Prediction and model evaluation for space-time data.

Evaluation metrics for prediction error, model selection and model averaging on space-time data are understudied and poorly understood. The absence of independent replication makes prediction ambiguous as a concept and renders evaluation procedures developed for independent data inappropriate for most space-time prediction problems. Motivated by air pollution data collected during California wildfires in 2008, this manuscript attempts a formalization of the true prediction error associated with spatial interpolation. We investigate a variety of cross-validation (CV) procedures employing both simulations and case studies to provide insight into the nature of the estimand targeted by alternative data partition strategies. Consistent with recent best practice, we find that location-based cross-validation is appropriate for estimating spatial interpolation error as in our analysis of the California wildfire data. Interestingly, commonly held notions of bias-variance trade-off of CV fold size do not trivially apply to dependent data, and we recommend leave-one-location-out (LOLO) CV as the preferred prediction error metric for spatial interpolation.

Effect modification of the association between fine particulate air pollution during a wildfire event and respiratory health by area-level measures of socio-economic status, race/ethnicity, and smoking prevalence.

Fine particulate air pollution (PM2.5) is decreasing in most areas of the United States, except for areas most affected by wildfires, where increasing trends in PM2.5 can be attributed to wildfire smoke. The frequency and duration of large wildfires and the length of the wildfire season have all increased in recent decades, partially due to climate change, and wildfire risk is projected to increase further in many regions including the western United States. Increasingly, empirical evidence suggests differential health effects from air pollution by class and race; however, few studies have investigated such differential health impacts from air pollution during a wildfire event. We investigated differential risk of respiratory health impacts during the 2008 northern California wildfires by a comprehensive list of socio-economic status (SES), race/ethnicity, and smoking prevalence variables. Regardless of SES level across nine measures of SES, we found significant associations between PM2.5 and asthma hospitalizations and emergency department (ED) visits during these wildfires. Differential respiratory health risk was found by SES for ED visits for chronic obstructive pulmonary disease where the highest risks were in ZIP codes with the lowest SES levels. Findings for differential effects by race/ethnicity were less consistent across health outcomes. We found that ZIP codes with higher prevalence of smokers had greater risk of ED visits for asthma and pneumonia. Our study suggests that public health efforts to decrease exposures to high levels of air pollution during wildfires should focus on lower SES communities.

Melan A and S100 protein immunohistochemistry in feline melanomas: 48 cases.

Immunohistochemistry, using a monoclonal antibody to Melan A and a polyclonal antibody to S100 protein, was applied to 48 formalin-fixed, paraffin-embedded specimens of feline melanoma. Forty-two cutaneous, three oral, one mucocutaneous, and two metastatic melanomas comprised the tumors. Thirty-two tumors (67%) were positive for Melan A and 42 (87.5%) were positive for S100. All but one of the tumors that were positive for Melan A were also positive for S100. S100 was detected in 11 of 16 tumors that were negative for Melan A. Seventy-five percent (9 of 12) of amelanotic melanomas were negative for Melan A. Normal adrenal cortex, the cerebellum, and the skin had cells that were positive for Melan A. Sebaceous adenoma was the only nonmelanocytic tumor examined that reacted with antibody to Melan A. Although less sensitive than S100 protein, Melan A is more specific for melanoma and is useful in differentiating feline cutaneous melanoma from the more common pigmented basal cell tumor.

Immunoreactivity of A103, an antibody to Melan A, in canine steroid-producing tissues and their tumors.

The monoclonal antibody A103 to the melanocytic differentiation antigen Melan A stains human steroid-producing cells and their tumors. A total of 200 formalin-fixed, paraffin-embedded canine normal tissues and hyperplastic and neoplastic lesions of the adrenal gland, testis, and ovary were immunohistochemically tested for Melan A with antibody A103. Leydig cell tumors (23/23, 100%), Sertoli cell tumors (14/15, 93%), and adrenocortical adenomas (12/13, 92%) were consistently positive. Adrenocortical carcinomas (23/35, 65%) and granulosa cell tumors (10/17, 59%) were less frequently positive. All pheochromocytomas, seminomas, and dysgerminomas were negative. The pattern of staining was cytoplasmic, but nuclear staining was also frequently seen in normal Leydig cells and their tumors. As in human tumors, immunohistochemistry for Melan A stains many canine steroid-producing tumors and can be used to distinguish these tumors from those of nonstereidogenic cells.

Regression of hypertrophic osteopathy in a cat after surgical excision of an adrenocortical carcinoma.

A 12-year-old, spayed, female domestic shorthair cat was diagnosed with severe and extensive hypertrophic osteopathy of the appendicular skeleton. Diagnostic ultrasound detected a mass lesion in the right adrenal gland. A right adrenalectomy was performed, and histopathological examination confirmed an adrenocortical carcinoma. No radiographic evidence of pulmonary metastasis was found on initial presentation or recheck thoracic radiographs taken 15 weeks later. Almost complete regression of periosteal new bone formation occurred 15 weeks following the successful surgical removal of the adrenal tumor.

Zinc toxicosis in a captive striped hyena (Hyaena hyaena).

An 11-yr-old captive-born female striped hyena (Hyaena hyaena) acutely developed lameness and swelling of the left front foot with anorexia, depression, and lethargy. Hematologic evaluation revealed regenerative anemia, azotemia, and other mild serum electrolyte and mineral abnormalities. Twenty radiographically visible coins and 10 coin fragments were removed by laparotomy and gastrotomy following unsuccessful medical therapy. The animal died during anesthetic recovery. Zinc serum levels were 41.0 ppm at first presentation and 36.0 ppm at the time of surgery, compared with concentrations of 1.78 ppm and 2.82 ppm for serum taken from this female and a male hyena 3 mo previously. Zinc toxicosis was diagnosed based on the similarity of clinical signs to those described in dogs, presence in the stomach of pennies minted after 1982 (when the zinc content of U.S. pennies was increased substantially), necropsy findings, and elevated serum and liver zinc values. The case highlights the risk posed by penny ingestion for subsequent zinc toxicosis in captive omnivores.

Growth hormone and insulin-like growth factor-I enhance beta-glucuronidase gene activation by androgen in mouse kidney.

Beta-glucuronidase (GUS) is a lysosomal enzyme that, in mouse kidney, is subject to control by multiple hormones: androgen, which increases GUS transcription; estrogen, which antagonizes androgen-mediated stimulation of GUS; and growth hormone (GH), which appears to be necessary for the full androgen effect. Neither estrogen nor GH affects GUS in the absence of androgen. In hypophysectomized or pituitary dwarf mice the reduced androgen stimulation of GUS can be partially restored with GH treatment. Androgen-induced GUS mRNA increased significantly with intermittent GH, compared to no GH or continuous GH. Intact mice subjected to continuous infusion of GH showed a depressed androgen effect on GUS similar to that seen in GH-deficient mice. Thus, pulsatile GH is required for the full androgen response. Insulin-like growth factor-I (IGF-I) also restored GUS induction by androgen in GH-deficient mice. We conclude that GH enhances the effect of androgen on the GUS gene via IGF-I. Using transgenic mice, we have also identified a genetic variant of the GUS gene that is insensitive to GH enhancement of the androgen effect.

Elimination of lysosomal storage in brains of MPS VII mice treated by intrathecal administration of an adeno-associated virus vector.

Mucopolysaccharidosis type VII (MPS VII) is an inherited lysosomal storage disease caused by insufficient beta-glucuronidase (GUS). To provide gene therapy in a mutant mouse model of this disease, we have used a recombinant adeno-associated virus (rAAV) vector to deliver GUS cDNA to a variety of tissues. Although intravenous administration of vector produced therapeutic levels of GUS in the liver, delivery to the brain was inadequate. To improve delivery to the brain intrathecal injection of the vector into the cerebrospinal fluid was employed. This route of administration to either neonatal or adult mutant mice resulted in therapeutic levels of GUS in the brain and the elimination of storage granules in brain tissue.

Disseminated Rhodococcus equi infection in two goats.

Rhodococcus equi infection was diagnosed in two goats from the same herd. At necropsy, numerous caseating granulomas were disseminated throughout the liver, lungs, abdominal lymph nodes, medulla of right humerus, and the right fifth rib of goat No. 1, and the liver of goat No. 2. Histopathologic examination confirmed the presence of multiple caseating granulomas in these organs. Numerous gram-positive and Giemsa-positive coccobacilli were identified within the cytoplasm of macrophages. Aerobic bacterial cultures of the liver and lung from both goats yielded a pure growth of R. equi. R. equi antigens were immunohistochemically identified in caseating granulomas from both goats. However, the 15- to 17-kd virulence antigens of R. equi were not detected, suggesting possible infection by an avirulent strain of this organism.

Intestinal multinodular A lambda-amyloid deposition associated with extramedullary plasmacytoma in three dogs: clinicopathological and immunohistochemical studies.

Intestinal extramedullary plasmacytomas (EMPs) are rare tumours in dogs. Three cases of canine intestinal EMP with amyloid deposits are described in this report. These tumours, which were located in the rectal submucosa, had variable numbers of well-differentiated plasma cells and fewer multinucleated giant cells of plasmacytoid and histiocytic morphology, admixed with abundant amyloid. Two cases had metaplastic cartilage and bone within the amyloid deposits. Immunohistochemically, the plasma cells of all three tumours reacted for lambda-light chains of immunoglobulins but not for kappa-chains, indicating monoclonality. Plasma cells of two tumours were also positive to CD79a antiserum. Amyloid deposits were labelled with an A lambda (amyloid of immunoglobulin lambda-light chain origin) antiserum but not with antisera against its precursor protein, the immunoglobulin lambda-light chains, indicating possible conformational changes of amyloidogenic proteins during their transformation into amyloid.

Actinomycotic splenitis and intestinal volvulus in an alpaca (Lama pacos).

Morphologic, microbiologic, and polymerase chain reaction amplification techniques were used to evaluate an alpaca (Lama pacos) with splenitis and intestinal volvulus. The intestinal volvulus produced a severe necrosuppurative typhlocolitis associated with vascular thrombosis and was most likely the cause of death of this animal. In addition, this animal had multiple coalescing abscesses affecting most of the splenic tissue. The isolation of Actinomyces spp. from the spleen and the morphology of the colonies when stained with Gram and Steiner stains support a diagnosis of splenic actinomycosis.

Prevalence of autoantibodies to thyroglobulin in dogs with nonthyroidal illness.

OBJECTIVE: To evaluate a thyroglobulin autoantibody (TgAA) assay and determine a diagnostic threshold. SAMPLE POPULATION: Serum samples from dogs with various endocrine abnormalities and from 30 obese adult female Beagles. PROCEDURE: TgAA were determined by use of the ELISA. Six experiments were done: 1, definition of positive results for TgAA using samples from normal and T3 autoantibody (T3AA) positive dogs; 2, establishment of prevalence of positive results in 91 clinically normal dogs; 3, evaluation of positive results for sera from dogs with nonthyroidal illnesses; 4, testing of samples from dogs with primary hypothyroidism but absence of T4AA or T3AA, or both; 5, determination of prevalence of false-negative results in dogs that are T4AA and/or T3AA positive, which were (18 dogs) or were not (22 dogs) receiving L-thyroxine replacement therapy; and 6, examination of thyroid biopsy specimens from 18 dogs (8 TgAA positive and 10 TgAA negative). RESULTS: Positive results were defined as at least twice (200%) the optical density of the negative-control sample. False-positive results were obtained for only 3.4% of 146 dogs with nonthyroidal illness. Thirty-seven percent of dogs with primary hypothyroidism, but no evidence of T4AA or T3AA, or both, were TgAA positive. False-negative results were found in 1 of 22 and 2 of 18 T3AA-positive dogs with and without thyroid replacement therapy, respectively. Thyroid biopsy specimens from 8 TgAA-positive dogs had evidence of lymphocytic thyroiditis, whereas those from 10 TgAA-negative dogs did not. CONCLUSION AND CLINICAL RELEVANCE: The assay is sensitive and specific for identification of lymphocytic autoimmune thyroiditis in dogs, and has potential for aiding early diagnosis of thyroiditis in dogs and identifying dogs likely to perpetuate hypothyroidism in breeding programs.

Characterization of natural occurring Pneumocystis carinii pneumonia in pigs by histopathology, electron microscopy, in situ hybridization and PCR amplification.

Macroscopic, histologic, ultrastructural, microbiologic, in situ hybridization (ISH) and PCR detection results in three 8-week-old pigs naturally infected with Pneumocystis carinii (PC) are described. All animals had a nonsuppurative interstitial pneumonia and intra-alveolar Pneumocystis organisms with foamy eosinophilic and PAS positive appearance. Ultrastructurally, PC trophozoites and cysts were observed in pigs No. 2 and No. 3, with the former being much more numerous. PC organisms were located on the alveolar surface or within the alveolar septa. Trophozoites had numerous filopodia and were thick-walled. Cysts had no or few filopodia, were thick-walled and contained intracystic bodies. Using non-isotopic ISH on formalin-fixed, paraffin-embedded lung tissue sections, PC DNA from pigs No. 2 and No. 3 hybridized with a probe specific for PC ribosomal RNA (rRNA). Using primers specific for mitochondrial rRNA gene (pAZ102-E/pAZ102-H), and for the internal transcriber spacers of ribosomal gene of PC, PCR methods amplified a product in the lung of pigs No. 2 and No. 3 using either frozen or formalin-fixed and paraffin-embedded lung tissue. DNA from Pig No. 1 samples did not amplify with any primer. This is the first time that molecular biology techniques (in situ hybridization and PCR) have been applied to the study of porcine pneumocystosis.

Treatment of lysosomal storage disease in MPS VII mice using a recombinant adeno-associated virus.

Mucopolysaccharidosis type VII (MPS VII) is a lysosomal storage disease caused by a genetic deficiency of beta-glucuronidase (GUS). We used a recombinant adeno-associated virus vector (AAV-GUS) to deliver GUS cDNA to MPS VII mice. The route of vector administration had a dramatic effect on the extent and distribution of GUS activity. Intramuscular injection of AAV-GUS resulted in high, localized production of GUS, while intravenous administration produced low GUS activity in several tissues. This latter treatment of MPS VII mice reduced glycosaminoglycan levels in the liver to normal and reduced storage granules dramatically. We show that a single administration of AAV-GUS can provide sustained expression of GUS in a variety of cell types and is sufficient to reverse the disease phenotype at least in the liver.

Intestinal extramedullary plasmacytoma associated with amyloid deposition in three dogs: an ultrastructural and immunoelectron microscopic study.

Samples from rectal plasmacytoma in three adult dogs that were diagnosed by light microscopy and immunohistochemistry were examined by electron microscopy. The most common cell type had typical plasmacytoid features. A second cell type was a plasmacytoid giant cell with single or multiple eccentric nuclei, irregular nuclear membrane, abundant and dilated rough endoplasmic reticulum, and numerous electron-dense granules. The third cell type was a histiocytic giant cell that intermingled with plasmacytoid cells. All three tumors had abundant amyloid, mainly in the interstitium but also within histiocytic cells and less commonly in plasma cells or plasmacytoid giant cells. Extracellular and intracellular amyloid fibrils and the contents of membrane-bound electron-dense bodies of plasma cells reacted with antibody to lambda-light chain of immunoglobulins by immunogold staining.

Sarcocystosis in mink (Mustela vison).

This report describes the clinical, microscopic, and ultrastructural findings in mink with muscular sarcocystosis. Three 2-3-mo-old mink were killed because they were ill with signs of progressive neurological disease. One mink had variable numbers of sarcocysts in multiple skeletal muscles. Sarcocysts were up to 300 microm in long and 20 microm wide. Ultrastructurally, the sarcocyst wall had numerous elongated 1.7-2.0-microm x 250-nm villar protrusions (VP). The VP had microtubules and irregularly distanced minute undulations. Both metrocytes and bradyzoites were present in sarcocysts. The mink with sarcocysts in muscles also had nonsuppurative meningoencephalitis and meningomyelitis. Similar brain lesions were found in other 2 mink from the same farm, but sarcocysts were not observed in the skeletal muscle of these animals. This is the first report of muscular sarcocystosis in mink.

Malignant cauda equina paraganglioma in a cat.

An 11-year-old spayed female domestic long-haired cat presented for surgical removal of a slowly growing and deeply invasive 2.5 x 3.5-cm mass cranial to the base of the tail. Light microscopic examination of surgical biopsy specimens revealed an encapsulated mass composed of packets of polygonal cells of various sizes separated by a delicate fibrovascular stroma. Gömöri's reticulum stain revealed a characteristic endocrine or "Zellballen" pattern. Tumor cells contained diffuse positive reactivity to synaptophysin and neuron-specific enolase, reactions consistent with a neuroendocrine neoplasm. S-100 protein-positive cells reminiscent of sustentacular (support) cells occurred singly or in small clusters within tumor packets. At postmortem examination 3 months later, a 9- x 5- x 4-cm multinodular raised tan mass involving the caudal pelvis, sacrum, and tail-head regions was found. The base of this neoplastic mass originated within the cauda equina region and involved approximately five caudal nerve roots. Numerous 1-3-mm metastatic nodules were identified disseminated throughout the pulmonary parenchyma. The tumor was diagnosed as a malignant paraganglioma of the cauda equina region with pulmonary metastasis.

Lymphosarcoma with plasmacytoid differentiation in a scarlet macaw (Ara macao).

A lymphosarcoma in a scarlet macaw (Ara macao) affecting periocular structures is described. Microscopically and ultrastructurally, many of the lymphoid cells had plasmacytoid features. Polymerase chain reaction amplification failed to detect exogenous avian retrovirus RAV-1 in the neoplastic mass.

Omeprazole in a dog with gastrinoma.

A 9-year-old male German Shepherd Dog was presented with the primary complaints of vomiting, profuse watery diarrhea, anorexia, and severe weight loss. The dog developed hematemesis and melena, which were unresponsive to treatment with an H2-receptor antagonist and a gastrointestinal protectant. A marked neutrophilia, panhypoproteinemia, hypokalemia, and mildly increased activities of alkaline phosphatase and alanine aminotransferase were the only relevant abnormalities found on a CBC, serum biochemical profile, and urinalysis. An exploratory laparotomy revealed several small nonresectable masses at the root of the mesentery, which were identified histologically as a neuroendocrine neoplasm. Immunohistochemical staining of the neoplasm was positive for gastrin and negative for insulin, glucagon, pancreatic polypeptide, and vasoactive intestinal polypeptide. Fasting serum gastrin concentrations were high. Zollinger-Ellison syndrome was diagnosed, and the dog was treated with omeprazole, an H+,K(+)-ATPase inhibitor. All clinical signs resolved, and the dog remains asymptomatic 2 years later. Omeprazole may be the gastric acid antisecretory drug of choice for dogs with gastrinoma.

Ceruminous gland tumors in dogs and cats: a review of 124 cases.

The histological features of 124 ceruminous gland tumors from canine and feline biopsy submissions were reviewed. The tissues, which represented submissions from private veterinary practices and a veterinary college, included ceruminous gland adenocarcinomas and adenomas as well as a single, mixed ceruminous gland adenocarcinoma. A majority of the masses from both dogs and cats were identified as malignant processes.