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.

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.

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.