Effects on Tissue Integration of Collagen Scaffolds Used for Local Delivery of Gentamicin in a Rat Mandible Defect Model.

Surgical site infections (SSIs) are a common complication following orthopedic surgery. SSIs may occur secondary to traumatic or contaminated wounds or may result from invasive procedures. The development of biofilms is often associated with implanted materials used to stabilize injuries and to facilitate healing. Regardless of the source, SSIs can be challenging to treat. This has led to the development of devices that act simultaneously as local antibiotic delivery vehicles and as scaffolds for tissue regeneration. The goal for the aforementioned devices is to increase local drug concentration in order to enhance bactericidal activity while reducing the risk of systemic side effects and toxicity from the administered drug. The aims of this study were to assess the effect of antibiotic loading of a collagen matrix on the tissue integration of the matrix using a rat mandibular defect model. We hypothesized that the collagen matrix could load and elute gentamicin, that the collagen matrix would be cytocompatible in vitro, and that the local delivery of a high dose of gentamicin via loaded collagen matrix would negatively impact the tissue-scaffold interface. The results indicate that the collagen matrix could load and elute the antimicrobial gentamicin and that it was cytocompatible in vitro with or without the presence of gentamicin and found no significant impact on the tissue-scaffold interface when the device was loaded with a high dose of gentamicin.

Development of a polymeric biomedical device platform with controlled disassembly and in vivo testing in a swine intestinal model.

The aim of this study was to create a surgical guide platform that maintains its integrity while the surgeon performs an intestinal anastomosis or another similar procedure, which then breaks apart and is eliminated from the body in a controlled manner. The device contains mixed polymeric structures that give it a controlled rate of disassembly that could meet the requirements of a specific surgical purpose. The intraluminal anastomotic guide was manufactured as a hollow cylinder composed of layers of porous polyurethane/PCL with polyvinylpyrrolidone as the binding agent similar to a "brick-mortar" architecture. This combination of polymeric structures is a promising manufacturing method from which a variety of tunable devices can be fabricated for specific medical procedures and site-specific indications. The guide was designed to rapidly disassemble within the intestinal lumen after use, reliably degrading while maintaining sufficient mechanical rigidity and stability to support manipulation during complex surgical procedures. The nature of the device's disassembly makes it suitable for use in hollow structures that discharge their contents, resulting in their elimination from the body. A swine model of intestinal anastomosis was utilized to validate the use and function of the device.

Pulmonary silicosis in 2 rock hyraxes, and literature review.

Two rock hyraxes (Procavia capensis), from the Chattanooga Zoo, were submitted separately for autopsy at the University of Tennessee Veterinary Medical Center. The first was a 4-y-old intact female that died without premonitory signs and the second was a 10-y-old intact male that was euthanized because of severe renal disease. Microscopically, the lungs of both hyraxes had multifocal-to-coalescing, <1-mm diameter aggregates of epithelioid macrophages separated by streams of fibrous tissue. Macrophages contained intracytoplasmic, clear, acicular, birefringent crystals. Transmission electron microscopy and energy-dispersive x-ray spectroscopy findings on the lung samples were consistent with silica crystal deposition. The hyraxes had been housed together on commercially sourced play sand composed of 99-99.5% quartz, a crystalline silica polymorph. The microscopic findings, transmission electron microscopy, and energy-dispersive x-ray spectroscopy of the intrahistiocytic crystals, in addition to the history of exposure to crystalline silica, were consistent with pulmonary silicosis. Pulmonary silicosis has not been reported previously in rock hyraxes, to our knowledge.

Influence of a novel scaffold composed of polyurethane, hydroxyapatite, and decellularized bone particles on the healing of fourth metacarpal defects in mares.

OBJECTIVE: To determine the effect of a novel scaffold, designed for use in bone regeneration, on healing of splint bone segmental defects in mares. STUDY DESIGN: In vivo experimental study. SAMPLE POPULATION: Five adult mares (4-10 years old; mean weight, 437.7 kg ± 29 kg). METHODS: Bilateral 2-cm full-thickness defects were created in the fourth metacarpal bones (MCIV) of each horse. Each defect was randomly assigned to either a novel scaffold treatment (n = 5) or an untreated control (n = 5). The scaffold was composed of polyurethane, hydroxyapatite, and decellularized bone particles. Bone healing was assessed for a period of 60 days by thermography, ultrasonography, radiography, and computed tomography (CT). Biopsies of each defect were performed 60 days after surgery for histological evaluation. RESULTS: On the basis of radiographic analysis, scaffold-treated defects had greater filling (67.42% ± 26.7%) compared with untreated defects (35.88% ± 32.7%; P = .006). After 60 days, CT revealed that the density of the defects treated with the scaffolds (807.80 ± 129.6 Hounsfield units [HU]) was greater than density of the untreated defects (464.80 ± 81.3 HU; P = .004). Evaluation of histology slides provided evidence of bone formation within an average of 9.43% ± 3.7% of the cross-sectional area of scaffolds in contrast to unfilled defects in which connective tissue was predominant throughout the biopsy specimens. CONCLUSION: The novel scaffold was biocompatible and supported bone formation within the MCIV segmental defects. CLINICAL SIGNIFICANCE: This novel scaffold offers an effective option for filling bone voids in horses when support of bone healing is indicated.

Development and in vivo Assessment of a Rapidly Collapsible Anastomotic Guide for Use in Anastomosis of the Small Intestine: A Pilot Study Using a Swine Model.

Various conditions in human and veterinary medicine require intestinal resection and anastomosis, and complications from these procedures are frequent. A rapidly collapsible anastomotic guide was developed for small intestinal end-to-end anastomosis and was investigated in order to assess its utility to improve the anastomotic process and to potentially reduce complication rates. A complex manufacturing method for building a polymeric device was established utilizing biocompatible and biodegradable polyvinylpyrrolidone and polyurethane. This combination of polymers would result in rapid collapse of the material. The guide was designed as a hollow cylinder composed of overlaying shingles that separate following exposure to moisture. An in vivo study was performed using commercial pigs, with each pig receiving one standard handsewn anastomosis and one guide-facilitated anastomosis. Pigs were sacrificed after 13 days, at which time burst pressure, maximum luminal diameter, and presence of adhesions were assessed. Burst pressures were not statistically different between treatment groups, but in vivo anastomoses performed with the guide withstood 10% greater luminal burst pressure and maintained 17% larger luminal diameter than those performed using the standard handsewn technique alone. Surgeons commented that the addition of a guide eased the performance of the anastomosis. Hence, a rapidly collapsible anastomotic guide may be beneficial to the performance of intestinal anastomosis.

Fatal pulmonary hemorrhage associated with vascular amyloid deposition in a cat.

CASE SUMMARY: An adult female spayed Siamese-cross cat of unknown age was presented for bilateral hemorrhagic otorrhea. Nasopharyngeal polyps were diagnosed by CT and biopsy; bilateral ventral bulla osteotomies were performed. Episodic epistaxis, otic hemorrhage and hemoptysis with respiratory distress progressed over 18 months. Systolic blood pressure, complete blood count, plasma biochemistries, prothrombin time, partial thromboplastin time and coagulation factor 12, 9 and 8 activities were normal. Serial thoracic radiographs revealed patchy interstitial to alveolar patterns. Airway hemorrhage prevented diagnostic bronchoscopy. Respiratory hemorrhage was ultimately fatal. Amyloid deposition was identified in pulmonary vasculature, bronchial wall, lymphoid tissues, nasal-pharyngeal tissue and tympanic bullae based on microscopic examination and confirmed by Congo red staining with green birefringence under polarized light. RELEVANCE AND NOVEL INFORMATION: Amyloidosis should be considered as a differential diagnosis in cats with spontaneous hemorrhage of the respiratory or otic tracts. Although systemic amyloidosis is associated with a grave prognosis, this case suggests that prolonged survival is possible after the initial onset of signs in cats with pulmonary amyloidosis.

Blastomyces dermatitidis pneumonia in a llama.

A 20-y-old female llama ( Lama glama) was euthanized after a history of chronic dyspnea and osteoarthritis. At autopsy, the lungs were covered by clear gelatinous material and expanded by firm, variably discrete, tan-white nodules up to 8 cm diameter containing tan-white, viscous material. The tracheobronchial lymph nodes were firm and enlarged up to 6 × 4 × 3 cm; the thoracic aorta and carotid arteries were lined by hard, tan-white, mineralized intimal plaques. Histologic examination of lung revealed numerous 10-20 µm diameter yeasts with clear 1-2 µm thick double-contoured walls, central basophilic nuclei, and frequent broad-based budding, consistent with Blastomyces dermatitidis. DNA sequencing confirmed the diagnosis. B. dermatitidis should be considered in the differential diagnosis of pulmonary disease in llamas.

Spontaneous neoplasms in captive Virginia opossums ( Didelphis virginiana): a retrospective case series (1989-2014) and review of the literature.

This retrospective project summarizes the types of neoplasms identified in Virginia opossums ( Didelphis virginiana) presented to the University of Tennessee, College of Veterinary Medicine (UTCVM) postmortem service in 1989-2014 and serves as a review of the literature. Of the 85 Virginia opossums identified from the UTCVM case database, there were 17 diagnoses of neoplasia from 12 cases (14%). These cases included 8 females, 2 males, and 2 neutered males. All opossums with known ages (11 of 12) were >2 y old. Pulmonary tumors, specifically minimally invasive or lepidic-predominant adenocarcinomas, were the most common diagnosis and accounted for 53% (9 of 17) of the neoplasms. Additional tumors included acute myeloid leukemia with eosinophil maturation, hepatic hemangiosarcoma, sarcoma (unknown origin), squamous cell carcinoma, disseminated mast cell tumor, trichoblastoma, thyroid adenoma, and an osteoma. These findings serve as a reference for the types of spontaneous neoplasms in Virginia opossums; based on these findings, neoplasia should be considered as a differential in mature captive Virginia opossums.

Temporal Development of Gut Microbiota in Triclocarban Exposed Pregnant and Neonatal Rats.

Alteration of gut microbial colonization process may influence susceptibility of the newborn/infant to infectious and chronic disease. Infectious disease risk leads to widespread use of non-prescription antimicrobials in household products such as Triclocarban (TCC), an antimicrobial compound in personal care products. TCC concentrates in and is transferred through the milk to suckling offspring. TCC exposure during gestation and lactation significantly reduced phylogenetic diversity (PD) among exposed dams and neonates. Among dams using weighted UniFrac distances, TCC induced significant dysbiosis of gut microbiota by gestational day (GD) 18, a trend that continued after delivery. Similarly, an overall restructuring of gut microbiota occurred in neonates. By postnatal day (PND) 12, communities separated based on exposure status and became significantly different at PND 16. The ability of TCC to drive microbial dysbiosis warrants future investigation to evaluate the safety of non-prescription antimicrobial use, including TCC, during critical exposure windows.

A novel approach to determine post mortem interval using neutron radiography.

One of the most difficult challenges in forensic research is to objectively determine the post-mortem interval (PMI). The accuracy of PMI is critical for determining the timeline of events surrounding a death. Most PMI techniques rely on gross morphological changes of cadavers that are highly sensitive to taphonomic factors. Recent studies have demonstrated that even exhumed individuals exposed to the same environmental conditions with similar PMIs can present different stages of decomposition. After death, tissue undergoes sequential changes consisting of organic and inorganic phase variations, as well as a gradual reduction of tissue water content. Hydrogen (H) is the primary contributor to neutron radiography (NR) contrast in biological specimens because (1) it is the most abundant element in biological tissues and (2) its nucleus scatters thermal and cold neutrons more strongly than any other atomic nucleus. These contrast differences can be advantageous in a forensic context to determine small changes in hydrogen concentrations. Neutron radiography of decaying canine tissues was performed to evaluate the PMI by measuring the changes in H content. In this study, dog cadavers were used as a model for human cadavers. Canine tissues and cadavers were exposed to controlled (laboratory settings, at the University of Tennessee, College of Veterinary Medicine) and uncontrolled (University of Tennessee Anthropology Research Facility) environmental conditions, respectively. Neutron radiographs were supplemented with photographs and histology data to assess the decompositional stages of cadavers. Results demonstrated that the increase in neutron transmission likely corresponded to a decrease in hydrogen content in the tissue, which was correlated with the decay time of the tissue. Tissues depleted in hydrogen were brighter in the neutron transmission radiographs of skeletal muscles, lung, and bone, under controlled conditions. Over a period of 10 days, changes in neutron transmission through lung and muscle were found to be higher than bone by 8.3%, 7.0%, and 2.0%, respectively. Results measured during uncontrolled conditions were more difficult to assess and further studies are necessary. In conclusion, neutron radiography may be used to detect changes in hydrogen abundance that can be correlated with the post-mortem interval.

Dorsal black skin necrosis in a Vietnamese pot-bellied pig.

BACKGROUND: Thermal burns are an uncommon cause of injury in large animals. CASE REPORT: A 10-month-old pet female black and white Vietnamese pot-bellied pig presented to the emergency service with fever and erythematous to purpuric skin lesions affecting the intermandibular space and hocks. One week prior to the emergency visit, she had appeared restless and in pain. Two weeks following the emergency visit, she again presented to the large animal clinic with sloughing of the pigmented skin on her head, face, dorsal and lateral trunk sparing the nonpigmented skin and pigmented ears. The affected skin constituted ~40% of her total skin. Histopathological findings for affected skin included full-thickness epidermal and partial to full-thickness dermal coagulative necrosis with follicular epithelial mineralization, while that from normal-appearing pigmented skin was within normal limits. A culture from a skin biopsy yielded meticillin-resistant Staphylococcus aureus (ST-72). Treatments included oral antibiotics, pain management, hyperbaric oxygen therapy and general anaesthesia to facilitate debridement. Healthy skin was often present when the necrotic skin was debrided, although in some areas the necrosis extended into the underlying fat. Complications that occurred during rehabilitation included intense pruritus that resulted in self-trauma and the formation of a nasal fistula, which was later surgically corrected. CONCLUSIONS AND CLINICAL IMPORTANCE: Cases of dorsal thermal necrosis in pot-bellied pigs are uncommon in the literature. Based on the clinical presentation and lack of another identifying cause, the lesions were attributed to a sun-induced thermal burn.

Nonsteroidal anti-inflammatory drug sulindac sulfide suppresses structural protein Nesprin-2 expression in colorectal cancer cells.

BACKGROUND: Nonsteroidal anti-inflammatory drugs (NSAIDs) are well known for treating inflammatory disease and have been reported to have anti-tumorigenic effects. Their mechanisms are not fully understood, but both cyclooxygenase (COX) dependent and independent pathways are involved. Our goal was to shed further light on COX-independent activity. METHODS: Human colorectal cancer cells were observed under differential interference contrast microscopy (DICM), fluorescent microscopy, and micro-impedance measurement. Microarray analysis was performed using HCT-116 cells treated with sulindac sulfide (SS). PCR and Western blots were performed to confirm the microarray data and immunohistochemistry was performed to screen for Nesprin-2 expression. Micro-impedance was repeating including Nesprin-2 knock-down by siRNA. RESULTS: HCT-116 cells treated with SS showed dramatic morphological changes under DICM and fluorescent microscopy, as well as weakened cellular adhesion as measured by micro-impedance. Nesprin-2 was selected from two independent microarrays, based on its novelty in relation to cancer and its role in cell organization. SS diminished Nesprin-2 mRNA expression as assessed by reverse transcriptase and real time PCR. Various other NSAIDs were also tested and demonstrated that inhibition of Nesprin-2 mRNA was not unique to SS. Additionally, immunohistochemistry showed higher levels of Nesprin-2 in many tumors in comparison with normal tissues. Further micro-impedance experiments on cells with reduced Nesprin-2 expression showed a proportional loss of cellular adhesion. CONCLUSIONS: Nesprin-2 is down-regulated by NSAIDs and highly expressed in many cancers. GENERAL SIGNIFICANCE: Our data suggest that Nesprin-2 may be a potential novel oncogene in human cancer cells and NSAIDs could decrease its expression.

Diagnosis and treatment of a dermal malignant melanoma in an African lion (Panthera leo).

A 13-yr-old intact male African lion (Panthera leo) presented with a 4-mo history of left maxillary lip swelling. On physical examination, a 10-cm-diameter, ulcerated, round, firm, and pigmented mass at the level of the left maxillary canine tooth was noticed. All other organ systems examined were within normal limits. Multiple biopsies of the mass were collected and fixed in 10% neutral buffered formalin. Histopathologic evaluation of the biopsies revealed a malignant dermal melanoma. Hematologic and plasma biochemical parameters were within normal reference ranges. Thoracic radiographs taken 3 days following initial presentation showed no evidence of metastasis of the tumor. Computed tomography of the skull and neck was performed to evaluate local tumor invasion and to plan for hypofractionated radiation therapy. Therapy included four weekly treatments of 8 gray external-beam hypofractionated radiation and four bimonthly immunotherapy treatments. Following this treatment regime, the tumor size was reduced by 50%, and surgical excision was performed. No major side effects associated with radiation or immunotherapy were seen. Six months after diagnosis, hematologic and plasma biochemical parameters were within normal limits, thoracic radiographs showed no evidence of metastasis, and the lion showed no clinical signs of disease. The lion will continue to receive immunotherapy every 6 mo for the rest of its life. To the authors' knowledge, this is the first report of a successful treatment of a malignant dermal melanoma with external-beam hypofractionated radiation, immunotherapy, and surgical excision in an African lion.

Effects of equine metabolic syndrome on inflammatory responses of horses to intravenous lipopolysaccharide infusion.

OBJECTIVE: To test the hypothesis that inflammatory responses to endotoxemia differ between healthy horses and horses with equine metabolic syndrome (EMS). Animals-6 healthy horses and 6 horses with EMS. PROCEDURES: Each horse randomly received an IV infusion of lipopolysaccharide (20 ng/kg [in 60 mL of sterile saline {0.9% NaCl} solution]) or saline solution, followed by the other treatment after a 7-day washout period. Baseline data were obtained 30 minutes before each infusion. After infusion, a physical examination was performed hourly for 9 hours and at 15 and 21 hours; a whole blood sample was collected at 30, 60, 90, 120, 180, and 240 minutes for assessment of inflammatory cytokine gene expression. Liver biopsy was performed between 240 and 360 minutes after infusion. Results-Following lipopolysaccharide infusion in healthy horses and horses with EMS, mean rectal temperature, heart rate, and respiratory rate increased, compared with baseline findings, as did whole blood gene expression of interleukin (IL)-1ß, IL-6, IL-8, IL-10, and tumor necrosis factor-α. The magnitude of blood cytokine responses did not differ between groups, but increased expression of IL-6, IL-8, IL-10, and tumor necrosis factor-α persisted for longer periods in EMS-affected horses. Lipopolysaccharide infusion increased liver tissue gene expressions of IL-6 in healthy horses and IL-8 in both healthy and EMS-affected horses, but these gene expressions did not differ between groups. CONCLUSIONS AND CLINICAL RELEVANCE: Results supported the hypothesis that EMS affects horses' inflammatory responses to endotoxin by prolonging cytokine expression in circulating leukocytes. These findings are relevant to the association between obesity and laminitis in horses with EMS.

Metal accumulation and health effects in raccoons (Procyon lotor) associated with coal fly ash exposure.

Approximately 5.4 million cubic yards of coal fly ash and water spilled into the Emory River embayment of Watts Bar Reservoir in east Tennessee on Dec 22, 2008. Raccoons were collected in 2009 and 2010 from the spill site (10/y) and unexposed areas (5/y) to determine whether metals and metalloids were accumulating in raccoons and if any negative health effects resulted from exposure to the spilled coal fly ash. Tissues were analyzed from each animal to determine the concentrations of 26 metals/metalloids. Complete blood cell counts (CBC), plasma biochemistry panels, and histopathology of select tissues also were performed. Results were analyzed by year and exposure status. Although significant differences were present in some tissues for some metals/metalloids, only arsenic in hair, iron in muscle, nickel in hair, selenium in hair and muscle, strontium in hair, and vanadium in hair and liver were increased in spill site animals (one or both years) compared with unexposed animals. No clinically important differences were observed between groups regarding CBC or plasma biochemistry analyses. Lesions were observed on histopathology in some tissues, but there was no difference in the prevalence of lesions between spill site and unexposed animals. There does not seem to be any important accumulation of metals/metalloids or negative health effects in raccoons associated with exposure to coal fly ash compared with unexposed animals.

Effects of a "two-hit" model of organ damage on the systemic inflammatory response and development of laminitis in horses.

The role of endotoxemia in the development of laminitis remains unclear. Although systemic inflammation is a risk factor for laminitis in hospitalized horses, experimental endotoxin administration fails to induce the disease. While not sufficient to cause laminitis by itself, endotoxemia might predispose laminar tissue to damage from other mediators during systemic inflammation. In "two-hit" models of organ damage, sequential exposure to inflammatory stimuli primes the immune system and causes exaggerated inflammatory responses during sepsis. Acute laminitis shares many characteristics with sepsis-associated organ failure, therefore an equine "two-hit" sepsis model was employed to test the hypothesis that laminitis develops with increased frequency and severity when repeated inflammatory events exacerbate systemic inflammation and organ damage. Twenty-four light breed mares (10) and geldings (14) with chronic disease conditions or behavioral abnormalities unrelated to laminitis that warranted euthanasia were obtained for the study. Horses were randomly assigned to receive an 8-h intravenous infusion of either lipopolysaccharide (5 ng/kg/h) or saline beginning at -24h, followed by oligofructose (OF; 5 g/kg) via nasogastric tube at 0 h. Euthanasia and tissue collection occurred at Obel grade 2 laminitis, or at 48 h if laminitis had not developed. Liver biopsies were performed at 24h in laminitis non-responders. Blood cytokine gene expression was measured throughout the study period. Lipopolysaccharide and OF administration independently increased mean rectal temperature (P<0.001), heart rate (P=0.003), respiratory rate (P<0.001), and blood interleukin (IL)-1ß gene expression (P<0.0016), but responses to OF were not exaggerated in endotoxin-pretreated horses. The laminitis induction rate did not differ between treatment groups and was 63% overall. When horses were classified as laminitis responders and non-responders, area under the blood IL-1ß expression curve (P=0.010) and liver and lung gene expression of IL-1ß, IL-6, IL-8, IL-10, and tumor necrosis factor-α (P<0.05) were higher in responders following OF administration. The results indicate that endotoxin pretreatment did not enhance responses to OF. However, systemic inflammation was more pronounced in laminitis responders compared to non-responders, and tissue-generated inflammatory mediators could pose a greater risk than those produced by circulating leukocytes.

Intervention of human breast cell carcinogenesis chronically induced by 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine.

More than 85% of breast cancers are sporadic and attributable to long-term exposure to environmental carcinogens, such as those in the diet, through a multistep disease process progressing from non-cancerous to premalignant and malignant stages. The chemical carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is one of the most abundant heterocyclic amines found in high-temperature cooked meats and is recognized as a mammary carcinogen. However, the PhIP's mechanism of action in breast cell carcinogenesis is not clear. Here, we demonstrated, for the first time, that cumulative exposures to PhIP at physiologically achievable, pico to nanomolar concentrations effectively induced progressive carcinogenesis of human breast epithelial MCF10A cells from a non-cancerous stage to premalignant and malignant stages in a dose- and exposure-dependent manner. Progressive carcinogenesis was measured by increasingly- acquired cancer-associated properties of reduced dependence on growth factors, anchorage-independent growth, acinar-conformational disruption, proliferation, migration, invasion, tumorigenicity with metastasis and increased stem-like cell populations. These biological changes were accompanied by biochemical and molecular changes, including upregulated H-Ras gene expression, extracellular signal-regulated kinase (ERK) pathway activation, Nox-1 expression, reactive oxygen species (ROS) elevation, increased HIF-1α, Sp1, tumor necrosis factor-α, matrix metalloproteinase (MMP)-2, MMP-9, aldehyde dehydrogenase activity and reduced E-cadherin. The Ras-ERK-Nox-ROS pathway played an important role in not only initiation but also maintenance of cellular carcinogenesis induced by PhIP. Using biological, biochemical and molecular changes as targeted endpoints, we identified that the green tea catechin components epicatechin-3-gallate and epigallocatechin-3-gallate, at non-cytotoxic doses, were capable of suppressing PhIP-induced cellular carcinogenesis and tumorigenicity.

Intracranial extension of retrobulbar blastomycosis (Blastomyces dermatitidis) in a dog.

Blastomycosis (Blastomyces dermatitidis) is a fungal disease that is endemic in the southern United States. This case report illustrates the clinical, MRI and histopathologic findings in a dog with invasion of a retrobulbar blastomycotic lesion into the calvarium. A 5-year-old intact female Weimaraner was referred for a 2-month history of change in behavior and recent onset of visual deficits. Magnetic resonance imaging (MRI) examination revealed a large (5.8 × 2.0 × 2.5 cm) mass extending from the left orbit through a circular defect in the left cranioventral aspect of the calvarium caudally to the level of the pituitary fossa and interthalamic adhesion. The mass was heterogeneously iso- to hypointense on T2-W images, slightly hypointense on T1-W images, did not attenuate on fluid attenuated inversion recovery (FLAIR) images, and did not show evidence of susceptibility artifact on T2*-W gradient recalled echo (GRE) images. Vasogenic edema and associated mass effect were noted. The mass showed strong homogeneous contrast enhancement with well-defined margins and had thickening of the adjacent meninges (dural tail sign). Based on MRI findings a malignant neoplastic process was considered most likely and the patient was placed on oral prednisone to decrease peri-tumoral inflammation. The dog initially improved but was euthanized 3 weeks later for worsening clinical signs. Histopathologic assessment of the mass revealed marked pyogranulomatous optic neuritis with intralesional fungal yeasts consistent with blastomycosis (Blastomyces dermatitidis). To our knowledge this is the first report of invasion of a retrobulbar blastomycotic lesion into the calvarium in a dog.

Lycoperdonosis in two dogs.

Lycoperdonosis is a rare respiratory disease that results from the inhalation of spores released from the Lycoperdon (puffball) mushroom. In the present study, 2 cases of confirmed canine lycoperdonosis are described. The first case presented to the Matthew J. Ryan Veterinary Hospital of the University of Pennsylvania, and the second case was submitted for postmortem examination to the University of Tennessee Veterinary Teaching Hospital. Both dogs presented in respiratory distress, and owners reported that the dogs had been playing or digging in areas with puffball mushrooms prior to the onset of clinical signs. In the initial case, thoracic radiographs revealed a diffuse interstitial and multifocal alveolar pulmonary pattern. Despite aggressive medical treatment and mechanical ventilation, the dog continued to worsen and was euthanized. Postmortem examination revealed firm lung lobes and enlarged tracheobronchial lymph nodes. Histologically, there was a severe diffuse histiocytic and pyogranulomatous bronchointerstitial pneumonia. Throughout the lung and lymph nodes, most commonly within macrophages, were round, 3-5 µm in diameter, Gomori methenamine silver-positive structures, consistent with Lycoperdon spores. An approximately 750-base pair DNA fragment was amplified from lung of both cases by polymerase chain reaction using primers specific to yeast ribosomal DNA, and the sequence of the fragment was determined to be most closely related to Lycoperdon pyriforme. Importantly, reexamination of an endotracheal wash from the initial case revealed intrahistiocytic spores, suggesting that airway sampling may assist in diagnosing lycoperdonosis.

Nonsteroidal anti-inflammatory drug-activated gene-1 expression inhibits urethane-induced pulmonary tumorigenesis in transgenic mice.

The expression of nonsteroidal anti-inflammatory drug-activated gene-1 (NAG-1) inhibits gastrointestinal tumorigenesis in NAG-1 transgenic mice (C57/BL6 background). In the present study, we investigated whether the NAG-1 protein would alter urethane-induced pulmonary lesions in NAG-1 transgenic mice on an FVB background (NAG-1(Tg+/FVB)). NAG-1(Tg+/FVB) mice had both decreased number and size of urethane-induced tumors, compared with control littermates (NAG-1(Tg+/FVB) = 16 +/- 4 per mouse versus control = 20 +/- 7 per mouse, P < 0.05). Urethane-induced pulmonary adenomas and adenocarcinomas were observed in control mice; however, only pulmonary adenomas were observed in NAG-1(Tg+/FVB) mice. Urethane-induced tumors from control littermates and NAG-1(Tg+/FVB) mice highly expressed proteins in the arachidonic acid pathway (cyclooxygenases 1/2, prostaglandin E synthase, and prostaglandin E(2) receptor) and highly activated several kinases (phospho-Raf-1 and phosphorylated extracellular signal-regulated kinase 1/2). However, only urethane-induced p38 mitogen-activated protein kinase (MAPK) phosphorylation was decreased in NAG-1(Tg+/FVB) mice. Furthermore, significantly increased apoptosis in tumors of NAG-1(Tg+/FVB) mice compared with control mice was observed as assessed by caspase-3/7 activity. In addition, fewer inflammatory cells were observed in the lung tissue isolated from urethane-treated NAG-1(Tg+/FVB) mice compared with control mice. These results paralleled in vitro assays using human A549 pulmonary carcinoma cells. Less phosphorylated p38 MAPK was observed in cells overexpressing NAG-1 compared with control cells. Overall, our study revealed for the first time that the NAG-1 protein inhibits urethane-induced tumor formation, probably mediated by the p38 MAPK pathway, and is a possible new target for lung cancer chemoprevention.