Comparison of abdominal ultrasound and magnetic resonance imaging for detection of abdominal lymphadenopathy in dogs with metastatic apocrine gland adenocarcinoma of the anal sac.

Imaging studies in humans with anal and rectal cancer indicate that magnetic resonance imaging (MRI) is a more sensitive technique than abdominal ultrasound (AUS) for the detection of abdominal lymphadenopathy. The purpose of this retrospective study was to directly compare the efficacy of these two techniques in detecting abdominal lymphadenopathy in dogs with apocrine gland adenocarcinoma of the anal sac (AGAAS). Six dogs with histologically confirmed AGAAS and histopathologic confirmation of metastasis to abdominal lymph nodes (LNs) had AUS and abdominal MRI. AUS identified lymphadenopathy in two of six dogs, whereas MRI identified lymphadenopathy in all the six dogs. Lymphadenopathy was predominantly sacral in location, with involvement of the medial iliac and hypogastric LNs in only two cases. These data suggest that MRI is more sensitive than AUS for detecting sacral abdominal lymphadenopathy in dogs with AGAAS. As such, MRI could be considered in any patient with AGAAS for initial staging of this disease.

Risk factor analysis of Histoplasma capsulatum fungemia.

The primary objective of the study was to investigate the risk factors for Histoplasma capsulatum fungemia. We conducted a retrospective case-control study among patients with histoplasmosis seen at Mayo Clinic in Rochester from 1 January 1991 through 31 December 2005. Blood cultures were prepared from specimens obtained from 111 patients with a diagnosis of histoplasmosis of which 55 had demonstrated H. capsulatum fungemia whereas the cultures of the remaining 56 patients were negative. The mean age of the patients was 56 years, of which 70% men and 95% were white. In univariate analysis, immunocompromised status (OR 2.9, P=0.008), peripheral leukocyte count (WBC)<3000 cells/mm(3) (OR 7.3, P<0.001), albumin<3.5 g/ dl (OR 3.1, P=0.018), and Charlson score of>4 (OR 2.9, P=0.022) were associated with H. capsulatum fungemia, but age>55 years was not (OR 1.4, P=0.38). In multivariable analysis, immunocompromised status (OR 2.4, P=0.043) and WBC<3000 cells/mm(3) (OR 6.5, P=0.001) remained significant factors associated with H. capsulatum fungemia. Immunocompromised status and WBC<3000 cells/ mm(3) are independent risk factors for the development of H. capsulatum fungemia.

Multicenter evaluation of a Candida albicans peptide nucleic acid fluorescent in situ hybridization probe for characterization of yeast isolates from blood cultures.

We evaluated aliquots from 244 clinical blood culture bottles that demonstrated yeasts on Gram stain using a Candida albicans peptide nucleic acid (PNA) fluorescent in situ hybridization (FISH) probe. The sensitivity, specificity, positive predictive value, and negative predictive value of the C. albicans PNA FISH test in this study were 99%, 100%, 100%, and 99.3%, respectively.

Influence of starch intake on growth and skeletal development of weanling horses.

Forty-four weanling horses were used in two experiments to evaluate the effect of starch intake on growth and skeletal development. In Exp. 1, the weanlings were fed either a grain-based, high-starch (31.1%, DM basis) concentrate or a by-product-based, low-starch (0.0%) concentrate with coastal bermudagrass (Cynodon dactylon) hay. Corn oil was used to equalize the energy concentration of the concentrates. The concentrate:hay ratio was 64:36 (as-fed basis), and intake was the same for both diets. Body weight gains were greater by the weanlings consuming the high-starch concentrate (0.81 vs. 0.67 kg/d; P = 0.01). Total body length gain also was greater for the weanlings consuming the high-starch concentrate (15.5 vs. 13.2 cm; P = 0.045). Other body measurements and bone mineral deposition were not influenced by diet or gender. At the end of the experiment, postprandial blood glucose concentrations suggested that the horses on the low-starch diet were less efficient in metabolizing blood glucose than were those that had been consuming the high-starch diets. In Exp. 2, the weanlings were fed either a high-starch (34.7%) or medium-starch (17.0%) concentrate plus coastal bermudagrass hay. Corn oil again was used to equalize the energy content of the medium-starch concentrate to that of the high-starch concentrate. The concentrate:hay ratio was 64:36 (as-fed basis), and the intake was the same for both diets. The diets did not influence rate of gain (0.75 kg/d; P = 0.98), body measurements (P = 0.11 to 0.93), or bone mineral deposition (P = 0.66). Animals on the medium-starch diet tended to have blood glucose concentrations that peaked earlier and were lower at later times than those consuming the high-starch concentrate. Bone osteochondrotic lesions were not related to the diet and were found to decrease during the course of the experiment for both the high-starch and the medium-starch diets (P = 0.006 and 0.016, respectively).

In situ hybridization for the differentiation of Aspergillus, Fusarium, and Pseudallescheria species in tissue section.

Identification of fungi in tissue sections can be difficult. In particular, species of Aspergillus, Fusarium, and Pseudallescheria all appear as septate, branched hyphae. However, their differentiation can have significant clinical implications, as the latter two groups are often resistant to commonly used antifungal agents. In situ hybridization may assist in rapidly distinguishing these organisms in the absence of available culture. Oligonucleotide DNA probes were directed against the 5S, 18S, or 28S rRNA sequences of three groups of fungi with a high degree of specificity for each. Probes were tested on 26 formalin-fixed, paraffin-embedded tissue specimens, each with culture-proven involvement by one of these organisms: Fusarium species, n = 12; Pseudallescheria boydii, n = 5; Aspergillus species, n = 9 ( probe set validated in an earlier study). Accuracy of both ISH and morphology was compared with culture. Morphologic examination (GMS and PAS) showed a greater sensitivity in detecting fungi (100%) as compared with in situ hybridization (84.6%). When detected, however, DNA probes allowed definitive identification of organisms. While there was no ability to distinguish between the three groups of organisms by morphologic features, ISH probes showed 100% positive predictive value (PPV, 19/19 organisms identified correctly). No cross-reactivity was observed when the probes were tested against other genera (100% specificity). Furthermore, the use of ISH allowed the detection of mixed fungal infections involving multiple organism types in two cases, demonstrating another advantage over morphology. In situ hybridization, directed against rRNA sequences, provides a rapid and accurate technique for distinguishing commonly encountered, nonpigmented filamentous fungi in histologic sections. While less sensitive than morphology, ISH is highly accurate and may help to distinguish between organisms that have similar or identical morphologic features by light microscopy.

In situ hybridization for the identification of filamentous fungi in tissue section.

Identification of fungi in tissue sections can be difficult because of limited biopsy tissue with only a few organisms present, or mycelial elements may be the only forms present, rendering common organism types indistinguishable from one another. In situ hybridization may assist in the rapid and accurate identification of such fungi. In this study, DNA probes were directed against the 5S or 18S ribosomal RNA sequences of three groups of fungi with a high degree of specificity for each. Two of the three, Aspergillus and Zygomycetes species, are usually seen in tissue purely in their hyphal forms. The third, Candida species is seen less commonly as predominantly mycelial elements. Probes were tested on 61 formalin-fixed, paraffin-embedded tissue specimens, each with culture-proven involvement by one of these organisms (Candida species, n = 21; Aspergillus species, n = 27; Zygomycetes, n = 13). Accuracy of both in situ hybridization (ISH) and morphology, based on the examination of Grocott methanamine silver (GMS)- and periodic acid-Schiff (PAS)-stained slides, was compared with culture. The results showed that morphologic examination (GMS and PAS) showed a slightly greater sensitivity in detecting the presence of fungi (98%) compared with in situ hybridization (95%). DNA probes, however, were more accurate in correctly identifying those organisms present. Although ISH specific probes showed 97% positive predictive value (PPV), examination of GMS-and PAS-stained slides had an 86% PPV when compared with culture-based identification methods. These results show that ISH, directed against ribosomal RNA, provides a rapid and accurate technique for the identification of mycelial fungal organisms in histologic tissue sections. Its primary use lies in the ability to accurately distinguish between organisms that have similar or identical morphologic features by light microscopy.

Quantitative analysis of chondroitin sulfate isomers in intervertebral disk chondrocyte culture using capillary electrophoresis.

Chondroitin sulfate (CS) isomers, 6-sulfate (CS6) and 4-sulfate (CS4), change their ratio to each other in cartilaginous tissues with aging. In this study, a quantitative measurement method of CS6 and CS4 was developed, using capillary electrophoresis (CE). Various buffer solutions, pH, and digestion times were studied, and the use of 0.1 M Tris-HCl at pH of 8.0 allowed the isolation of CS6 and CS4 from CS most efficiently when combined with chondrotinase ABC at a concentration of 1 mU/microg of the substrate during a 3 hr digestion period. Amounts of newly synthesized CS6 and CS4 in the intervertebral disk chondrocyte three-dimensional culture were quantified by this method after the proteoglycans were extracted by equilibrium density centrifugation.

Computed radiography.

CR is a useful tool for the equine veterinarian and has many advantages. Its only major disadvantage is the initial cost, in dollars and time, for setup in one's practice. CR is already in use at several universities and private practices in the United States and around the world. In the future, as is the case with any computer-based technology, this modality should become more affordable and readily available to smaller practices. The potential of CR in veterinary medicine combined with saturation of the human market is driving the development of specialized software and algorithms for veterinary medicine.

Functional interaction between fluorodeoxyuridine-induced cellular alterations and replication of a ribonucleotide reductase-negative herpes simplex virus.

G207 is an oncolytic herpes simplex virus (HSV) which is attenuated by inactivation of viral ribonucleotide reductase (RR) and deletion of both gamma(1)34.5 genes. The cellular counterparts that can functionally substitute for viral RR and the carboxyl-terminal domain of ICP34.5 are cellular RR and the corresponding homologous domain of the growth arrest and DNA damage protein 34 (GADD34), respectively. Because the thymidylate synthetase (TS) inhibitor fluorodeoxyuridine (FUdR) can alter expression of cellular RR and GADD34, we examined the effect of FUdR on G207 bioactivity with the hypothesis that FUdR-induced cellular changes will alter viral proliferation and cytotoxicity. Replication of wild-type HSV-1 was impaired in the presence of 10 nM FUdR, whereas G207 demonstrated increased replication under the same conditions. Combined use of FUdR and G207 resulted in synergistic cytotoxicity. FUdR exposure caused elevation of RR activity at 10 and 100 nM, whereas GADD34 was induced only at 100 nM. The effect of enhanced viral replication by FUdR was suppressed by hydroxyurea, a known inhibitor of RR. These results demonstrate that the growth advantage of G207 in FUdR-treated cells is primarily based on an RR-dependent mechanism. Although our findings show that TS inhibition impairs viral replication, the FUdR-induced RR elevation may overcome this disadvantage, resulting in enhanced replication of G207. These data provide the cellular basis for the combined use of RR-negative HSV mutants and TS inhibitors in the treatment of cancer.

In situ hybridization for the identification of yeastlike organisms in tissue section.

The identification of yeast and yeastlike organisms in tissue sections can be very difficult. Biopsy tissues may be limited, with only occasional organisms present. In addition, several common species have overlapping histologic features. Deoxyribonucleic acid probes were designed to detect both the 18S and 28S ribosomal ribonucleic acid sequences of five fungal organisms with a high degree of specificity for each fungus. Each of these organisms--Blastomyces dermatitidis, Coccidioides immitis, Cryptococcus neoformans, Histoplasma capsulatum, and Sporothrix schenckii--can be manifested histologically as round, yeastlike structures, often within a similar size range. Probes were tested against 98 archived, formalin-fixed, paraffin-embedded tissue specimens, each of which had culture-proved involvement by one of these organisms. Assessment of accuracy was based on the presence of yeastlike organisms in consecutive Grocott's methanemine silver (GMS)-stained tissue sections, and agreement with culture results. The results indicated that GMS had a greater overall sensitivity in detecting fungal organisms (95.9%) compared with in situ hybridization (ISH; 82.7%). ISH with oligonucleotide deoxyribonucleic acid probes, however, was more specific, with all species-specific probes yielding 100% specificity (compared with 96.2-100% specificity based on morphology alone). ISH also had a higher positive predictive value (100% in all cases) compared with GMS (83.3-100%). In addition, four cases with rare organisms present (4.1% of cases tested) were detected by ISH but not by GMS staining. These results show that ISH, directed against ribosomal ribonucleic acid, provides a rapid, accurate technique for the identification of yeastlike organisms in histologic tissue sections. Its primary strength lies in the ability to speciate organisms accurately that are too few or atypical to identify based solely on morphologic features.

Ramichloridium mackenziei brain abscess: report of two cases and review of the literature.

We report two cases of brain abscesses caused by Ramichloridium mackenziei, a neurotropic dematiaceous fungus that seems to be geographically restricted to the Middle East. One of the patients had chronic myelomonocytic leukemia but did not receive any chemotherapeutic agents. The other patient was a normal host. Both cases had a fatal outcome despite aggressive antifungal therapy and surgical intervention. Herein, we review all previously described cases in the literature, and discuss the epidemiology, mycology and histopathology of this life-threatening organism.

Quantitative hepatobiliary scintigraphy in normal cats and in cats with experimental cholangiohepatitis.

Quantitative hepatobiliary scintigraphy using 99mTc-mebrofenin was performed on eight normal cats and on the same cats after induction of experimental cholangiohepatitis by infection with the liver fluke Platynosomum concinnum. Hepatobiliary scintigraphy was performed 3 times at 10 weeks, 4 months and 6 months after infection. In addition, routine biochemical tests, hepatic ultrasound and ultrasound guided hepatic biopsy samples were obtained at the same time points, and the results compared with hepatobiliary scintigraphy. The normal hepatic extraction fraction was determined to be 85%, and the normal hepatic excretion half time (T 1/2) was 14 minutes. There was no significant change in scintigraphic parameters compared to pre-infection values at any time following infection with the liver fluke. No correlation between scintigraphic parameters and histologic scores was found; however, significant correlation was identified between parasite burden and histologic scores 6 months following infection. Despite the presence of severe multifocal histologic abnormalities, minimal clinical, biochemical and scintigraphic derangements were identified using this model of cholangiohepatitis. Based on this study, hepatobiliary scintigraphy appears to be an insensitive test for structural hepatobiliary abnormalities. The role of hepatobiliary scintigraphy in functional hepatobiliary abnormalities of the feline liver has not been determined.

Sporotrichosis masquerading as pyoderma gangrenosum: case report and review of 19 cases of sporotrichosis.

We present the case of a 59-year-old woman who had large ulcerations on her right leg that were diagnosed initially as pyoderma gangrenosum and treated with three immunosuppressive agents (cyclosporin, prednisone and azathioprine) for 6 months. Results of a biopsy at 6 months showed numerous cigar-shaped bodies consistent with Sporothrix schenckii; identification was confirmed by tissue culture. A retrospective review was performed for all cases diagnosed as sporotrichosis from tissue culture or biopsy specimens at the Mayo Clinic. Nineteen cases were identified. The present case was the only one in which fungal organisms were visible on histological examination. The present case emphasizes the importance of making a definitive histological diagnosis in unusual ulcer cases or in suspected cases of pyoderma gangrenosum before the initiation of immunosuppressive therapy. The large number of cigar-shaped bodies in the tissue is a rare finding in sporothrix infection and has been reported in only two cases previously.

Performance of five agar media for recovery of fungi from isolator blood cultures.

We studied the recovery of 1,270 fungal isolates from 176,144 Isolator blood cultures (0.72% positive) on bacterial and fungal media, under routine and differing incubation conditions. Except with Histoplasma capsulatum, chocolate agar incubated for only 3 days proved to be an excellent medium for the recovery of fungi from the Isolator system.

Thoracic vertebral chordoma in a domestic ferret (Mustela putorius furo).

A castrated adult male 1.0-kg domestic ferret (Mustela putorius furo) was referred for evaluation of a mass of >5 mo duration that was associated with the thoracic spine. The ferret had motor dysfunction and no conscious proprioception or pain perception in either hind limb. Segmental reflexes were present. Survey radiography, myelography, and magnetic resonance imaging showed a mass involving the ninth and 10 thoracic vertebrae and compressing the spinal cord. Decompressive surgery was performed, but clinical signs persisted. A diagnosis of chordoma was made using histopathology and immunohistochemistry. This is the first reported case of a chordoma involving the thoracic vertebrae in a domestic ferret.

Esophageal transit of capsules in clinically normal cats.

OBJECTIVE: To evaluate the esophageal passage of capsules in clinically normal cats and determine the incidence of prolonged transit or entrapment. ANIMALS: 12 clinically normal adult cats. PROCEDURE: Esophageal transit of barium sulfate-filled capsules was evaluated fluoroscopically. Each cat was examined 3 times (36 examinations). Esophageal transit times were classified as normal (< or = 30 seconds) or prolonged (> 30 but < or = 240 seconds). Capsules were considered entrapped when transit times were > 240 seconds. RESULTS: Transit times were normal in 10 of the 36 (27.8%) examinations, whereas times were prolonged in 7 (19.4%) examinations. Capsules became entrapped in the midcervical region of the esophagus during 19 (52.8%) examinations. Following termination of each examination, cats with entrapped capsules were fed a small amount (0.5 to 1 ounce) of food; this resulted in passage of the capsule to the stomach. CONCLUSIONS AND CLINICAL RELEVANCE: The possibility of medication-induced esophagitis should be considered when orally administering ulcerogenic drugs to cats. It is recommended that a small volume of food be given following medications to ensure complete esophageal clearance.

Radiographic and ultrasonographic evaluation of the normal feline postpartum uterus.

The radiographic and ultrasonographic appearance of the normal involution process of the feline postpartum uterus has not been previously described. Six queens were examined to determine the normal radiographic and ultrasonographic appearance of the involuting postpartum uterus. Radiographic and ultrasonographic examinations were performed daily from days one through ten, then on days 12, 14, 18, 24, and 28 postpartum. Radiographically the mean total uterine thickness was 16.5 mm at day one postpartum. By day 14 the mean total uterine thickness was 10.5 mm and by day 24 postpartum the uterus was not radiographically visible. Ultrasonographically at day one postpartum the mean total uterine thickness was 16.6 mm and the mean uterine wall thickness was 2.7 mm. At day 14 postpartum the mean total uterine thickness was 6.2 mm and the mean wall thickness was 2.1 mm. At day 28 postpartum the uterus could still be identified ultrasonographically however individual wall layers were not discernable.

Ultrasonographic features of canine gastrointestinal pythiosis.

Pythiosis is a chronic pyogranulomatous infection of the gastrointestinal tract or skin caused by the water borne pathogen Pythium insidiosum. The ultrasonographic features of nine dogs with gastrointestinal pythiosis are reported. The stomach, duodenum, jejunum or colon were affected. All dogs had thickening of the gastrointestinal wall and areas with obliteration of the normal layered appearance. In one dog an eccentric mass was found arising from the serosal surface of the wall of the colon with mild diffuse wall thickening. Regional lymph node enlargement was seen in seven of the nine dogs. One dog had invasion of the pancreas and signs compatible with extrahepatic biliary obstruction. When compared to previous reports of gastrointestinal neoplasia, the features of wall thickening, loss of layering and regional lymphadenopathy are not considered specific for gastrointestinal pythiosis. Histological examination of tissue specimens is required for diagnosis.

Dynamic magnetic resonance imaging of the normal canine pituitary gland.

The pituitary glands of six normal dogs were evaluated using dynamic magnetic resonance imaging. T1 weighted images were obtained every 13 seconds for three minutes of three contiguous slices through the pituitary gland following a bolus intravenous injection of gadolinium-DTPA. Contrast enhancement was seen initially in the region of the pituitary stalk at 52-65 seconds followed by uniform enhancement at 104-143 seconds post injection. This pattern of enhancement was seen in all subjects and is similar to that reported in humans.