No correlation found between palpation and ultrasound for evaluation of effusion in the medial femorotibial and femoropatellar joint compartments of horses.

OBJECTIVE: To compare palpation and ultrasound scores of effusion of the medial femorotibial and femoropatellar joints of horses. ANIMALS: 40 horses (80 stifles) were evaluated over a 12-week period. METHODS: Horses > 1 year of age without history of stifle disease were enrolled from September to December 2022. Palpation of right and left medial femorotibial and femoropatellar joint compartments was performed. Amount of effusion was scored by a board-certified large animal surgeon, a third-year large animal surgery resident, and an equine sports medicine intern. Effusion of right and left medial femorotibial and femoropatellar joints was quantified with ultrasound by a board-certified equine sports medicine and rehabilitation clinician. Amount of effusion on palpation and ultrasound was graded as none-mild (1), moderate (2), or severe (3). A 2-way intraclass correlation coefficient evaluated interrater reliability of palpation scores. The Spearman rank correlation determined association between palpation and ultrasound scores. RESULTS: Interrater reliability for palpation of effusion was poor between all observers for all joint compartments. No significant correlation was identified between palpation and ultrasound scores for any joint compartment for any observer. CLINICAL RELEVANCE: Clinicians often rely on palpation of joint effusion as an indication of stifle pathology. We found interrater reliability to be poor for palpation scores, indicating low agreement for palpation of joint effusion between clinicians within our group. No correlation was found between palpation and ultrasound scores for joint effusion, indicating that clinicians should not rely on palpation alone to quantify joint effusion of the medial femorotibial and femoropatellar joints.

Histologic characterization of the major duodenal papilla and association with concurrent biliary, pancreatic, and intestinal pathology in cats.

Conjoining of the major pancreatic duct and common bile duct at the major duodenal papilla (MDP) is suspected to predispose cats to the clinical syndrome of "triaditis." However, microanatomy of the MDP or presence of lesions at the MDP has not been assessed in cats with or without triaditis. The aims of this study were to characterize feline MDP histomorphology and to identify associations between MDP anatomy/disease and the presence of biliary, pancreatic, or intestinal inflammation or neoplasia. Histologic assessment was prospectively performed on the MDP, duodenum, jejunum, ileum, liver, and pancreas from 124 client-owned cats undergoing postmortem examination. The majority of cats (104/124, 84%) had a complex ductular network at the MDP, with no distinction between pancreatic and common bile ducts. Lymphoid aggregates at the MDP were common (63/124, 51%). Inflammation of the MDP (MDPitis) was present in 35 of 124 cats (28%) and was often concurrent with cholangitis, pancreatitis, or enteritis (32/35, 91%), but was only associated with enteritis (19/35, 54%, P < .05). Triaditis was less common (19/124, 15%), but was associated with both conjoined MDP anatomy (19/19, 100%, P < .05) and MDPitis (12/19, 63%, P < .05). Neoplasia was present in 37 of 124 cats (29%), with lymphoma (28/37, 78%) predominating. Enteropathy-associated T-cell lymphoma type 2 (EATL2) was most common (n = 16/37, 43%) and was associated with triaditis and MDPitis (P < .05). These findings suggest that anatomy, immune activation, and/or inflammation of the MDP may play a role in the pathogenesis of triaditis. Further studies are needed to elucidate the relationships between triaditis, MDPitis, and EATL2.

Ex vivo analysis of ultraviolet radiation transmission through ocular media and retina in select species.

The aim of this study was to assess the transmission of the ultraviolet (UV) radiation (200-400 nm) through intact enucleated globes of different species (dogs, cats, pigs, rabbits, horses, and humans) using spectrophotometry. Globes of cats (n = 6), dogs (n = 18), pigs (n = 10), rabbits (n = 6), horses (n = 10), and humans (n = 4) were analyzed. A 5-10 mm circular area of sclera and choroid from the posterior aspect of the globe was removed under a surgical microscope, leaving the retina intact in all species except the horse. Glass coverslips were added in horses and rabbits due to retinal and globe fragility. The %T of wavelengths from 200 to 800 nm were measured through the ocular media (cornea, aqueous humor, lens, and vitreous humor) and retina, and compared between species. The globes of cats and dogs allowed the most amount of UV radiation transmission, while those of pigs and humans allowed the least amount of UV radiation transmission. A small amount of UV radiation transmission through the ocular media was detected in the rabbit and horse. Results from this study will support further vision research that may be used to train companion, working, and service animals.

Whole exome sequencing analysis of canine urothelial carcinomas without BRAF V595E mutation: Short in-frame deletions in BRAF and MAP2K1 suggest alternative mechanisms for MAPK pathway disruption.

Molecular profiling studies have shown that 85% of canine urothelial carcinomas (UC) harbor an activating BRAF V595E mutation, which is orthologous to the V600E variant found in several human cancer subtypes. In dogs, this mutation provides both a powerful diagnostic marker and a potential therapeutic target; however, due to their relative infrequency, the remaining 15% of cases remain understudied at the molecular level. We performed whole exome sequencing analysis of 28 canine urine sediments exhibiting the characteristic DNA copy number signatures of canine UC, in which the BRAF V595E mutation was undetected (UDV595E specimens). Among these we identified 13 specimens (46%) harboring short in-frame deletions within either BRAF exon 12 (7/28 cases) or MAP2K1 exons 2 or 3 (6/28 cases). Orthologous variants occur in several human cancer subtypes and confer structural changes to the protein product that are predictive of response to different classes of small molecule MAPK pathway inhibitors. DNA damage response and repair genes, and chromatin modifiers were also recurrently mutated in UDV595E specimens, as were genes that are positive predictors of immunotherapy response in human cancers. Our findings suggest that short in-frame deletions within BRAF exon 12 and MAP2K1 exons 2 and 3 in UDV595E cases are alternative MAPK-pathway activating events that may have significant therapeutic implications for selecting first-line treatment for canine UC. We developed a simple, cost-effective capillary electrophoresis genotyping assay for detection of these deletions in parallel with the BRAF V595E mutation. The identification of these deletion events in dogs offers a compelling cross-species platform in which to study the relationship between somatic alteration, protein conformation, and therapeutic sensitivity.

Hurdles of Cataract Surgery: Veterinary Ophthalmology Resident's Perspective (Part B).

The purpose of this study was to describe veterinary ophthalmology residents' perceived preparedness for performing cataract surgery who are currently enrolled in, or recently graduated from, veterinary academic or private practice institutions. A descriptive survey was distributed online to 127 residents at academic and private practice training programs in the United States. The survey included items about educational resources available for residents and techniques commonly taught during cataract surgery. Residents were asked to describe their perceived preparedness in performing various surgical steps or techniques, difficulty of each surgical step, and the available educational resources. Thirty-five (27.5%) residents completed the survey and were included in this study. Residents who had access to wet labs gained surgical competency in creating a clear corneal incision, capsulorhexis, and wound closure. They reported sculpting with the phacoemulsification handpiece, quadrant or cortical removal, and capsulorhexis as most difficult and were not as prepared or a little prepared in performing capsulorhexis and sculpting during active phacoemulsification. When comparing residents' perceived competency before and after their first surgical experience, there was a significant change in their ability to perform all surgical steps except hydrodissection (p < .05). Cataract surgery is one of the more advanced surgical skills obtained during residency training. Supervised wet lab time improves a resident's preparedness for executing certain surgical steps. However, further research is needed to determine whether educational resources such as structured curriculum or virtual simulation may improve residents' preparedness for executing surgical steps not easily replicated in a wet lab.

Equine pectinate ligament descemetization is associated with age.

PURPOSE: To evaluate the correlation between equine pectinate ligament descemetization and ocular disease. METHODS: The pathology database of the North Carolina State University Veterinary Medical Center was searched from 2010-2021 for all equine globes. Disease status was then assigned as affected by glaucoma, uveitis, or "other" based upon clinical records. The iridocorneal angles (ICA) of each globe were evaluated for the presence of pectinate ligament descemetization, the length of descemetization, as well as for the degree of angle collapse and the extent of cellular infiltrate or proteinaceous debris. One slide from each eye was evaluated by two separate, blinded investigators (HW & TS). RESULTS: A total of 66 eyes from 61 horses were identified, with a total of 124 sections of ICA of sufficient quality to review. 16 horses were affected by uveitis, 8 by glaucoma, 7 by both glaucoma and uveitis, and 30 horses by other ocular disease, most commonly ocular surface disease or neoplasia, which served as controls. Pectinate ligament descemetization was most prevalent in the control group compared to the glaucoma and uveitis groups. Pectinate ligament descemetization length was positively correlated with age, with an increase of 13.5 µm per year of age (p = .016). Infiltrate scores and angle closure scores were higher in both the glaucoma and uveitis group compared to the control group (p < .001). CONCLUSIONS: Equine pectinate ligament descemetization appears to be correlated with increased age and should not be used as a histologic marker for the presence of glaucoma.

Effects of Intramuscular Alfaxalone and Midazolam Compared With Midazolam and Butorphanol in Rhode Island Red Hens (Gallus gallus domesticus).

Chickens (Gallus gallus domesticus) often undergo veterinary procedures requiring sedation; however, there is little published research evaluating the efficacy of sedation protocols in this species. The objective of this study was to assess the effects of intramuscular alfaxalone and midazolam compared with intramuscular butorphanol and midazolam in chickens. In a complete crossover study, 11 healthy adult hens were randomly administered midazolam 2.5 mg/kg IM combined with either alfaxalone 15 mg/kg IM (AM, n = 11) or butorphanol 3 mg/kg IM (BM, n = 11), with a 35-day washout period between groups. Time to first effects, recumbency, standing, and recovery were recorded. Physiologic parameters and sedation scores were recorded every 5 minutes by 2 blinded investigators. Fifteen minutes after injection, positioning for sham whole body radiographs was attempted. At 30 minutes, flumazenil 0.05 mg/kg IM was administered to all hens. Peak total sedation score was significantly higher for AM compared with BM (P < 0.001). Mean ± SD or median (range) time to initial effects, recumbency, standing, and recovery in AM and BM were 1.9 ± 0.6 and 2.6 ± 0.9 (P = 0.02), 3.5 (1.6-7.6) and 4.8 (2.2-13.0) (P = 0.10), 40.3 (28.0-77.8) and 33.2 (5.2-41.3) (P = 0.15), and 71.2 (45.7-202.3) and 39.9 (35.9-45.9) minutes (P = 0.05), respectively. Radiographic positioning was successful in 6 of 11 (54.5%) and 0 of 11 (0%) birds in the AM and BM groups at 15 minutes, respectively. Heart and respiratory rates remained within acceptable clinical limits for all birds. Intramuscular AM resulted in significantly faster onset of sedative effects, significantly longer duration of recumbency, significantly higher peak sedation, and improved success of radiographic positioning compared with intramuscular BM. Intramuscular AM produces clinically effective sedation in chickens without clinically significant cardiorespiratory effects.

Bilateral lysis of aortic saddle thrombus with early tissue plasminogen activator (BLASTT): a prospective, randomized, placebo-controlled study in feline acute aortic thromboembolism.

OBJECTIVES: The aim of this study was to investigate the impact of tissue plasminogen activator (TPA) on the treatment of feline aortic thromboembolism (FATE). METHODS: Cats diagnosed with FATE involving ⩾2 limbs were enrolled in a prospective, multicenter, double-blinded, randomized, placebo-controlled study within 6 h of an event. Diagnosis was made by clinical findings and one confirmatory criterion. Cats received placebo or TPA (1 mg/kg/h with the first 10% by bolus). All cats received pain control and thromboprophylaxis. The primary outcome was a change from baseline in a published limb score at 48 h. Secondary outcomes included 48 h survival, survival to discharge and complication proportions. Statistical analyses included pattern-mixture models, logistic regression and Fisher's exact, Student's t- and Mann-Whitney-Wilcoxon tests. RESULTS: Based on a power analysis, 40 cats were enrolled; however, only 20 survived to 48 h (TPA, n = 12; placebo, n = 8 [P = 0.34]). There was a statistically significant improvement in limb scores compared with baseline for both groups (P <0.001). Limb score at 48 h was 1 point lower (better) in the TPA group (P = 0.19). Thrombolysis had no statistically significant effect on 48 h survival (P = 0.22). Lower affected limb lactate was associated with better 48 h survival (odds ratio 1.53, 95% confidence interval 1.08-2.17; P = 0.02). The survival to discharge rates were 45% (TPA) and 30% (placebo; P = 0.51). Complications in the TPA and placebo groups included acute kidney injury (22% and 19%, respectively; P = 1.00) and/or reperfusion injuries (33% and 19%, respectively; P = 0.45). CONCLUSIONS AND RELEVANCE: Survival and complication rates of acute FATE were not different with or without thrombolysis. High in-hospital mortality decreased the statistical power to detect a statistically significant difference between treatments with regard to our primary outcome.

Point-of-care viscoelastic coagulation assessment in healthy dogs during the perianesthetic period.

BACKGROUND: The viscoelastic coagulation monitor (VCM Vet) is a novel, portable device that provides a global assessment of hemostasis. The study aims were to evaluate serial viscoelastic analysis during the perianesthetic period in healthy dogs and to compare the agreement between two VCM Vet devices. Twenty healthy dogs undergoing orthopedic surgery were enrolled. Whole blood samples were collected from an intravenous catheter at four time points: baseline, 15 min after premedication, 60 min after inhalant initiation, and 60 min after inhalant termination. Viscoelastic tests were performed in duplicate on different devices, providing: clot time (CT; seconds), clot formation time (CFT; seconds), alpha angle (α; degrees), amplitude (units) at 10 (A10) and 20 (A20) minutes post clot time, maximum clot firmness (MCF; units), and lysis index (%) at 30 (Li30) and 45 (Li45) minutes post maximum clot formation. RESULTS: One hundred sixty samples were analyzed. The speed of CT and CFT significantly decreased an average of 25.5 s (95% confidence interval [CI]15.9-35.0) and 6.9 s (95% CI 3.1-10.7) per time point, respectively. There were no significant changes in clot strength or lysis variables. The Bland-Altman style plot shows an acceptable rate of agreement for all variables with intra-class correlation ranging from 0.64-0.94. CONCLUSION: The rate of clot formation (CT and CFT) decreased over the perianesthetic period in healthy dogs undergoing surgery. These changes were small and occurred without changes in clot strength or fibrinolysis rate, thus were not clinically relevant. There was clinically acceptable consistency between devices.

Vaccines based on the replication-deficient simian adenoviral vector ChAdOx1: Standardized template with key considerations for a risk/benefit assessment.

Replication-deficient adenoviral vectors have been under investigation as a platform technology for vaccine development for several years and have recently been successfully deployed as an effective COVID-19 counter measure. A replication-deficient adenoviral vector based on the simian adenovirus type Y25 and named ChAdOx1 has been evaluated in several clinical trials since 2012. The Brighton Collaboration Benefit-Risk Assessment of VAccines by TechnolOgy (BRAVATO) was formed to evaluate the safety and other key features of new platform technology vaccines. This manuscript reviews key features of the ChAdOx1-vectored vaccines. The simian adenovirus Y25 was chosen as a strategy to circumvent pre-existing immunity to common human adenovirus serotypes which could impair immune responses induced by adenoviral vectored vaccines. Deletion of the E1 gene renders the ChAdOx1 vector replication incompetent and further genetic engineering of the E3 and E4 genes allows for increased insertional capability and optimizes vaccine manufacturing processes. ChAdOx1 vectored vaccines can be manufactured in E1 complementing cell lines at scale and are thermostable. The first ChAdOx1 vectored vaccines approved for human use, against SARS-CoV-2, received emergency use authorization in the UK on 30th December 2020, and is now approved in more than 180 countries. Safety data were compiled from phase I-III clinical trials of ChAdOx1 vectored vaccines expressing different antigens (influenza, tuberculosis, malaria, meningococcal B, prostate cancer, MERS-CoV, Chikungunya, Zika and SARS-CoV-2), conducted by the University of Oxford, as well as post marketing surveillance data for the COVID-19 Oxford-AstraZeneca vaccine. Overall, ChAdOx1 vectored vaccines have been well tolerated. Very rarely, thrombosis with thrombocytopenia syndrome (TTS), capillary leak syndrome (CLS), immune thrombocytopenia (ITP), and Guillain-Barre syndrome (GBS) have been reported following mass administration of the COVID-19 Oxford-AstraZeneca vaccine. The benefits of this COVID-19 vaccination have outweighed the risks of serious adverse events in most settings, especially with mitigation of risks when possible. Extensive immunogenicity clinical evaluation of ChAdOx1 vectored vaccines reveal strong, durable humoral and cellular immune responses to date; studies to refine the COVID-19 protection (e.g., via homologous/heterologous booster, fractional dose) are also underway. New prophylactic and therapeutic vaccines based on the ChAdOx1 vector are currently undergoing pre-clinical and clinical assessment, including vaccines against viral hemorrhagic fevers, Nipah virus, HIV, Hepatitis B, amongst others.

Refinement of the Feline Musculoskeletal Pain Index (FMPI) and development of the short-form FMPI.

OBJECTIVES: The aim of this study was to investigate the reliability and responsiveness of the Feline Musculoskeletal Pain Index (FMPI) using the collective results of multiple clinical studies and iteratively refine the FMPI for future use. METHODS: Data were compiled from previously conducted studies involving client-owned cats with degenerative joint disease (DJD) and which used the FMPI. The reliability of the FMPI was assessed using the data from the initial visits of those studies. For the assessment of responsiveness of the FMPI, only placebo-controlled studies that used analgesic treatments were included. Treatment groups from each study were combined and categorized as 'placebo' group and 'analgesic' group. Then, the mean change from baseline in score of each FMPI item and across all items within and between these groups were assessed. Based on the results of the reliability and responsiveness of the FMPI, stepwise elimination was used to remove the items that were least able to distinguish between the placebo and analgesic groups. Finally, after the stepwise elimination, a proposed new FMPI-short form (FMPI-sf) was constructed and its reliability was reassessed using the data sets described above. Individual and combined data sets of the studies were also used to compare the responsiveness of the original FMPI and the FMPI-sf. RESULTS: The data from 180 cats from four studies were included. The original FMPI had a reasonable reliability, but low/no responsiveness. The elimination process of FMPI items refined the responsiveness of the instrument while maintaining its reliability. When the responsiveness was compared between the original FMPI (17 items) and the FMPI-sf (nine items), the treatment effect between groups was always greater when the FMPI-sf was used. CONCLUSIONS AND RELEVANCE: The proposed FMPI-sf may be able to better distinguish between placebo and analgesic effects in cats with DJD.

HOPX+ injury-resistant intestinal stem cells drive epithelial recovery after severe intestinal ischemia.

Intestinal ischemia is a life-threatening emergency with mortality rates of 50%-80% due to epithelial cell death and resultant barrier loss. Loss of the epithelial barrier occurs in conditions including intestinal volvulus and neonatal necrotizing enterocolitis. Survival depends on effective epithelial repair; crypt-based intestinal epithelial stem cells (ISCs) are the source of epithelial renewal in homeostasis and after injury. Two ISC populations have been described: 1) active ISC [aISC; highly proliferative; leucine-rich-repeat-containing G protein-coupled receptor 5 (LGR5+)-positive or sex-determining region Y-box 9 -antigen Ki67-positive (SOX9+Ki67+)] and 2) reserve ISC [rISC; less proliferative; homeodomain-only protein X positive (HOPX+)]. The contributions of these ISCs have been evaluated both in vivo and in vitro using a porcine model of mesenteric vascular occlusion to understand mechanisms that modulate ISC recovery responses following ischemic injury. In our previously published work, we observed that rISC conversion to an activated state was associated with decreased HOPX expression during in vitro recovery. In the present study, we wanted to evaluate the direct role of HOPX on cellular proliferation during recovery after injury. Our data demonstrated that during early in vivo recovery, injury-resistant HOPX+ cells maintain quiescence. Subsequent early regeneration within the intestinal crypt occurs around 2 days after injury, a period in which HOPX expression decreased. When HOPX was silenced in vitro, cellular proliferation of injured cells was promoted during recovery. This suggests that HOPX may serve a functional role in ISC-mediated regeneration after injury and could be a target to control ISC proliferation.NEW & NOTEWORTHY This paper supports that rISCs are resistant to ischemic injury and likely an important source of cellular renewal following near-complete epithelial loss. Furthermore, we have evidence that HOPX controls ISC activity state and may be a critical signaling pathway during ISC-mediated repair. Finally, we use multiple novel methods to evaluate ISCs in a translationally relevant large animal model of severe intestinal injury and provide evidence for the potential role of rISCs as therapeutic targets.

Irradiation of the Normal Murine Tongue Causes Upregulation and Activation of Transient Receptor Potential (TRP) Ion Channels.

Signal transduction at sensory neurons occurs via transmembrane flux of cations, which is largely governed by the transient receptor potential (TRP) family of ion channels. It is unknown whether TRP channel activation contributes to the pain that accompanies radiation-induced oral mucositis. This study sought to characterize changes in TRP channel expression and function that occur in the locally irradiated tissues and afferent neurons of mice. Female CD-1 mice received single high-dose (27 Gy) tongue irradiation, or sham irradiation. Animals were euthanized either before overt glossitis developed (days 1 and 5 postirradiation), when glossitis was severe (day 11), or after mice had recovered (days 21 and 45). Tongue irradiation caused upregulation of the Trpv1 gene in trigeminal ganglia (TG) neurons. Other TRP genes (Trpv2, Trpv4, Trpa1, Trpm8) and Gfrα3 (which acts upstream of several TRP channels) were also upregulated in TGs and/or tongue tissue, in response to radiation. Ex vivo calcium imaging experiments demonstrated that the proportions of TG neurons responding to histamine (an activator of TRPV1, TRPV4 and TRPA1), TNF-α (an activator of TRPV1, TRPV2 and TRPV4), and capsaicin (a TRPV1 agonist), were increased as early as one day after tongue irradiation; these changes persisted for at least 21 days. In a subsequent experiment, we found that genetic deletion of TRPV1 mitigated weight loss (a surrogate marker of pain severity) in mice with severe glossitis. The results intimate that various TRP channels, and TRPV1 in particular, should be explored as analgesic targets for patients experiencing pain after oral irradiation.

Leveraging the potential of nature to meet net zero greenhouse gas emissions in Washington State.

The State of Washington, USA, has set a goal to reach net zero greenhouse gas emissions by 2050, the year around which the Intergovernmental Panel on Climate Change (IPCC) recommended we must limit global warming to 1.5 °C above that of pre-industrial times or face catastrophic changes. We employed existing approaches to calculate the potential for a suite of Natural Climate Solution (NCS) pathways to reduce Washington's net emissions under three implementation scenarios: Limited, Moderate, and Ambitious. We found that NCS could reduce emissions between 4.3 and 8.8 MMT CO2eyr-1 in thirty-one years, accounting for 4% to 9% of the State's net zero goal. These potential reductions largely rely on changing forest management practices on portions of private and public timber lands. We also mapped the distribution of each pathway's Ambitious potential emissions reductions by county, revealing spatial clustering of high potential reductions in three regions closely tied to major business sectors: private industrial forestry in southwestern coastal forests, cropland agriculture in the Columbia Basin, and urban and rural development in the Puget Trough. Overall, potential emissions reductions are provided largely by a single pathway, Extended Timber Harvest Rotations, which mostly clusters in southwestern counties. However, mapping distribution of each of the other pathways reveals wider distribution of each pathway's unique geographic relevance to support fair, just, and efficient deployment. Although the relative potential for a single pathway to contribute to statewide emissions reductions may be small, they could provide co-benefits to people, communities, economies, and nature for adaptation and resiliency across the state.

Vaccines based on replication incompetent Ad26 viral vectors: Standardized template with key considerations for a risk/benefit assessment.

Replication-incompetent adenoviral vectors have been under investigation as a platform to carry a variety of transgenes, and express them as a basis for vaccine development. A replication-incompetent adenoviral vector based on human adenovirus type 26 (Ad26) has been evaluated in several clinical trials. The Brighton Collaboration Viral Vector Vaccines Safety Working Group (V3SWG) was formed to evaluate the safety and features of recombinant viral vector vaccines. This paper reviews features of the Ad26 vectors, including tabulation of safety and risk assessment characteristics of Ad26-based vaccines. In the Ad26 vector, deletion of E1 gene rendering the vector replication incompetent is combined with additional genetic engineering for vaccine manufacturability and transgene expression optimization. These vaccines can be manufactured in mammalian cell lines at scale providing an effective, flexible system for high-yield manufacturing. Ad26 vector vaccines have favorable thermostability profiles, compatible with vaccine supply chains. Safety data are compiled in the Ad26 vaccine safety database version 4.0, with unblinded data from 23 ongoing and completed clinical studies for 3912 participants in five different Ad26-based vaccine programs. Overall, Ad26-based vaccines have been well tolerated, with no significant safety issues identified. Evaluation of Ad26-based vaccines is continuing, with >114,000 participants vaccinated as of 4th September 2020. Extensive evaluation of immunogenicity in humans shows strong, durable humoral and cellular immune responses. Clinical trials have not revealed impact of pre-existing immunity to Ad26 on vaccine immunogenicity, even in the presence of Ad26 neutralizing antibody titers or Ad26-targeting T cell responses at baseline. The first Ad26-based vaccine, against Ebola virus, received marketing authorization from EC on 1st July 2020, as part of the Ad26.ZEBOV, MVA-BN-Filo vaccine regimen. New developments based on Ad26 vectors are underway, including a COVID-19 vaccine, which is currently in phase 3 of clinical evaluation.

TGF-ß2 Reduces the Cell-Mediated Immunogenicity of Equine MHC-Mismatched Bone Marrow-Derived Mesenchymal Stem Cells Without Altering Immunomodulatory Properties.

Allogeneic mesenchymal stem cells (MSCs) are a promising cell therapy for treating numerous diseases, but major histocompatibility complex (MHC)-mismatched MSCs can be rejected by the recipient's immune system. Pre-treating MSCs with transforming growth factor-ß2 (TGF-ß2) to downregulate surface expression of MHC molecules may enhance the ability of allogeneic MSCs to evade immune responses. We used lymphocyte proliferation assays and ELISAs to analyze the immunomodulatory potential of TGF-ß2-treated equine bone marrow-derived MSCs. T cell activation and cytotoxicity assays were then used to measure the in vitro cell-mediated immunogenicity. Similar to untreated MSCs, TGF-ß2-treated MSCs inhibited T cell proliferation and did not stimulate MHC-mismatched T cells to proliferate. Additionally, similar quantities of prostaglandin E2 and TGF-ß1 were detected in assays with untreated and TGF-ß2-treated MSCs supporting that TGF-ß2-treated MSCs retain their strong immunomodulatory properties in vitro. Compared to untreated MSCs, TGF-ß2-treated MSCs induced less T cell activation and had reduced cell-mediated cytotoxicity in vitro. These results indicate that treating MSCs with TGF-ß2 is a promising strategy to reduce the cell-mediated immunogenicity of MHC-mismatched MSCs and facilitate allogeneic MSC therapy.

Outcomes and prognostic indicators in 59 paraplegic medium to large breed dogs with extensive epidural hemorrhage secondary to thoracolumbar disc extrusion.

OBJECTIVE: To evaluate outcomes and prognostic factors after decompressive hemilaminectomy in paraplegic medium to large breed dogs with extensive epidural hemorrhage (DEEH) and thoracolumbar intervertebral disc extrusion (TL-IVDE). STUDY DESIGN: Retrospective, cohort, descriptive study. ANIMALS: Fifty-nine client-owned dogs. METHODS: Medical records and advanced imaging were reviewed for paraplegic dogs with DEEH. Ambulatory status 6 months after surgery and postoperative complications were recorded. Multiple logistic regression models were constructed to explore prognostic factors. RESULTS: Records of 22 dogs with and 37 dogs without pelvic limb pain perception at presentation were included. Median age of dogs was 5 years (interquartile range, 4-7), and mean weight was 26.9 kg (SD, ±9.71). Labradors and Labrador mixes were most common (17/59 [28.8%]). Recovery of ambulation occurred in 17 of 22 (77.3%) dogs with and in 14 of 37 (37.8%) dogs without pain perception prior to surgery. Progressive myelomalacia was recorded in three of 59 (5.1%) dogs, one with pain perception and two without pain perception at presentation. Postoperative complications (14/59 [23.7%]) were common. Factors independently associated with outcome included clinical severity (odds ratio [OR] 0.179, P = .005), number of vertebrae with signal interruption in half Fourier single-shot turbo spin-echo sequences (HASTEi; OR, 0.738; P = .035), and ratio of vertebral sites decompressed to HASTEi (OR, 53.79; P = .03). CONCLUSION: Paraplegic medium to large breed dogs with DEEH have a less favorable outcome after surgical decompression than paraplegic dogs with TL-IVDE. CLINICAL SIGNIFICANCE: Dogs with DEEH can have severe postoperative complications. Loss of pain perception and increased HASTEi are associated with a poor outcome, while more extensive decompression improves outcome.

Proteinuria in dogs with gallbladder mucocele formation: A retrospective case control study.

BACKGROUND: Proteinuria is an independent risk factor for morbidity and mortality in dogs. An association between proteinuria and gallbladder mucocele formation in dogs is unknown. OBJECTIVE: Determine if gallbladder mucocele formation or clinicopathologic comorbidities are associated with proteinuria. ANIMALS: Twenty-five dogs with mucocele formation and 25 breed and age-matched control dogs from a prior study. METHODS: Retrospective case control study. Proteinuria defined by calculated urine dipstick protein concentration (mg/mL) to urine specific gravity (USG) ratio. Clinicopathologic findings, postcosyntropin cortisol concentration, thyroid function profile, and illness severity score were recorded. RESULTS: Median urine dipstick protein concentration to USG ratio and number of dogs having a ratio ≥1.5 were significantly higher for dogs with mucocele formation compared to control dogs. Proteinuria was not significantly associated with CBC or serum biochemistry profile abnormalities but increased in relation to severity of illness. CONCLUSIONS AND CLINICAL IMPORTANCE: Gallbladder mucocele formation is significantly associated with proteinuria in dogs. Diagnosis and treatment of proteinuria in dogs with mucocele formation might minimize long term kidney morbidity in these patients.

Neutropenia in dogs receiving vincristine for treatment of presumptive immune-mediated thrombocytopenia.

BACKGROUND: Neutropenia is an adverse effect of vincristine when used in multidrug chemotherapy protocols. OBJECTIVE: To determine the incidence of neutropenia, identify potential risk factors for neutropenia, and determine the effect of neutropenia on outcome, in dogs receiving vincristine for treatment of immune-mediated thrombocytopenia (ITP). ANIMALS: One hundred twenty-seven client-owned dogs presumptively diagnosed with ITP. METHODS: In this retrospective cohort study, medical records were reviewed to identify dogs presumptively diagnosed with ITP, and treated with vincristine, over a 15-year period. Logistic regression was used to identify risk factors for the development of neutropenia in dogs receiving vincristine. Time to platelet count ≥40 000 platelets/µL, survival, and duration of hospitalization were compared between neutropenic and non-neutropenic dogs. RESULTS: Vincristine was administered to 127 dogs with presumptive ITP; 19 became neutropenic. Administration of cyclosporine was significantly (P < .001) associated with the development of neutropenia (odds ratio: 12.97, 95% confidence interval: 4.17, 40.35). There was no difference in median time to ≥40 000 platelets/µL between neutropenic dogs (4 days; range, 1-14 days) and non-neutropenic dogs (3 days; range, 0-48 days). Percentage survival to discharge was 95% in both groups, but median duration of hospitalization was significantly longer in neutropenic dogs (6 days; range, 3-22 days) compared to non-neutropenic dogs (4 days; range, 2-15 days). CONCLUSIONS AND CLINICAL IMPORTANCE: Cyclosporine administration was associated with the development of neutropenia in dogs receiving vincristine, which might be related to effects on metabolism of vincristine. Neutrophil counts should be monitored in dogs receiving vincristine treatment for ITP, particularly if administered in conjunction with cyclosporine.

Early radiation-induced oral pain signaling responses are reduced with pentoxifylline treatment.

Radiation-induced acute oral mucositis is associated with inflammation and pain. In other realms of pain research, nociceptors are known to be activated by inflammatory cytokines; for example, tumor necrosis factor alpha (TNF-α) can activate transient receptor potential ion channels on sensory neurons. But there is an unclear relationship between inflammatory cytokines and molecular mediators of pain in radiation-induced mucositis (RIM) and radiation-associated pain (RAP). In this prospective, analytical, experimental pilot study, a common drug (pentoxifylline [PTX]) was used with the goal of inhibiting TNF-α signaling in mice that underwent lingual irradiation to induce severe acute oral RIM/RAP. Body weight and glossitis scores were recorded daily. Eye wiping behaviors were assayed as a surrogate measure of oral discomfort (which is possible due to cross-sensitization of the mandibular and ophthalmic branches of the trigeminal nerve). Quantitative real-time reverse transcription polymerase chain reaction was performed on irradiated tongue tissue to measure changes in expression of TNF-α, its receptor, nuclear factor kappa-light-chain-enhancer of activated B cells, transient receptor potential vanilloid type 1 (TRPV1), and transient receptor potential vanilloid type 4 (TRPV4). Responsiveness of afferent sensory trigeminal neurons to TNF-α, a TRPV1 agonist (capsaicin), and a partial TRPV4 agonist (histamine) was measured via calcium imaging. Although PTX treatment did not reduce glossitis severity or mitigate weight loss in mice with RIM/RAP, it did inhibit the upregulation of TNF-α's receptor that normally accompanies RIM, and it also reduced neuronal responsiveness to each of the aforementioned chemical stimuli. These results provide provisional evidence that inhibition of TNF-α signaling with PTX treatment may serve as a useful tool for reducing pain in head and neck cancer patients.