circulating concentrations of trypsin-like immunoreactivity and activities of lipase and amylase after pancreatic duct ligation in dogs.

The possibility that assay of circulating trypsin-like immunoreactivity (TLI) could assist in the diagnosis of acute pancreatitis in dogs has been examined by assaying plasma TLI concentrations after pancreatic duct ligation and comparing the results with plasma activities of lipase and amylase. Venous blood samples were obtained from 8 dogs before surgery, then daily for 5 days and at 14 days after ligation of pancreatic ducts. Plasma concentrations of TLI increased within 24 hours and tended to peak before and to decrease more rapidly than activities of lipase and amylase, remaining greater than the control range for 5 days in all but 2 dogs. Plasma lipase and amylase activities increased together and remained greater than the control range in all dogs for 5 days after surgery. Regression analysis of all postoperative data indicated significant correlations between concentration of TLI and lipase activity (r = 0.67, P less than 0.001), concentration of TLI and amylase activity (r = 0.53, P less than 0.001), and between lipase and amylase activities (r = 0.74, P less than 0.001). These findings suggested that assay of TLI may provide an early indication of acute pancreatitis in dogs. Because TLI is specifically pancreatic in origin, high plasma TLI concentration may prove a more reliable indicator of clinical pancreatitis than high activities of amylase or lipase, which may be derived from extrapancreatic tissues.

Amylases↗
PubMed1989

Biochemical changes in the jejunal mucosa of dogs with exocrine pancreatic insufficiency following pancreatic duct ligation.

The effects of exocrine pancreatic insufficiency on the small intestinal mucosa were examined in dogs following pancreatic duct ligation. There were no significant changes either in villus architecture or enterocyte height after duct ligation, but numbers of bacteria in duodenal juice increased then subsequently decreased following treatment with exogenous pancreatic enzymes. Pancreatic insufficiency resulted in a considerable increase in the proportion of microvillar membrane proteins of molecular mass over 200 kDa from 3.3 +/- 4 per cent (mean +/- SEM) to 13.6 +/- 7.2 per cent, and this decreased to 6.9 +/- 5.2 per cent following pancreatic enzyme supplementation. However, anticipated increases in activities of maltase and sucrase were not observed following duct ligation, and there was a reduction in lactase activity which was reversed by pancreatic supplementation. Activities of marker enzymes for the other subcellular organelles showed relatively minor or no changes throughout the study. These findings are consistent with a specific role for pancreatic enzymes in the post-translational processing of intestinal microvillar membrane proteins, and suggest that reduced degradation of brush border proteins in the absence of pancreatic secretions may be masked by quantitative and qualitative changes in the intestinal microflora.

Alkaline Phosphatase↗
PubMed1989

Effect of exocrine pancreatic insufficiency on cobalamin absorption in dogs.

The possibility that the canine pancreas might have an important role in the physiologic absorption of cobalamin (vitamin B12) has been explored by determining the effect of exocrine pancreatic insufficiency on cobalamin absorption and by examining the subsequent influence of bovine pancreatic enzymes and canine pancreatic juice. Exocrine pancreatic insufficiency was induced by ligation of pancreatic ducts and confirmed by indirect assessment of exocrine pancreatic function. Cobalamin absorption was determined by oral administration of cyano[58Co]cobalamin and quantitation of radioactivity in blood, urine, and feces during 48 hours. Pancreatic duct ligation resulted in a significant (P less than 0.001) decrease in cobalamin absorption, which was not restored by oral administration of bovine pancreatic enzymes, despite considerable improvements in steatorrhea and in vivo proteolytic activities. In marked contrast, malabsorption of cobalamin was significantly (P less than 0.05) reversed by oral administration of canine pancreatic juice. These results indicate that pancreatic secretions have an important role in the normal absorption of cobalamin in the dog, a role that does not appear to be attributable to pancreatic enzymes, but is consistent with the existence of a pancreatic intrinsic factor in this species.

Animals↗
PubMed1988

Experimental epikeratophakia with biological adhesive.

Successful experimental epikeratophakia grafting was done with the use of a biological adhesive. The use of an adhesive eliminated the need for any suturing of the corneal lenticule to the host cornea, and this method reduced the length of the operative procedure to only one third of the time that is taken when sutures are used. Tisseel (Immuno AG, Vienna), a commercially available two-component adhesive system based on human fibrinogen, which is activated by thrombin, was used. The glue was used in combination with an antifibrinolytic agent. With alteration of the operative technique and lenticule design, 70% of glued epikeratophakia grafts in a rabbit model were retained, compared with a 50% success rate with grafts applied without the use of an adhesive.

Animals↗
PubMed1988

Fresh frozen plasma therapy in acute pancreatitis: an experimental study.

Fresh frozen plasma therapy was assessed in a rat model of acute haemorrhagic pancreatitis incorporating the facility for continuous intravenous infusion in the unrestrained animal. Infusions including fresh frozen plasma were shown to significantly improve 72 h survival in the model compared to crystalloid (P less than 0.001) and colloid (P less than 0.05) control groups. The mechanism of this improved survival has yet to be elucidated, but this work supports the establishment of a controlled clinical trial of fresh frozen plasma therapy in this disease.

Acute Disease↗
PubMed1988

Quantitative assessment of human proteinases as agents for chemonucleolysis.

A rabbit model system is described. It allows accurate measurement of the dose-dependent loss of glycosaminoglycan from the nucleus pulposus of lumbar intervertebral discs after injection of a proteinase. At the dose equivalent to that of chymopapain used in human chemonucleolysis, two human serine proteinases, cathepsin G and chymotrypsin, were as effective as chymopapain in removing up to 80% of the glycosaminoglycan from the disc. A cysteine proteinase, cathepsin B released no more than 45% of glycosaminoglycan. X-ray films clearly showed narrowing of the disc space when 30-40% of glycosaminoglycan was removed. The degradation of the nucleus pulposus was seen histologically as loss of toluidine blue metachromasia.

Animals↗
PubMed1988

Experimental epikeratophakia using tissue lathed at room temperature.

This report presents for the first time the results of carrying out epikeratophakia with tissue lathed at room temperature. Using an experimental model of epikeratophakia in the rabbit, we evaluated tissue handling techniques for the preparation of donor lenticules. Details of the technique are described and the in-vivo and histopathological findings reported.

Animals↗
PubMed1988

The EGPs: the eclosion hormone and cyclic GMP-regulated phosphoproteins. I. Appearance and partial characterization in the CNS of Manduca sexta.

The present study describes 2 phosphoproteins, both with an apparent molecular weight of 54 kDa, in the CNS of the tobacco hornworm, Manduca sexta. Their phosphorylation is regulated by a neuropeptide, eclosion hormone (EH), and the second messenger cGMP, which thus have been named the EGPs (eclosion hormone- and cGMP-regulated phosphoproteins). Although cAMP was more effective than cGMP at stimulating the phosphorylation of the EGPs in CNS homogenates, in the intact CNS cGMP was more effective. Since cGMP mediates the action of EH, this strongly suggests that cGMP is the second messenger that stimulates the phosphorylation of the EGPs in vivo. The EGPs can only be phosphorylated in vitro during discrete time periods during the life of Manduca. During the larval and pupal molts, the EGPs can first be phosphorylated just prior to ecdysis. Their ability to be phosphorylated is correlated with the time when the insect is sensitive to EH. This close temporal correlation suggests that the ability to phosphorylate the EGPs determines when the insect can first respond to EH. During adult development, the EGPs first appeared on fluorograms 6 d before sensitivity to EH, suggesting that at this stage other factors may also be involved in the regulation of sensitivity. For the ecdyses of all 3 stages, EH appeared to stimulate the phosphorylation of the EGPs at ecdysis. The EGPs were found in all regions of the prepupal nervous system that were investigated, but only in the abdominal and pterothoracic ganglia of the developing adult. Fractionation of nervous system homogenates by ultracentrifugation revealed that one of the EGPs was present only in the pellet fraction, whereas the other was approximately equally distributed between pellet and supernatant. Furthermore, the EGPs in the pellet fraction could be partially solubilized with detergents and high salt concentrations.

Animals↗
PubMed1988

The EGPs: the eclosion hormone and cyclic GMP-regulated phosphoproteins. II. Regulation of appearance by the steroid hormone 20-hydroxyecdysone in Manduca sexta.

Two proteins (the EGPs) in the CNS of the tobacco hornworm Manduca sexta are phosphorylated by the action of the neuropeptide eclosion hormone (EH), which triggers ecdysis behavior (Morton and Truman, 1986, 1988). The onset of sensitivity to EH requires prior exposure to the steroid hormone 20-hydroxyecdysone (20-HE). A series of steroid removal and replacement experiments indicates that the EGPs are also regulated by 20-HE with a time course that is similar to that seen for the 20-HE regulation of behavioral sensitivity to EH. This suggests that the steroid regulation of EH sensitivity is due to the regulation of the EGPs. The appearance of the EGPs requires not only the presence of 20-HE, but also its subsequent removal. The appearance of the EGPs can be blocked in vivo and in vitro by maintaining artificially elevated levels of 20-HE, but only up to a particular time in development. The ending of this steroid-sensitive period occurs 4 hr before the normal appearance of the EGPs, consistent with the hypothesis that the EGPs are synthesized de novo in response to the removal of 20-HE.

Animals↗
PubMed1987

Reactions of hemocytes of immune and non-immune Galleria mellonella larvae to Proteus mirabilis.

Immune larval Galleria mellonella removed live Proteus mirabilis from the hemolymph less effectively than did non-immune larvae. This was attributed to a decline in total hemocyte counts, levels of plasmatocytes and granulocytes and hemocyte adhesion capacity. Immune serum possessed factors which reduced bacterial adhesion to the hemocytes. This was not due to altering bacterial surfaces but rather to irreversible suppression of hemocyte activity.

Animals↗
PubMed1987

DNA fingerprints of dogs and cats.

Human minisatellite probes consisting of tandem repeats of the 'core' sequence, a putative recombination signal in human DNA, cross-hybridize to multiple polymorphic fragments in dog and cat DNA to produce individual-specific DNA 'fingerprints'. Pedigree analysis shows that most of the DNA fragments detected in an individual are heterozygous, and that these fragments are derived from multiple dispersed autosomal loci. DNA fingerprints of cats and dogs should prove suitable for individual identification and for establishing family relationships. They are also suitable for rapid marker generation in large pedigrees and could be applied to linkage analysis in these animals.

Animals↗
PubMed1986

Some failings of pulsatility index and damping factor.

Pulsatility index (PI) is a commonly used method of objective assessment of the Doppler waveform. PI falls with increasing proximal stenosis and is raised by increasing peripheral resistance. Damping factor (DF) for an arterial segment is calculated by dividing the proximal by the distal PI. DF rises with increasing severity of disease of the arterial segment. DF is not, however, sufficiently accurate to be used alone but is usually combined with transit time measurements to provide information of diagnostic use. Both PI and DF have been examined in a canine model of combined segment disease. With increasing stenosis, distal PI falls as expected but so also does proximal PI. Such a stenosis is, in effect, a flow-throttling resistance so that although the characteristics of blood flow are altered by its presence, similar changes are observed both above and below the stenosis. The reduction of PI by a stenosis distal to the insonation site may result in the false interpretation of a low PI as indicating disease proximal to the insonation site. The observed similarity between PI proximal and distal to a stenosis reduces the usefulness of pulsatility index damping factor, particularly in the assessment of the femoro-popliteal segment in combined segment disease.

Animals↗
PubMed1985

Guidelines on the recognition of pain, distress and discomfort in experimental animals and an hypothesis for assessment.

Under the 1876 Cruelty to Animals Act it is necessary to recognise pain so that an assessment may be made to determine if it is 'an experiment calculated to give pain' and 'to prevent the animal feeling pain'. Under the conditions of the licence it is also necessary to recognise 'severe pain which is likely to endure' and 'suffering considerable pain'. In the White Paper May 1983 (Command 8883) it is stated that: 'in the application of controls the concept of pain should be applied in a wide sense' and 'the Home Secretary's practice has been to interpret the concept of pain to include disease, other disturbances of normal health, adverse change in physiology, discomfort and distress'. The draft European Convention for the Protection of Vertebrate Animals used for Experimental and other Purposes, aims to control, subject to specific exceptions, any experimental or other scientific procedure which 'may cause pain, suffering, distress or lasting harm'. (The White Paper states that UK control will be stricter than the Council of Europe proposals.) Thus, there is a considerable onus on the experimenter to recognise pain (not to define it) and to alleviate it. It is intended that this article should be of help, not only to newcomers inexperienced in the recognition of pain, but also possibly to those relatively experienced workers who may be called upon to evaluate the pain involved in a new model or an individual animal.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1985

Steroid regulation of the peptide-mediated increase in cyclic GMP in the nervous system of the hawkmoth, Manduca sexta.

The action of the peptide, eclosion hormone (EH) on the CNS of Manduca sexta appears to be mediated via the second messenger cGMP. Injections of EH or release of endogenous EH cause a rapid increase in cGMP in the CNS. Cyclic GMP, 8-bromo-cGMP and the phosphodiesterase inhibitors IBMX and theophylline mimic the action of EH in triggering premature ecdysis behavior. The CNS is only sensitive to EH just before ecdysis, both in triggering ecdysis and increasing endogenous cGMP levels. The development of the ability to increase cGMP levels occurs earlier than the behavioral sensitivity and the relative timing of these events is discussed in terms of the likely site for the block in behavioral sensitivity. The steroid hormone 20-hydroxyecdysone is shown to regulate the ability of EH to elevate cGMP levels in the CNS.

1-Methyl-3-isobutylxanthine↗