[Magnetic resonance imaging and ultrasonography in dogs and cats with ocular and orbital diseases. Part 1: Ocular diseases]. / Kernspintomographische und sonographische Untersuchungen bei Hunden und Katzen mit intraokulären und orbitalen Erkrankungen. Teil 1: Intraokuläre Erkrankungen.

OBJECTIVE: This two-part study describes the clinical usefulness and value of ultrasound and magnetic resonance imaging (MRI) in dogs and cats with ocular (n=30) and orbital diseases (n=31). MRI and ultrasonography characteristics are described in single cases with ocular and orbital disease. MATERIAL AND METHODS: Ultrasonography and MRI were performed in 15 dogs and 15 cats with intraocular neoplasia or intraocular inflammatory disease. RESULTS: In all patients with intraocular neoplasia, sonography revealed masses with increased echogenicity and fairly uniform echotexture, thus allowing the tentative diagnosis of an intraocular tumour. In these cases, MRI often proved to be a valuable diagnostic tool in showing the complete extent of intraocular lesion. An additional benefit of MRI was seen in the tissue characterization of tumours based on MRI signal characteristics and pattern of contrast enhancement. Discreet intraocular inflammatory alterations, in particular to the anterior and posterior segment of the eyeball, were more clearly shown by ultrasound than by MRI. Neoplasia could be excluded and inflammatory disease was successfully diagnosed using MRI due to the different image sequences with or without contrast medium administration. Traumatic ruptures of the lens capsule and the globe after trauma were depicted more clearly with MRI. CONCLUSIONS AND CLINICAL RELEVANCE: When opacity of the anterior eye segment is present, various intraocular changes can be quickly diagnosed by ultrasound with high accuracy, without requiring anaesthesia of the patient. MRI of the globe allows differentiation of diverse pathologies, gives detailed information of infiltration in orbital structures and the exact degree of ocular lesions after trauma. This additional evidence often makes it easier to predict the correct prognosis and choose the best therapy.

[Two cases of disseminated toxoplasmosis]. / Zwei Fallberichte einer disseminierten Toxoplasmose.

Two cases of disseminated fatal toxoplasmosis of a pet mouse and a red panda are described. The pet mouse had a private owner; the red panda lived in a zoological garden in Saxony. At necropsy, both animals suffered from a systemic toxoplasmosis. A severe necrotizing hepatitis was the main histological feature in both animals. Parasitic cysts could be abundantly found in the liver, moderately in the brain and in a low number in other organs. With the PAS-reaction, cysts showed a weak staining in the mouse and a strong staining in the red panda. Diagnosis was confirmed by PCR and immunohistochemistry.

Immunohistochemical study of angiogenesis and angiogenic factors in equine granulosa cell tumours.

The first part of our study (Müller et al., 2009) characterized angiogenesis in the equine cycling ovary through histomorphological and immunohistochemical examinations (vascular endothelial growth factors A and B [VEGF A, VEGF B], vascular endothelial growth factor receptors 1 and 2 [VEGF-R1, VEGF-R2], vascular angiopoietins 1 and 2 [Ang1, Ang2], angiopoietin receptor [Tie2], and von Willebrand Factor). Since angiogenesis plays an important role in development and growth of numerous tumours, the second part of our study involved a similar examination of 70 equine granulosa cell tumours (GCTt). The results of the second study were compared with those of the normal equine ovary. Certain similarities in the expression pattern could be detected between normal, cyclical ovaries (Müller et al., 2009) and GCTt. The immunoreactivity of granulosa cells and Leydig-like cells in GCTt resembles granulosa cells and luteinized thecal cells in periovulatory cycling ovaries. The neoplastic cells support circulation, supply and growth of GCTt by contributing to angiogenesis.

Morpho-functional studies regarding the fertility prognosis of mares suffering from equine endometrosis.

The aim of the present study was to characterize the morpho-functional features of endometrosis in barren and foaling mares, using both conventional histopathological and immunohistochemical methods. Endometrial biopsy samples were collected during the physiological breeding season from 159 estrous, clinically healthy mares (mean age 12 years), and the quality and degree of endometrosis was histomorphologically defined. The mares were bred and those that foaled were put in the foaling group whereas those that did not foal were placed in the barren group. Foaling mares were then compared with barren mares. Sixty-four percent (101/159) of uterine samples showed varying degrees of endometrosis and were used for this study. The sample population consisted of 51 barren and 50 foaling mares suffering from endometrosis. Expression of steroid hormone receptors (estrogen receptor, progesterone receptor) and endometrial protein secretion patterns (uteroglobin [UG], uterocalin [UC], calbindin(D9k) [CAL], uteroferrin [UF]) was evaluated by immunohistochemistry (barren mares N = 51, foaling mares N = 31). In comparison with unaffected glands, fibrotic glands generally showed a cycle-asynchronous, partially patchy protein expression pattern which is interpreted as a sign of endometrial maldifferentiation within fibrotic areas. In barren mares (N = 51) more than half of biopsy samples (27/51) showed a destructive mostly moderate (20/27) type of endometrosis. In affected glands, staining for UG (17/21) was decreased (P < 0.001). Foaling mares (N = 50) frequently showed a mild, nondestructive endometrosis (35/50). Compared with barren mares, foaling mares had statistically (P < 0.05) more often a cycle-synchronous or increased UG expression pattern within fibrotic glands. Obvious deviations of either UG or UC rarely occurred. Within fibrotic foci, UF often demonstrated a cycle-synchronous or more intense expression pattern in both foaling (28/31) and barren mares (41/51), compared with healthy glands. Mares of both groups showed a cycle-asynchronous staining for estrogen receptor and progesterone receptor in the stromal cells in areas of periglandular fibrosis and the glandular epithelia. These findings indicate that affected areas become independent of the uterine control mechanisms and exhibit specific differentiation dynamics. Immunohistochemical investigations showed that the secretory patterns differ between barren and foaling mares. The findings in this study should be considered as a useful addition to the "classical" Kenney categorization.

Fungal contamination of paraffin wax blocks in a pathology archive.

While searching for paraffin wax blocks for research purposes in our archive we detected numerous larval and some dead adult moths. Some wax blocks were riddled with a white-brown crumbling substance. The entire archive was checked and profoundly-infested blocks were separated from unaffected blocks. Mycological and parasitological investigations were performed. Fungi were identified by culture and polymerase chain reaction, which revealed high sequence homology to six different fungal species. The moths were determined to be Nemapogon personellus. A total of 8,484 wax blocks had to be removed from the archive and destroyed. Pathologists should be alerted to the importance of checking the humidity of the air where archival material is stored.

Histomorphological and immunohistochemical study of angiogenesis and angiogenic factors in the ovary of the mare.

Cyclical ovaries of 18 mares were examined histologically and immunohistochemically for vascular endothelial growth factor A and B (VEGF A; VEGF B), angiopoietin1 and 2 (Ang1; Ang2), vascular endothelial growth factor receptor 1 and 2 (VEGF-R1; VEGF-R2), angiopoietin receptor (Tie2) and von Willebrand factor. The most intensive coexpression of the examined factors and receptors was detected in the periovulatory period, when a distinctive ovarian angiogenesis takes place, being essential for tertiary follicle maturation and for the endocrine function of the Corpus luteum. Based on the immunohistochemical results, VEGF A, Ang2, VEGF-R2 and Tie2 in particular seem to play a significant role on angiogenesis during follicular and luteal development in the mare, while Ang1 supports vessel stabilisation. The findings of luteal regression and follicular atresia showed that, in the absence of VEGF A, Ang2 and its receptor Tie2 contribute substantially to vessel regression and therefore to luteolysis and follicular atresia.

Histological and immunohistochemical characterization of equine anovulatory haemorrhagic follicles (AHFs).

Anovulatory haemorrhagic follicles (AHFs) are often the reason for ovulation failure in the mare. As the underlying factors that lead to AHF development are not well understood, it was of interest to investigate the vascularization of AHFs compared with normal follicles and corpora lutea (controls). In the present study, the ovarian cell populations investigated immunohistochemically included granulosa and luteal cells as well as various vascular structures. None of these cell types showed differences in the expression of vascular endothelial growth factor A (VEGF-A) between control ovaries containing normal follicles and corpora lutea and ovaries with AHFs. In contrast, a considerable reduction in the proportion of Flk-1-expressing cells, together with a decreased intensity of staining, was apparent in the AHFs. This greatly reduced expression of Flk-1 in the luteinized cells and the vascular structures of AHFs may lead to a distinct decrease in the potential pro-angiogenic activity of VEGF-A in these structures compared with the situation in normal follicles and corpora lutea. Furthermore, the authors suspect that the distinct expression of angiopoietin2 and VEGF-A seen in the cells within the inner fibrous layers of the AHFs was caused by hypoxia resulting from deficient vascularization, as suggested by the irregularity of the capillaries present in the luteinized wall of the AHF. In addition, whereas LH-receptor (LH-R) expression occurred uniformly in all stages of development of the corpora lutea in normal control ovaries, there was highly variable labelling for LH-R in all the AHFs examined, thereby indicating a possible numerical deficiency of LH-receptors in AHFs. The authors concluded that, despite the apparent expression of sufficient VEGF-A in the AHFs allows ovulation and corpus luteum formation, a relative lack of receptor, Flk-1, effects the pro-angiogenic activity of VEGF-A which could be a reason for ovulation failure associated with AHF formation.

Immunohistochemical and histochemical identification of proteins and carbohydrates in the equine endometrium Expression patterns for mares suffering from endometrosis.

Although alterations in patterns of protein secretion revealed in uterine flushings from mares suffering from endometrosis have been described, little is known about alterations at the cellular level. Hence, the aim of this study was to characterize deviations in patterns of uterine gland secretion patterns using endometrial biopsies, histochemical and newly established immunohistochemical methods. Forty-eight endometrial biopsies were obtained from mares suffering from various types of endometrosis (active and inactive, destructive and non-destructive) and degree (mild to severe) were analyzed for expression of the proteins uteroglobin, uteroferrin, calbindinD9k and uterocalin as representatives of endometrial proteins detectable by immunohistochemistry using polyclonal antibodies. Glycogen was identified using the PAS-reaction and mucopolysaccharides were stained with alcian blue. Uterine glandular epithelia within fibrotic foci mostly revealed a protein and carbohydrate pattern of expression which was independent of hormonal changes during the estrous cycle. In comparison to non-affected glands, most epithelial cells within periglandular fibrosis exhibited decreased immunostaining intensity for proteins, especially when there was destructive endometrosis. However, uteroferrin staining intensity was strong within areas of severe destructive endometrosis. Moreover, only few basal glandular epithelial cells, especially those in cystic glands, stained for mucopolysaccharides that are typically seen within the luminal epithelia. Usually a single fibrotic focus caused only slight alterations in glandular proteins and carbohydrate reaction patterns, so that only more severe endometrosis lead to deviations which were detectable in uterine flushings. The highly sensitive methods used in the present study allow studies of uterine secretion patterns in the context of routine assessment of endometrial biopsies.

Immunolocalisation of the uterine secretory proteins uterocalin, uteroferrin and uteroglobin in the mare's uterus and placenta throughout pregnancy.

Previous studies have shown that the equine uterus produces many progesterone-dependent proteins throughout gestation. In particular, uterocalin and uteroferrin are detectable using electrophoresis or blot analyses but information regarding the immunohistochemical placental distribution of these two proteins is rare and information regarding uteroglobin is still lacking. The aim of the present study was to co-immunolocalise these three secretory proteins in the mare's uterus throughout gestation in an effort to understand their functional role in the maintenance of pregnancy. Therefore, endometrial biopsy samples were obtained from 20 pregnant mares between 16 and 309 days of gestation and labelled immunohistochemically for uteroglobin, uteroferrin and uterocalin. Uteroferrin remained detectable in almost every endometrial gland at all stages but with an increase in staining intensity as gestation advanced. The most progesterone-dependent protein, uterocalin, showed variable staining throughout gestation with the most intense labelling in early pregnancy and during the period of endometrial cup reaction. Uteroglobin secretion was only detectable in traces and only in individual glands throughout gestation. The results indicate that uterocalin and uteroferrin, but not uteroglobin, may play important roles in supplying nutrients for the conceptus, thereby contributing to the maintenance of pregnancy. However, further investigations are necessary to understand the role of uteroglobin during gestation.

Effects of methylene blue on propofol requirement during anaesthesia induction and surgery.

Delayed emergence from anaesthesia and neurological disturbances have been reported in patients undergoing parathyroidectomy who received methylene blue (MB) pre-operatively. We hypothesised that MB would decrease propofol requirements. The Bispectral index (BIS) and a target-controlled infusion of propofol were used in two groups of 11 matched patients. Patients in one group were pretreated with MB. During induction, clinical sedation scores and BIS values were significantly lower at the predicted effect-site propofol concentration of 2 microg x ml(-1) in the MB compared with the control group. Intra-operatively, although similar BIS values were achieved in the two groups, patients pretreated with MB required a mean 50% lower dose of propofol compared with controls. In view of these findings, care should be taken to ensure an adequate depth of anaesthesia by titrating the administration of anaesthetic agents whenever MB is infused peri-operatively.

Mycobacterium pinnipedii: transmission from South American sea lion (Otaria byronia) to Bactrian camel (Camelus bactrianus bactrianus) and Malayan tapirs (Tapirus indicus).

Tuberculosis infections caused by Mycobacterium (M.) pinnipedii in a South American sea lion, Bactrian camel, and Malayan tapirs kept in two zoological gardens spanning a time period of 5 years are reported. The zoos were linked by the transfer of one tapir. Conventional bacteriological and molecular methods were applied to detect the pathogen. Spoligotyping and MIRU/VNTR-typing performed to assess the genetic similarity revealed identical molecular characteristics of the isolates from all animals involved. Anti-tuberculosis antibodies were detected using ELISA and a recently developed serological rapid test. The study shows that: (i) using molecular methods, the assessment of the genetic relationship of infectious agents helps to confirm the routes of infection, and that (ii) immunological tests may help to detect tuberculosis infections ante mortem more reliably and early. This would prevent the transfer of tuberculosis by asymptomatic animals.

Histomorphological and immunohistochemical characterization of equine granulosa cell tumours.

Benign (n=33) and malignant metastasizing (n=1) granulosa cell tumours (GCTs) from 34 mares aged 3-21 years, and normal (control) ovaries from nine mares aged 3-10 years, were examined histologically and immunohistochemically (for inhibin alpha, glutathione S-transferase alpha [GSTalpha], c-erbB-2 oncoprotein [cerb], cytokeratin, vimentin, desmin and alpha-actin), the results being related where appropriate to clinical signs and endocrinological data. Availability permitting, serum samples from GCT-affected mares before and several weeks after ovariectomy were examined for the following hormones: oestradiol, progesterone and testosterone (by radioimmunoassay); and inhibin B (by a cross-reactive ELISA). Histological examination revealed that the GCTs were predominantly well differentiated neoplasms. The metastasizing GCT differed immunohistochemically from the benign GCTs in respect of the expression patterns of vimentin, cerb and GSTalpha in the granulosa cells. A notable feature was the presence of Leydig-like cells in mares with stallion-like behaviour or elevated serum testosterone, or both. GSTalpha immunolabelling indicated that the Leydig-like cells were potential producers of steroid hormone. From the immunohistochemical and endocrinological findings it was concluded that GCTs produce abnormally high concentrations of inhibin, which reduce the release of follicle-stimulating hormone, leading to atrophy of the contralateral ovary-a finding in 27 of the mares.

Primary and secondary heart tumours in dogs and cats.

Primary and secondary neoplasms of the canine and feline heart are uncommon. During a 2-year period, 83 dogs suffering from primary cardiac (n=11), extracardiac benign (n=6) or malignant (n=66) tumours and 30 cats with primary cardiac (n=1) or extracardiac (n=29) malignant tumours were examined. Echocardiography revealed four cases of primary cardiac neoplasms in dogs, but secondary heart tumours were not detected. After necropsy, tissue samples from the heart and tumours were examined histologically and immunohistochemically. In dogs, primary neoplasms included seven haemangiosarcomas, two chemodectomas, one rhabdomyosarcoma, and one neurofibrosarcoma. In 24 of 66 dogs examined, metastases of extracardiac neoplasms were found in the heart (15 carcinomas, six malignant lymphomas, three haemangiosarcomas). In cats, one case of primary haemangiosarcoma of the pericardium and five cases of secondary cardiac tumours (two malignant lymphomas, three carcinomas) occurred. Cardiac neoplasms in cats were not identified clinically but were detected by detailed gross sectioning of the heart (n=2) or histopathological examinations (n=3). This study showed an unexpectedly high number (36%) of dogs with cardiac metastases.

Excitation spectrum of two correlated electrons in a lateral quantum dot with negligible Zeeman splitting.

The excitation spectrum of a two-electron quantum dot is investigated by tunneling spectroscopy in conjunction with theoretical calculations. The dot made from a material with negligible Zeeman splitting has a moderate spatial anisotropy leading to a splitting of the two lowest triplet states at zero magnetic field. In addition to the well-known triplet excitation at zero magnetic field, two additional excited states are found at finite magnetic field. The lower one is identified as the second excited singlet state on the basis of an avoided crossing with the first excited singlet state at finite fields. The measured spectra are in remarkable agreement with exact-diagonalization calculations. The results prove the significance of electron correlations and suggest the formation of a state with Wigner-molecular properties at low magnetic fields.

Cowpox virus infection associated with a streptococcal septicaemia in a foal.

Cowpox virus infection associated with a streptococcal septicaemia was diagnosed in a weak German Warmblood filly, born 29 days prematurely, and humanely destroyed on the sixth day of life. At necropsy, ulcerative lesions in the alimentary tract, colitis, polyarthritis and nephritis were observed. Transmission electron microscopical examination of specimens from ulcerative lesions revealed typical orthopox virions. Cowpox virus was unequivocally identified by virological and molecular-biological methods.

Functional pathology of the ovaries and uteri of premature female piglets exposed to distinct amounts of zearalenone.

The objective of this study was to investigate the effects of four different concentrations (groups II-V) of orally administered zearalenone (ZON) over a feeding period of 35 days on parts of the genital tract of 75 day-old female piglets in comparison to a control group (I). In order to determine possible dose related characteristics for a ZON intoxication in ovaries and uteri, histomorphological (HE and Azan stained slides) and immunohistochemical methods [expression patterns of estrogen (ER) and progesterone (PR) receptor, proliferating cell nuclear antigen (PCNA)] were carried out. Using these methods, there were no obvious (immuno-) histomorphological differences within the treatment groups or in comparison to the control group. Nevertheless, compared to the group I, statistical analyses of all parameters investigated revealed significance only in the increased mitotic activity of muscle cells in the uterinelamina muscularis in treatment group V.

Cardiovascular response to acute normovolaemic haemodilution in patients with severe aortic stenosis: assessment with transoesophageal echocardiography.

Using multiplane transoesophageal echocardiography (TOE), we investigated the haemodynamic response to acute normovolaemic haemodilution (ANH) in anaesthetised patients with critical aortic stenosis. Twenty-eight patients were randomly assigned to ANH or control groups. In the control group, haemodynamic data remained unchanged over a 20-min period. In the ANH group, haemoglobin levels decreased from a mean (SD) of 134 (7) to 91 (9) g x l(-1) (p < 0.001) whereas stroke volume, central venous pressure and left ventricular (LV) end-diastolic area all increased significantly (mean (SD) +15 (6) ml; +2.0 (1.1) mmHg; +2.1 (0.8) cm2, respectively). During ANH, the accelerated blood flow through the stenotic valve caused an increased loss (SD) in LV stroke work: from 24 (8)% to 30 (10)%), (p < 0.01). Hence, lowering viscosity with ANH resulted in improved venous return, higher cardiac preload and increased stroke volume. However, this adaptive haemodynamic response was limited by less efficient LV stroke work due to dissipation of fluid kinetic energy.

[Uncommon mycobacterial infections in domestic and zoo animals: four cases with special emphasis on pathology]. / Ungewöhnliche Mykobakterien-Infektionen bei Haus- und Zootieren: eine Kasuistik unter besonderer Berücksichtigung der Pathologie.

Infections caused by classical tubercle bacilli are rare during the last years. Nevertheless, diseases caused by other mycobacteria have to be considered clinically and in diagnostic pathology especially in cases of immunosuppression and due to their potential zoonosis risk. An infection by mycobacteria was diagnosed in four animals (Mayotte Maki, Blue-headed Parrot, Patagonian sealion, Beagle) necropsied between 1995 and 2002 in the Institute of Veterinary-Pathology of the University of Leipzig. The Maki, the blue-headed parrot and the dog showed a disseminated character of the disease caused by Mycobacterium genavense (monkey and bird) resp. Mycobacterium avium (dog), while an open chronical tuberculosis of the lungs due to a pathogenic member of Mycobacterium tuberculosis complex was observed in the seal. All these bacteria are potential causes of zoonoses. So, if granulomatous or disseminated histiocytic alterations are detected in diagnostic pathology, mycobacterial infections should always be included in differential diagnoses and require careful aetiological investigations by histopathological and bacteriological methods.

The effect of vasodilators on aspirin-induced antagonism of t-PA thrombolysis.

Although i.v. t-PA has proven successful in reducing neurologic deficits in acute ischemic stroke, the disadvantages of a narrow therapeutic time window and the failure of thrombolysis in more than 50% of patients treated have necessitated an examination of adjuvant therapies to improve the rate of thrombolysis. Experimentally, the combination of aspirin therapy with t-PA has resulted in a paradoxical antagonism of thrombolysis. Reversal of this antagonism with nitric oxide (NO) donors suggested that aspirin may inhibit/ antagonize NO-related mechanisms. Using this rabbit model of thromboembolic stroke, this hypothesis is now expanded to compare two clinically relevant anti-hypertensive agents, atenolol (NO-dependent) and hydralazine (NO-independent), for their ability to improve t-PA-mediated clot lysis following aspirin pre-treatment. Thirty rabbits (10 per group) were pre-treated with aspirin (20mg kg(-1), i.v.) and then randomized to receive either vehicle, atenolol (20 microg kg(-1) h(-1), i.v.) or hydralazine (10 microg kg(-1) min(-1), i.v.) beginning 30 min following autologous clot embolization. All rabbits then received t-PA (6.3 mg kg(-1), i.v.) beginning 1 h after embolization, with completion of the protocol 4 h after embolization. Aspirin therapy reduced regional cerebral blood flow (rCBF) from 82.8m +/- 4.7 to 62.5 +/- 6.6 (n = 30; p = 0.0005). In the aspirin control group only 30% (3 of 10) rabbits demonstrated complete clot lysis, whereas the combined atenolol (60%) and hydralazine (70%) groups experienced a clot lysis rate of 65% (13 of 20 rabbits), similar to clot lysis rates previously observed with t-PA alone. In a separate series of experiments, all agents able to reverse aspirin antagonism of thrombolysis demonstrated an improvement in rCBF, suggesting a common mechanism for this diverse group of agents in reversing aspirin's antagonism of thrombolysis.