Oclacitinib (APOQUEL®) is a selective Janus kinase 1 inhibitor with efficacy in a canine model of flea allergic dermatitis.

Oclacitinib is a novel Janus kinase (JAK) inhibitor that potently inhibits JAK1-dependent cytokines involved in allergy, inflammation, and pruritus (IL-2, IL-4, IL-6, IL-13, and IL-31). Oclacitinib (Apoquel®, Zoetis Inc, Parsippany, NJ) is approved for the treatment/control of pruritus associated with allergic dermatitis and treatment/control of clinical manifestations of atopic dermatitis in dogs at least 12 months of age. To evaluate the effectiveness of oclacitinib in dogs with flea allergy dermatitis, the JAK1 selective inhibitor was tested in a placebo-controlled, masked, single-dose (0.4 mg/kg) or repeat-dose (0.4 mg/kg, twice daily for 2 weeks) study. Pruritic behaviors were quantitated by video recording, and erythema and skin lesions were assessed using a 10-cm visual analog scale (VAS). Results showed that oclacitinib reduced pruritus by 61% as early as 1.5 h after a single oral dose compared to placebo, with an average reduction (compared to placebo) of 85% 1-5 h after dosing (0.4 mg/kg; p < .0001). Oclacitinib also significantly reduced erythema (p < .0001) and skin lesion (p < .0005) VAS scores on Day 14 compared to placebo in a repeat dose study. No adverse events were noted during the conduct of these studies. IL-31 concentrations were elevated in the majority of dogs after flea infestation, suggesting JAK1-dependent cytokines may drive clinical signs of flea allergy dermatitis. These findings show that oclacitinib, an inhibitor of JAK1-dependent cytokines involved in allergy and inflammation can rapidly reduce clinical signs associated with flea allergic dermatitis in dogs.

Comparison of intradermal and serum testing for environmental allergen-specific immunoglobulin E determination in a laboratory colony of cats with naturally acquired atopic syndrome.

BACKGROUND: Allergen testing is used to select antigens included in the desensitisation vaccine. Intradermal skin test (IDT) is the gold standard in cats, yet allergen-specific immunoglobulin (Ig)E serological testing (ASIS) is often used. Feline data are lacking regarding the agreement between IDT and ASIS results. HYPOTHESIS/OBJECTIVES: The first objective of the study was to establish a colony of cats with naturally acquired feline atopic syndrome (FAS). Further objectives were to define their hypersensitivity disorder to detail the allergen tests results, and to assess similarity between the allergen tests. ANIMALS: Thirty-five cats with FAS and 10 control cats. MATERIALS AND METHODS: Enrolled cats went through a five phase-screening and quarantine process before joining the colony. An elimination diet trial was performed on all FAS cats. ASIS and IDT were consecutively performed on all cats under sedation. RESULTS: Reactions to 34 allergens were compiled for the 45 cats. Global sensitivity and specificity of ASIS were 34.7% and 78.9%, respectively. Only flea (ICC = 0.26, p = 0.040) and Dermatophagoides pteronyssinus (ICC = 0.48, p < 0.001) allergens had a significant intraclass correlation (weak agreement). Two FAS cats had negative tests including one cat with a concomitant food allergy. CONCLUSIONS AND CLINICAL RELEVANCE: This study depicts the first reported colony of cats with naturally acquired FAS. This is the first feline study to compare and show the poor agreement between allergen tests with a panel of 34 allergens. This colony also harbours two cats with FAS with negative allergen tests. These may represent the first described cats with an intrinsic form of atopic syndrome.

Cytokine transcriptome profiling in acute experimental canine atopic dermatitis skin lesions after IL-31 inhibition with lokivetmab.

BACKGROUND: The caninised monoclonal antibody lokivetmab (LKV), directed at interleukin (IL)-31, is very effective at controlling pruritus in most dogs with atopic dermatitis (AD). However, evidence exists that IL-31 is not required for the induction of acute allergic skin inflammation, which might explain why this treatment is less efficacious in some dogs with AD. HYPOTHESIS/OBJECTIVES: To compare the comprehensive transcriptome analysis of house dust mite (HDM)-sensitised dogs with and without treatment with LKV to attest our hypothesis that LKV does not majorly affect acute cytokine/chemokine production. ANIMALS: Six HDM-sensitised atopic Maltese-beagle dogs. MATERIALS AND METHODS: In this cross-over study, the cytokine profiling of acute AD skin lesions was compared by RNA sequencing (RNA-Seq), with or without LKV-induced inhibition of IL-31. Skin biopsies were obtained from each dog at 0, 6, 12, 24, 48, and 96 h after epicutaneous HDM allergen provocation. RESULTS: Macroscopic and microscopic skin lesion scores were not significantly different between the LKV- and nontreatment groups at any time points. Likewise, the results of RNA-Seq analysis revealed no significant difference in the messenger (m)RNA expression of the major cytokines between these two groups. In LKV-treated dogs, IL6, IL9, IL13, IL33, CCL17, and CCL22 were significantly upregulated compared to their baseline expression levels, suggesting that these cytokines are unaffected by IL-31 inhibition. CONCLUSIONS AND CLINICAL RELEVANCE: IL-31 inhibition is insufficient to prevent the expression of other proinflammatory mediators in acute AD and these could be considered as other potential therapeutic targets.

Speed of onset of a new chewable formulation of oclacitinib maleate (Apoquel®) in a canine model of IL-31-induced pruritus.

Oclacitinib maleate (Apoquel®, Zoetis Inc.) is commonly used around the world for the control/treatment of pruritus associated with allergic dermatitis and the control/treatment of atopic dermatitis in dogs at least 12 months of age. A new flavored chewable formulation of oclacitinib has been developed where more than 90% of doses offered to dogs were freely accepted when tested in clinical trials. The objective of this study was to determine whether the new chewable formulation of oclacitinib has a similar onset of anti-pruritic activity as the original oclacitinib film-coated tablets (FCT). Twenty-one laboratory beagle dogs were randomized to treatment and received placebo, 0.4-0.6 mg/kg oclacitinib FCT or 0.4-0.6 mg/kg flavored chewable oclacitinib tablet (n = 7/group). Efficacy was measured by assessing reduction in pruritus 1-3 h post-administration of treatments. Pruritus was induced by injecting canine IL-31, intravenously (2.5 µg/kg), approximately 15 min prior to the pruritus observation window. Results from this study demonstrated both oclacitinib FCT and the flavored chewable oclacitinib tablet significantly reduced IL-31-induced pruritus within 1-3 h post-dosing compared to placebo (p = .0069 and .0113, respectively), suggesting the new formulation of oclacitinib chewable tablets works as quickly to reduce pruritus in dogs as the oclacitinib FCT.

Onset and duration of action of lokivetmab in a canine model of IL-31 induced pruritus.

BACKGROUND: Interleukin (IL)-31 is a cytokine involved in allergic inflammation which induces pruritus across species including dogs. Using recombinant canine IL-31 we have developed a model of pruritus in the dog to evaluate onset of action and duration of effect of therapeutic drugs. OBJECTIVE: To assess the onset of action and duration of effect of lokivetmab (Cytopoint) in the IL-31-induced pruritus model. ANIMALS: Twenty-four purpose-bred beagle dogs (neutered males, spayed and intact females) 1.5-4.7 years old and weighing between 6 and14 kg. METHODS AND MATERIALS: Randomized, blinded, placebo-controlled studies were designed to evaluate the antipruritic properties of lokivetmab. Laboratory beagle dogs were given either placebo, 0.125, 0.5 or 2.0 mg/kg lokivetmab, subcutaneously. IL-31 then was administered to evaluate pruritus 3-5 h post-placebo or -lokivetmab administration as well as one, seven, 14, 28, 42 and 56 days post-dosing. Pruritus was evaluated over a 2 h window in animals by video monitoring and scored using a categorical scoring system. RESULTS: When animals were given 2.0 mg/kg lokivetmab, a significant reduction in pruritus was observed at 3-4, 4-5 and 3-5 h post-treatment (P ≤ 0.0001). When animals were given either 0.125, 0.5 or 2 mg/kg lokivetmab, the duration of effect was dose-dependent and statistically significant for 14, 28 and 42 days, respectively (P ≤ 0.0288). CONCLUSION: These data indicate that a single subcutaneous injection of 2 mg/kg lokivetmab produces a significant suppression of pruritus starting 3 h post-treatment that can be sustained for 42 days.

Cytokine expression in feline allergic dermatitis and feline asthma.

BACKGROUND: The pathogenesis of feline allergic dermatitis (FAD) is unclear, with several differences from allergic dermatitis in dogs and humans. HYPOTHESIS/OBJECTIVES: To survey cytokine expression levels in healthy cats and cats affected with allergic dermatitis or asthma. ANIMALS: Formalin-fixed, paraffin-embedded skin biopsies from 22 cats with allergic dermatitis and 21 cats without allergic dermatitis were used for cutaneous assays. Serum was obtained from 17 healthy cats, 18 cats with allergic dermatitis, and 18 cats with a presumptive diagnosis of asthma. METHODS AND MATERIALS: Cutaneous mRNA expression was evaluated with quantitative PCR [interleukin (IL)-31 and IL-31 Receptor A] and RNA in situ hybridisation (ISH) [IL-5, IL-31, IL-31RA, IL-33 and Oncostatin M receptor (OSMR)-ß]. IL-31 protein concentrations were evaluated in serum with an enzyme-linked immunosorbent assay. Serum levels of 19 additional cytokines were evaluated using a Luminex panel. RESULTS: IL-31, IL-31RA, IL-5 and IL-33 mRNA expression were either expressed in low quantities or undetectable in most samples. By contrast, OSMR-ß expression was significantly higher in the skin of allergic versus healthy cats (P < 0.0001). Although serum IL-31 was detected in a larger number of cats with allergic dermatitis than healthy cats, and concentrations appeared to be higher in cats with allergies, this difference was not statistically significant. Cats affected by asthma also exhibited insignificantly higher concentrations of IL-31 in the serum. CONCLUSIONS AND CLINICAL RELEVANCE: Our results suggest that feline allergic diseases may exhibit different pathomechanisms from allergic diseases affecting other species. These findings are useful in guiding further therapeutic development toward targets that may have a role in the pathogenesis of feline allergic skin disease.

Serum concentrations of IL-31 in dogs with nonpruritic mast cell tumours or lymphoma.

BACKGROUND: The aim of this study was to compare serum interleukin (IL)-31 concentrations in dogs with lymphoma and mast cell tumours (MCT) without pruritus to those of healthy dogs. HYPOTHESIS/OBJECTIVES: To determine if IL-31 plays a role in tumour pathogenesis and if IL-31 could be a biological marker for disease progression. ANIMALS: Forty-eight healthy dogs and 36 dogs with neoplasia [multicentric lymphoma (14), MCT (15) and cutaneous lymphoma (7)] were included in the study. METHODS AND MATERIALS: Dogs with neoplasia were assigned to three different groups. Group 1 consisted of patients with multicentric lymphoma, which were diagnosed by cytological, histopathological and clonality investigations. Thoracic radiographs, ultrasound examination of the abdominal cavity, and fine-needle aspirates from liver and spleen were used to determine the lymphoma stage Patients with cutaneous lymphoma, diagnosed by cytological and histopathological findings, were included in Group 2. Patients with MCT, diagnosed by cytological and histopathological findings, were included in Group 3. Serum was frozen at -80ºC before measuring the concentration of IL-31 via a Simoa ultra-sensitive, fully automated two-step immunoassay. RESULTS: Serum concentrations of IL-31, regardless of the disease and its staging, were within the normal range in all patients; there was no difference between any of the different tumour groups and healthy dogs. CONCLUSIONS AND CLINICAL IMPORTANCE: IL-31 is not likely to be involved in the pathogenesis of canine MCT or lymphoma without pruritus.

Type 2 inhibitor leads of human tropomyosin receptor kinase (hTrkA).

In silico virtual screening using the ligand-based ROCS approach and the commercially purchasable compound collection from the ZINC database resulted in the identification of distinctly different and novel acetamide core frameworks with series representatives 1a and 2a exhibiting nanomolar affinity in the kinase domain only hTrkA HTRF biochemical assay. Additional experimental validation using the Caliper technology with either the active or inactive kinase conditions demonstrated the leads, 1a and 2a, to preferentially bind the kinase inactive state. X-ray structural analysis of the kinase domain of hTrkA…1a/2a complexes confirmed the kinase, bind the inhibitor leads in the inactive state and to exhibit a type 2 binding mode with the DFG-out and αC-helix out conformation. The leads also demonstrated sub-micromolar activity in the full length hTrkA cell-based assay and selectivity against the closely related hTrkB isoform. However, the poor microsomal stability and permeability of the leads is suggestive of a multiparametric lead optimization effort requirement for further progression.

Lead identification and characterization of hTrkA type 2 inhibitors.

Virtual in silico structure-guided modeling, followed by in vitro biochemical screening of a subset of commercially purchasable compound collection resulted in the identification of several human tropomyosin receptor kinase A (hTrkA) inhibitors that bind the orthosteric ATP site and exhibit binding preference for the inactive kinase conformation. The type 2 binding mode with the DFG-out and αC-helix out hTrkA kinase domain conformation was confirmed from X-ray crystallographic solution of a representative inhibitor analog, 1b. Additional hTrkA and hTrkB (selectivity) assays in recombinant cells, neurite outgrowth inhibition using rat PC12 cells, early ADME profiling, and preliminary pharmacokinetic evaluation in rodents guided the lead inhibitor progression in the discovery screening funnel.

IL-31-induced pruritus in dogs: a novel experimental model to evaluate anti-pruritic effects of canine therapeutics.

BACKGROUND: Pruritus is a characteristic clinical sign of allergic skin conditions including atopic dermatitis (AD) in the dog. IL-31 is a cytokine found in the serum of some dogs with AD and can induce pruritic behaviours in laboratory beagle dogs. HYPOTHESIS/OBJECTIVES: The objectives were to characterize an IL-31-induced pruritus model by evaluating the efficacy of prednisolone, dexamethasone and oclacitinib, and to compare the speed of anti-pruritic effects of oclacitinib against those of prednisolone and dexamethasone. ANIMALS: Purpose-bred beagle dogs were used in all studies. METHODS: Randomized, blinded, placebo-controlled studies were designed to evaluate and compare the anti-pruritic properties of prednisolone, dexamethasone and oclacitinib following a single intravenous injection of recombinant canine IL-31. Video surveillance was used to monitor and score pruritic behaviours in study animals. RESULTS: Prednisolone [0.5 mg/kg, per os (p.o.)] reduced IL-31-induced pruritus when given 10 h prior to observation. When the time interval between drug treatment and observation was shortened to 1 h, dexamethasone (0.2 mg/kg, intramuscular) but not prednisolone (0.25 or 0.5 mg/kg, p.o.) reduced IL-31-induced pruritus. Oclacitinib (0.4 mg/kg, p.o.) reduced pruritus when given 1, 6, 11 and 16 h prior to the observation period, and the anti-pruritic activity of oclacitinib was greater when compared to prednisolone and dexamethasone at all time points assessed. CONCLUSION AND CLINICAL IMPORTANCE: The efficacy of prednisolone, dexamethasone and oclacitinib in the IL-31-induced pruritus model gives confidence that this may be a relevant model for acute pruritus associated with allergic dermatitis including AD and can be used to evaluate novel compounds or formulations.

Possible Spreading of SARS-CoV-2 from Humans to Captive Non-Human Primates in the Peruvian Amazon.

Human-to-animal transmission events of SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2) have been reported in both domestic and wild species worldwide. Despite the high rates of contagion and mortality during the COVID-19 (Coronavirus Diseases 2019) pandemic in Peru, no instances of natural virus infection have been documented in wild animals, particularly in the Amazonian regions where human-wildlife interactions are prevalent. In this study, we conducted a surveillance investigation using viral RNA sequencing of fecal samples collected from 76 captive and semi-captive non-human primates (NHPs) in the Loreto, Ucayali, and Madre de Dios regions between August 2022 and February 2023. We detected a segment of the RNA-dependent RNA polymerase (RdRp) gene of SARS-CoV-2 by metagenomic sequencing in a pooled fecal sample from captive white-fronted capuchins (Cebus unicolor) at a rescue center in Bello Horizonte, Ucayali. Phylogenetic analysis further confirmed that the retrieved partial sequence of the RdRp gene matched the SARS-CoV-2 genome. This study represents the first documented instance of molecular SARS-CoV-2 detection in NHPs in the Peruvian Amazon, underscoring the adverse impact of anthropic activities on the human-NHP interface and emphasizing the importance of ongoing surveillance for early detection and prediction of future emergence of new SARS-CoV-2 variants in animals.

Interleukin-31: its role in canine pruritus and naturally occurring canine atopic dermatitis.

BACKGROUND: Interleukin-31 (IL-31) is a member of the gp130/interleukin-6 cytokine family that is produced by cell types such as T helper 2 lymphocytes and cutaneous lymphocyte antigen positive skin homing T cells. When overexpressed in transgenic mice, IL-31 induces severe pruritus, alopecia and skin lesions. In humans, IL-31 serum levels correlate with the severity of atopic dermatitis in adults and children. HYPOTHESIS/OBJECTIVE: To determine the role of IL-31 in canine pruritus and naturally occurring canine atopic dermatitis (AD). ANIMALS: Purpose-bred beagle dogs were used for laboratory studies. Serum samples were obtained from laboratory animals, nondiseased client-owned dogs and client-owned dogs diagnosed with naturally occurring AD. METHODS: Purpose-bred beagle dogs were administered canine interleukin-31 (cIL-31) via several routes (intravenous, subcutaneous or intradermal), and pruritic behaviour was observed/quantified via video monitoring. Quantitative immunoassay techniques were employed to measure serum levels of cIL-31 in dogs. RESULTS: Injection of cIL-31 into laboratory beagle dogs caused transient episodes of pruritic behaviour regardless of the route of administration. When evaluated over a 2 h period, dogs receiving cIL-31 exhibited a significant increase in pruritic behaviour compared with dogs that received placebo. In addition, cIL-31 levels were detectable in 57% of dogs with naturally occurring AD (≥ 13 pg/mL) but were below limits of quantification (<13 pg/mL) in normal, nondiseased laboratory or client-owned animals. CONCLUSIONS: Canine IL-31 induced pruritic behaviours in dogs. Canine IL-31 was detected in the majority of dogs with naturally occurring AD, suggesting that this cytokine may play an important role in pruritic allergic skin conditions, such as atopic dermatitis, in this species.

Oclacitinib 10 years later: lessons learned and directions for the future.

Oclacitinib was approved in the United States 10 years ago for the management of atopic dermatitis (AD) and allergic skin disease in dogs. Many studies and case reports have been published in the past 10 years on the efficacy and safety of this medication, both at labeled doses to treat allergic dogs and off label to treat other diseases and given to other species. Concerns and confusion have occurred for both clinicians and owners regarding the long-term safety of this drug. The purpose of this review is to present the current knowledge on the efficacy, speed of action, effects on the immune system, and clinical safety of oclacitinib, based on evidence and published literature. We also aim to summarize the lessons learned in the past 10 years and to propose directions for the future.

Lack of Interference of Oclacitinib with the results of intradermal testing or allergen-specific IgE serology in Dermatophagoides farinae-sensitized beagle dogs.

Canine atopic dermatitis (AD) is associated with increased levels of allergen-specific IgE due to hyper-sensitization to environmental allergens. Intradermal testing (IDT) and allergen-specific IgE serology testing are often used to determine the allergens which elicit an IgE response in animals with a diagnosis of AD. The objective of this study was to determine the effects of oclacitinib on IDT and allergen-specific IgE serology testing using a laboratory model of house-dust mite sensitized Beagle dogs. Twenty-four (24) normal, healthy purpose-bred Beagle dogs were sensitized to house dust mites (HDM, Dermatophagoides farinae) and randomly assigned to placebo-, oclacitinib- (0.4 mg/kg/dose PO), or prednisolone-treated (0.5 mg/kg/dose PO) groups. After 14 days of twice daily dosing, the effects of prednisolone and oclacitinib were compared to placebo using baseline and post-dose IDT and allergen-specific IgE serum measurements. Sensitized dogs had increased circulating HDM-specific IgE for at least two weeks post-sensitization. Prednisolone significantly inhibited the measurable sensitivity of IDT, while oclacitinib did not. Neither prednisolone nor oclacitinib imposed significant effects on allergen-specific IgE serum levels, suggesting oclacitinib may have potential to be used in dogs concurrently undergoing intradermal skin testing and/or allergen-specific IgE serology testing without interference with test results.

Dynamic monitoring of service members to quantify blast exposure levels during combat training using BlackBox Biometrics Blast Gauges: explosive breaching, shoulder-fired weapons, artillery, mortars, and 0.50 caliber guns.

CONQUER is a pilot blast monitoring program that monitors, quantifies, and reports to military units the training-related blast overpressure exposure of their service members. Overpressure exposure data are collected using the BlackBox Biometrics (B3) Blast Gauge System (BGS, generation 7) sensors mounted on the body during training. To date, the CONQUER program has recorded 450,000 gauge triggers on monitored service members. The subset of data presented here has been collected from 202 service members undergoing training with explosive breaching charges, shoulder-fired weapons, artillery, mortars, and 0.50 caliber guns. Over 12,000 waveforms were recorded by the sensors worn by these subjects. A maximum peak overpressure of 90.3 kPa (13.1 psi) was recorded during shoulder-fired weapon training. The largest overpressure impulse (a measure of blast energy) was 82.0 kPa-ms (11.9 psi-ms) and it was recorded during explosive breaching with a large wall charge. Operators of 0.50 caliber machine guns have the lowest peak overpressure impulse (as low as 0.62 kPa-ms or 0.09 psi-ms) of the blast sources considered. The data provides information on the accumulation of blast overpressure on service members over an extended period of time. The cumulative peak overpressure, peak overpressure impulse, or timing between exposures is all available in the exposure data.

Significant Mitigation of Blast Overpressure Exposure During Training by Adjustment of Body Position as Demonstrated With Field Data.

INTRODUCTION: During training and deployment, service members (SMs) experience blast exposure, which may potentially negatively impact brain health in the short and long term. This article explores if blast exposure mitigation can be effectively achieved for four different weapon training scenarios that are being monitored as part of the CONQUER (COmbat and traiNing QUeryable Exposure/event Repository) program. The training scenarios considered here are a detonating cord linear (det linear) breaching charge, a water breaching charge, a shoulder-fired weapon, and a 120-mm mortar. MATERIALS AND METHODS: This article focuses on the efficacy of modification of position and standoff distance on SMs' exposure to blast overpressure. Blast overpressure exposures were measured using BlackBox Biometrics (B3) Blast Gauge System (BGS) sensors worn by SMs during normal training. The BGS involves the use of three gauges/sensors, which are worn on the head, chest, and nondominant shoulder to record surface pressures at multiple locations on the SM. For the breaching charges, we compared the level of exposure when the SMs were directly in front of the blast with a breaching blanket to a modified standoff position around a corner from the charge without a breaching blanket. For the shoulder-fired weapon training, the modified approach simply increased the standoff distance of the SM. Finally, for mortars, blast overpressure exposures were compared for different levels of their ducking height (body position) below the mortar tube at the time of firing. RESULTS: Modification of the position of SMs during training with the det linear breaching charge had the highest measured blast exposure percent reduction, at 79%. Both the water breaching charge and shoulder-fired weapon showed lowered peak overpressures on all gauges. The measured percent reduction for the 120-mm mortar was 35%. When the blast gauges did not trigger at the modified standoff distance, the percent reduction was calculated with the assumption that the new overpressures were below ∼3.4 kPa (0.5 psi) (the lowest trigger threshold for the gauges). A figure summarizes the percent reduction for each subject in the training scenarios. CONCLUSIONS: Results show that the modification of the SMs' position effectively mitigated blast exposures for all considered weapon scenarios. There was at least a 50% overpressure reduction from the initial to modified standoff distances and a 35% reduction from the change in SM body posture. Based on these observations, new locations and body positioning of SMs during training have been suggested for blast mitigation.

Laboratory safety evaluation of lokivetmab, a canine anti-interleukin-31 monoclonal antibody, in dogs.

Lokivetmab (Cytopoint®, Zoetis) is a canine monoclonal antibody that specifically binds and neutralizes interleukin (IL)-31. Lokivetmab is approved for use in dogs for the treatment of atopic dermatitis (AD) and allergic dermatitis. The laboratory safety of lokivetmab was evaluated in 2 studies by adapting the science-based, case-by-case approach used for preclinical and early clinical safety evaluation of human biopharmaceuticals. The main objectives were to demonstrate the safety of lokivetmab in healthy laboratory Beagle dogs by using integrated clinical, morphologic, and functional evaluations. In Study 1, dogs were treated s.c. with saline or lokivetmab at 3.3 mg/kg (1X, label dose) or 10 mg/kg (3X intended dose) for 7 consecutive monthly doses, with terminal pathology and histology assessments. In Study 2, the functional immune response was demonstrated in naïve dogs using the T-cell dependent antibody response (TDAR) test with 2 different dose levels of unadjuvanted keyhole limpet hemocyanin (KLH) as the model immunogen. The primary endpoint was anti-KLH IgG antibody titer, and secondary endpoints were ex vivo IL-2 enzyme-linked immunospot (ELISpot) and peripheral blood mononuclear cell lymphoproliferation assays. Both studies included monitoring general health, periodic veterinary clinical evaluations, serial clinical pathology and toxicokinetics, and monitoring for anti-drug antibodies. In both studies, the health of dogs receiving lokivetmab was similar to controls, with no treatment-related changes uncovered. Extensive pathology evaluations of immune tissues (Study 1) revealed no lokivetmab-related morphologic changes, and in dogs treated at 10 mg/kg lokivetmab, immunization with the model antigen KLH did not impair the functional antibody or T-cell recall responses. There were no immunogenicity-related or hypersensitivity-related responses observed in either study. These studies in healthy laboratory dogs showed that lokivetmab was well-tolerated, did not produce any treatment-related effects, and had no effect on immune system morphology or its functional response. These studies also demonstrated the utility of a science-based case-by-case approach to the safety evaluation of a veterinary biopharmaceutical product.

Development of a Fast-Running Algorithm to Approximate Incident Blast Parameters Using Body-Mounted Sensor Measurements.

INTRODUCTION: The Office of Naval Research sponsored the Blast Load Assessment-Sense and Test program to develop a rapid, in-field solution that could be used by team leaders, commanders, and medical personnel to make science-based stand-down decisions for service members exposed to blast overpressure. However, a critical challenge to this goal was the reliable interpretation of surface pressure data collected by body-worn blast sensors in both combat and combat training scenarios. Without an appropriate standardized metric, exposures from different blast events cannot be compared and accumulated in a service member's unique blast exposure profile. In response to these challenges, we developed the Fast Automated Signal Transformation, or FAST, algorithm to automate the processing of large amounts of pressure-time data collected by blast sensors and provide a rapid, reliable approximation of the incident blast parameters without user intervention. This paper describes the performance of the FAST algorithms developed to approximate incident blast metrics from high-explosive sources using only data from body-mounted blast sensors. METHODS AND MATERIALS: Incident pressure was chosen as the standardized output metric because it provides a physiologically relevant estimate of the exposure to blast that can be compared across multiple events. In addition, incident pressure serves as an ideal metric because it is not directionally dependent or affected by the orientation of the operator. The FAST algorithms also preprocess data and automatically flag "not real" traces that might not be from blasts events (false positives). Elimination of any "not real" blast waveforms is essential to avoid skewing the results of subsequent analyses. To evaluate the performance of the FAST algorithms, the FAST results were compared to (1) experimentally measured pressures and (2) results from high-fidelity numerical simulations for three representative real-world events. RESULTS: The FAST results were in good agreement with both experimental data and high-fidelity simulations for the three case studies analyzed. The first case study evaluated the performance of FAST with respect to body shielding. The predicted incident pressure by FAST for a surrogate facing the charge, side on to charge, and facing away from the charge was examined. The second case study evaluated the performance of FAST with respect to an irregular charge compared to both pressure probes and results from high-fidelity simulations. The third case study demonstrated the utility of FAST for detonations inside structures where reflections from nearby surfaces can significantly alter the incident pressure. Overall, FAST predictions accounted for the reflections, providing a pressure estimate typically within 20% of the anticipated value. CONCLUSIONS: This paper presents a standardized approach-the FAST algorithms-to analyze body-mounted blast sensor data. FAST algorithms account for the effects of shock interactions with the body to produce an estimate of incident blast conditions, allowing for direct comparison of individual exposure from different blast events. The continuing development of FAST algorithms will include heavy weapons, providing a singular capability to rapidly interpret body-worn sensor data, and provide standard output for analysis of an individual's unique blast exposure profile.

In Pursuit of an Allosteric Human Tropomyosin Kinase A (hTrkA) Inhibitor for Chronic Pain.

Human ß-nerve growth factor (ß-NGF) and its associated receptor, human tropomyosin receptor kinase A (hTrkA), have been demonstrated to be key factors in the perception of pain. However, efficacious small molecule therapies targeting the intracellularly located hTrkA kinase have not been explored thoroughly for pain management. Herein, we report the pharmacological properties of a selective hTrkA allosteric inhibitor, 1. 1 was shown to be active against the full length hTrkA, showing preferential binding for the inactive kinase, and was confirmed through the X-ray of hTrkA···1 bound complex. 1 was also found to inhibit ß-NGF induced neurite outgrowth in rat PC12 cells. Daily oral administration of 1 improved the joint compression threshold of rats injected intra-articularly with monoiodoacetate over a 14-day period. The efficacy of 1 in a relevant chronic pain model of osteoarthritis coupled with in vitro confirmation of target mediation makes allosteric hTrkA inhibitors potential candidates for modulating pain.