Development of a Pain Scoring System for Use in Sheep Surgically Implanted with Ventricular Assist Devices.

Purpose/Aim: In prey species, such as sheep, clinical signs of postoperative pain can manifest in subtle ways or may be concealed entirely. Previous publications describing pain assessment in ruminants focus on lameness and flock behavior, often in a farm environment. These indicators of pain may be difficult to assess in sheep housed in biomedical research settings. We have developed a novel pain scoring system for sheep undergoing thoracotomy for implantation of ventricular assist devices that are permanently housed in modified stanchions. Materials and Methods: The pain scoring system includes ruminant-specific behavioral signs of pain in addition to objective measurements that can be readily evaluated in a biomedical research setting. A numerical score is generated by the evaluator for each category. A decision tree is utilized to help guide further action following the generation of a cumulative score by the evaluator. A total score of 0-2 requires no intervention, 3-9 requires the consideration of additional analgesic administration, and a pain score ≥ 10 warrants the consideration of additional multimodal analgesia. Results: A novel pain scoring system and decision tree specifically designed for sheep undergoing thoracotomy in a biomedical research environment was developed and successfully utilized. Out of 102 postoperative pain scores measured, 86 scores were <2. There were 17/102 postoperative pain scores ≥3, which typically resulted in the administration of supplemental rescue analgesia in the immediate postoperative period. Conclusions: A novel pain scoring system was developed and utilized in a biomedical research environment for evaluating postoperative pain in sheep undergoing thoracotomy for implantation of a ventricular assist device. Further studies are necessary to validate the reliability of this novel pain scoring system.

Sex differences in antinociceptive response to Δ-9-tetrahydrocannabinol and CP 55,940 in the mouse formalin test.

Cannabinoids have shown promise for the treatment of intractable pain states and may represent an alternative pharmacotherapy for pain management. A growing body of clinical evidence suggests a role for sex in pain perception and in cannabinoid response. We examined cannabinoid sensitivity and tolerance in male and female mice expressing a desensitization-resistant form (S426A/S430A) of the cannabinoid type 1 receptor (CB1R). Mice were assessed for acute and inflammatory nociceptive behaviors in the formalin test following pretreatment with either vehicle or mixed CB1R/CB2R agonists, Δ-9-tetrahydrocannabinol ([INCREMENT]-THC) (1-6 mg/kg) or CP 55,940 (0.06-0.2 mg/kg). Tolerance to the effects of 6 mg/kg [INCREMENT]-THC or 0.1 mg/kg CP 55,940 was examined by the formalin test following chronic daily dosing. Female mice showed decreased sensitivity to the effects of [INCREMENT]-THC and CP 55,940 compared with male mice. The S426A/S430A mutation increased the attenuation of nociceptive behaviors for both agonists in both sexes. Female mice displayed delayed tolerance to [INCREMENT]-THC compared with male mice, whereas the S426A/S430A mutation conferred a delay in tolerance to [INCREMENT]-THC in both sexes. Male S426A/S430A mutant mice also display resistance to tolerance to CP 55,940 compared with wild-type controls. This study demonstrates sex and genotype differences in response for two different cannabinoid agonists. The results underscore the importance of including both male and female mice in preclinical studies of pain and cannabinoid pharmacology.

Cucurbitacin I inhibits cell motility by indirectly interfering with actin dynamics.

BACKGROUND: Cucurbitacins are plant natural products that inhibit activation of the Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) pathway by an unknown mechanism. They are also known to cause changes in the organization of the actin cytoskeleton. METHODOLOGY/PRINCIPAL FINDINGS: We show that cucurbitacin I potently inhibits the migration of Madin-Darby canine kidney (MDCK) cell sheets during wound closure, as well as the random motility of B16-F1 mouse melanoma cells, but has no effect on movement of Dictyostelium discoideum amoebae. Upon treatment of MDCK or B16-F1 cells with cucurbitacin I, there is a very rapid cessation of motility and gradual accumulation of filamentous actin aggregates. The cellular effect of the compound is similar to that observed when cells are treated with the actin filament-stabilizing agent jasplakinolide. However, we found that, unlike jasplakinolide or phallacidin, cucurbitacin I does not directly stabilize actin filaments. In in vitro actin depolymerization experiments, cucurbitacin I had no effect on the rate of actin filament disassembly at the nanomolar concentrations that inhibit cell migration. At elevated concentrations, the depolymerization rate was also unaffected, although there was a delay in the initiation of depolymerization. Therefore, cucurbitacin I targets some factor involved in cellular actin dynamics other than actin itself. Two candidate proteins that play roles in actin depolymerization are the actin-severing proteins cofilin and gelsolin. Cucurbitacin I possesses electrophilic reactivity that may lead to chemical modification of its target protein, as suggested by structure-activity relationship data. However, mass spectrometry revealed no evidence for modification of purified cofilin or gelsolin by cucurbitacin I. CONCLUSIONS/SIGNIFICANCE: Cucurbitacin I results in accumulation of actin filaments in cells by a unique indirect mechanism. Furthermore, the proximal target of cucurbitacin I relevant to cell migration is unlikely to be the same one involved in activation of the JAK2/STAT3 pathway.