Fentanyl and methadone used as adjuncts to bupivacaine for lumbosacral epidural analgesia in sheep.

Six healthy, female, mixed-breed 18-24-month-old sheep weighing 30-48 kg were submitted to lumbosacral epidural bupivacaine in combination with either methadone or fentanyl. Epidural catheters were placed in six sheep that were given three treatments: (Bup) bupivacaine (0.5 mg/kg) alone; (BupMet) bupivacaine (0.25 mg/kg) plus methadone (0.3 mg/kg); and (BupFent) bupivacaine (0.25 mg/kg) plus fentanyl (0.002 mg/kg). Haemodynamic variables, respiratory rate, rectal temperature, analgesia by applying a standard painful stimulus, motor block and sedative scores were compared among the three treatments. These parameters were determined before epidural administration and at 5, 10, 20, 30, 60, 90, 120 minutes after treatment administration, and then every 60 minutes thereafter until the end of analgesic effect. Parametrical data were analysed by proc glimmix (SAS) for repeated measures on time and means tested by ls-means. Non-parametrical data were analysed by Fisher's exact test. Duration of analgesia was longer with BupMet (240 minutes) compared with BupFent (180 minutes; P=0.0127), but BupMet was similar to Bup (240 minutes). Both treatments with opioids produced moderate motor blockade. BupMet and BupFent produced mild sedation. Only treatment with bupivacaine alone induced cardiovascular and respiratory rate changes that stayed within acceptable limits.

Effects of intra-articular injection of mesenchymal stem cells associated with platelet-rich plasma in a rabbit model of osteoarthritis.

The current study aims to evaluate the macroscopic and histological effects of autologous mesenchymal stem cells (MSC) and platelet-rich plasma on knee articular cartilage regeneration in an experimental model of osteoarthritis. Twenty-four rabbits were randomly divided into four groups: control group, platelet-rich plasma group, autologous MSC undifferentiated group, and autologous MSC differentiated into chondrocyte group. Collagenase solution was used to induce osteoarthritis, and treatments were applied to each group at 6 weeks following osteoarthritis induction. After 60 days of therapy, the animals were euthanized and the articular surfaces were subjected to macroscopic and histological evaluations. The adipogenic, chondrogenic, and osteogenic differentiation potentials of MSCs were evaluated. Macroscopic and histological examinations revealed improved tissue repair in the MSC-treated groups. However, no difference was found between MSC-differentiated and undifferentiated chondrocytes. We found that MSCs derived from adipose tissue and platelet-rich plasma were associated with beneficial effects in articular cartilage regeneration during experimental osteoarthritis.

Comparison of analgesic and systemic effects of bupivacaine, methadone, or bupivacaine/methadone administered epidurally in conscious sheep.

OBJECTIVE: The aim of this study was to evaluate the combination of bupivacaine and methadone administered epidurally in sheep. METHODS: Six healthy female mixed-breed sheep weighing 35-46 kg and aged 12-18 months were included. Each sheep was assigned to receive three treatments: 0.5 mg/kg 0.25% bupivacaine (BP), 0.3 mg/kg 1% methadone (MT) or 0.25 mg/kg bupivacaine and 0.15 mg/kg methadone (BPMT). All drugs were injected into the lumbosacral space through an epidural catheter. Each animal received each treatment at random. Heart rate, arterial blood pressure (systolic, diastolic and mean), respiratory rate, rectal temperature, analgesia, sedation and motor block were determined before treatment and at predetermined intervals. RESULTS: The duration of analgesia was 240, 220, and 180 min for BP, MT and BPMT, respectively (P < 0.05). Motor block for all agents was mild to moderate. None or the treatments significantly altered the heart rate, blood pressure or respiratory rate. CONCLUSION: Our findings suggest that lumbosacral epidural administration of bupivacaine, methadone or a combination of the two drugs can provide perioperative analgesia in sheep as part of their management for surgical procedures in the flank and hindlimbs.

Evaluation of pH effects on genomic integrity in adipose-derived mesenchymal stem cells using the comet assay.

The use of mesenchymal stem cells (MSCs) in experimental, clinical, and therapeutic trials has grown in recent years. However, the issue remains of whether these procedures are completely safe for transplant patients. Therefore, this study was designed and carried out with the aim of evaluating two different comet assay protocols for genomic damage pattern analysis in MSCs derived from adipose tissue. The analyzed and interpreted results suggest that genetic testing is needed to support clonal expansion safety in cell therapy procedures with MSCs. Furthermore, they also suggest that if the comet assay technique would be used as a genomic integrity screening assay, the protocol performed at pH = 12 (that yielded a frequency of damaged cells: tail intensity = 9.50 ± 0.60, tail moment = 0.0122 ± 0.0007; results are reported as means ± standard deviation) would be indicated as genomic damage, and that subsequent single-strand breaks occur at pH > 13 (frequency of damaged cells: tail intensity = 30.71 ± 4.23, tail moment = 0.0447 ± 0.0073). Our study demonstrates that, in the era of regenerative medicine, it is necessary to standardize and establish a battery of tests in order to identify genomic damage prior to MSC transplantation.

Efficacy of epidural lidocaine combined with tramadol or neostigmine on perineal analgesia in the horse.

REASONS FOR PERFORMING STUDY: Short duration of analgesia is among the limitations of a single epidural injection with lidocaine in horses. OBJECTIVES: To evaluate the effectiveness and safety of epidural lidocaine in combination with either tramadol or neostigmine for perineal analgesia in horses. METHODS: Epidural catheters were placed in 6 saddle horses that then were given 3 treatments: 2% lidocaine (0.2 mg/kg bwt) alone, 2% lidocaine (0.2 mg/kg bwt) plus tramadol (0.5 mg/kg bwt), and 2% lidocaine (0.2 mg/kg bwt) plus neostigmine (1.0 µg/kg bwt). The order of treatments was randomised. Haemodynamic variables, respiratory rate, rectal temperature, analgesia, motor block and behaviour scores were compared among the 3 treatments. These parameters were determined before drug administration (baseline), at 5, 10, 15, 30, 45, 60, 75 and 90 min, and every 30 min thereafter until loss of analgesia. RESULTS: Duration of analgesia was longer with lidocaine plus tramadol (210 ± 12 min) compared with lidocaine plus neostigmine (150 ± 35 min) or lidocaine alone (70 ± 12 min; P<0.05). All treatments produced mild or moderate motor block without behavioural changes. Other adverse effects were not observed in any of the horses. CONCLUSION AND POTENTIAL RELEVANCE: Further studies are required to demonstrate whether tramadol or neostigmine have a role in the management of post operative pain when coadministered with lidocaine.

Hyaluronidase shortens levobupivacaine lumbosacral epidural anaesthesia in dogs.

OBJECTIVES: The aim of this study was to evaluate the effects of hyaluronidase added to levobupivacaine in lumbosacral epidural blockade in dogs. METHODS: Six adult mixed breed dogs (two males and four females) weighing 7 to 14 kg (10.5 ±1.5 kg) and aged two to five years were used. Each dog received both treatments in random order: levobupivacaine alone (LBA; n=6) or levobupivacaine plus hyaluronidase (LBH; n=6) administered in the lumbosacral epidural space. Systemic effects, spread and duration of anaesthesia and motor block were determined before treatment and at predetermined intervals. RESULTS: The duration of local anaesthesia was 90 ±10 minutes (P=0.001) for LBH treatment and 150 ±15 minutes for LBA treatment. In the LBH treatment, anaesthesia reached the T12 to T13 dermatome and in the LBA treatment it reached the T11 to T12 dermatome in all animals in 5 and 15 minutes, respectively. Complete motor blockade was 75 ±12 minutes (P=0.01) and 120 ±15 minutes for LBH and LBA treatments, respectively. CLINICAL SIGNIFICANCE: Hyaluronidase added to levobupivacaine significantly shortens the duration of epidural anaesthesia with the same dermatome spread into the epidural space in dogs.

Lignocaine versus bupivacaine for flank anaesthesia using multiport catheters via a caudal epidural approach in cattle.

OBJECTIVE: To determine the anaesthetic and systemic effects of dorsolumbar epidural anaesthesia using non-stylet multiport catheters via the caudal approach to administer hypertonic 5% lignocaine (HL) or hypertonic 0.5% bupivacaine (HB) to the flank in standing cattle. MATERIALS AND METHODS: Six healthy adult cattle weighing 310-455 kg received 0.2 mg/kg HL or 0.025 mg/kg of HB; control animals received 0.9% saline solution. All drugs were injected into the dorsolumbar epidural space via a caudal approach through a non-stylet multiport catheter. Each animal received each treatment at random. Evaluations of anaesthesia, ataxia, heart rate, arterial pressures, respiratory rate and rectal temperature were obtained at 0 (basal), 5, 10, 15, 30, 45, 60, 75, and 90 min after epidural injection and then at 30-min intervals until loss of anaesthesia. All animals received a standard noxious stimulus and a 4-point scale was used to score the response. A second scale was used to score ataxia. RESULTS: The duration of anaesthesia in the upper and lower flanks in cattle was 68 +/- 12 and 110 +/- 15 min (mean +/- SD) after dorsolumbar epidural HL or HB, respectively. Both hypertonic local anaesthetics produced a mild ataxia. The systemic changes were within acceptable limits in these clinically healthy cattle. CONCLUSION: In standing cattle the dorsolumbar epidural injection of hypertonic lignocaine provided faster onset of anaesthesia and fewer cardiovascular effects, but had a shorter duration of anaesthesia than hypertonic bupivacaine.

Effects of lumbosacral epidural ketamine and lidocaine in xylazine-sedated cats.

In order to determine the analgesic and cardiovascular effects of the combination of epidural ketamine and lidocaine, 6 sedated cats were studied. Six healthy, young cats were used in a prospective randomised study. Each cat underwent 3 treatments, at least 1 week apart, via epidural injection: (1) ketamine (2.5 mg/kg), (2) lidocaine (4.0 mg/kg), and (3) ketamine (2.5 mg/kg) plus lidocaine (4.0 mg/kg). Epidural injections were administered through the lumbosacral space. Analgesia, motor block, sedation, heart rate, arterial blood pressure, respiratory rate and arterial oxygen saturation were measured. Rectal temperature was compared before and after sedation as well as after epidural administration of the drugs. Epidural administration of the ketamine/lidocaine combination induced prolonged analgesia extending from the coccygeal to the T13-L1 dermatomes, leading to severe ataxia. Cardiovascular effects were significant in all treatments: heart rate decreased, but there was a minimal reduction in arterial pressure. It was concluded that adding a dose of ketamine to epidural lidocaine in cats is feasible and effective.

Clinical evaluation of clonidine added to lidocaine solution for subarachnoid analgesia in sheep.

Clonidine (CL) is a alpha2-adrenergic agonist that produces analgesia in animals and humans by a non-opiate alpha2-adrenergic action in the spinal cord dorsal horn. The objective of this prospective randomized study was to investigate the clinical effects of CL/lidocaine (LD) combination administered by the subarachnoid route in sheep. Each sheep received each of three treatments, at no shorter than weekly intervals. Treatments consisted of 0.003 mg/kg CL, 1.2 mg/kg LD and a combination of CL (0.003 mg/kg) and LD (1.2 mg/kg) (CLLD). Subarachnoid injections were given in all animals between the last lumbar and first sacral vertebra. Heart rate (HR), arterial pressures, respiratory rate, rectal temperature, analgesia, sedation, and motor blockade were determined before drug administration (basal) and 5, 10, 15 and 30 min after drug administration, and at 30-min intervals until loss of analgesia occurred. The duration of analgesia after subarachnoid CLLD administration was 187 +/- 24 min (mean +/- SD), i.e. more than twice of that obtained with CL (99 +/- 19 min) or LD (55 +/- 4.4 min) alone. In all sheep, CL, administered either alone or with LD, induced moderate sedation. After subarachnoid administration of three treatments, all sheep had ataxia and subsequent sternal recumbency. The CL treatment causes decreases in blood pressure (diastolic arterial pressure and mean arterial pressure) and HR. Data suggest that the CLLD combination could be used subarachnoidally in sheep requiring prolonged surgery.

Analgesic and systemic effects of xylazine, lidocaine and their combination after subarachnoid administration in goats.

The objective of the study was to determine the analgesic and systemic effects of subarachnoid administration of xylazine hydrochloride (XY), lidocaine hydrochloride (LI) and their combination (XYLI) in goats. Six healthy goats were used in a prospective randomised study. Three treatments were administered to each goat, with 1-week intervals between each treatment. Treatments consisted of 0.1 mg/kg xylazine, 2.5 mg/kg lidocaine and a combination of xylazine 0.05 (mg/kg) and lidocaine (1.25 mg/kg). Analgesia, ataxic, sedative, cardiovascular and respiratory effects, and rectal temperature were evaluated before (baseline) and at 5, 10, 15, and 30 min after subarachnoid injection, and then at 30-min intervals until loss of analgesia occurred. Lidocaine induced analgesia in 3.1 +/- 1 min (mean +/- SD), which lasted for 66 +/- 31 min. Heart and respiratory rates and blood pressure remained unchanged after lidocaine-induced analgesia. Xylazine induced analgesia in 9.5 +/- 2.6 min and xylazine-lidocaine in 3.2 +/- 1.2 min. Xylazine-lidocaine-induced analgesia lasted longer (178.3 +/- 37 min) than that induced by xylazine (88.3 +/- 15 min). The XYLI treatment induced prolonged motor blocking (115 min), more than the XY (80 min) and LI (90 min) treatments. Both xylazine and xylazine-lidocaine caused significant decreases in the heart and respiratory rates, but not in blood pressure. The combination of xylazine (0.05 mg/kg) and lidocaine (1.25 mg/kg) can be administered subarachnoidally (between last lumbar vertebra and 1st sacral vertebra) to produce prolonged (> 2.5 h) analgesia of the tail, perineum, hind limbs, flanks and caudodorsal rib areas in goats. Despite the prolonged analgesia, using this combination is desirable for relieving postoperative pain, but it may be a disadvantage due to a motor block when dealing with goats.

L-Bupivacaine 0.5% vs. racemic 0.5% bupivacaine for caudal epidural analgesia in horses.

Bupivacaine is available as a racemic mixture of its enantiomers, d-bupivacaine and l-bupivacaine (LB). The aim of this randomized, double-blind study was to investigate the clinical efficacy and safety of S(-)-bupivacaine compared with standard racemic bupivacaine (RB) in horses under caudal epidural analgesia. Two treatments were administered to each horse, with a 2-week interval between subsequent treatments. Treatment 1 consisted of 0.5% LB at a dose of 0.06 mg/kg of body weight, and treatment 2 consisted of 0.5% RB at a dose of 0.06 mg/kg of body weight. Epidural injections were given in all animals between the first and second coccygeal vertebra. Heart rate (HR), arterial pressures, respiratory rate (RR), rectal temperature (RT), analgesia, and motor blocking were determined before drug administration (basal) and 5, 10, 15 and 30 min after drug administration, and at 30 min intervals thereafter. There were no significant differences between the two treatments in the quality of sensory and motor block. The duration of analgesia was 320 +/- 30 min (mean +/- SD) for RB and 360 +/- 42 min for LB. HRs and RRs, arterial pressures and RT did not change (P < 0.05) significantly from basal values after epidural administration of LB or RB. This study supports that 0.5% LB is an effective alternative to RB in caudal epidural analgesia in conscious, standing horses. The use of LB vs. RB warrants further investigation, particularly for long-lasting surgery in the perineal region.

Sperm capacitation and the acrosome reaction are compromised in teratospermic domestic cats.

The efficiency of sperm capacitation and of the acrosome reaction was studied in the teratospermic domestic cat to evaluate further the etiology of compromised zona pellucida penetration and oocyte fertilization. Specific objectives were to compare normospermic and teratospermic cat ejaculates for 1) the kinetics and timing of sperm capacitation in vitro as determined by an ionophore-induced acrosome reaction; 2) the incidence of spontaneous acrosomal loss; 3) the ability of capacitated, swim-up processed sperm to acrosome-react in response to chemical (calcium ionophore) or physiological (solubilized zonae pellucidae) inducers; and 4) differences in acrosomal ultrastructure by use of transmission electron microscopy (TEM). Acrosomal status was determined with the fluorescent probe Arachis hypogaea (peanut) agglutinin. The timing of in vitro capacitation differed (p < 0.05) between cat populations. Normospermic samples were capacitated at 2.0 h postcentrifugation, whereas teratospermic samples required 2.5 h to become capacitated. At 2.5 h, sperm from teratospermic males were less capable (p < 0.05) of completing the acrosome reaction after ionophore exposure (49.3 +/- 8.0%) than sperm from normospermic males (73.3 +/- 3.8%). Levels of spontaneous acrosomal loss/reaction over time were similar (p > 0.05) between cat groups (range, 7.6-17.8%). In swim-up separated sperm from normospermic cats, ionophore A23187 was a more potent inducer (p < 0.05) of the acrosome reaction (70.1 +/- 6.5%) than solubilized zonae pellucidae (31.1 +/- 1.2%). Swim-up separated sperm from teratospermic cats, however, were compromised in the ability to acrosome react, regardless of inducer (ionophore, 23.9 +/- 3.3%; solubilized zonae pellucidae, 23.9 +/- 4.7%; p > 0.05). Sperm motility patterns over time indicated that differences in acrosomal status were not influenced by cell death. The frequency of abnormal acrosomes detected by TEM was higher (p < 0.05) in teratospermic (30.0 +/- 3.9%) than in normospermic (3.1 +/- 1.3%) samples. Swim-up separation failed to reduce (p > 0.05) the proportion of sperm cells with malformed acrosomes (swim-up, 33.5 +/- 3.5%; washed, 26.6 +/- 4.6%). These results indicate that sperm from teratospermic cats exhibit a high incidence of malformed acrosomes detectable only at the ultrastructural level. Nevertheless, acrosomal dysfunction is not related exclusively to structural defects because > 40.0% of swim-up separated sperm with structurally normal acrosomes still are incapable of completing the acrosome reaction. This suggests that compromised capacitation and acrosomal dysfunction may be responsible for low fertilization success in the teratospermic domestic cat.

Role of needle biopsy in thyroid nodules.

The role of fine-needle aspiration cytology of thyroid nodules is analyzed in 116 cases. Forty-six percent of the patients with "cold" solid nodules were operated on with a 1% known false-negative and a 5% false-positive result. Ninety-four percent of the aspirations provided adequate cytologic material for evaluation. A new approach to the assessment and management of the thyroid nodule with fine-needle aspiration is proposed as the initial step in the evaluation.