Analgesic, cardiorespiratory effects and motor block characteristics of epidural levobupivacaine alone or in combination with methadone or dexmedetomidine in bitches undergoing unilateral total mastectomy.

The aim of this study was to evaluate the latency, extent of analgesia, and duration of motor block of levobupivacaine alone and combined with methadone or dexmedetomidine after epidural administration during and after mastectomy in dogs. Twenty-four mature, mixed-breed female dogs were randomly divided into three experimental groups with eight animals each, according to the agents used in lumbosacral epidural analgesia: levobupivacaine 0.75% alone (1.5mg/kg - control group), levobupivacaine 0.75% (1.5 mg/kg) + methadone 1% (0.3 mg/kg), or levobupivacaine 0.75% (1.5 mg/kg) + dexmedetomidine 0.05% (3 µg/kg). During surgery, cardiorespiratory parameters were evaluated. Rescue analgesia was given when there were signs of nociception and was necessary in all three treatment groups. Since all animals received rescue analgesia during the surgery and immediately post-surgery, the duration of the sensitive block were not evaluated. The extent of sensory block was between the 12º and 13º thoracic vertebrae for the control group, 7º thoracic vertebra to 5º lumbar vertebra (methadone group), and 8º thoracic vertebra to 4º lumbar vertebra for the dexmedetomidine group. Methadone or dexmedetomidine combined with levobupivacaine increased the extent of the sensory block and the duration of the motor block in bitches when administered via the epidural route.

Comparison of lidocaine, levobupivacaine or ropivacaine for distal paravertebral thoracolumbar anesthesia in ewes.

OBJECTIVE: To compare the effects of lidocaine, levobupivacaine, or ropivacaine on the onset time and duration of anesthesia of the flank of ewes, using the distal paravertebral thoracolumbar approach. STUDY DESIGN: Randomized experimental study. ANIMALS: Twenty-six healthy mixed-breed ewes (46 ± 3.1 kg). METHODS: Thoracolumbar paravertebral nerve blocks were performed using the distal approach in sheep for ruminal fistulation. The 13th thoracic (T13), first lumbar (L1) and second lumbar (L2) nerves were infiltrated with 2% lidocaine (group GLI, n = 9), 0.5% levobupivacaine (group GLE, n = 8) or 0.5% ropivacaine (group GRO, n = 9); 1.5 mL on the dorsal branch and 2.5 mL on the ventral branch, total volume of 12 mL per ewe. Anesthesia onset time and duration were assessed by application of superficial and deep pin pricks, and skin clamping with a hemostat. Heart rate, respiratory rate, rectal temperature and systemic arterial pressures were recorded prior to nerve block (T0), after the anesthetic agent injection and onset time (T1) and predetermined time points during the surgical procedure (T2-T6). RESULTS: Incomplete nerve blocks were present in five of the 26 ewes enrolled in the study and they were not included in the statistical analyzes. Onset times in GLI, GLE and GRO were 1.5 ± 0.5, 3.1 ± 1.5 and 2.1 ± 0.8 minutes, respectively, with GLE significantly longer than GLI. The durations of anesthesia for GLI, GLE and GRO were 80 ± 27, 649 ± 68 and 590 ± 40 minutes, respectively, with the duration of GLI significantly shorter than GLE and GRO. There were no clinically important changes in cardiopulmonary variables. CONCLUSION AND CLINICAL RELEVANCE: Administration of levobupivacaine and ropivacaine at the distal paravertebral site to block nerves T13, L1 and L2 produced a longer duration of anesthesia of the ewe's flanks compared with lidocaine.

Triethanolamine stabilization of methotrexate-ß-cyclodextrin interactions in ternary complexes.

The interaction of methotrexate (MTX) with beta-cyclodextrin (ß-CD) in the presence of triethanolamine (TEA) was investigated with the aim to elucidate the mechanism whereby self-assembly cyclodextrin systems work in association with this third component. Solubility diagram studies showed synergic increment of the MTX solubility to be about thirty-fold. Experiments using 2D ROESY and molecular modeling studies revealed the inclusion of aromatic ring III of the drug into ß-CD cavity, in which TEA contributes by intensifying MTX interaction with ß-CD and stabilizes MTX:ß-CD:TEA ternary complex by electrostatic interaction. The maintenance of these interactions in solid phase was also studied in ternary MTX:ß-CD:TEA and comparisons were made with freeze dried binary MTX:ß-CD and physical mixtures. FTIR studies evidenced that MTX-ß-CD interaction remained in solid ternary complexes, which was also supported by thermal (differential scanning calorimetry (DSC), thermogravimetric analysis (TG)/first derivative of TG analysis (DTG) and C,N,H elementary analysis) and structural (X-ray diffraction analysis, (XRD)) studies, mainly regarding the increment of drug stability. The efficient in vitro drug dissolution studies successfully demonstrated the contribution of ternary complexes, which highlights the importance of this possible new raw material for further applications in drug delivery systems.

Validation of the Portuguese Version of the Short-Form Glasgow Composite Measure Pain Scale (CMPS-SF) According to COSMIN and GRADE Guidelines.

We aimed to validate the CMPS-SF according to COSMIN and GRADE guidelines. Four trained evaluators assessed 208 videos (pre-operative-M1, peak of pain-M2, 1 h after the peak of pain and analgesia (rescue)-M3, and 24 h post-extubation-M4) of 52 dogs, divided into negative controls (n = 10), soft tissue surgeries (n = 22), and orthopedic surgeries (n = 20). The videos were randomized and blinded as to when they were filmed, and were evaluated in two stages, 21 days apart. According to confirmatory analysis, the CMPS-SF is a unidimensional scale. Intra-observer reliability was between 0.80 and 0.99 and inter-observer reliability between 0.73 and 0.86. Criterion validity was confirmed by the correlation between the CMPS-SF and other unidimensional scales (≥0.7). The differences between the scores were M2 ≥ M3 > M4 > M1 (responsiveness), and the scale presented construct validity (higher postoperative pain scores in dogs undergoing surgery versus control). Internal consistency was 0.7 (Cronbach's α) and 0.77 (McDonald's ω), and the item-total correlation was between 0.3 and 0.7, except for "A(ii)-Attention to wound". Specificity and sensitivity were 78-87% and 74-83%, respectively. The cut-off point for rescue analgesia was ≥5 or ≥4 excluding item B(iii) mobility, and the GRADE classification was high, confirming the validity of the scale.

Laminin database: a tool to retrieve high-throughput and curated data for studies on laminins.

The Laminin(LM)-database, hosted at http://www.lm.lncc.br, is the first database focusing a non-collagenous extracellular matrix protein family, the LMs. Part of the knowledge available in this website is automatically retrieved, whereas a significant amount of information is curated and annotated, thus placing LM-database beyond a simple repository of data. In its home page, an overview of the rationale for the database is seen and readers can access a tutorial to facilitate navigation in the website, which in turn is presented with tabs subdivided into LMs, receptors, extracellular binding and other related proteins. Each tab opens into a given LM or LM-related molecule, where the reader finds a series of further tabs for 'protein', 'gene structure', 'gene expression' and 'tissue distribution' and 'therapy'. Data are separated as a function of species, comprising Homo sapiens, Mus musculus and Rattus novergicus. Furthermore, there is specific tab displaying the LM nomenclatures. In another tab, a direct link to PubMed, which can be then consulted in a specific way, in terms of the biological functions of each molecule, knockout animals and genetic diseases, immune response and lymphomas/leukemias. LM-database will hopefully be a relevant tool for retrieving information concerning LMs in health and disease, particularly regarding the hemopoietic system.

Reliability and Validity of UNESP-Botucatu Cattle Pain Scale and Cow Pain Scale in Bos taurus and Bos indicus Bulls to Assess Postoperative Pain of Surgical Orchiectomy.

Pain assessment guides decision-making in pain management and improves animal welfare. We aimed to investigate the reliability and validity of the UNESP-Botucatu cattle pain scale (UCAPS) and the cow pain scale (CPS) for postoperative pain assessment in Bos taurus (Angus) and Bos indicus (Nelore) bulls after castration. METHODS: Ten Nelore and nine Angus bulls were anaesthetised with xylazine-ketamine-diazepam-isoflurane-flunixin meglumine. Three-minute videos were recorded at -48 h, preoperative, after surgery, after rescue analgesia and at 24 h. Two evaluators assessed 95 randomised videos twice one month apart. RESULTS: There were no significant differences in the pain scores between breeds. Intra and inter-rater reliability varied from good (>0.70) to very good (>0.81) for all scales. The criterion validity showed a strong correlation (0.76-0.78) between the numerical rating scale and VAS versus UCAPS and CPS, and between UCAPS and CPS (0.76). The UCAPS and CPS were responsive; all items and total scores increased after surgery. Both scales were specific (81-85%) and sensitive (82-87%). The cut-off point for rescue analgesia was >4 for UCAPS and >3 for CPS. CONCLUSIONS: The UCAPS and CPS are valid and reliable to assess postoperative pain in Bos taurus and Bos indicus bulls.

Clinical validation of the short and long UNESP-Botucatu scales for feline pain assessment.

BACKGROUND: The UNESP-Botucatu multidimensional feline pain assessment scale (UFEPS) is a valid and reliable instrument for acute pain assessment in cats. However, its limitations are that responsiveness was not tested using a negative control group, it was validated only for ovariohysterectomy, and it can be time-consuming. We aimed to evaluate the construct and criterion validity, reliability, sensitivity, and specificity of the UFEPS and its novel short form (SF) in various clinical or painful surgical conditions. METHODS: Ten client-owned healthy controls (CG) and 40 client-owned cats requiring pain management for clinical or surgical care (20 clinical and 20 surgery group (12 orthopedic and eight soft tissue surgeries) were recruited. Three evaluators assessed pain, in real-time, in clinical cases before and 20 min after rescue analgesia and in surgical cases before and up to 6.5 hours postoperatively, by using the visual analog, numerical ratio, and a simple descriptive scale, in this order, followed by the UFEPS-SF, UFEPS and Glasgow multidimensional feline pain (Glasgow CMPS-Feline) in random order. For the surgical group, rescue analgesia (methadone 0.2 mg/kg IM or IV and/or dipyrone 12.5 mg/kg IV) was performed when the UFEPS-SF score was ≥4 or exceptionally according to clinical judgement. If a third interventional analgesia was required, methadone (0.1-0.2 mg/kg IM) and ketamine (1 mg/kg IM) were administered. For the clinical group, all cats received rescue analgesia (methadone 0.1-0.2 mg/kg IM or IV or nalbuphine 0.5 mg/kg IM or IV), according to the clinician in charge, regardless of pain scores. Construct (1-comparison of scores in cats undergoing pain vs pain-free control cats by unpaired Wilcoxon-test and 2-responsiveness to analgesia by paired Wilcoxon test) and concurrent criterion validity (Spearman correlation of the total score among scales), inter-rater reliability, specificity and sensitivity were calculated for each scale (α = 0.05). RESULTS: Reliability ranged between moderate and good for the UFEPS and UFEPS-SF (confidence intervals of intraclass coefficients = 0.73-0.86 and 0.63-0.82 respectively). The Spearman correlation between UFEPS and UFEPS-SF was 0.85, and their correlation with Glasgow CMPS-Feline was strong (0.79 and 0.78 respectively), confirming criterion validity. All scales showed construct validity or responsiveness (higher scores of cats with clinical and postoperative pain vs healthy controls, and the reduction in scores after rescue analgesia). The sensitivity and specificity of the UFEPS, UFEPS-SF and Glasgow CMPS-Feline were moderate (sensitivity 83.25, 78.60% and 74.28%; specificity 72.00, 84.67 and 70.00%, respectively). CONCLUSIONS: Both UFEPS and UFEPS-SF showed appropriate concurrent validity, responsiveness, reliability, sensitivity, and specificity for feline acute pain assessment in cats with various clinical and orthopedic and soft tissue surgical conditions.

Monitoring structural features, biocompatibility and biological efficacy of gamma-irradiated methotrexate-loaded spray-dried microparticles.

In this study, biodegradable and biocompatible gamma irradiated poly-(dl-lactide-co-glycolide) (PLGA) spray-dried microparticles were prepared aiming to improve the efficacy of methotrexate (MTX). The experimental design included three formulations of microparticles containing distinct drug amount (9%, 18%, and 27% w/w) and three distinct gamma irradiation dose (15kGy, 25kGy, and 30kGy). The physicochemical and drug release properties of the microparticles supported their biocompatibility and biological efficacy studies in different cell lines. The irradiation induced slight changes in the spherical shape of the microparticles and the formation of free radicals was dependent on the drug loading. However, the amorphous character, particle size, drug loading, and drug release rate of the microparticles were preserved. The drug release data from all microparticles formulation were evaluated by using four drug kinetic models and by comparison of their similarity factor (f2). The gamma irradiation did not induce changes in the biocompatibility of PLGA microparticles and in the biological activity of the MTX-loaded microparticles. Finally, the spray-dried MTX-loaded PLGA microparticles enhanced the efficacy of the drug in the human cervical cancer cells (SiHa cell line). This study demonstrated the feasibility of the gamma irradiated spray dried PLGA microparticles for prolonged release of MTX, supporting a promising antitumor-drug delivery system for parenteral (subcutaneous) or pulmonary use.

Molecular characterization of Caulobacter crescentus mutator strains.

Mutator strains were identified by screening random Tn5 insertion clones of Caulobacter crescentus. We identified clones with robust increases in mutation rates with Tn5 insertions in the mutY, mutS, mutL and uvrD genes, known to act in mutation-preventing pathways in Escherichia coli. Analysis of mutations in the rpoB gene revealed that in both the parental strain and mismatch repair-deficient mutants, A:T→G:C transitions predominate by a large margin over C:G→T:A. We have also investigated the role of the error-prone polymerase encoded by imuC (dnaE2) in spontaneous mutagenesis, and found that a imuC mutant strain shows mutation rates and sequences comparable to the parental strain. Our study characterizes for the first time mutator strains in a member of the alphaproteobacteria group. In spite of the limitations of using a single marker, possible reasons for the observed mutational bias are discussed in the light of the repertoire of DNA repair genes in this bacterium.

Evaluation of cardiopulmonary parameters and recovery from anesthesia in cougars (Puma concolor) anesthetized with detomidine/ketamine and isoflurane or sevoflurane / Avaliação de parâmetros cardiorrespiratórios e recuperação da anestesia em pumas (Puma concolor) anestesiados com detomidina/cetamina e isoflurano ou sevoflurano

The aim of this study was to assess the cardiopulmonary effects, the onset time after the administration of a detomidine/ketamine combination, and the recovery from anesthesia of cougars (Puma concolor) anesthetized with detomidine/ketamine and isoflurane or sevoflurane for abdominal ultrasound imaging. Fourteen animals were randomly allocated into two experimental groups: GISO (n=7) and GSEVO (n=7). Chemical restraint was performed using 0.15mg/kg detomidine combined with 5mg/kg ketamine intramuscularly; anesthesia induction was achieved using 2mg/kg propofol intravenously and maintenance with isoflurane (GISO) or sevoflurane (GSEVO). The following parameters were assessed: heart rate, respiratory rate, systolic and diastolic arterial blood pressure, mean arterial blood pressure, oxyhemoglobin saturation, rectal temperature, central venous pressure, and end-tidal carbon dioxide. The time to sternal recumbency (TSR) and time to standing position (TSP) were also determined. There was not statistically significant difference for the cardiopulmonary variables or TSP whereas TSR was significantly shorter in GSEVO. The time to onset of anesthesia was 11.1±1.2 minutes and 11.3±1.8 minutes for GISO and GSEVO, respectively. The anesthesia of cougars with detomidine/ketamine and isoflurane or sevoflurane was conducted with safety, cardiopulmonary stability, and increased time to sternal recumbency in the GISO group.

O objetivo do presente estudo foi avaliar os efeitos cardiorrespiratórios e a recuperação anestésica de onças-pardas (Puma concolor) submetidas à anestesia com detomidina/cetamina e isofluorano ou sevofluorano para avaliação ultrassonográfica abdominal. Para isso, foram utilizados 14 animais divididos aleatoriamente em dois grupos experimentais GISO (n=7) e GSEVO (n=7). Foram submetidos à contenção química com detomidina 0,15mg/kg associada à cetamina 5mg/kg pela via intramuscular, induzidos com propofol 2mg/kg pela via intravenosa e mantidos com isofluorano (GISO) ou sevofluorano. Foram avaliados os parâmetros: frequência cardíaca e respiratória, pressão arterial sistólica, média e diastólica saturação de oxihemoglobina, temperatura retal, pressão venosa central e fração expirada de dióxido de carbono. O tempo para adoção de decúbito esternal e posição quadrupedal também foram avaliados. Não houve diferença estatística para as variáveis cardiorrespiratórias e no tempo para adoção da posição quadrupedal. O tempo para adoção de decúbito esternal foi significativamente menor no GSEVO em relação ao GISO. Concluiu-se que a anestesia de onças pardas com detomidina/cetamina e isoflurano ou sevoflurano foi realizada de maneira segura, com estabilidade cardiorrespiratória e com aumento no tempo para adoção de decúbito esternal no GISO.