The Role of Targeted Osmotic Lysis in the Treatment of Advanced Carcinoma in Companion Animals: A Case Series.

Background: Targeted osmotic lysis (TOL) is a novel technology that involves concomitant stimulation of voltage-gated sodium channels (VGSCs) and the pharmacological blockade of Na+, K+-ATPase causing lysis of highly malignant cancer cells. Hypothesis/Objectives. TOL offers an option for treating advanced carcinomas in companion animals. Animals. Two cats and 2 dogs that presented to veterinary hospitals for evaluation and treatment of one of several forms of carcinoma. Methods: Digoxin was administered to achieve steady-state, therapeutic concentrations. The animals were then exposed to pulsed electric field stimulation. Pre- and posttreatment assessments of tumor size and quality of life were compared. The treatment frequency and survivability varied, based on the patient's premorbid functioning and response to treatment. Results: Regardless of cancer type, TOL consistently increased survival beyond expected, often improving, but without compromising of quality of life. Conclusions and Clinical Importance. TOL warrants consideration as an option for managing advanced carcinomas.

Ocular teratoid medulloepithelioma in a northern red-shouldered macaw: case report and literature review.

A 4-mo-old northern red-shouldered macaw (Diopsittaca nobilis) was admitted to the veterinary hospital of the Arruda Câmara Zoo, in the State of Paraiba, Brazil, for investigation of an orbital mass. Given rapid progression and lack of response to treatment, the bird was euthanized, and an autopsy was performed. Histologically, the mass consisted of a retrobulbar invasive tumor characterized by tubular and rosette-like structures, with interspersed heteroplastic tissues, such as aggregates of neuroglial cells and islands of hyaline cartilage. The tumor was immunopositive for pancytokeratin, GFAP, NSE, and S100. These findings were compatible with an ocular teratoid medulloepithelioma, a neoplasm best described in humans but also reported rarely in young cockatiels and African Grey parrots.

Severe Concomitant Physaloptera sp., Dirofilaria immitis, Toxocara cati, Dipylidium caninum, Ancylostoma sp. and Taenia taeniaeformis Infection in a Cat.

Here we describe an unusual and severe mixed parasitic infection in a cat that died during routine surgery. Gastric Physaloptera sp., cardiac Dirofilaria immitis, and intestinal Toxocara cati, Dipylidium caninum, Ancylostoma sp. and Taenia taeniaeformis were observed. Histologic lesions included chronic proliferative pulmonary endarteritis, mild increase of mucosal intestinal white cells, and terminal aspiration of gastric content. The severe dirofilariasis may have contributed to this patient death during anesthesia.

Novel Oncolytic Herpes Simplex Virus 1 VC2 Promotes Long-Lasting, Systemic Anti-melanoma Tumor Immune Responses and Increased Survival in an Immunocompetent B16F10-Derived Mouse Melanoma Model.

Oncolytic virotherapy (OVT) is now understood to be an immunotherapy that uses viral infection to liberate tumor antigens in an immunogenic context to promote the development of antitumor immune responses. The only currently FDA-approved oncolytic virotherapy, T-Vec, is a modified type 1 herpes simplex virus (HSV-1). While T-Vec is associated with limited response rates, its modest efficacy supports the continued development of novel OVT viruses. Herein, we test the efficacy of a recombinant HSV-1, VC2, as an OVT in a syngeneic B16F10-derived mouse model of melanoma. VC2 possesses mutations that block its ability to enter neurons via axonal termini. This greatly enhances its safety profile by precluding the ability of the virus to establish latent infection. VC2 has been shown to be a safe, effective vaccine against both HSV-1 and HSV-2 infection in mice, guinea pigs, and nonhuman primates. We found that VC2 slows tumor growth rates and that VC2 treatment significantly enhances survival of tumor-engrafted, VC2-treated mice over control treatments. VC2-treated mice that survived initial tumor engraftment were resistant to a second engraftment as well as colonization of lungs by intravenous introduction of tumor cells. We found that VC2 treatment induced substantial increases in intratumoral T cells and a decrease in immunosuppressive regulatory T cells. This immunity was critically dependent on CD8+ T cells and less dependent on CD4+ T cells. Our data provide significant support for the continued development of VC2 as an OVT for the treatment of human and animal cancers.IMPORTANCE Current oncolytic virotherapies possess limited response rates. However, when certain patient selection criteria are used, oncolytic virotherapy response rates have been shown to increase. This, in addition to the increased response rates of oncolytic virotherapy in combination with other immunotherapies, suggests that oncolytic viruses possess significant therapeutic potential for the treatment of cancer. As such, it is important to continue to develop novel oncolytic viruses as well as support basic research into their mechanisms of efficacy. Our data demonstrate significant clinical potential for VC2, a novel type 1 oncolytic herpes simplex virus. Additionally, due to the high rates of survival and the dependence on CD8+ T cells for efficacy, our model will enable study of the immunological correlates of protection for VC2 oncolytic virotherapy and oncolytic virotherapy in general. Understanding the mechanisms of efficacious oncolytic virotherapy will inform the rational design of improved oncolytic virotherapies.

Detection of SARS-CoV-2 by RNAscope® in situ hybridization and immunohistochemistry techniques.

In situ hybridization (ISH) and immunohistochemistry (IHC) are essential tools to characterize SARS-CoV-2 infection and tropism in naturally and experimentally infected animals and also for diagnostic purposes. Here, we describe three RNAscope®-based ISH assays targeting the ORF1ab, spike, and nucleocapsid genes and IHC assays targeting the spike and nucleocapsid proteins of SARS-CoV-2.

Cytotoxic Activity of the Mesoionic Compound MIH 2.4Bl in Breast Cancer Cell Lines.

In this work, we report the synthesis of a new 1,3-thiazolium-5-thiolate derivative of a mesoionic compound (MIH 2.4Bl) and the characterization of its selective cytotoxicity on a panel of breast cancer cells lines. The cytotoxic effect of MIH 2.4Bl on breast cancer cell lines was determined by XTT and crystal violet assays, flow cytometry analysis, electron microscopy characterization, and terminal deoxynucleotidyl transferase (TdT) deoxyuridine triphosphate (dUTP) nick end labeling (TUNEL) apoptosis assays. As determined using XTT cell growth and survival assays, MIH 2.4Bl exhibited growth inhibition activity on most breast cancer cell lines tested, compared with normal human mammary epithelial cells. Three breast cancer cell lines (MCF-7, T-47D, and ZR-75-1) showed a more potent sensitivity index to growth inhibition by MIH 2.4Bl than the other breast cancer cell lines. Interestingly, these 3 cell lines were derived from tumors of Luminal A origin and have ER (estrogen receptor), PR (progesterone receptor), and HER2 (human epidermal growth factor receptor 2) positive expression. Additional analysis of cytotoxicity mediated by MIH 2.4Bl was performed using the MCF-7 cell line. MCF-7 cells displayed both time- and dose-dependent decreases in cell growth and survival, with a maximum cytotoxic effect observed at 72 and 96 hours. The MCF-7 cells were also characterized for cell cycle changes upon treatment with MIH 2.4Bl. Using flow cytometry analysis of cell cycle distribution, a treatment-dependent effect was observed; treatment of cells with MIH 2.4Bl increased the G2/M population to 34.2% compared with 0.1% in untreated (control) cells. Ultrastructural analysis of MFC-7 cells treated with MIH 2.4Bl at 2 different concentrations (37.5 and 75 µM) was performed by transmission electron microscopy. Cells treated with 37.5 µM MIH 2.4Bl showed morphologic changes beginning at 6 hours after treatment, while cells treated with 75 µM showed changes beginning at 3 hours after treatment. These changes were characterized by an alteration of nuclear morphology and mitochondrial degeneration consistent with apoptotic cell death. Results of a TUNEL assay performed on cells treated for 96 hours with MIH 2.4Bl supported the observation of apoptosis. Together, these results suggest that MIH 2.4Bl is a promising candidate for treating breast cancer and support further in vitro and in vivo investigation.

A retrospective histologic study of 140 cases of clinically significant equine ocular disorders.

Ocular diseases are an important category in equine medicine; however, most articles regarding histologic ocular lesions in horses are exclusive to a specific condition and do not provide a complete review of clinically significant ocular disease frequency in a diagnostic laboratory. We reviewed sections of equine eyes from 140 cases (98 enucleations [biopsies] and 42 autopsies) with clinically relevant ocular alterations at 2 diagnostic centers in the United States. The most common primary conditions were non-traumatic keratitis (36), equine recurrent uveitis (ERU; 31), traumatic injuries (22), ocular and periocular neoplasms (19), and uveitis and/or endophthalmitis resulting from sepsis (18). Congenital anomalies (3) and retinal atrophy and detachment alone (3) were infrequent. Non-traumatic keratitis was frequently accompanied by anterior uveitis (22), corneal rupture (16), pre-iridal fibrovascular membrane formation (13), and secondary mycotic infection (11). ERU was the second and third most prevalent disease in autopsies and enucleations, respectively. This condition was commonly associated with glaucoma (15). Glaucoma (25) and cataract (20) were the most prevalent secondary alterations in the evaluated cases. Keratitis (20) and corneal rupture (16) were among the most prevalent consequences of trauma. Information presented herein may guide clinicians and pathologists, contributing to the early diagnosis of potentially vision-impairing conditions and raising the chances of successful treatment and cure.

Bovine papillomavirus E5 oncoprotein expression and its association with an interactor network in aggresome-autophagy pathway.

E5 protein, the major oncoprotein of bovine Deltapapillomavirus (BPV), was found to be expressed in 18 of 21 examined urothelial cancers of cattle. E5 oncoprotein was found to interact with p62 which was degraded through the autophagosome-lysosome pathway as well as LC3-II and appeared to be involved in the phosphorylation of the α-subunit of eukaryotic initiation factor 2 (eIF2α). Autophagy was morphologically documented by transmission electron microscope (TEM) through the detection of double-membrane autophagosomes and autolysosomes. Overexpression of Bag3 known to mediate selective autophagy was also demonstrated. Furthermore, Bag3 and BPV E5 oncoprotein were seen to co-localize with dynein and 14-3-3γ, which suggested that Bag3 could be involved in inducing the retrograde transport of BPV E5 along microtubules to aggresomes, perinuclear sites with high autophagic flux. Electron dense perinuclear structures consistent with aggresomes were also documented by TEM in urothelial cancer cells. Finally, Bag3 was found to also interact with synaptopodin 2 (Synpo2), which would seem to contribute to cargo degradation as it has been shown to facilitate autophagosome formation. This study provides mechanistic insights into the potential role(s) of autophagy in BPV disease, which can help to develop future treatment and control measures for BPV infection. Activation of autophagy correlates positively with BPV infection and may play a role in biological behavior of bladder cancer as urothelial carcinomas of cattle are known to be characterized by a relatively low rate of metastasis.

Liver tumorigenesis is promoted by a high saturated fat diet specifically in male mice and is associated with hepatic expression of the proto-oncogene Agap2 and enrichment of the intestinal microbiome with Coprococcus.

Liver cancer results in a high degree of mortality, especially among men. As fatty liver disease is a risk factor for development of hepatocellular carcinoma, we investigated the role of dietary fat type in tumor promotion by high-fat diets in mice after initiation with the chemical carcinogen diethyl nitrosamine. Tumor incidence and multiplicity were significantly greater in males than those in females. In males, fat type had complex effects on tumorigenesis. Preneoplastic foci were most prevalent in mice fed a polyunsaturated fat diet enriched in docosahexaenoic acid, whereas carcinomas and large visible liver tumors were significantly greater in mice fed a saturated fat diet made with cocoa butter relative to mice fed mono- or polyunsaturated fats. Different mechanisms thus seemed involved in early and late tumor promotion. The hepatic transcriptome and gut microbiome were assessed for traits associated with tumorigenesis. Hepatic expression of more than 20% of all genes was affected by sex, whereas fat type affected fewer genes. In males, the saturated fat diet induced expression of the proto-oncogene Agap2 and affected the expression of several cytochrome P450 genes, and genes involved in lipid, bile acid and fatty acid metabolism. The gut microbiome had a higher level of genus Akkermansia and a lower level of Firmicutes in females than in males. Males fed saturated fat had an altered microbiome, including an enrichment of the genus Coprococcus. In conclusion, sex and the dietary fat type affect the gut microbiome, the hepatic transcriptome and ultimately hepatic tumor growth.

Investigation of a Cyprinid Herpesvirus 1 Disease Episode in a Group of Pond-Reared Koi.

Fifteen adult koi (a variant of Common Carp Cyprinus carpio) simultaneously developed white cutaneous proliferations affecting up to 30% of their bodies. The onset of these lesions (in September 2014) was associated with their return to a remodeled backyard water garden after temporarily being maintained in a plastic swimming pool. A single water temperature taken during the outbreak read 21°C on November 17, 2014. The water garden had no extrinsic heat source, with average ambient temperatures ranging from 9.4 to 26.4°C during the outbreak (September 2014-January 2015). Representative skin biopsies were obtained from two fish; the histologic features included severe epidermal hyperplasia, dysplasia, keratinocyte apoptosis, decreased and haphazardly distributed goblet cells with the absence of club cells, keratinocyte hydropic degeneration, and moderate infiltration by lymphocytes and eosinophilic granular cells. Ultrastructural findings included intranuclear nonenveloped hexagonal nucleocapsids and abundant cytoplasmic-enveloped virions morphologically consistent with the Alloherpesviridae family. Polymerase chain reaction was performed on formalin-fixed, paraffin-embedded shavings from the two biopsied koi targeting the thymidine kinase gene of cyprinid herpesvirus 1 (CyHV-1). Together with the aforementioned findings, these results are consistent with an outbreak of CyHV-1 in a population of adult koi.

Effect of LysM+ macrophage depletion on lung pathology in mice with chronic bronchitis.

Macrophages (MΦ) are key sentinels of respiratory exposure to inhaled environmental stimuli. In normal "healthy" tissues, MΦ are believed to be a dormant cell type that, upon exposure to stress-causing stimuli, may get activated to exhibit pro- or anti-inflammatory roles. To test whether stress present in chronic bronchitic (CB) airways triggers MΦ to manifest protective or detrimental responses, the DTA+ (LysM-regulated Diphtheria Toxin A expressing) strain with partial MΦ-deficiency was crossed with the Scnn1b-Tg mouse model of CB and the progenies were studied at 4-5 weeks of age. Compared with DTA- littermates, the DTA+ mice had ~50% reduction in bronchoalveolar lavage (BAL) MΦ, and the recovered MΦ were immature, phenotypically distinct, and functionally defective. DTA+/Scnn1b-Tg mice exhibited a similar depletion of LysM+ MΦ offset by a significant increase in LysM- MΦ in the BAL. In DTA+/Scnn1b-Tg mice, lung disease was more severe than in DTA-/Scnn1b-Tg littermates, as indicated by an increased incidence of mucus plugging, mucous cells, airway inflammation, higher levels of cytokines/chemokines (KC, TNF-α, MIP-2, M-CSF, IL-5, and IL-17), and worsened alveolar airspace enlargement. DTA+/Scnn1b-Tg mice exhibited increased occurrence of lymphoid nodules, which was concomitant with elevated levels of immunoglobulins in BAL. Collectively, these data indicate that numerical deficiency of MΦ in stressed airspaces is responded via compensatory increase in the recruitment of immature MΦ and altered non-MΦ effector cell-centered responses, for example, mucus production and adaptive immune defense. Overall, these data identify dynamic roles of MΦ in moderating, rather than exacerbating, the severity of lung disease in a model of CB.

Intramuscular vaccination of guinea pigs with the live-attenuated human herpes simplex vaccine VC2 stimulates a transcriptional profile of vaginal Th17 and regulatory Tr1 responses.

Herpes simplex virus is a common causative agent of oral and genital diseases. Novel vaccines and therapeutics are needed to combat herpes infections especially after the failure of subunit vaccines in human clinical trials. We have shown that the live-attenuated HSV-1 VC2 vaccine strain is unable to establish latency in vaccinated animals and produces a robust immune response capable of completely protecting mice against lethal vaginal HSV-1 or HSV-2 infections. The guinea pig represents the best small animal model of genital HSV-2 disease. Reported here, twenty-one female Hartley guinea pigs received intramuscular injection with either the VC2 vaccine, or equal volume of conditioned tissue culture media. Animals received 2 booster vaccinations at 21 day intervals following the initial vaccination. After vaccination, animals were challenged with the highly virulent HSV-2 (G) strain. Histologically, VC2 vaccinated animals had little to no apparent inflammation/disease following challenge. Unvaccinated animals developed moderate to severe erosive and ulcerative vaginitis. Quantitative reverse-transcriptase PCR analysis in VC2 vaccinated and challenged animals identified transcriptional signatures of Th17 and regulatory Tr1 cells associated with the inflammatory response primed by VC2 vaccination. Treatment of cultured human vaginal epithelial cells (VK2 cells) with a combination of IL-17A and IL-22 resulted in the significant induction of beta-defensin 3 expression. Further, treatment of VK2 cells with IL-17A, IL-22, IL-36 or beta-defensin 3 resulted in diminished HSV-2 replication. Overall, these results suggest that intramuscular vaccination with the live-attenuated vaccine VC2 primes a mucosal immune response predisposing the adaptive expression of transcripts associated with a Th17 response to challenge and these responses contribute to antiviral immunity.

Intramuscular Immunization of Mice with the Live-Attenuated Herpes Simplex Virus 1 Vaccine Strain VC2 Expressing Equine Herpesvirus 1 (EHV-1) Glycoprotein D Generates Anti-EHV-1 Immune Responses in Mice.

Vaccination remains the best option to combat equine herpesvirus 1 (EHV-1) infection, and several different strategies of vaccination have been investigated and developed over the past few decades. Herein, we report that the live-attenuated herpes simplex virus 1 (HSV-1) VC2 vaccine strain, which has been shown to be unable to enter into neurons and establish latency in mice, can be utilized as a vector for the heterologous expression of EHV-1 glycoprotein D (gD) and that the intramuscular immunization of mice results in strong antiviral humoral and cellular immune responses. The VC2-EHV-1-gD recombinant virus was constructed by inserting an EHV-1 gD expression cassette under the control of the cytomegalovirus immediate early promoter into the VC2 vector in place of the HSV-1 thymidine kinase (UL23) gene. The vaccines were introduced into mice through intramuscular injection. Vaccination with both the VC2-EHV-1-gD vaccine and the commercially available vaccine Vetera EHVXP 1/4 (Vetera; Boehringer Ingelheim Vetmedica) resulted in the production of neutralizing antibodies, the levels of which were significantly higher in comparison to those in VC2- and mock-vaccinated animals (P < 0.01 or P < 0.001). Analysis of EHV-1-reactive IgG subtypes demonstrated that vaccination with the VC2-EHV-1-gD vaccine stimulated robust IgG1 and IgG2a antibodies after three vaccinations (P < 0.001). Interestingly, Vetera-vaccinated mice produced significantly higher levels of IgM than mice in the other groups before and after challenge (P < 0.01 or P < 0.05). Vaccination with VC2-EHV-1-gD stimulated strong cellular immune responses, characterized by the upregulation of both interferon- and tumor necrosis factor-positive CD4+ T cells and CD8+ T cells. Overall, the data suggest that the HSV-1 VC2 vaccine strain may be used as a viral vector for the vaccination of horses as well as, potentially, for the vaccination of other economically important animals.IMPORTANCE A novel virus-vectored VC2-EHV-1-gD vaccine was constructed using the live-attenuated HSV-1 VC2 vaccine strain. This vaccine stimulated strong humoral and cellular immune responses in mice, suggesting that it could protect horses against EHV-1 infection.

Evaluation of the genes encoding CD39/NTPDase-1 and CD39L1/NTPDase-2 in horses with and without abnormal hemorrhage and in horses with pathologic evidence of exercise-induced pulmonary hemorrhage.

BACKGROUND: Exercise-induced pulmonary hemorrhage (EIPH) is a common disorder of equine athletes. The role of polymorphisms in genes encoding hemostasis-regulatory proteins in horses with abnormal hemorrhage is unknown. OBJECTIVES: The goal of this study was to evaluate the genes encoding 2 ectonucleotidases, CD39/NTPDase-1 and CD39L1/NTPDase-2, and one ecto-5' nucleotidase, CD73, in horses with abnormal hemorrhage or pathologic changes consistent with EIPH. METHODS: Twenty-three horses with histories of abnormal hemorrhage, 8 horses with gastrointestinal signs, and 45 healthy horses were evaluated using polymerase chain reaction-based techniques. Formalin-fixed tissues from 21 horses with pathologic changes consistent with EIPH were also evaluated. RESULTS: Three single nucleotide polymorphisms (SNPs) were identified in the gene encoding CD39 and one SNP was identified in the gene encoding CD39L1. No SNPs were identified in the gene encoding CD73. CD39 SNPs were identified in 19 of 20 (95%) horses with unexplained hemorrhage and 20 of 21 (95%) horses with pathologic features consistent with EIPH. CD39L1 SNPs were identified in 6 of 20 (30%) horses with unexplained hemorrhage and 8 of 21 (38%) horses with pathologic features consistent with EIPH. CD39 and CD39L1 SNPs were identified in 5 of 8 (62.5%) and one of 8 (12.5%) horses, respectively, presenting with colic or weight loss. CD39 and CD39L1 SNPs were identified in 28 of 45 (62%) and 13 of 45 (28.8%) healthy horses, respectively. CONCLUSIONS: CD39 and CD39L1 are critically important in maintaining normal hemostasis and limiting inflammation. Further studies are needed to evaluate their role in the pathogenesis of equine EIPH.

Evaluation of bone regeneration potential of dental follicle stem cells for treatment of craniofacial defects.

BACKGROUND AIMS: Stem cell-based tissue regeneration offers potential for treatment of craniofacial bone defects. The dental follicle, a loose connective tissue surrounding the unerupted tooth, has been shown to contain progenitor/stem cells. Dental follicle stem cells (DFSCs) have strong osteogenesis capability, which makes them suitable for repairing skeletal defects. The objective of this study was to evaluate bone regeneration capability of DFSCs loaded into polycaprolactone (PCL) scaffold for treatment of craniofacial defects. METHODS: DFSCs were isolated from the first mandibular molars of postnatal Sprague-Dawley rats and seeded into the PCL scaffold. Cell attachment and cell viability on the scaffold were examined with the use of scanning electron microscopy and alamar blue reduction assay. For in vivo transplantation, critical-size defects were created on the skulls of 5-month-old immunocompetent rats, and the cell-scaffold constructs were transplanted into the defects. RESULTS: Skulls were collected at 4 and 8 weeks after transplantation, and bone regeneration in the defects was evaluated with the use of micro-computed tomography and histological analysis. Scanning electron microscopy and Alamar blue assay demonstrated attachment and proliferation of DFSCs in the PCL scaffold. Bone regeneration was observed in the defects treated with DFSC transplantation but not in the controls without DFSC transplant. Transplanting DFSC-PCL with or without osteogenic induction before transplantation achieved approximately 50% bone regeneration at 8 weeks. Formation of woven bone was observed in the DFSC-PCL treatment group. Similar results were seen when osteogenic-induced DFSC-PCL was transplanted to the critical-size defects. CONCLUSIONS: This study demonstrated that transplantation of DFSCs seeded into PCL scaffolds can be used to repair craniofacial defects.

miR-196a expression in human and canine osteosarcomas: a comparative study.

Osteosarcoma (OS) is the most common primary malignant bone tumour in dogs and humans. MicroRNAs are short non-coding RNA molecules involved in post-transcriptional gene expression. Here, we compared the effects of miR-196a deregulation in human and canine OS cells after having observed a more uniform distribution and stronger down-expression in the human specimens. Cell response to miR-196a transfection was different in human and canine OS. A decreased proliferation rate was seen in human MG63 and 143B OS cells, while no appreciable changes occurred in canine DAN cells. Transient decrease of motility was highly remarkable and longer in MG63, concomitant with decreased levels of annexin1, a target of miR-196a promoting cell migration and invasion. In conclusion, the effects of miR-196a over-expression on tumour cell response may be strictly related to species and cell type. Further studies are needed to define the impact of miRNA deregulation on OS development.

First report of Cylicospirura felineus in a feral domestic shorthair cat in North America.

CASE SUMMARY: A feral domestic shorthair cat was euthanized owing to acute onset and progression of neurological signs attributed to ethylene glycol toxicity. At post-mortem examination two nodules were identified within the fundus of the stomach. Examination of the gastric nodules revealed an intact mucosal surface, each with multiple red slender nematodes extending through an individual central pore. Histopathological evaluation of the nodules highlighted unique reactive fibroplasia, mimicking feline gastrointestinal eosinophilic sclerosing fibroplasia (FGESF), encasing numerous nematodes with females possessing gravid uteri containing abundant larvated eggs. The latter findings were highly suggestive of the Cylicospirura genus, further supported by an en face evaluation of the buccal cavity, highlighting a distinctive trifid tooth appearance. Together, these findings are consistent with Cylicospirura felineus. PCR for the COX-1 gene was unsuccessful on formalin-fixed specimens, attributed to nucleic acid and protein crosslinking. RELEVANCE AND NOVEL INFORMATION: This represents the first documented case of Cylicospirura species in a feral domestic shorthair cat in North America. This particular cat lived in the highly urban environment of New Orleans, Louisiana. Identification of this case demonstrates the potential for feral cats to serve as reservoir hosts and ultimately support transmission of Cylicospirura species into domesticated cat populations. Gastric cylicospiruriasis may present clinically as a firm abdominal mass, potentially with a history of chronic vomiting. The latter emphasizes the importance of differentiating this condition from a neoplastic process such as alimentary lymphoma and adenocarcinoma. Histologically, the unique thick anastomosing collagenous cords encasing nematodes represent a stereotypical response observed in a broad array of gastrointestinal inflammation in felines, including intralesional bacteria, fungal hyphae, foreign bodies and, in this case, gastric nematodes that closely resemble FGESF. Additionally, these unique histological lesions have previously been misinterpreted as neoplastic conditions, including sclerosing mast cell tumor and extraosseous osteosarcoma.

In vitro and in vivo characterization of pentaerythritol triacrylate-co-trimethylolpropane nanocomposite scaffolds as potential bone augments and grafts.

A thiol-acrylate-based copolymer synthesized via an amine-catalyzed Michael addition was studied in vitro and in vivo to assess its potential as an in situ polymerizing graft or augment in bone defect repair. The blends of hydroxyapatite (HA) with pentaerythritol triacrylate-co-trimethylolpropane (PETA), cast as solids or gas foamed as porous scaffolds, were evaluated in an effort to create a biodegradable osteogenic material for use as a bone-void-filling augment. Osteogenesis experiments were conducted with human adipose-derived mesenchymal stromal cells (hASCs) to determine the ability of the material to serve as an osteoinductive substrate. Poly(ɛ-caprolactone) (PCL) composites PCL:HA (80:20) (wt/wt%) served as the control scaffold, while the experimental scaffolds included PETA:HA (100:0), (85:15), (80:20), and (75:25) composites (wt/wt%). The results indicate that PETA:HA (80:20) foam composites had higher mechanical strength than the corresponding porous PCL:HA (80:20) scaffolds made by thermo-precipitation method, and in the case of foamed composites, increasing HA content directly correlated with increased yield strength. For cytotoxicity and osteogenesis experiments, hASCs cultured for 21 days on PETA:HA scaffolds in stromal medium displayed the greatest number of live cells compared with PCL:HA composites. Moreover, hASCs cultured on foamed PETA:HA (80:20) scaffolds resulted in the greatest mineralization, increased alkaline phosphatase (ALP) expression, and the highest osteocalcin (OCN) expression after 21 days. Overall, the PETA:HA (80:20) and PETA:HA (85:15) scaffolds, with 66.38% and 72.02% porosity, respectively, had higher mechanical strength and cytocompatibility compared with the PCL:HA control. The results of the 6-week in vivo biocompatibility study using a posterior lumbar spinal fusion model demonstrate that PETA:HA can be foamed in vivo without serious adverse effects at the surgical site. Additionally, it was demonstrated that cells migrate into the interconnected pore volume and are found within centers of ossification.