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A comparison of several hypertonic solutions for resuscitation of bled sheep.

Small volumes (4 ml/kg) of 2400 mOsm NaCl restore cardiac output and mean arterial pressure to 80% of baseline after hemorrhage (65% of blood volume) in unanesthetized sheep. An equal volume of normal saline is less effective. To identify an optimal hypertonic solution, we screened six 2400 mOsm solutions in 18 randomized experiments in 8 sheep: NaCl, NaHCO3, NaCl/sodium acetate, NaCl/mannitol, NaCl/6% Dextran 70, and glucose. Cardiovascular function, as determined by cardiac output and mean arterial pressure, was restored best with NaCl, NaCl/NaAc, and NaCl/Dex. These three solutions were then evaluated using 18 sheep in 36 experiments. Following a 1-hr baseline period, the sheep were bled to a mean arterial pressure of 50 mm Hg for 2 hr. One of the solutions was then given in a volume of 4 ml/kg over 2 min and the sheep were monitored for 3 hr. Within 3 min of the infusion, cardiac output increased to greater than 100% of baseline for all three solutions. The NaCl-Dex solution sustained a significantly higher cardiac output over the 3-hr observation period than the other solutions. Plasma volume increased for all solutions following infusion. NaCl-Dex maintained plasma volume significantly better than the other solutions. As a further control, an isotonic solution of 6% Dextran 70 in normal saline was studied. It was not as effective as the hypertonic NaCl-Dex in maintaining cardiac output, mean arterial pressure, or plasma volume. Osmolality increased 10% (309 to 326 mOsm/kg H2O), plasma [NA] increased 7% (151 to 161 meq/liter), and plasma [K] decreased from 3.9 to 2.6 meq/liter following the hypertonic infusions. The sheep appeared to tolerate these electrolyte changes well. We conclude that a single bolus infusion of 2400 mOsm NaCl with 6% Dextran 70 best resuscitates sheep that have been subjected to a moderate degree of hemorrhagic shock compared to several other solutions. Its beneficial effects are caused in part by a sustained reestablishment of plasma volume. More studies are needed to document the safety of dextran in the clinical setting of hemorrhagic shock. Small volumes of hypertonic solutions may be valuable in the initial fluid resuscitation of patients in hemorrhagic shock.

Animals↗

Food aversion conditioned in anesthetized sheep.

We discovered that a food aversion could be conditioned in anesthetized sheep. Sheep were allowed to eat a familiar food (alfalfa-grain pellets) for 30 min, and 90 min later they were given either an intraruminal (IR) injection of water (C), an IR injection of LiCl (L), anesthesia followed by an IR injection of water (A), or anesthesia followed by an IR injection of LiCl (A+L). Induction of anesthesia was by an intravenous injection of pentobarbitone sodium, and maintenance of deep anesthesia was by halothane. Sheep were maintained in deep anesthesia for 2 h to ensure that the effects of LiCl on the acquisition of a food aversion, which occur within about 1 h, were completed before they awakened. When tested 5 days later, sheep that received LiCl (treatments L and A+L) consumed less alfalfa-grain pellets than sheep that did not receive LiCl (treatments C and A) (241 g vs. 306 g; p = 0.057). Intake of sheep that were anesthetized (treatments A and A+L) did not differ from that of sheep that were not anesthetized (treatments C and L) (295 g vs. 252 g; p = 0.183). Nor was there an interaction between LiCl and anesthesia (p = 0.423). Thus, we conclude that changes in preferences for foods caused by postingestive feedback occur automatically every time food is ingested (i.e., they are noncognitive), and the kind and amount of feedback is a function of the match between the food's chemical characteristics and its ability to meet the animal's current demands for nutrients.

Anesthesia, General↗

Airway effects of inhaled bradykinin, substance P, and neurokinin A in sheep.

The effects of inhaled bradykinin (BK), substance P (SP), and neurokinin A (NKA) on pulmonary resistance and airway responsiveness to carbachol were studied in conscious allergic sheep. Inhaled BK (20 breaths, 0.1 to 5.0 mg.ml-1) caused dose-dependent increases in pulmonary resistance. Neither inhaled SP nor NKA (20 breaths, 0.1 to 1.0 mg.ml-1) produced significant bronchoconstriction in allergic sheep. However, the response to SP could be enhanced (p less than 0.05) by pretreatment with the neutral endopeptidase inhibitor, thiorphan (40 breaths, 1 mg.ml-1). Sheep that were allergic to Ascaris suum antigen were 5.9 times (p less than 0.05) more sensitive to the constrictor effects of BK than nonallergic sheep. BK-induced bronchoconstriction was blocked in a dose-dependent fashion by the BK beta 2-receptor antagonist, NPC 567 (D-arginine[hydroxyproline3,D-phenylalanine7]BK). Atropine (0.2 mg.kg-1, intravenously) and nedocromil sodium (1 mg.kg-1 in 3 ml of saline, aerosolized) significantly inhibited the BK-induced bronchoconstriction by 97% and 43%, respectively. Chlorpheniramine (2 mg.kg-1, intravenously) had no effect. NKA caused a transient increase in airway responsiveness in allergic sheep, producing a mean 1.9-fold leftward shift in dose-response curves to aerosolized carbachol (p less than 0.05). This hyperresponsiveness was not evident 24 hours after NKA challenge. Neither SP nor BK changed airway responsiveness. Thus, in allergic sheep, inhaled BK caused a more pronounced bronchoconstriction than that observed in nonallergic sheep. The bronchoconstriction was blocked by a BK-receptor antagonist and appeared to be partially mediated via cholinergic reflexes.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Inhalation↗

T lymphocyte subset alterations following bluetongue virus infection in sheep and cattle.

To determine potential mechanisms of differential disease expression in ruminants infected with bluetongue virus (BTV), clinically normal, BTV-seronegative, yearling sheep and cattle were infected subcutaneously with a standardized insect-source inoculum of BTV serotype 17 (BTV-17) (three infected and one contact control each) or animal adapted BTV serotype 10 (BTV-10) (three sheep only). BTV was isolated from peripheral blood cell components of infected sheep and cattle and all infected animals showed evidence of seroconversion by 14 days post infection (PI). Sheep infected with both serotypes of BTV developed pyrexia, oral lesions, and leukopenia which were most severe on days 7-8 PI. Analysis of peripheral blood mononuclear leukocytes with specific monoclonal antibodies and flow cytometry revealed panlymphocytopenia on day 7 PI. This response was further characterized by an increase in the CD4/CD8 ratio (greater than 3) resultant from a greater decrease in absolute numbers of circulating SBU-T8(CD8+) ("cytotoxic/suppressor") lymphocytes compared to SBU-T4 (CD4)+ ("helper") lymphocytes. SBU-T19+ lymphocytes were also decreased below baseline values on days 5-14 post infection. On day 14 PI there were increased CD8+ lymphocytes and decreased CD4/CD8 ratios (approximately 0.6) in these sheep. Clinical and hematologic changes in cattle infected with BTV-17 were minimal and consisted of mild pyrexia (rectal temperature 103 degrees F) on day 9 PI in two of three infected animals and mild leukopenia on several days PI in one animal. This leukopenia was the result of a pan T lymphocytopenia with CD4/CD8 ratios in the expected range (1-2). Similar to infected sheep, infected cattle did have a shift (decrease, approximately 0.8) in the peripheral CD4/CD8 ratio associated with an increase in circulating BoT8 (CD8)+ lymphocytes on day 14 post infection. Lymphocytes in the peripheral blood of all sheep and cattle infected with BTV-17 proliferated in vitro in response to purified BTV-17. These results confirm and extend those of previous studies that indicate species differences in the hematologic response to an equivalent BTV infection in domestic ruminants.

Animals↗

An effective peptide vaccine to eliminate bovine leukaemia virus (BLV) infected cells in carrier sheep.

Protective effects of the gp51 of bovine leukaemia virus (BLV) expressed by a recombinant baculovirus (rgp51) and synthetic multiple antigenic peptides (MAP) of T-helper, T-cytotoxic, and B-cell epitopes of gp51 were investigated against BLV challenge. Two and three sheep were immunized with rgp51 and a mixture of peptides with Freund's complete adjuvant, respectively. BLV was detected from all the immunized sheep at 2 weeks and showed peak levels at 4 weeks after the challenge. However, in two sheep immunized with the mixed peptides, the titer of BLV gradually decreased and one sheep eliminated BLV completely at 28 weeks after the challenge. These two sheep showed higher lymphocyte proliferative responses against the immunized peptides than the other sheep. One of the sheep also showed the specific cytotoxic lymphocyte activity against the BLV gp51-expressing target in vitro. These results suggest the possibility of the peptide vaccine for elimination of BLV in carrier animals in vivo.

Amino Acid Sequence↗

Receptor changes in the spinal cord of sheep associated with exposure to chronic pain.

There is evidence that post-injury hypersensitivity is partly due to changes in the central nervous system. Sheep with foot rot were used to investigate the effect of chronic pain on some receptors thought to be involved in spinal nociceptive processing systems (alpha 2 adrenoceptor and mu and delta opioid receptors). Saturation binding studies showed a variable distribution of [3H] clonidine (alpha 2 adrenoceptor agonist) in the spinal cord of normal sheep. The number of receptors (Bmax) present in areas thought to be involved in nociceptive processing, laminae I and II and lamina X, increased to 131% and 169% of control sheep values respectively in animals exposed to chronic pain. The affinity of the receptors (KD), however, remained unchanged at approximately 2 nM. There was less [3H]DAGO (mu opioid agonist) and [3H]DPDPE (delta opioid agonist) binding in the sheep spinal cord. Both opioid receptor types being mainly located in the superficial dorsal horn. The [3H]DPDPE binding was unchanged in the sheep with foot rot, whilst the number (Bmax), but not the affinity, of the [3H]DAGO binding sites increased in laminae I and II in lame animals to 130% of the control sheep values. Hence, in animals in chronic pain, the number of alpha 2 adrenoceptors and mu opioid receptors increased mainly in areas of the sheep spinal cord associated with nociception.

Animals↗

Evidence for multiple anthelmintic resistance in sheep and goats reared under the same management in coastal Kenya.

Four experiments, two with sheep and two with goats, were carried out to determine the efficacy of ivermectin, fenbendazole, levamisole, closantel and some of their combinations by faecal egg count reduction tests. In the first experiment, injectable ivermectin, oral ivermectin, fenbendazole and levamisole were tested in 6-month-old lambs, and their reduction percentages were 77%, 13%, 42% and 92%, respectively. In the second experiment, with yearling sheep, the reduction percentages were 35% for injectable ivermectin, 32% for fenbendazole, 99% for levamisole, 48% for closantel, 92% for injectable ivermectin combined with fenbendazole, 99% for injectable ivermectin combined with levamisole, and 100% for fenbendazole combined with levamisole. In the study with 18-month-old goats given the same dose rates as those recommended for sheep, the reduction percentages were 73% for injectable ivermectin, 25% for fenbendazole, and 78% for levamisole. Another group of 14-month-old goats was treated with dose rates 1.5 times those recommended for sheep and the reduction percentages were 93% for levamisole, 92% for injectable ivermectin, and 97% for a combination of levamisole and ivermectin. In all experiments with sheep and goats the gastrointestinal nematode parasites identified by larval cultures were Haemonchus contortus, Trichostrongylus spp. and Oesophagostomum spp. The gastrointestinal nematodes of both sheep and goats on this farm are resistant to ivermectin and fenbendazole, whereas levamisole is still effective in sheep, but not in goats. The results are discussed in relation to the farm as a source of breeding stock to smallholder farmers and its potential to spread anthelmintic resistance.

Animal Husbandry↗

Fatty acid synthesis from amino acids in sheep adipose tissue.

The rates of incorporation of 14C from 14C labelled acetate, glucose, alanine, leucine, isoleucine and valine into fatty acids has been measured in perirenal adipose tissue from foetal lambs and 8-month-old sheep, and into both fatty acids and acylglycerol glycerol in adipose tissue from 3-year-old sheep and 220-240 g female rats. Rates of incorporation of 14C from amino acids into fatty acids were much lower in adipose tissue from sheep (at all three ages) than from rats, whereas rates of incorporation of 14C into acylglycerol glycerol were either greater in sheep adipose tissue or the same as in rat adipose tissue. The rate of incorporation of 14C from amino acids into fatty acids decreased in the order leucine greater than alanine greater than isoleucine greater than valine in adipose tissue from rats and foetal lambs, and in the order leucine greater than alanine = isoleucine greater than valine in adipose tissue from 8-month- and 3-year-old sheep. Amino acids make a very small contribution to fatty acid synthesis in adipose tissue from sheep at all stages of development examined while fatty acids are a minor product of amino acid metabolism in sheep adipose tissue. The study provides further evidence for an important role for ATP-citrate lyase in restricting the utilization of acetyl-CoA generated in the mitochondria for fatty acid synthesis.

Acetates↗

Distinct pulmonary and hepatic forms of flavin-containing monooxygenase in sheep.

1. Flavin-containing monooxygenase (FMO) in pulmonary and hepatic microsomes from sheep was analyzed by western blotting by probing with antibodies raised against FMO purified from rabbit lung and pig liver. 2. Pulmonary microsomes from sheep contain a single major protein which cross-reacts with the antibody to rabbit lung FMO, but no band can be observed when probed with the antibody to the pig liver enzyme. Likewise, sheep liver microsomes contain a protein which cross-reacts with the antibody to pig liver FMO, but no significant staining is observed following incubation with antibody to the lung enzyme. 3. Sheep pulmonary and hepatic microsomal FMO also display a difference in activity toward chlorpromazine and n-dodecylamine. 4. Preliminary evidence suggests that sheep FMO may be induced (liver) or repressed (lung) during pregnancy. 5. Sheep are similar to rodents (rat, mouse, guinea pig, hamster and rabbit) in having distinct forms of pulmonary and hepatic FMO. The immunochemical and catalytic difference between sheep liver and lung FMO is similar to that of rabbit.

Amines↗

Temporal development of bluetongue virus protein-specific antibody in sheep following natural infection.

Humoral immune responses of sheep to natural bluetongue virus (BTV) infection were studied on a temporal basis. The temporal development of viral protein-specific IgG was determined by western immunoblotting; virus neutralization and agar gel immunodiffusion (AGID) were conducted for comparative purposes. Prior to the emergence of the arthropod vector and the associated transmission of BTV, virus-neutralizing antibody was absent from all sentinel sheep; 3 sheep had pre-existing AGID antibody and all sheep had IgG, specific for 4 viral proteins, as determined by immunoblotting. Following emergence of the BTV vector, 9 of 11 sheep became infected, as determined by virus isolation, with BTV. All sheep developed virus-neutralizing and AGID antibody. However, only those sheep with a demonstrable viremia experienced an increase in viral protein-specific antibody. Development of viral protein-specific IgG varied with the individual animal and no obvious correlation between a specific response and protective immunity or viral clearance was noted. From a diagnostic viewpoint, the immunoblotting procedure was superior in identifying past exposure to BTV, as compared with neutralization and AGID. In addition, the application of immunoblotting to paired serum samples appeared to be a sensitive indicator of viremia.

Animals↗

Evaluation of enzyme-linked immunosorbent assay (ELISA) and enzyme-linked immunoelectrotransfer blot (EITB) for immunodiagnosis of hydatid diseases in sheep.

This study was undertaken to investigate antigenic characteristics of hydatid cyst fluid in sheep by SDS-PAGE method, to evaluate sensitivity and specificity of Enzyme-linked immunosorbent assay (ELISA) and Enzyme-linked immunoelectrotransfer blot (EITB) assay for diagnosis of sheep hydatidosis, and to determine seroprevalance of hydatidosis in sheep population in Elazig, Turkey. SDS-PAGE analysis of hydatid cyst fluids indicated that 6 specific-protein bands were detected at molecular weights of 29, 45, 58, 68, 98 and 116 kDa. EITB analysis showed presence of 29, 38, 42, 58, 62, 68, 98, 116, 120, 150 and 205 kDa bands in positive sheep sera, while 38, 58, 62, 68, 116 and 205 kDa bands were detected in negative sheep sera. Therefore, it was concluded that the 116 kDa band was specific for diagnosis of sheep hydatid disease by EITB assay. Sensitivity and specificity of EITB assay were determined as 88% and 84%, respectively, whereas corresponding rates for ELISA were 60% and 94%, respectively. Sensitivity of ELISA was 47.3% in hepatic cysts, 60% in pulmonary cysts, 69.2% in hepato-pulmonar cysts. Sensitivity ratios of ELISA were 67.8%, 75%, and 38.4% for fertile, sterile, and under-growth cysts, respectively. Sensitivity of EITB was found 84.2% in hepatic cysts, 80% in pulmonary cysts, and 92.3% in hepato-pulmonar cysts. Corresponding ratios for sensitivity of EITB for fertile, sterile, calcified, and under growth cysts were 92.8%, 75%, 100%, and 84.6%, respectively. In addition, seroprevalance of hydatidosis in sheep was found as 62% by ELISA and 66.4% by EITB in Elazig, Turkey and seroprevalance increased by age.

Animals↗

Trabecular structure of the condyle of the jaw joint in young and mature sheep: a comparative histomorphometric reference.

Much of the literature regarding arthrotic changes in the temporomandibular joint (TMJ) is based on the assumption, rather than the demonstration, that joint degeneration is pathologically and biochemically similar to that which has been described for other arthrodial joints. Understanding such changes is axiomatic of an understanding of the specific histomorphometric structure of the normal TMJ, in particular the condyle. Unfortunately, very little has been established about the trabecular bone patterns in the mandibular condyle as it develops. As a consequence of the obvious practical difficulties in investigations of the human TMJ, the sheep has been variously used as an animal model. In order to augment a fuller characterisation of this animal model, this study focuses on the quantitative histomorphometries of the trabeculae in the mandibular condyles of young and mature sheep. Quantitative histomorphometric analyses of condylar trabeculae were performed on histological sections prepared from mature and young sheep condyles. Lateral, central and medial sagittal sections, and anterior and posterior coronal sections of the condyle were analysed using a Quantimet 500MC image analysis system that had been programmed to provide structural index values of trabecular bone volume, surface, thickness, separation and number. Analysis of histoquantitation data revealed a significant concordance in bone structural index values between lateral, central and medial regions in young and mature sheep as well as anterior and posterior regions in young sheep. Moreover, there was little significant variation between similar regions in the respective age groups. This study provides the first comparative histomorphometric quantitative analysis of the trabeculae in the mandibular condyle of both young and mature sheep. The findings of this study reinforce the appropriateness of the sheep TMJ as a model in previous experimental studies of the bony architecture of the condyle.

Aging↗

Evaluation of an absorbed ELISA and an agar-gel immuno-diffusion test for ovine paratuberculosis in sheep in Australia.

The sensitivities and specificities of an absorbed enzyme-linked immunosorbent assay (ELISA) and an agar-gel immuno-diffusion (AGID) test for the detection of Johne's disease in sheep were estimated using data from six known infected and 12 assumed uninfected sheep flocks. Sensitivities were estimated for all histologically positive sheep, as well as by histological lesion score, lesion type (paucibacillary or multibacillary) and sheep body-condition score, with ELISA sensitivities estimated at 95 and 99% specificity. Logistic-regression analysis was used to test for significant effects of lesion score and condition score, with flock included in the model as a random effect. Estimated specificities were 95% (95% CI: 93.4, 95.6%) and 99% (98.4, 99.4%) for ELISA cut-point ratios of 2.4 and 3.6, respectively, and 100% (99.7, 100.0%) for the AGID. Estimated sensitivities for all infected sheep were 41.5% (35.0, 48.3%), 21.9% (16.6, 27.9%) and 24.6% (19.1, 30.7%) for ELISA cut-point ratios of 2.4 and 3.6 and for AGID, respectively. Sensitivities of all tests and cut-points varied significantly between flocks and between categories of lesion score and condition score. Sensitivity ranged from 25 to 73, 10 to 47 and 9.2 to 63% between flocks, for the ELISA with cut-points of 2.4 and 3.6, and for the AGID, respectively. Sensitivity was highest in thin sheep and in sheep with multibacillary lesions. The effects of lesion type and condition score on test sensitivity were significant in the logistic regressions for the AGID and ELISA at both cut-points and the flock effect was significant for the AGID but not for the ELISA at either cut-point.

Animals↗

Blockade of haem oxygenase and nitric oxide synthetase causes cortical dysfunction in sheep exposed to carbon monoxide.

Twenty adult ewes underwent common surgery and following recovery were exposed to 1% carbon monoxide (CO) for 2 h. Ten of these sheep were randomly selected for treatment with haem oxygenase (HO) and nitric oxide synthetase (NOS) blockers. All sheep were killed 5 days later. The CO exposure was narcotic and EEG frequency was suppressed. The EEG recovery was rapid in the control sheep and both slow and incomplete in the treated sheep. This difference was statistically significant (P<0.05). For the first time in our CO studies in sheep, one showed multiple cortical infarcts. This sheep was blocked for HO and NOS function. No significant differences were seen in peri-ventricular white matter infarction distribution and frequency. We have previously shown brain protection against CO- and inert diluent-hypoxemia by way of an increase in brain blood flow (BBF) that maintains adequate brain O2 uptake, and by an increase in circulating red blood cells. From this study, we propose that the induction of neuronal and glial HO and NOS in sheep exposed to CO is protective, especially for the cortex. We intend to study this further by both selective and collective enzyme blockade and by measuring regional BBF changes.

Animals↗

Lack of a critical cardiac output and critical systemic oxygen delivery during low cardiac output in the third trimester in the pregnant sheep.

OBJECTIVE: We sought to determine whether a critical cardiac output and oxygen delivery exist in normal pregnancy. We also sought to determine the role of fetoplacental oxygen demand on maternal oxygen transport variables in response to decreased maternal cardiac output. STUDY DESIGN: We studied 10 adult female sheep, 5 nonpregnant and 5 pregnant. We placed a flow-directed thermodilution catheter in the pulmonary artery and a balloon-tipped catheter in the right atrium of the sheep. We also placed a catheter for pressure monitoring and blood sampling in the descending thoracic aorta in both the mother and fetus. We decreased maternal cardiac output by incremental inflation of the right atrial balloon. We measured maternal cardiac output by intravenous bolus thermodilution technique. We also measured maternal and fetal acid-base status and serum lactate concentrations. We calculated a variety of maternal cardiorespiratory variables, including systemic oxygen delivery, systemic oxygen consumption, and fractional whole body tissue oxygen extraction. RESULT: The nonpregnant sheep displayed a critical cardiac output below which there was an abrupt decrease in oxygen consumption. In contrast, there was no critical level of cardiac output in the pregnant sheep. Maternal oxygen consumption was linearly dependent on cardiac output. Maximum fractional oxygen extraction was significantly lower in the pregnant sheep than in the nonpregnant sheep. CONCLUSION: States of low cardiac output in the pregnant sheep are associated with a lack of a critical cardiac output; the flow-dependent oxygen consumption observed is the result of either an impairment in tissue oxygen extraction or some degree of metabolic arrest or a combination of both. If this unique cardiac output-oxygen consumption relationship is seen in human pregnancy, it could have significant implications in the care of the critically ill obstetric patient.

Animals↗

Ischemic preconditioning protection against stunning in conscious diabetic sheep: role of glucose, insulin, sarcolemmal and mitochondrial KATP channels.

INTRODUCTION: Sarcolemmal and mitochondrial ATP-sensitive potassium (KATP) channels have been postulated to participate in preconditioning protection against infarction and stunning. However, these structures appear to be altered in diabetes and thus, it would be possible that preconditioning does not develop in diabetic hearts. OBJECTIVE: The purpose of this study was to know whether early (EP) and late (LP) ischemic preconditioning against stunning develop in conscious diabetic (D) sheep and whether diabetes affects KATP channel function. METHODS: Male castrated sheep received alloxan monohydrate (1 g) and were ascribed to three experimental groups: control [DC, 12 min of ischemia (i) followed by 2 h of reperfusion (r)], early preconditioning (DEP, six 5 min i-5 min r periods were performed 45 min before the 12 min i) and late preconditioning (DLP, same as DEP except that the preconditioning stimulus was performed 24 h before the 12 min i). Regional mechanics during reperfusion was evaluated by wall thickening fraction (%WTH) and expressed as percentage of basal values (100%), and KATP channel behavior was indirectly assessed by monophasic action potential duration (MAPD) in relation to its sensitivity to glibenclamide blockade (0.1 and 0.4 mg/kg). The results were compared to those obtained in normal (N) sheep. The effects of sarcolemmal and mitochondrial KATP channel blockade on recovery from stunning were assessed by administration of glibenclamide (0.1 and 0.4 mg/kg) and 5-hydroxydecanoate (5-HD, 5 mg/kg i.v.) and/or diazoxide (10 microg/kg/min over 90 min). Whether acute hyperglycemia (H) in normal animals and insulin (I) treatment in diabetic sheep affected preconditioning protection and KATP channel behavior were also evaluated. RESULTS: Results expressed as mean % recovery of %WTH showed that preconditioning protected against stunning in normal sheep (NC=65+/-3.5, NLP=82+/-6**, NEP=76+/-4*, *P<0.05 and **P<0.01 against NC) while this did not occur in diabetic ones, where DLP (58+/-7.6) afforded a similar recovery to DC (54+/-5) and DEP worsened instead of improving mechanical function (37+/-9, P<0.01 against DC). Acute hyperglycemia did not affect preconditioning development (NEPH=72+/-3 and NLPH=80+/-4) and insulin treatment reverted the lack of early and late preconditioning protection in diabetic hearts (DEPI=72+/-4* and DLPI=76+/-3*, P<0.05 against DC). Sarcolemmal KATP channel behavior appeared altered in diabetic hearts as shown by MAPD in normal sheep (276+10 ms) compared to diabetic ones (365+9 ms, P<0.05) and by the sensitivity to glibenclamide [0.1 mg/kg completely blocked KATP channels in diabetic (P<0.05) but not in normal hearts]. Insulin also restored MAPD in diabetic heart. Mitochondrial KATP channels appeared not to account for the reported results in diabetes, since glibenclamide (%WTH=40+/-4, P<0.01 vs. NC), but not 5HD nor diazoxide affected myocardial functional recovery during reperfusion. CONCLUSIONS: Sarcolemmal KATP channel dysfunction due to the lack of insulin affords a primary approach to explain the absence of preconditioning protection against stunning in diabetic sheep hearts.

Action Potentials↗

Responses of sheep and kitten isolated coronary arteries to some vasoactive agents.

In the present study, the effects of a range of vasoactive agents were assessed in sheep isolated coronary artery strips and in kitten isolated perfused coronary arteries. Both preparations contained muscarinic cholinoceptors. However, these receptors mediated contraction of sheep coronaries while dilatation was seen in the kitten. In sheep arteries, 5-hydroxytryptamine caused increases in resting tone and relaxation of acetylcholine-induced tone, while vasoconstriction was the predominant response in the kitten. Angiotensin increased tone in both preparations, although tachyphylaxis was evident. Conversely, adenosine and ATP caused reductions in tone in both preparations. Alpha-adrenoceptor agonists caused vasodilatation in kitten coronaries and decreased acetylcholine-induced tone in sheep arteries. However, weak contractions were observed in sheep coronaries in the absence of induced tone. Results also indicate that sympatomimetic amines mediate relaxation of sheep coronary arteries via beta 1-adrenoceptors. Sheep coronaries are readily and inexpensively obtained and provide a useful tool for the further study of coronary vascular smooth muscle.

Acetylcholine↗

The distribution of scrapie-associated fibrils in neural and non-neural tissues of advanced clinical cases of natural scrapie in sheep.

The distribution of scrapie-associated fibrils (SAFs) throughout four brain regions, the pituitary gland, along the whole length of the spinal cord and in the sciatic nerve was assessed in 10 sheep terminally affected by scrapie and in four control sheep. Tonsils, retropharyngeal, broncho-mediastinal and mesenteric lymph nodes, the distal ileum, proximal colon and spleen were also examined for fibrils in all 14 sheep. Fibrils were detected in all four brain regions and throughout the length of the spinal cord in nine of the scrapie affected sheep. SAFs were not detectable in any of the sciatic nerve samples tested. In one of the 10 clinically affected sheep only minimal lesions were found by histopathology and fibrils were detected only from the cerebrum and one spinal cord region (taken at the C1 C2 vertebrae). Fibrils were not detected in the tonsils or retropharyngeal lymph nodes but were detected in other non-neural tissues of some of the scrapie-affected sheep. These tissues included pituitary gland, broncho-mediastinal and mesenteric portal lymph nodes, distal ileum, proximal colon and spleen. Fibrils could not be detected in any of the tissues taken from the four control sheep.

Animals↗