Adipose tissue insulin receptor and glucose transporter 4 expression, and blood glucose and insulin responses during glucose tolerance tests in transition Holstein cows with different body condition.

Glucose uptake in tissues is mediated by insulin receptor (INSR) and glucose transporter 4 (GLUT4). The aim of this study was to examine the effect of body condition during the dry period on adipose tissue mRNA and protein expression of INSR and GLUT4, and on the dynamics of glucose and insulin following the i.v. glucose tolerance test in Holstein cows 21 d before (d -21) and after (d 21) calving. Cows were grouped as body condition score (BCS) ≤3.0 (thin, T; n = 14), BCS = 3.25 to 3.5 (optimal, O; n = 14), and BCS ≥3.75 (overconditioned, OC; n = 14). Blood was analyzed for glucose, insulin, fatty acids, and ß-hydroxybutyrate concentrations. Adipose tissue was analyzed for INSR and GLUT4 mRNA and protein concentrations. During the glucose tolerance test 0.15 g/kg of body weight glucose was infused; blood was collected at -5, 5, 10, 20, 30, 40, 50, and 60 min, and analyzed for glucose and insulin. On d -21 the area under the curve (AUC) of glucose was smallest in group T (1,512 ± 33.9 mg/dL × min) and largest in group OC (1,783 ± 33.9 mg/dL × min), and different between all groups. Basal insulin on d -21 was lowest in group T (13.9 ± 2.32 µU/mL), which was different from group OC (24.9 ± 2.32 µU/mL. On d -21 the smallest AUC 5-60 of insulin in group T (5,308 ± 1,214 µU/mL × min) differed from the largest AUC in group OC (10,867 ± 1,215 µU/mL × min). Time to reach basal concentration of insulin in group OC (113 ± 14.1 min) was longer compared with group T (45 ± 14.1). The INSR mRNA abundance on d 21 was higher compared with d -21 in groups T (d -21: 3.3 ± 0.44; d 21: 5.9 ± 0.44) and O (d -21: 3.7 ± 0.45; d 21: 4.7 ± 0.45). The extent of INSR protein expression on d -21 was highest in group T (7.3 ± 0.74 ng/mL), differing from group O (4.6 ± 0.73 ng/mL), which had the lowest expression. The amount of GLUT4 protein on d -21 was lowest in group OC (1.2 ± 0.14 ng/mL), different from group O (1.8 ± 0.14 ng/mL), which had the highest amount, and from group T (1.5 ± 0.14 ng/mL). From d -21 to 21, a decrease occurred in the GLUT4 protein levels in both groups T (d -21: 1.5 ± 0.14 ng/mL; d 21: 0.8 ± 0.14 ng/mL) and O (d -21: 1.8 ± 0.14 ng/mL; d 21: 0.8 ± 0.14 ng/mL). These results demonstrate that in obese cows adipose tissue insulin resistance develops prepartum and is related to reduced GLUT4 protein synthesis. Regarding glucose metabolism, body condition did not affect adipose tissue insulin resistance postpartum.

Field trial on progesterone cycles, metabolic profiles, body condition score and their relation to fertility in Estonian Holstein dairy cows.

Resumption of luteal activity postpartum and fertility were investigated in an Estonian Holstein high milk production and good fertility dairy herd. Body condition was scored after every 10 days in 54 multiparous dairy cows (71 lactations) calving inside from December to March during 4-year period. Blood samples were taken 1-14 days before calving and 1-14, 28-42 and 63-77 days after calving: analytes estimated were serum aspartate aminotransferase (AST), glucose, ketone bodies, total cholesterol, non-esterified fatty acids and triglycerides. The general linear mixed model was used to compare the data for cows with different characteristics in luteal activity postpartum based on their milk progesterone profiles. Forty-five per cent of cases had abnormal profiles; delayed resumption of ovarian cyclicity postpartum (DC) was the most prevalent abnormality. There was no difference in body condition scores between the groups. The DC and prolonged luteal phase groups had higher serum AST activity (p < 0.01) 1-14 days postpartum compared with normal group. The DC group also had higher cholesterol and triglyceride values (p < 0.05) 28-42 days postpartum and higher milk fat/protein ratio (p < 0.01) on the first month of lactation compared with normal profile group. Despite long post-calving anoestrous period (71 +/- 5.0 days; mean +/- SEM) DC group had 64.7% first service pregnancy rate (normal group 48.6% and PLP group 37.5%). This study did not find any detrimental effect of prolonged anovulatory period postpartum on subsequent fertility.

Field trial on the relationship of blood metabolites and body condition score with the recurrence of luteal activity in Estonian Holstein cows.

Associations of body condition scores and blood metabolites, measured before calving and at different periods during early lactation, with recurrence of luteal activity were investigated in a 250-head commercial dairy farm during a 4-year period (1999-2002). The study was conducted on 48 dairy cows (60 lactations) with average 305-day milk yield of 8149 kg per cow. Blood samples taken 1-14 days before calving and 1-14, 28-42 and 63-77 days after calving were analysed for aspartate aminotransferase, glucose, ketone bodies, triglycerides, non-esterified fatty acids and cholesterol. Milk progesterone (P(4)) profiles (samples collected twice a week, P(4) levels measured in whole milk by enzyme immunoassay) were used to evaluate the interval from calving to first luteal response, P(4) >5 ng/ml, and the interval from calving to first normal cycle. The MIXED procedure of the sas system was used to study the association of investigated parameters. A higher concentration of ketone bodies before calving was associated with shorter interval to recurrence of first normal cycle (P = 0.007) and tended to be related to shorter interval from calving to first luteal response (P = 0.071). A lower prepartum aminotransferase activity showed a tendency to be associated with shorter interval from calving to first luteal response (P = 0.084). Results suggest metabolic status up to 2 weeks prepartum to be related to the resumption of postpartum luteal activity in Estonian Holstein dairy cows.

Intrathecal and oral clonidine as prophylaxis for postoperative alcohol withdrawal syndrome: a randomized double-blinded study.

UNLABELLED: In this study, we evaluated the effect of intrathecal and oral clonidine as supplements to spinal anesthesia with lidocaine in patients at risk of postoperative alcohol withdrawal syndrome (AWS). We hypothesized that clonidine would have a prophylactic effect on postoperative AWS. Forty-five alcohol-dependent patients (daily ethanol intake >60 g) scheduled for transurethral resection of the prostate were double-blindly randomized into three groups. All patients received hyperbaric lidocaine 100 mg intrathecally. The diazepam group (DiazG) was premedicated with diazepam 10 mg orally; the intrathecal clonidine group (Clon(i/t)G) received a placebo (saline) tablet and clonidine 150 microg intrathecally; and the oral clonidine group (Clon(p/o)G) received clonidine 150 microg orally. For patients diagnosed with AWS, the Clinical Institute Withdrawal Assessment for Alcohol, revised scale, was used. Twelve patients in the DiazG had symptoms of AWS, compared with two in the Clon(i/t)G and one in the Clon(p/o)G. The median Clinical Institute Withdrawal Assessment for Alcohol, revised scale, score was 12 in the DiazG versus 1 in the clonidine-treated groups. Two patients in the DiazG had severe delirium. Patients receiving oral clonidine had a slightly decreased mean arterial blood pressure 6-12 h after spinal anesthesia (P < 0.05); patients in the DiazG had a hyperdynamic circulatory reaction 24-72 h after surgery. In conclusion, preoperative clonidine 150 microg, intrathecally or orally, prevented significant postoperative AWS in ethanol-dependent patients. IMPLICATIONS: In this randomized, double-blinded study, clonidine 150 microg both intrathecally and orally prevented postoperative alcohol-withdrawal symptoms in alcohol-dependent men. The effect was superior to that with a single dose of diazepam 10 mg orally.

Postoperative pain relief after laparoscopic cholecystectomy: a randomised prospective double-blind clinical trial.

BACKGROUND AND AIMS: The clinical value of infiltration of wounds with local anaesthetics (LA) and their intraperitoneal application for treating pain after laparoscopic cholecystectomy (LC) still remain controversial. In this study the use of intraincisional and intraperitoneal LA was evaluated. MATERIAL AND METHODS: Eighty patients were prospectively randomised into four groups. In the control group (G1) LA was not used. In G2 all wounds were infiltrated with 80 ml of 0.125 % Bupivacaine containing 5 mg of Phenylephrine. In G3 the wounds were infiltrated with 80 ml of 0.9 % NaCl. In G4, in addition to wound infiltration with Bupivacaine/Phenylephrine, 200 ml of normal saline, containing 0.15 % of Lidocaine, was left intraperitoneally under the right diaphragm. Postoperative abdominal and shoulder pain scores were recorded on a visual analogue scale (VAS) during 24 hours after LC. Narcotic analgesic consumption was also recorded. RESULTS: The mean abdominal pain scores were significantly lower in G2, compared with G3, 3 to 24 hours after operation, compared with G4, 3 to 6 hours and compared with G1, 3 to 24 hours (except at hour 12) after surgery. The incidence of shoulder pain was 30 %. There were no significant differences in the mean shoulder pain scores between the groups. The mean dosage and the total amount of Pethidine at 24 hours were significantly lower in G2 compared with G1. CONCLUSIONS: Intraincisional infiltration with a Bupivacaine/Phenylephrine mixture reduces significantly abdominal postoperative pain (for up to 24 h) and narcotic analgesic consumption after LC. An intraperitoneal subdiaphragmatic dilute solution of Lidocaine was not effective in reducing overall pain and shoulder pain after LC.

Postoperative pain relief following intrathecal bupivacaine combined with intrathecal or oral clonidine.

BACKGROUND: The purpose of the present study was to evaluate the postoperative analgesic and adverse effects of equal doses of oral or intrathecal clonidine in spinal anaesthesia with bupivacaine plain. METHODS: Forty-five ASA I-III orthopaedic patients scheduled for osteosynthesis of a traumatic femur fracture were randomised in a double-blind fashion to one of 3 groups. Patients received 15 mg of plain bupivacaine intrathecally (group B) or an intrathecal mixture of bupivacaine 15 mg and clonidine 150 mg (group CIT). In group CPO oral clonidine 150 mg was administered 60 min before intrathecal injection of bupivacaine 15 mg. RESULTS: Oral and intrathecal clonidine prolonged the time until the first request for analgesics, 313 +/- 29 and 337 +/- 29 min, respectively, vs. 236 +/- 27 min in group B (P < 0.01). The total 24- h PCA morphine dose was significantly lower in group CIT(19.3 +/- 1.3 mg) compared to groups B and CPO(33.4 +/- 2.0 and 31.2 +/- 3.1 mg). MAP was decreased significantly during the first hour after intrathecal clonidine(14%) and during the first 5 h after oral clonidine(14-19%). HR decreased in CIT during the 5th and 6th postoperative hours(7-9%) and during the first 2 h(9%) in CPO (P < 0.01). The degree of sedation was more pronounced in group CPO during the first 3 h. Four patients had pruritus in group B. CONCLUSIONS: Addition of intrathecal clonidine prolonged analgesia and decreased morphine consumption postoperatively more than oral clonidine. Hypotension was more pronounced after oral than after intrathecal clonidine. Intrathecal clonidine is therefore recommended.

Enhancement of intrathecal lidocaine by addition of local and systemic clonidine.

BACKGROUND: Enhancement of local anesthetic-produced regional blocks by clonidine seems well established. There are insufficient data about dose-effect relationship of combinations of clonidine with individual agents, efficiency of local versus systemic administration of clonidine, and comparative evaluation of clonidine with vasoconstrictors. Because of unavailability of long-acting local anaesthetics at the time of study, our aim was to evaluate augmentation of lidocaine spinal block with local or systemic clonidine and to compare the results with the efficacy of intrathecal phenylephrine. METHODS: Ninety pts of age 50-72 yrs with ASA 1-4 physical status, scheduled for open prostatectomies, hysterectomies or ostheosynthesis of fractured hip were randomized to one of 6 treatment groups, 15 pts in each. Patients received intrathecally (L3-L4) either 100 mg of plain lidocaine (group L100); or a mixture of lidocaine 40 and 80 mg with clonidine 100 micrograms (groups L40-C100 and L80-C100); or a combination of lidocaine 40 and 80 mg with clonidine 300 micrograms orally 60 min before spinal puncture (L40-C300 and L80-C300). Addition of intrathecal phenylephrine 5 mg to 80 mg of lidocaine was also investigated (L80-P5). RESULTS: There were no significant intergroup differences concerning demographic data or type of surgery. All operations (duration up to 150 min) were completed without need for analgesic supplementation. The addition of clonidine resulted in a significant reduction of the onset time of spinal block and prolongation of the duration of sensory and motor blocks compared to plain lidocaine or lidocaine with phenylephrine. In spite of the well-known hypotensive action of alpha 2-agonists, haemodynamic depression only in group L80-C300 was significantly more pronounced than in L100 and L80-P5 groups. The least decrease of BP and minimal need of rescue ephedrine among all patients studied were recorded in the group receiving low dosage of lidocaine with intrathecal clonidine (L40-C100). Sedation occurred in most patients receiving clonidine. CONCLUSION: Our results indicate that addition of clonidine to lidocaine, irrespective of the route of administration, prolongs the duration of spinal block and permits a reduction of the lidocaine dose needed for a given duration of block. Addition of phenylephrine results in a less pronounced statistically significant prolongation of anaesthesia. The regression of sensory block before restoration of motor function seems to be a specific (and unfortunate) effect of both clonidine and phenylephrine.

Distribution of local anesthetic solution in retromediastinal block. Preliminary experimental results.

BACKGROUND AND OBJECTIVES: Interpleural anesthesia blocks pain perception from the thoracoabdominal wall without impairment of leg function. Bilateral interpleural anesthesia is not recommended because of possible bilateral impairment of respiratory function. Infiltration of the retromediastinum with local anesthetic might cause bilateral thoracoabdominal somatic block and block of sympathetic afferents from the abdominal cavity without impairing respiration. METHOD: Distribution of stained fluid was studied after injection into the retromediastinum through a catheter placed about 10 cm cephalad to the diaphragm via the esophageal hiatus in three human cadavers of normal size and in six anesthetized pigs of 20-30 kg. In the pigs serum levels of bupivacaine were measured after injection of 10 mL of 0.5% bupivacaine stained with 1 mL of methylene blue. RESULTS: The injected Dye stained intercostal nerves 6-11 in cadavers and 5-12 in pigs symmetrically on both sides, along with the adjacent parts of the sympathetic chain and both vagal nerves but not the phrenic nerves. During the sampling period of 50-60 minutes, bupivacaine serum concentrations rose slowly to a maximum of 4.2 micrograms/mL. CONCLUSIONS: Block of pain perception from the abdominal wall and cavity is possible by injection of local anesthetic into the retromediastinum via a catheter introduced through the esophageal diaphragm hiatus. The block would not be expected to impair respiratory or leg function. Its efficacy and safety have yet to be established.

The evidence of oxidative stress in cardiac surgery and septic patients: a comparative study.

In present study, lipid peroxidation products (thiobarbituric acid reactive substances (TBARS) and diene conjugates (DC)) and markers of blood antioxidant status (serum antioxidative capacity (AOC) and red blood cells glutathione (RBC-GSH)) were measured to compare the extent of oxidative stress in 12 cardiac surgery and 10 septic general surgery patients. In heart surgery, arterial TBARS were significantly increased 15 min after the start and 15 min after cessation of cardiopulmonary bypass, while AOC at these times was decreased. Eighteen hours after surgery all parameters, except antioxidative capacity, had returned to preoperative levels. In septic patients, the preoperative level of lipid peroxidation was significantly higher and antioxidative capacity lower than in heart surgery patients. Surgery had no influence on oxidative stress markers in this group of patients. Increase in lipid peroxidation and reduction in blood antioxidant capacity, induced either by sepsis or cardiopulmonary bypass, were of comparable extent.

Time course of oxidative stress during open-heart surgery.

Oxidative stress and subsequent lipid peroxidation have been suggested as pathogenetically important for postischaemic reperfusion injury. We studied the time-course of oxidative stress in 14 adults undergoing cardiac surgery, evaluating serum levels of lipid peroxidation products--diene conjugates (DC) and basal and Fe-stimulated thiobarbituric acid reactive substances (TBARS, FeTEBARS)--as well as markers of blood antioxidant status--serum antioxidative capacity (AOC) and red blood cell glutathione (RBC-GSH) at 6 perioperative time-points. Arterial TBARS were significantly increased 15 minutes after start of cardiopulmonary bypass, 5 minutes after release of aortic cross-clamp and 15 minutes after cessation of bypass, compared with the preoperative levels (respective means 20.8, 38.5, 34.8 vs 7.5 nmol/g protein, p < 0.05). AOC had decreased at these times (means 21.3, 18.1, 23.2 vs 34.9%, p < 0.05). The TBARS changes correlated with AOC decrease (r = 0.30, p < 0.001). Changes in serum DC and RBC-GSH were not statistically significant. All lipid peroxidation parameters had returned to preoperative levels on the following morning, while antioxidative capacity remained suppressed (28.1%, p < 0.05). These data demonstrate a definite time-course of oxidative stress markers in arterial blood during open-heart surgery.

Atrial versus ventricular pacing in sick sinus syndrome: the role of retrograde ventriculoatrial conduction.

This study compares the acute haemodynamic effect of atrial versus ventricular pacing, and ventricular pacing in the presence versus absence of retrograde ventriculoatrial conduction. Twenty-two patients with symptomatic sick sinus syndrome were tested: 12 with ventriculoatrial conduction (Group 1) and 10 without it (Group 2). Cardiac haemodynamics were measured by thermodilution technique at spontaneous rhythm as baseline and during atrial versus ventricular pacing at two different pacing rates. Atrial pacing offered an average augmentation for 20% in cardiac index from spontaneous rhythm in both groups (P < 0.05). The presence of ventriculoatrial conduction was the crucial determinant of cardiac performance during ventricular pacing. The change in cardiac index from the baseline was towards decrease in Group 1 and towards increase in Group 2. The difference between the groups was significant (P < 0.05). We conclude that atrial pacing is superior to ventricular and should be preferred in symptomatic sick sinus syndrome patients without atrioventricular conduction defects. However, ventricular pacing can be considered for preventing clinical symptoms associated with attacks of severe bradycardia or temporary cardiac arrest in patients without ventriculoatrial conduction.