A caprine experimental model for studies on inflammation

A new experimental model forin vivo studies on local inflammation in the goat is presented. The teat and udder cisterns were separated by a surgical procedure, resulting in the teat cistern being an isolated pouch which is easily accessible through the teat canal and suitable for experimental studies. The surgery was consistently successful in closing the passage and no post-surgical complications were observed.The model was applied to a study of the inflammatory response induced by infusion ofSalmonella endotoxin. A marked response was observed as measured by the accumulation of leukocytes, serum albumin andN-acetyl--d-glucosaminidase (NAGase) in the test cistern. An initial increase in serum albumin, indicating an increase in the epithelial permeability, was observed from 1.5 h after endotoxin infusion. Approximately 0.5 h later, the cell count started to increase, reaching its peak level 3 h after infusion. The NAGase concentration was closely correlated with the cell count.The model provides new possibilities forin vivo studies on local inflammation and fulfils many of the requirements of an inflammatory model; for example, it allows non-traumatic repeated samplings from the same animal. The goat is a suitable experimental animal for many studies and, as each goat has two teats, intra-goat comparisons can be performeed.Abbreviations LPS lipopolysaccharide - FreeNAG NAGase in cell-free sample - NAGase N-acetyl--d-glucosaminidase; saline, sterile 0.9% (w/v) NaCl - TotNAG NAGase in cell-containing samples     

Characterization of the complex carbohydrates in the zona pellucida of mammalian oocytes using lectin histochemistry

The objective of this study was to characterize the glycoconjugates present in the zona pellucida of the follicular oocytes in sheep, goats and pigs. The zona pellucida was stained with periodic acid-Schiff, low iron diamine, high iron diamine, and nine different lectin horseradish conjugates: Con-A, SBA, DBA, PNA, RCA-I, GSA-II, WGA, LTA and UEA-I. Staining with DBA, PNA, SBA and RCA-I was performed with and without saponification with KOH and sialidase digestion. The results showed the presence of neutral and acidic glycoconjugates with different terminal sugars and also sialic acid radicals in the zona pellucida of all the animals studied. In particular, the positive staining with WGA, SBA, PNA and RCA-I suggests the presence of oligosaccharides with N-acetyl-d-glucosamine and sialic acid linked to the penultimate -N-acetyl-d-galactosamine and to the disaccharide galactosyl-(1-3)-N-acetyl-d-galactosamine. The terminal trisaccharide sialic acid galactosyl-(1-4)-N-acetyl-d-glucosamine was identified only in the zona pellucida of ovine and procine oocytes. Thus, the zona pellucida exhibited species-specific variations in the content and distribution of lectin-binding patterns that may reflect the species specificity of gamete interaction.Abbreviations HID high iron diamine - HRP horseradish peroxidase - LID low iron diamine - PAS periodic acid-Schiff - PBS phosphate-buffered saline; see also Tables I and II     

Nephrotoxicity in lambs apparently caused by experimental feeding withNarthecium ossifragum

Seven lambs were fed a mixture of bog plants containing 15 g (wet matter)Narthecium ossifragum per kg live weight for 10 consecutive days. Their serum creatinine concentration increased from day 1 to 4 but then fell to normal by day 6 after feeding started. In the same animals, the serum magnesium concentration was increased on days 3, 4 and 5. Histopathological examination of the kidneys of 2 lambs killed after being fed 25 gN. ossifragum per kg live weight for one day revealed tubular epithelial cell degeneration and necrosis. There were no signs of disease or distress in the experimental animals and their appetite remained normal throughout the experimental period. It was concluded thatN. ossifragum is probably nephrotoxic to lambs.Abbreviations ALP alkaline phosphatase - ASAT aspartate aminotransferase - -HBA -hydroxybutyric acid - CK creatine kinase - d.m. dry matter - -GT -glutamyltransferase - GLDH glutamate dehydrogenase - MCV mean cell volume - RBC red blood cells - SEM standard error of the mean - WBC white blood cells - w.m. wet matter     

Preliminary findings of altered follicular activity in Holstein cows with coagulation factor XI deficiency

Factor XI (F XI) deficiency is an autosomal recessive coagulopathy found in Holstein cattle. Affected animals have a 50% greater prevalence of repeat breeding. Therefore, several parameters describing ovarian function were studied. Daily blood sampling revealed that progesterone concentrations were slower to decline from a peak at day 16 (p<0.01) to values less than 3 nmol/L in F XI-deficient cows (5.14±0.69 days (mean ± SD) versus 4.05±0.63 days in control animals), resulting in an oestrous cycle length of 24.7±2.1 days compared to 22.9±3.0 days, respectively. This was not due to an alteration in the availability of prostaglandin F₂ (PGF₂) or oxytocin (OT) involved in luteolysis. No significant differences (p>0.05) were seen between normal (n=7) and F XI-deficient (n=7) cows in the peak values or the area under the curve for the pulse in 13,14-dihydro-15-keto PGF₂ in response to OT challenge or in the parameters describing the pulse of ovarian OT secretion after PGF₂ injection (n=7 for each) between days 12 and 14. Ovulatory follicular development was assessed by ultrasound monitoring and plasma 17-oestradiol values at 8-h intervals after a luteolytic injection of cloprostenol (n=6 for each). Follicular diameter was smaller (p<0.05) and accompanied by lower peak oestradiol values near the time of ovulation in F XI-deficient cows. The results suggest that the oestrous cycle in F XI-deficient cows is characterized by a slower process of luteolysis that may be associated with smaller follicular development.Abbreviations F XI factor XI - OT oxytocin - PGF₂ prostaglandin F₂ - PGFM 13,14-dihydro-15-keto-prostaglandin F₂ - i.m. intramuscularly     

The use of divalent cation chelating agents (EDTA/EGTA) to reduce non-specific serum protein interaction in enzyme immunoassay

An indirect enzyme immunoassay (ELISA) for detection of bovine antibody toBrucella abortus was modified by the addition of divalent chelating agents to the serum diluent. This addition resulted in an increase in specificity from 96.0% in the regular assay to 99.4% in the modified procedure. Of the 15 715 sera initially tested by the indirect ELISA, 691 that had given positive reactions were selected for retesting in the indirect ELISA with EDTA/EGTA added. The buffered plate antigen test (BPAT) correctly identified 98.6% of the samples as negative. The addition of chelating agents did not alter the sensitivity of the indirect ELISA, which correctly classified 609 sera from animals from whichB. abortus had been isolated as positive. The sensitivity of the BPAT was 97.8%.Abbreviations ABTS 2,2-azinobis(3-ethylbenzthiazoline sulphonic acid) - BPAT buffered plate antigen test - EDTA ethylenediaminetetraacetic acid disodium salt - EGTA ethylene glycol-bis(-aminoethyl ether)-N,N,N,N-tetraacetic acid - ELISA enzyme-linked immunosorbent assay - IgG₁ immuno- globulin G₁ - IgM immunoglobulin M - Tris tris(hydroxymethyl)aminomethane     

Effect of PCB on some MFO enzyme activities in pregnant and virgin rabbits

Fenclor 64 (a PCB's commercial mixture) was administered twice i. p. to virgin and pregnant New Zealand rabbits at the dose of 100 mg/Kg to evaluate its inducing properties on the following hepatic microsomal MFO activities: p-nitro-anisole-O-demethylase, amino-pyrine-N-demethylase, acetanilide-hydroxylase and arylhydrocarbons-hydroxylase.The results indicate that Fenclor 64 evoked a mixed type induction in virgin as well as in pregnant rabbits since it increased liver weight, cytochrome P-450 levels, as well as p-nitro-anisole-O-demethylase and acetanilide-hydroxylase. Pregnancy by itself affects only amino-pyrine-N-demethylase levels.Work supported by a grant from M. P. I. 40% (1982)     

The effect of induced fever on the biokinetics of norfloxacin and its interaction with probenecid in goats

The kinetic profiles of norfloxacin were evaluated in afebrile, febrile and probenecid pre-treated (70 mg/kg orally) febrile goats after a single intravenous (i.v) dose (5 mg/kg). Fever was induced and maintained for 12 h by injecting Escherichia coli endotoxin (0.2 g/kg, i.v.) and repeating it in half the dose (0.1 g/kg) 5 h later. The plasma pharmacokinetic values for norfloxacin were best represented using a two-compartment open model. The peak norfloxacin plasma level of 90.52±3.18 g/ml attained in the probenecid pre-treated febrile goats was higher than that in the febrile (75.46±0.72 g/ml) or afebrile goats (62.25±1.23 g/ml). Cl_B and K _el values were significantly (p<0.01) decreased in febrile compared with afebrile goats. These values were further reduced in febrile goats after probenecid pre-treatment. However, t _1/2 was not affected by the fever-probenecid interaction. Norfloxacin may be used as an infusion with probenecid in caprine diseases where very high plasma levels are required to combat resistant organisms such as Bacteroides.Abbreviations MIC minimum inhibitory concentration - LPS lipopolysaccharide - i.v. intravenous(ly)     

The disposition kinetics and residues of fenvalerate in tissues following a single dermal application to black bengal goats

The disposition kinetics of fenvalerate were studied in goats after dermal application of 100 ml of 0.25% (w/v) solution. The insecticide persisted in the blood for 72 h. The mean (±SEM) V _d(area) and apparent t _1/2 () were 9.92±1.44 L/kg and 17.51±2.65 h, while the AUC and Cl_B values were respectively 82.15±7.40 g h/ml and 0.56±0.05 L/(kg h). Four days after the dermal application, the highest concentration of fenvalerate residues was found in the adrenal gland, followed by the biceps muscle, omental fat, liver, kidney, lung and cerebrum in that order. Fenvalerate caused hyperglycaemia but had no effect on serum protein and cholesterol levels. Serum acetylcholinesterase activities were increased after 24 h but were below the initial values from 48 to 120 h.Abbreviations Ache acetylcholinestase - AUC total area under the blood insecticide concentration-versus-time curve - Cl_B total body clearance - GLC gas-liquid chromatography - t _1/2() apparent elimination half-life - V _d(area) apparent volume of insecticide distribution based on area method     

Nephrotoxicity and hepatotoxicity in calves apparently caused by experimental feeding withNarthecium ossifragum

Seven calves were fed a mixture of bog plants containing 15 g (wet matter)Narthecium ossifragum per kg live weight for two consecutive days. Their serum creatinine, urea and magnesium concentrations increased, whereas the serum calcium concentration decreased. Histopathological examination of the kidneys of the 5 calves that were killed revealed tubular epithelial cell degeneration and necrosis. There were signs of liver dysfunction in all the calves including increased aspartate aminotransferase (ASAT), glutamate dehydrogenase (GLDH) and -glutamyltransferase activities.All the calves refused to ingestN. ossifragum after 2 days feeding, and their appetite for hay and concentrate was also reduced. It can be concluded thatN. ossifragum is nephrotoxic and hepatotoxic to calves.Abbreviations ALP alkaline phosphatase - ASAT aspartate aminotransferase - -HBA -hydroxybutyric acid - CK creatine kinase - d.m. dry matter - -GT -glutamyltransferase - GFR glomerular filtration rate - GLDH glutamate dehydrogenase - MCV mean cell volume - RBC red blood cells - SEM standard error of the mean - WBC white blood cells - w.m. wet matter     

Preliminary Studies on Hepatic Carnitine Palmitoyltransferase in Dairy Cattle with or without Fatty Liver

The hepatic mitochondrial carnitine palmitoyltransferase (CPT) activity was measured by fluorimetric assay in dairy cows with or without fatty liver. CPT activities in 13 lactating cattle and in 6 non-lactating cows were 304.4±86.6 mol CoA/min per g protein and 169.3±84.8 mol CoA/min per g protein, respectively. This difference was significant (<0.05). CPT activities in early lactation (0–110 days after calving), mid-lactation (111–220 days after calving) and late lactation (over 220 days after calving) were 278.9±68.0, 312.4±124.1 and 320±59.3 mol CoA/min per g protein, respectively. There was no significant difference between the values at different stages of lactation. The CPT activity in 10 lactating cows with fatty liver unrelated to calving was 201.3±80.0 mol CoA/min per g protein. CPT activity in 10 cattle with fatty liver was significantly lower than that in normal lactating cattle. Based on these findings, clinical fatty liver unrelated to calving appears to be associated with a decrease in hepatic CPT activity.     

Pharmacokinetic data on doxycycline and its distribution in different biological fluids in female goats

A pharmacokinetic study of doxycycline after intravenous administration at 5 mg/kg body weight in goats revealed that a concentration of 0.5 g/ml was maintained for 5 min-2 h, 4–12 h, 2–12 h and 5 min- >48 h in plasma, interstitial fluid, milk and urine respectively. The low t1/2 of 0.73±0.11 h and high t1/2 of 16.63±1.58 h show that the drug is rapidly distributed but slowly eliminated from the body. The tissue:plasma concentration of 4.86±1.06 during the elimination phase [K12/(K21-)] indicates a high expected tissue concentration, which is supported by similarly increased drug concentration in interstitial fluid and milk. The high Vd_area of 9.78±0.86 L/kg observed denotes that, apart from its wide distribution, the drug may be stored in fat depots as it is known to be highly lipophilic. As the drug maintained a therapeutic concentration for a shorter time in plasma, and the calculated dose rate for maintaining a minimal plasma concentration of 0.5–1.5 g/ml is relatively high, it may not be of much use in treating septicaemia in this species. Since the observed tissue:plasma concentration was higher and a therapeutic concentration was maintained in interstitial fluid and milk for longer, the drug can be used for other systemic infections at a lower dose rate than that required for treating septicaemia. As the drug maintained a very high concentration in urine, it may be of particular value in treating urinary tract infections caused by sensitive micro-organisms.     

Feed intake and rumen motility in dwarf goats. Effects of some α-adrenergic agonists,prostaglandins and posterior pituitary hormones

The purpose of the present investigation was to test the hypothesis that drug-induced changes in rumen contractions influence feed intake in dwarf goats. Intravenous (i.v.) administration of clonidine (1 g kg^-1 min^-1 for 10 min), xylazine (1 g kg^-1 min^-1 for 10 min), and PGF-2 (10 g kg^-1 min^-1 for 15 min) caused bradycardia and inhibition of rumen contractions. However, no appetite-stimulating effect of these drugs was observed. Other clinical changes induced by the ₂-adrenergic agonists included slight sedation and a decrease in body temperature; all clinical effects of clonidine and xylazine were partly antagonized by tolazoline pretreatment (10 g kg^-1 min^-1 for 30 min). These results suggest that the CNS control of feeding differs in ruminants and monogastric species.In dwarf goats fasted for 2 h, i.v. administration of oxytocin (0.01 IU kg^-1 min^-1 for 15 min), vasopressin (0.01 IU kg^-1 min^-1 for 15 min), octapressin (0.003 IU kg^-1 min^-1 for 15 min) or PGE (0.8 g kg^-1 min^-1 for 15 min) did not change feeding behaviour during the two observation periods (0–30 min and 180–210 min after drug infusion, respectively). In previous studies, similar doses of these drugs induced changes in heart rate and inhibition of rumen contraction in goats. These findings demonstrate that drug-induced changes in forestomach contractions do not simply cause changes in feeding behaviour. The i.v. infusion of the PGF₂ analogues etiproston (10 g kg^-1 min^-1 for 15 min), luprostiol (30 g kg^-1 min^-1 for 15 min), cloprostenol (1 g kg^-1 min^-1 for 15 min) and tiaprost (1 g kg^-1 min^-1 for 15 min) induced hypophagic effects and stimulated intestinal propulsion.     

Pharmacokinetics and dosing regimen of aminosidine in the dog

The kinetic behaviour of the aminoglycoside aminosidine, given at 15 mg/kg intravenously, intramuscularly and subcutaneously, was studied in 5 dogs to determine the appropriate dosage schedule. The pharmacokinetic behaviour of aminosidine in dogs was similar to that in other species, except that it was eliminated more slowly (=0.007±0.0003 min^-1). Intramuscular and subcutaneous administration produced peak serum concentrations (C _max[im]=32±6.4 g/ml; C _max[sc]=36±3.4 /ml) and times to peak concentration (T _max=60 min for both) that did not differ significantly; and neither compartmental nor non-compartmental analysis revealed any significant differences between any of the kinetic parameters obtained for these two extravenous routes of administration. Comparison of these results with previously published data suggests that aminosidine given once daily at 15 mg/kg would be as effective as, and safer than, the two or three daily administrations commonly employed in dogs.Abbreviations ALAT alanine aminotransferase - ASAT aspartate aminotransferase - AUC area under the curve - BUN blood urea nitrogen - Cl_b body clearance - C _max peak plasma concentration - CV coefficient of variation - i.m. intramuscular(ly) - i.v. intravenous(ly) - LDH lactate dehydrogenase - MIC minimal inhibitory concentration - MRT mean residence time - PAE post-antibiotic effect - PCV packed cell volume - RBC red blood cell count - s.c. subcutaneous(ly) - SD standard deviation - WBC white blood cell count - V _d(ss) distribution volume at steady state     

The use of cultured hepatocytes from goats and cattle to investigate xenobiotic oxidative metabolism

The oxidative metabolism of aldicarb (ALD), a carbamate pesticide, and fenbendazole (FBZ), an anthelmintic, was studied using cultured hepatocytes obtained from 4 goats and a bullock and incubated with ALD (50 mol/L) and FBZ (10 mol/L). The parent compounds and the metabolites were measured by HPLC. Both compounds are metabolized at the sulphur atom via two sequential oxidations, first to the sulphoxide (aldicarb sulphoxide and oxfendazole, respectively) and then to the sulphone. Oxfendazole and fenbendazole sulphone from FBZ, and aldicarb sulphoxide from ALD were found in both species. Aldicarb sulphone was not produced by the hepatocyte preparations from the bullock. The good correlation obtained comparing the in vitro results of FBZ metabolism with published in vivo dat obtained on FBZ kinetics in the same species confirmed the usefulness of in vitro models for predictive analysis of in vivo xenobiotic biotransformations.Abbreviations ALD aldicarb - ALDSON aldicarb sulphone - ALDSOX aldicarb sulphoxide - BSA bovine serum albumin - ID internal diameter - EGTA ethylene glycol bis(-aminoethyl ether) N,N,N,N-tetraacetic acid - FBZ fenbendazole - FBZSON fenbendazole sulphone - HBSS Hanks' balanced saline solution - HPLC high-pressure liquid chromatography - LDH lactate dehydrogenase - MFO mixed function oxidase - NCS newborn calf serum - OXF oxfendazole - WME Williams' Medium E     

Dose-dependent Response to an Intratesticular Injection of Calcium Chloride for Induction of Chemosterilization in Adult Albino Rats

This study concerned the minimum and optimum effective doses of calcium chloride needed for induction of chemosterilization in male albino rats, 30 days after a single intratesticular injection of calcium chloride (CaCl₂.2H₂O) solution at 2.5, 5, 10 or 20 mg per 100 g body weight per testis. There was a significant diminution in the relative wet weight of the sex organs (p<0.01), epididymal sperm count (p<0.001), plasma concentration of testosterone (p<0.01), testicular activities of D,3-hydroxy steroid dehydrogenase (D,3-HSD), 17-hydroxy steroid dehydrogenase (17-HSD) (p<0.01), glutathione S-transferase (GST) (p<0.01), superoxide dismutase (SOD) (p<0.01), and peroxidase (p<0.01), significant elevations in testicular content of malondialdehyde (MDA) and conjugated dienes (p<0.01), along with derangement of seminiferous tubular architecture and degeneration of the Leydig cells in the testis and elevations in the concentrations in the plasma of LH and FSH (p<0.01), commencing at a dose of 5 mg, with the greatest effects at a dose of 20 mg. No significant alterations in these factors occurred at the dose of 2.5 mg in comparison to the control that received only the vehicle. There was no significant alteration in the plasma concentrations of prolactin (p>0.05), corticosterone (p>0.05) or fasting blood glucose or in the rectal temperature (p>0.05) at any of the doses relative to the control group, suggesting that this chemosterilizing procedure did not exert any chronic stress on the experimental animals. From these observations, it may be suggested that 5 mg should be considered as the minimum dose, and 10 mg or 20 mg as the optimum dose, whereas 2.5 mg was ineffective for induction of chemosterilization. There would seem to be little point in using more than 20 mg of calcium chloride for this purpose. Intratesticular injection of calcium chloride at an effective dose may be considered as an alternative to surgical castration.     

Milk whey as a serum supplement for the growth of FMD virus in BHK-21 cells

Two batches of experimental media containing 10% milk whey (sweet or sour) and 1% bovine serum supported the growth of BHK-21 cells in serial passages. The cell yield was greater than 2×10⁶/ml at 48 h. The cells were susceptible to inoculation with foot and mouth disease (FMD) virus type A and the virus titres obtained were comparable with those in Eagle's MEM-G with 10% serum. The use of milk whey could reduce the consumption of serum required for the growth of FMD virus in BHK-21 cells by up to 90%.     

Pharmacokinetics of benzydamine in dairy cows following intravenous or intramuscular administration

Five lactating cows were given benzydamine hydrochloride by rapid intravenous (0.45 mg/kg) and by intramuscular (0.45 and 1.2 mg/kg) injection in a crossover design. The bioavailability, pharmacokinetic parameters and excretion in milk of benzydamine were evaluated. After intravenous administration, the disposition kinetics of benzydamine was best described using a two-compartment open model. Drug disposition and elimination were fast (t _1/2: 11.13±3.76 min;t _1/2: 71.98±24.75 min; MRT 70.69±11.97 min). Benzydamine was widely distributed in the body fluids and tissues (V _d(area): 3.549±1.301 L/kg) and characterized by a high value for body clearance (33.00±5.54 ml/kg per min). After intramuscular administration the serum concentration-time curves fitted a one-compartment open model. Following a dose of 0.45 mg/kg, theC _max value was 38.13±4.2 ng/ml at at _max of 67.13±4.00 min; MAT and MRT were 207.33±22.64 min and 278.01±12.22 min, respectively. Benzydamine bioavailability was very high (92.07%±7.08%). An increased intramuscular dose (1.2 mg/kg) resulted in longer serum persistence (MRT 420.34±86.39 min) of the drug, which was also detectable in milk samples collected from both the first and second milking after treatment.Abbreviations HPLC high-pressure liquid chromatography - IC50 concentration to inhibit the activity of an organism by 50% - IM intramuscular(ly) - IV intravenous(ly) - NSAID non-steroidal antiinflammatory drugs - pK _a negative logarithm of the ionization constant (K _a) of a drug; other abbreviations are listed in footnotes to tables     

Identification of Functional α-Adrenoceptor Subtypes in the Bovine Female Genital Tract During Different Phases of the Oestrous Cycle

The concentration and functionality of the -adrenoceptor (-AR) subtypes in the genital tract of cyclic heifers were investigated. In each tissue sample, a single class of ₁-ARs was observed, whereas two distinct classes of ₂-ARs were discriminated: low-affinity (LA) and high-affinity (HA) ₂-ARs. Statistical analysis showed the presence of significantly (p < 0.05) higher concentrations of all -AR subtypes in the follicle than in the corpus luteum. No significant differences were found in the ovary or myometrium between the luteal and follicular phases. In the ovary, the density of ₁-ARs was significantly (p < 0.05) higher than that of ₂-ARs. By contrast, there were significantly (p < 0.05) more ₂-ARs than ₁-ARs in the myometrium. As far as ₂-ARs are concerned, LA ₂-ARs were significantly (p < 0.05) higher than HA ₂-ARs in all tested tissues. Competition studies suggested that the rank order of potency of antagonists for ₁-ARs was prazosin > phentolamine > yohimbine, whereas for ₂-ARs the order of potency was yohimbine phentolamine > prazosin. Functional assays performed on myometrium showed that noradrenaline, phenylephrine and clonidine elicited concentration-dependent contractions only in dioestrus and pro-oestrus preparations and that clonidine was more effective than phenylephrine as a contractile agent. It appeared that there were no significant modifications in -AR affinity or concentration during the different stages of bovine oestrous cycle, whereas the uterine spontaneous activity and the responsiveness to -adrenergic stimulation was strongly influenced by hormonal levels. The modifications of uterine contractility observed during the oestrous cycle may be related to modifications induced in the transductional mechanisms of -ARs.     

The influence of ageing on muscarinic receptors, β-adrenoceptors and adenylate cyclase activity in the bovine lung

Muscarinic and -adrenoceptors were identified in airway epithelium, smooth muscle and lung parenchyma from Holstein-Friesian calves and cows and were characterized with [³H]quinuclidinyl benzilate and [³H]dihydroalprenolol, respectively. The muscarinic receptor density in the smooth muscle of cows (B _max=4803±245 fmol/mg protein) was 33% greater (p<0.01) than in calves. Low receptor numbers were detected in the epithelium and parenchyma. In both calves and cows, the density of epithelial -adrenoceptors was twice as high as in smooth muscle and parenchyma. The quantity of -adrenoceptors in the tracheal epithelium (B _max=994±83 fmol/mg protein) and smooth muscle (B _max=492±41 fmol/mg protein) in cows was respectively 37% (p<0.001) and 35% (p<0.01) lower than in calves. Adenylate cyclase (AC) assays indicated that the basal and the (–)-isopropylnoradrenaline- (ISO-) stimulated cAMP production were not significantly different between the calves and cows. After stimulation with NaF, significantly higher cAMP production was found in all tissues from cows. Significant correlations were found between absolute AC responses to NaF and -adrenoceptor density in epithelium (r=–0.75,p<0.001) and smooth muscle (r=–0.63,p<0.01). It seems that, in older animals, the production of cAMP is independent of the number of receptors, indicating the presence of fully active compensatory mechanisms.Abbreviations AC adenylate cyclase - cAMP intra-cellular adenosine monophosphate - G-protein GTP binding protein - GTP guanosine 5-triphosphate - ISO isopropylnoradrenaline - M muscarine - QNB quinuclidinyl benzilate     

The effects of glibenclamide,a blocker of K+ ATP-sensitive potassium channels,on diaphragmatic fatigue during endotoxaemia in pigs

An in vivo porcine model of endotoxaemia was used to study the effects of glibenclamide, a K⁺ ATP-sensitive potassium channel blocker. Escherichia coli lipopolysaccharides (LPS, 70 g/kg, i.v., as a bolus) were infused into anaesthetized, mechanically ventilated, indomethacin-treated pigs. After 120 min of endotoxaemia, glibenclamide was administered (10 mg/kg, i.v., over 5 min) to half the pigs. The steength at different frequencies of stimulation (10, 20, 30, 50 Hz, 20 V, 1 s) and the endurance capacity (10 Hz, 20 V, 30 s) of the diaphragm were evaluated after 120 min of endotoxaemia and 5, 10, 20 and 30 min after drug infusion. Glibenclamide transiently increased the blood pressure without changing the decreased cardiac output and at the same time further impaired the diaphragmatic activity. The reduced ability of the diaphragm to generate force in response to different electrical stimulations was shown by a significant reduction in strength. The endurance index decreased 5 min after glibenclamide infusion, returning to the pre-glibenclamide values by 150 min. These results indicate that glibenclamide modifies the activity of vascular smooth muscle and of the diaphragm.Abbreviations BP blood pressure - CO cardiac output - LPS lipopolysaccharides - Glib glibenclamide - i.m. intramuscular - i.v. intravenous - SEM standard error of the mean - v/v volume/volume P _ab, abdominal pressure - P _di transdiaphragmatic pressure - P _oes oesophageal pressure