Description and validation of a new descriptive and multiparametric numeric rating scale to assess sedation in cats.

The objective of this study was to design and assess the validity and reliability of a new feline multiparametric sedation scale (FMSS). A total of 89 household cats were recruited, enabling a total of 534 sedation assessments. Every assessment was performed by 3 blinded observers with varying expertise levels (Level 1: Student; Level 2: RVT; Level 3: ACVAA diplomate or senior resident). For comparison purposes, a visual analogue scale (VAS) and a Simple Qualitative Scale (SQS) were also used concurrently, with the VAS considered the gold standard. The new scale had excellent inter-observer agreement among experience groups with weighted Kappa scores of 0.84 (Levels 1 versus 2), 0.82 (Levels 2 versus 3), and 0.84 (Levels 1 versus 3), with P < 0.0001 for all comparisons. There was a high degree of association between FMSS and VAS (r = 0.90, P < 0.0001) and between FMSS and SQS (r = 0.89, P < 0.0001). Final FMSS numerical values were paired with levels of sedation with None = 0 (0 to 5), Mild = 4 (1 to 7), Moderate = 6 (2 to 10), and Profound = 12 (7 to 12); furthermore, differences were detected between pre- and post-sedation evaluations (P = 0.001). This scale demonstrated internal consistency and sensitivity even when evaluating drugs or doses with minimal sedative effects and there was very strong interrater reliability, independent of experience level. Based on this clinical study, we concluded that the use of this sedation scale is appropriate when objective numerical sedation quantification is required, in either a clinical or research setting.

Description et validation d'une nouvelle échelle d'évaluation numérique descriptive et multiparamétrique pour évaluer la sédation chez le chat. L'objectif de cette étude était de concevoir et d'évaluer la validité et la fiabilité d'une nouvelle échelle de sédation multiparamétrique féline (FMSS). Un total de 89 chats domestiques a été recruté, permettant un total de 534 évaluations de sédation. Chaque évaluation a été effectuée par trois observateurs en aveugle avec différents niveaux d'expertise (Niveau 1 : étudiant; Niveau 2 : RVT; Niveau 3 : diplomate de l'ACVAA ou résident senior). À des fins de comparaison, une échelle visuelle analogique (VAS) et une échelle qualitative simple (SQS) ont également été utilisées simultanément, VAS étant considérée comme l'étalon. La nouvelle échelle présentait un excellent accord inter-observateurs parmi les groupes d'expérience avec des scores Kappa pondérés de 0,84 (niveaux 1 versus 2), 0,82 (niveaux 2 versus 3) et 0,84 (niveaux 1 versus 3), avec P < 0,0001 pour toutes les comparaisons. Il y avait un degré élevé d'association entre FMSS et VAS (r = 0,90, P < 0,0001) et entre FMSS et SQS (r = 0,89, P < 0,0001). Les valeurs numériques FMSS finales ont été appariées avec les niveaux de sédation avec Aucun = 0 (0 à 5), Léger = 4 (1 à 7), Modéré = 6 (2 à 10) et Profond = 12 (7 à 12); en outre, des différences ont été détectées entre les évaluations pré- et post-sédation (P = 0,001). Cette échelle a démontré une cohérence interne et une sensibilité même lors de l'évaluation de médicaments ou de doses avec des effets sédatifs minimes et il y avait une très forte fiabilité inter-évaluateur, indépendamment du niveau d'expérience. Sur la base de cette étude clinique, nous avons conclu que l'utilisation de cette échelle de sédation est appropriée lorsqu'une quantification numérique objective de la sédation est requise, dans un cadre clinique ou de recherche.(Traduit par Dr Serge Messier).

Biosafety Level 2 cabinet UV-C light exposure of sports antidoping human urine samples does not affect the stability of selected prohibited substances.

The current study examined the stability of several antidoping prohibited substances analytes in urine after 15-min exposure to UV-C light in a Biosafety Level 2 cabinet. The urine matrices were exposed within the original antidoping bottles with the aim to destroy DNA/RNA and possible SARS CoV-2. The analytes small molecules Phase I and Phase II metabolites and peptides, in total 444, endogenous, internal standards, and prohibited substances, pH, and specific gravity in urine were studied. The accredited analytical methods were used by Anti-Doping Laboratory Qatar for the comparison of data of the same urine samples analyzed with and without UV-C exposure. In the study conditions, no problems of stability were detected in the substances spiked in the urine samples exposed in the UV-C irradiation.

Dose and cardiopulmonary effects of propofol alone or with midazolam for induction of anesthesia in critically ill dogs.

OBJECTIVE: To determine the dose and cardiopulmonary effects of propofol alone or with midazolam for induction of anesthesia in American Society of Anesthesiologists status ≥III dogs requiring emergency abdominal surgery. STUDY DESIGN: Prospective, randomized, blinded, clinical trial. ANIMALS: A total of 19 client-owned dogs. METHODS: Dogs were sedated with fentanyl (2 µg kg-1) intravenously (IV) for instrumentation for measurement of heart rate, arterial blood pressure, cardiac index, systemic vascular resistance index, arterial blood gases, respiratory rate and rectal temperature. After additional IV fentanyl (3 µg kg-1), the quality of sedation was scored and cardiopulmonary variables recorded. Induction of anesthesia was with IV propofol (1 mg kg-1) and saline (0.06 mL kg-1; group PS; nine dogs) or midazolam (0.3 mg kg-1; group PM; 10 dogs), with additional propofol (0.25 mg kg-1) IV every 6 seconds until endotracheal intubation. Induction/intubation quality was scored, and anesthesia was maintained with isoflurane. Variables were recorded for 5 minutes with the dog in lateral recumbency, breathing spontaneously, and then in dorsal recumbency with mechanical ventilation for the next 15 minutes. A general linear mixed model was used with post hoc analysis for multiple comparisons between groups (p < 0.05). RESULTS: There were no differences in group demographics, temperature and cardiopulmonary variables between groups or within groups before or after induction. The propofol doses for induction of anesthesia were significantly different between groups, 1.9 ± 0.5 and 1.1 ± 0.5 mg kg-1 for groups PS and PM, respectively, and the induction/intubation score was significantly better for group PM. CONCLUSIONS AND CLINICAL RELEVANCE: Midazolam co-induction reduced the propofol induction dose and improved the quality of induction in critically ill dogs without an improvement in cardiopulmonary variables, when compared with a higher dose of propofol alone.

Population reference ranges of urinary endogenous sulfate steroids concentrations and ratios as complement to the steroid profile in sports antidoping.

The population based Steroid Profile (SP) ratio of testosterone (T) and epitestosterone (E) has been considered as a biomarker approach to detect testosterone abuse in '80s. The contemporary Antidoping Laboratories apply the World Antidoping Agency (WADA) Technical Document (TD) for Endogenous Androgenic Anabolic Steroids (EAAS) in the analysis of SP during their screening. The SP Athlete Biological Passport (ABP) adaptive model uses the concentrations of the total of free and glucuronide conjugated forms of six EAASs concentrations and ratios measured by GC/MS. In the Antidoping Lab Qatar (ADLQ), the routine LC/MS screening method was used to quantitatively estimate the sulfate conjugated EAAS in the same analytical run as for the rest qualitative analytes. Seven sulfate EAAS were quantified for a number of routine antidoping male and female urine samples during screening. Concentrations, statistical parameters and selected ratios for the 6 EAAS, the 6 sulfate EAAS and 29 proposed ratios of concentrations from both EAAS and sulfate EAAS, which potentially used as SP ABP biomarkers, population reference limits and distributions have been estimated after the GC/MSMS analysis for EAAS and LC/Orbitrap/MS analysis for sulfate EAAS.

Comparison of 3 blind brachial plexus block techniques during maintenance of anesthesia and postoperative pain scores in dogs undergoing surgical procedures of the thoracic limb.

The degree of analgesia provided by blind techniques for brachial plexus blocks (BPBs) has not been compared in clinical cases undergoing surgery of the thoracic limb. The objective of this study was to evaluate the anesthetic conditions and postoperative analgesia resulting from 3 different BPB local anesthetic techniques in canine patients undergoing such surgery. Twenty-four client-owned dogs received a standardized premedication/induction protocol (hydromorphone and acepromazine/propofol), maintained with isoflurane in oxygen using mechanical ventilation, in a prospective, randomized, blinded clinical trial. Before surgery, dogs received 1 of 3 anatomical BPB techniques: traditional, perpendicular, or axillary, with 0.2 mL/kg body weight (BW) of bupivacaine 0.5%. Cardiorespiratory variables and isoflurane end-tidal concentrations were recorded throughout anesthesia. Scores for anesthetic maintenance (0-best to 4-responsive), recovery quality (0-not responsive to 3-responsive), and Glasgow pain scale were recorded for up to 24 h postoperatively. All dogs recovered uneventfully from anesthesia and no differences in the measured variables or scores were noted among groups, during and after anesthesia. When thoracic limb amputations in each of the 3 groups (n = 9; 4 in traditional, 3 in perpendicular, 2 in axillary) were compared to the other surgical procedures (n = 15); however, scores for anesthetic maintenance were higher [0 (0 to 1) versus 0 (0 to 0); median (interquartile range)], recovery [1 (0 to 2) versus 0 (0 to 0)], and pain [2.4 (2.4 to 3.0) versus 1.6 (1.4 to 2.2)] in the first 3 h post-extubation. Surgery times were also longer with amputations [115 min (100 to 138 min) versus 50 min (41 to 90 min)]. The 3 BPB techniques provided similar anesthesia and postoperative pain scores. Despite higher pain scores in thoracic limb amputations than in less invasive surgeries, the BPB appeared to provide significant comfort.

Le degré d'analgésie fournie par les blocs à l'aveugle du plexus brachial (BPBs) n'a pas été comparé lors de cas cliniques soumis à une chirurgie du membre thoracique. La présente étude visait à évaluer les conditions anesthésiques et l'analgésie post-opératoire résultant de trois techniques différentes d'anesthésie locale par BPB chez des patients canins soumis à une chirurgie. Vingt-quatre chiens appartenant à des propriétaires ont reçu une prémédication/induction standardisée (hydromorphone et acépromazine/propofol), avec maintien à l'isoflurane dans de l'oxygène en utilisant une ventilation mécanique, dans une étude clinique prospective, randomisée, et à l'aveugle. Avant la chirurgie, les chiens ont reçu un BPB par une des trois techniques anatomiques : traditionnelle, perpendiculaire, ou axillaire, avec 0,2 mL/kg de poids corporel de bupivacaïne 0,5 %. Les variables cardiorespiratoires et les concentrations d'isoflurane en fin d'expiration furent enregistrées tout au long de l'anesthésie. Les pointages pour le maintien de l'anesthésie (0-meilleur à 4-réactif), la qualité du rétablissement (0-non réactif à 3-réactif), et l'échelle de la douleur de Glasgow ont été notés jusqu'à 24 h post-opération. Tous les chiens ont récupéré sans problème de l'anesthésie et aucune différence mesurable dans les variables mesurées ou les pointages ne fut notée parmi les groupes, durant ou après l'anesthésie. Toutefois, lorsque les amputations du membre thoracique dans chacun des trois groupes (n = 9; 4 dans la traditionnelle, 3 dans la perpendiculaire, 2 dans l'axillaire) furent comparées aux autres procédures chirurgicales (n = 15), les pointages pour le maintien de l'anesthésie étaient plus élevés [0 (0 à 1) versus 0 (0 à 0); médiane (écart interquartile)], rétablissement [1 (0 à 2) versus 0 (0 à 0)], et douleur [2,4 (2,4 à 3,0) versus 1,6 (1,4 à 2,2)] dans les 3 h suivant l'extubation. La durée des chirurgies étaient également plus longues lors des amputations [115 min (100 à 138 min) versus 50 min (41 à 90 min)]. Les trois techniques de BPB produisaient des pointages similaires d'anesthésie et de douleur postopératoire. Malgré des pointages de douleur plus élevés lors d'amputations du membre thoracique que lors de chirurgies moins invasives, les BPB semblent fournir un confort significatif.(Traduit par Docteur Serge Messier).

High resolution full scan liquid chromatography mass spectrometry comprehensive screening in sports antidoping urine analysis.

The aim of this paper is to present the development and validation of a high-resolution full scan (HR-FS) electrospray ionization (ESI) liquid chromatography coupled to quadrupole Orbitrap mass spectrometer (LC/Q/Orbitrap MS) platform for the screening of prohibited substances in human urine according to World Antidoping Agency (WADA) requirements. The method was also validated for quantitative analysis of six endogenous steroids (epitestosterone, testosterone, 5α-dihydrotestosterone, dehydroepiandrosterone, androsterone and etiocholanolone) in their intact sulfates form. The sample preparation comprised a combination of a hydrolyzed urine liquid-liquid extraction and the dilute & shoot addition of original urine in the extracted aliquot. The HR-FS MS acquisition mode with Polarity Switching was applied in combination of the Quadrupole-Orbitrap mass filter. The HR-FS acquisition of analytical signal, for known and unknown small molecules, allows the inclusion of all analytes detectable with LC-MS for antidoping investigations to identify the use of known or novel prohibited substances and metabolites after electronic data files' reprocessing. The method has been validated to be fit-for-purpose for the antidoping analysis.

Evaluation of longitudinal steroid profiles from male football players in UEFA competitions between 2008 and 2013.

Testosterone and related compounds are the most recurrent doping substances. The steroid profile, consisting of the quantification of testosterone and its metabolites, has been described as the most significant biomarker to detect doping with pseudo-endogenous anabolic steroids. The steroidal module of the Athlete Biological Passport (ABP) was launched by the World Anti-Doping Agency (WADA) in 2014. To assess the value of introducing the module to its anti-doping programme, the Union of European Football Associations (UEFA) decided to analyze retrospectively the steroid profile data of 4195 urine samples, collected from 879 male football players and analyzed in 12 WADA-accredited laboratories between 2008 and mid-2013. This study focused on the evaluation of T/E ratios. The coefficient of variation (CV) and the adaptive model were the two statistical models used to study the longitudinal follow-up. A CV of 46% was determined to be the maximal natural intra-individual variation of the T/E when the sequence consisted of single data points analyzed in different laboratories. The adaptive model showed some profiles with an atypical T/E sequence and also enabled an estimate of the prevalence of external factors impacting the T/E sequences. Despite the limitations of this retrospective study, it clearly showed that the longitudinal and individual follow-up of the T/E biomarker of the players is a good tool for target testing in football. UEFA has therefore decided to implement the steroidal module of the ABP from the start of the next European football season in September 2015. Copyright © 2015 John Wiley & Sons, Ltd.

Involvement of spinal cord BDNF in the generation and maintenance of chronic neuropathic pain in rats.

Brain-derived neurotrophic factor (BDNF) is involved in neuronal survival and synaptic plasticity of the central and peripheral nervous system. In chronic pain, plastic changes in dorsal horn neurons contribute to a phenomenon of hypersensitivity to pain sensation that is maintained over time, known as central sensitization. This process is accompanied by BDNF overexpression, but the role of BDNF in the generation and maintenance of the hyperalgesic phenomenon is still unclear. The present study was aimed to investigate if exogenous BDNF administered to the rat spinal cord, in addition to trigger pain, participates in the maintenance of the central sensitization process (i.e., pain persistence) and to determine if the pain generated is comparable to that observed in a neuropathic pain model. Results showed that a single intrathecal injection of 0.003 ng of BDNF was able to decrease the nociceptive threshold (Randall-Selitto test) in normal rats, for at least a 42-day period. Furthermore, the hyperalgesia generated was comparable to that observed in rats with a 42-day history of mononeuropathy. Increasing the dose or administering additional doses of BDNF resulted neither in additional effectiveness in reducing the pain threshold nor in the prolongation of the hyperalgesic effect, thus showing that central sensitization induced by BDNF is a dose-independent, all-or-none process. It is concluded that BDNF alone is sufficient for generating a long-lasting neural excitability change in the spinal cord via tyrosine kinase B receptor signaling, similar to that observed in chronic pain models such as neuropathy.

The influence of diet on isotope ratio mass spectrometry values.

OBJECTIVES: Athletes have increasingly used testosterone (T) and other endogenous anabolic steroids that cannot be detected by conventional gas chromatography-mass spectrometry. This led to gas chromatography-combustion-isotope ratio mass spectrometry(GC/C/IRMS), which measures the relative amount of 13C in urinary steroids. Because exogenous testosterone is relatively low in 13C content, this study will determine if consuming a diet low in 13C plants, such as soy, can be confused with a GC/C/IRMS-positive test for exogenous testosterone. DESIGN: Cross-sectional study in which 22 vegetarians known to consume a diet depleted of 13C isotope were compared with a geographic control group of 14 subjects consuming a normal diet. SETTING: Two distinct subject populations with respect to diet. SUBJECTS: Subjects were recruited from a soy-based cooperative and control volunteers. Twenty-two of 24 research subjects completed the protocol compared with 14 of 22 control subjects. INTERVENTIONS: Independent variables were delta13C IRMS values,urinary steroid profile, and isoflavone analysis. MAIN OUTCOME MEASURES: Comparisons were made with respect to dietary analysis, isoflavones, and urinary steroid measurements using GC-C-IRMS. RESULTS: The delta13C values for 2 major metabolites of T (androsterone and etiocholanolone) were lower for the vegetarians than the controls (P = 0.005). The vegetarians excreted a median of 23 micromol/d of total isoflavones compared with 2.7 micromol/d for the control group (P =0.0002). CONCLUSIONS: The carbon isotope ratios of urinary testosterone metabolites of vegetarians consuming a diet that is markedly depleted of 13C content were lower than that of control subjects, but not low enough to result in World Anti-Doping Agency criteria for a positive IRMS analysis.

Drug testing data from the 2007 Pan American Games: delta13C values of urinary androsterone, etiocholanolone and androstanediols determined by GC/C/IRMS.

The main purpose of this article is to show the application of the CG/C/IRMS in real time during competition in the steroid confirmation analysis. For this reason, this paper summarizes the results obtained from the doping control analysis during the period of the 2007 Pan American Games held in Rio de Janeiro, Brazil. Approximately 5600 athletes from 42 different countries competed in the games. Testing was performed in accordance to World Anti-Doping Agency (WADA) technical note for prohibited substances. This paper reports data where abnormal urinary steroid profiles, have been found with the screening procedures. One 8 mL urine sample was used for the analysis of five steroid metabolites with two separate analyses by gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS). Urine samples were submitted to GC/C/IRMS for confirmation analysis to determine the (13)C/(12)C ratio of selected steroids. Fifty-seven urine samples were analyzed by GC/C/IRMS and the delta(13)C values ( per thousand) of androsterone, etiocholanolone, 5beta-androstane-3alpha, 17beta-diol (5beta-diol), 5alpha-androstane-3alpha, 17beta-diol (5alpha-diol) and 5beta-pregnane-3alpha, 20alpha-diol (5beta-pdiol), the endogenous reference compound are presented. One urine sample with a testosterone/epitestosterone (T/E) ratio of 4.7 was confirmed to be positive of doping by GC/C/IRMS analysis. The delta values of 5beta-diol and 5alpha-diol were 3.8 and 10.8, respectively, compared to the endogenous reference compound 5beta-pdiol, which exceeded the WADA limit of 3 per thousand. The results obtained by CG/C/IRMS confirmation analyses, in suspicious samples, were conclusive in deciding whether or not a doping steroid violation had occurred.