Characterization of chymotrypsin activity during early ontogeny of larval red drum (Sciaenops ocellatus)

The temporal evolution of chymotrypsin activity during early ontogeny of laboratory reared red drum larvae was accomplished using a combination of biochemical assays and electrophoretic methods (substrate SDS-PAGE). Optimal functional conditions for chymotrypsin were also determined. Chymotrypsin activity was first detected prior to the onset of exogenous feeding. Total chymotrypsin activity increased with age and standard length. Specific activity was greatest on day 10 post-hatch. Maximal chymotrypsin activity was observed at 50 °C, pH 7.8, and Ca²⁺ concentration of 25 mM. Using substrate gel electrophoresis and specific inhibitors the molecular weight of red drum chymotrypsin was estimated to be 26–27 kD. Our results indicate that the digestive system of red drum larvae is capable of alkaline proteolysis before first feeding and suggest that chymotrypsin may have potential as an indicator of nutritional condition.     

Recovery from neuromuscular block in dogs: restoration of spontaneous ventilation does not exclude residual blockade

ObjectiveTo evaluate if return of spontaneous ventilation to pre-relaxation values indicates complete recovery from neuromuscular blockade.Study designProspective, with each individual acting as its own control.AnimalsTen healthy adult female Beagle dogs weighing 6.2–9.4 kg.MethodsDogs were anesthetized with propofol, dexemedetomidine and isoflurane. Spontaneous ventilation was assessed by measuring end-tidal CO₂, expired tidal volume, peak inspiratory flow, respiratory rate and minute ventilation. Vecuronium 25 μg kg^?1 IV was administered and neuromuscular block was evaluated by measuring the train-of-four (TOF) ratio with acceleromyography in the hind limb. During spontaneous recovery from neuromuscular block, the TOF ratio when each ventilatory variable returned to baseline was recorded.ResultsThis dose of vecuronium produced moderate neuromuscular block in all dogs, with TOF ratio values of 0–18% at maximal block. Expired tidal volume, peak inspiratory flow and minute ventilation returned to pre-relaxation values when the median TOF ratio was ≤ 20%. The median TOF ratio was 42% when the end-tidal CO₂ returned to pre-relaxation values.Conclusions and clinical relevanceSignificant residual neuromuscular block could be measured at the hind limb with acceleromyography when ventilation had spontaneously returned to pre-vecuronium values. Monitoring spontaneous ventilation, including end-tidal CO₂, expired tidal volume, peak inspiratory flow or minute ventilation cannot be used as a surrogate for objective neuromuscular monitoring, and this practice may increase the risk of postoperative residual paralysis.     

Egtved virus: temperature-dependent immune response of trout to infection with low-virulence virus

Abstract. The present report deals with experiments concerning the theoretical basis for development of a live, attenuated vaccine against viral haemorrhagic septicaemia (VHS) of rainbow trout, Salmo gairdneri Richardson, on the basis of a virus strain derived from the Egtved virus reference strain F, the pathogenicity of which has been strongly reduced by in vitro passage. It is shown that the low pathogenicity of the virus (designated the Reva strain) is a genetically stable feature and that the protection against VHS induced by infection with the virus is due to an immune response. Following immunological priming at 10°C partial protection developed at 5° 10° and 15° but not at 20°C, whereas neutralizing antibody was produced at all four temperatures by some of the fish. The duration of virus persistence in trout was found to be inversely proportional to the water temperature.     

Ultrasonographic measurement of normal renal parameters

Both kidneys were imaged ultrasono-graphically in each of 100 normal adult dogs. Small but significant differences were found in the dimensions of the left and right kidneys and in renal dimensions of male and female dogs of similar bodyweight. There was a statistically significant correlation between renal length and bodyweight (r = 0–84, P<0–001). The relationship between renal length and bodyweight was linear except at very low or very high bodyweights. There was also a statistically significant correlation between renal volume and bodyweight (r = 0–91, P<0–001), and the relationship remained linear even at extremes of bodyweight. Graphs could therefore be constructed for renal lengthy bodyweight and renal volume/bodyweight with 95 per cent confidence limits.     

Subcellular distribution of copper in the liver of fetal deer in the last month of gestation

Fetal deer, in the last month of gestation, accumulate high concentrations of copper in the liver. Livers from fetal deer in late gestation were homogenised and fractionated by continuous sucrose density gradient centrifugation. The distribution of copper closely followed that of DNA; approximately two thirds of the metal was localised to the nuclear fractions with the remaining third in the cytosolic fractions. The fractionation procedure was repeated with digitonin, a lysosomal perturbant: lysosomes were disrupted and the marker enzyme, N-acetyl-beta-glucosaminidase, shifted from the particulate fractions to the cytosolic fractions; the distribution of copper was unaffected. Differential centrifugation of homogenate confirmed that approximately two thirds of the copper was associated with the nuclear fraction. Further confirmation of a nuclear localisation of copper was provided by X-ray microanalysis of purified nuclei.     

Recovery characteristics following maintenance of anaesthesia with sevoflurane or isoflurane in dogs premedicated with acepromazine

A standard anaesthetic protocol was used to anaesthetise 40 dogs for intravenous urography and a retrograde urethrogram or vaginourethrogram. The dogs were allocated by blocked randomisation to receive either isoflurane or sevoflurane for maintenance of anaesthesia after they had been premedicated with acepromazine and pethidine, and anaesthesia induced with propofol. An observer who was unaware of which agent had been used assessed ataxia 30 and 60 minutes after discontinuation of administration of the anaesthetic and assigned an overall recovery score. No complications occurred during anaesthesia of either group of dogs. The scores for ataxia were significantly lower after 60 minutes than after 30 minutes, but there was no significant difference between the groups. The quality of recovery was significantly better in the dogs that received sevoflurane than in those that received isoflurane, but the recovery times were similar.     

Regulation of protein secretion through controlled aggregation in the endoplasmic reticulum

A system for direct pharmacologic control of protein secretion was developed to allow rapid and pulsatile delivery of therapeutic proteins. A protein was engineered so that it accumulated as aggregates in the endoplasmic reticulum. Secretion was then stimulated by a synthetic small-molecule drug that induces protein disaggregation. Rapid and transient secretion of growth hormone and insulin was achieved in vitro and in vivo. A regulated pulse of insulin secretion resulted in a transient correction of serum glucose concentrations in a mouse model of hyperglycemia. This approach may make gene therapy a viable method for delivery of polypeptides that require rapid and regulated delivery.