Prolactin and growth hormone immunoactivity in canine mammary adenomas and adenocarcinomas.

It is now widely accepted in human medicine that prolactin (PRL) and growth hormone (GH) function in the mammary gland in an autocrine and paracrine manner in tumour formation. The aim of this study was to compare PRL and GH immunoactivity in canine mammary tumours submitted for histopathologic evaluation. Formalin-fixed specimens from spontaneously occurring mammary adenomas and adenocarcinomas from 24 female client-owned dogs were used. Information pertaining to the reproductive status of the patient at the time of mammary tumour diagnosis was obtained from each of the submitting veterinarians. Tissues were paraffin-embedded and sectioned (5 µm) onto charged slides. All slides were deparaffinized and rehydrated. Endogenous peroxidase activity was inactivated with 3% H2 O2, and non-specific binding was blocked. Polyclonal rabbit antihuman PRL (DAKO A0569) and GH antibody (DAKO A0570) were applied at a 1:250 and 1:200 dilutions, respectively. A universal rabbit negative control (DAKO N1699) was used. Slides were then reacted with anti-rabbit horseradish peroxidase followed by Nova Red Peroxidase substrate. Slides were counter-stained with haematoxylin, dehydrated and mounted. Tumour type and reproductive status at time of tumour diagnosis were compared individually between tumours that were negative or positive for PRL and GH using a two-tailed analysis of variance. Significance was defined as p < .05. There was no significant relationship between tumour type and PRL and GH presence. In addition, reproductive status at the time of tumour removal was found to be not significant. These results vary from previous reports in canine mammary tumours and warrant further investigation.

One hundred two tumors in 100 goats (1987-2011).

Of 1146 caprine necropsy or biopsy specimens submitted from 1987 through 2011 to the Veterinary Diagnostic Laboratory at Oregon State University, 100 goats (8.7%) had 102 tumors. Detailed records were available for 89 cases. Fifty-five goats were female, 17 were castrated males, and 12 were intact males. Breeds included 21 Nubian, 16 Pygmy, 10 Pygora, 8 Alpine, 4 Angora, 4 Saanen, 2 Toggenburg, and 9 crossbred goats. Dwarf, Nubian, and Saanen goats were overrepresented and Alpine and Boer goats underrepresented among cases with neoplastic disease in comparison to submissions overall. Age ranged from 7 months to 19 years (median, 7 years). Histopathology was performed on 97 tumors. Lymphoma (n = 17) was the most common tumor, followed by cutaneous squamous cell carcinoma (n = 10) and thymoma (n = 9). Most lymphomas were multicentric. All 7 mammary neoplasms were adenocarcinomas. Five of 7 vascular proliferations were hemangiosarcomas. All 4 melanocytic tumors were classified as (malignant) melanoma. Rarely reported caprine tumors included a choroid plexus carcinoma, 2 rhabdomyosarcomas, and 3 pheochromocytomas. Cutaneous round cell tumors were provisionally diagnosed as 2 histiocytomas and 5 mast cell tumors. Single cases of previously unreported caprine tumors included amyloid-producing odontogenic tumor, myxosarcoma, sebaceous carcinoma, apocrine sweat gland adenoma, and thyroid carcinoma. Nonneoplastic entities included 2 cases of mammary fibroadenomatous hyperplasia and single cases of vascular hamartoma, cervical adenomatous hyperplasia, and cervical leiomyofibromatosis. The results of this 25-year retrospective study indicate that lymphoma in particular and tumors in general are common in goats.

Characterization of the microanatomy and histopathology of placentas from aborted, stillborn, and normally delivered alpacas (Vicugna pacos) and llamas (Lama glama).

From 2002 to 2007, 101 camelid abortions and stillbirths were submitted to the Veterinary Diagnostic Laboratory at Oregon State University (84 alpacas [Vicugna pacos], 13 llamas [Lama glama], 4 unknown). For most cases (n = 67), a cause was not determined by routine testing. Eighty-five submissions included placenta for microscopic examination, of which 55 were from abortions to unknown causes (idiopathic). Microscopic features of placentas from abortion/stillbirth were compared with those from 19 camelids delivered normally (6 alpacas, 12 llamas, 1 unknown) and with those from 4 alpaca fetuses of known gestational age collected during the dam's necropsy. The most common microscopic findings in abortion/stillbirth placentas were mineralization (n = 57) and mucinous edema (n = 27) of the chorioallantoic stroma. One or more of these features were also observed in 22 of 23 placentas from normal pregnancies/deliveries and therefore interpreted as incidental findings. The comparison of alpaca placentas after matching for gestational parameters (crown-rump length, weight, days of gestation; n = 41) revealed hypoplasia of placental villi in 5 of 22 idiopathic abortions and in 1 abortion due to umbilical torsion; hypoplasia was further suspected in an additional 6 abortions of unknown cause and 2 abortions of known cause. The identified villous hypoplasia is assumed to have resulted in placental insufficiency. When placental insufficiency is included as cause, idiopathic abortions are reduced from 66.2 to 47.9% of alpaca cases with histopathologic examination of placenta and from 66.3 to 52.5% of alpaca and llama abortions overall. This study also permitted the generation of a linear regression curve correlating alpaca fetal crown-rump length with fetal age.

Mesenteric lymphangitis and sepsis due to RTX toxin-producing Actinobacillus spp in 2 foals with hypothyroidism-dysmaturity syndrome.

Actinobacillus suis-like organisms (ASLOs) have been isolated from the genital, respiratory, and digestive tracts of healthy adult horses, horses with respiratory disease, and septic foals. Two foals with congenital hypothyroidism-dysmaturity syndrome from separate farms developed ASLO infection. At necropsy, both had contracted carpal flexor tendons, thyroid hyperplasia, and thrombotic and necrotizing mesenteric lymphangitis and lymphadenitis; one foal also had mandibular prognathism. Numerous ASLOs were isolated from tissues from both foals, including intestine. Biochemical testing and mass spectrometric analysis of the two Actinobacillus isolates did not allow unequivocal identification. Comparative genetic analysis was done on these and similar isolates, including phylogeny based on 16S rRNA, rpoB and recN genes, as well as RTX (repeat in toxin) toxin typing of apxIA-apxIVA and aqxA genes. One isolate was identified as Actinobacillus suis sensu stricto, based on the presence of apxIA and apxIIA but not aqxA, whereas the other isolate had aqxA but neither apxIA nor apxIIA, consistent with A equuli ssp haemolyticus. Based on genotypic analysis of the isolates included for comparison, 3 of 3 equine ASLOs and 2 of 5 A equuli isolates were reclassified as A equuli subsp haemolyticus, emphasizing the importance of toxin genotyping in accurate classification of actinobacilli.

Pathology and viral antigen distribution of lethal pneumonia in domestic cats due to pandemic (H1N1) 2009 influenza A virus.

A novel swine-origin H1N1 influenza A virus has been identified as the cause of the 2009 influenza pandemic in humans. Since then, infections with the pandemic (H1N1) 2009 influenza virus have been documented in a number of animal species. The first known cases of lethal respiratory disease associated with pandemic (H1N1) 2009 influenza virus infection in house pets occurred in domestic cats in Oregon. A 10-year-old neutered domestic shorthair and an 8-year-old spayed domestic shorthair died shortly after developing severe respiratory disease. Grossly, lung lobes of both cats were diffusely firm and incompletely collapsed. Histologically, moderate to severe necrotizing to pyonecrotizing bronchointerstitial pneumonia was accompanied by serofibrinous exudation and hyaline membranes in the alveolar spaces. Influenza A virus was isolated from nasal secretions of the male cat and from lung homogenate of the female cat. Both isolates were confirmed as pandemic (H1N1) 2009 influenza virus by real-time reverse transcriptase polymerase chain reaction. With immunohistochemistry, influenza A viral antigen was demonstrated in bronchiolar epithelial cells, pneumocytes, and alveolar macrophages in pneumonic areas. The most likely sources of infection were people in the household with influenza-like illness or confirmed pandemic (H1N1) 2009 influenza. The 2 cases reported here provide, to the best of the authors' knowledge, the first description of the pathology and viral antigen distribution of lethal respiratory disease in domestic cats after natural pandemic (H1N1) 2009 influenza virus infection, probably transmitted from humans.

Preferential transport and metabolism of glucose in Bergmann glia over Purkinje cells: a multiphoton study of cerebellar slices.

Knowing how different cell types handle glucose should help to decipher how energy supply is adjusted to energy demand in the brain. Previously, the uptake of glucose by cultured brain cells was studied in real-time using fluorescent tracers and confocal microscopy. Here, we have adapted this technique to acute slices prepared from the rat cerebellum by means of multiphoton microscopy. The transport of the fluorescent glucose analogs 2NBDG and 6NBDG was several-fold faster in the molecular layer of the cerebellar cortex than in Purkinje cell somata and granule cells. After washout of free tracer, it became apparent that most phosphorylated tracer was located in Bergmann glia, which was confirmed by counterstaining with the glial marker sulforhodamine 101. The effective recovery of fluorescence after photobleaching showed that 2NBDG-P can diffuse horizontally across the molecular layer, presumably through gap junctions between Bergmann glial cells. Our main conclusion is that in acute cerebellar slices, the glucose transport capacity and glycolytic rate of Bergmann glia are several-fold higher than those of Purkinje cells. Given that the cerebellum is largely fueled by glucose and Purkinje neurons are estimated to spend more energy than Bergmann glial cells, these results suggest substantial shuttling of an energy-rich metabolite like lactate between glial cells and neurons.

Prevention of feeding tube dislodgement with the Wiesbaden rein: a case series.

We describe initial experience with the use of a new fixation method (Wiesbaden rein), which has been developed to prevent dislodgement of feeding tubes in the gastrointestinal tract. The Wiesbaden rein has been used in three patients without complication. In none of the patients was dislodgement or malfunction of the feeding tube observed. Therefore, the use of the Wiesbaden rein might prevent dislodgement of feeding tubes. Clinical trials are required before this new method can be recommended for general use.

Diagnostic accuracy, validity, and reliability of Tensiomyography to assess muscle function and exercise-induced fatigue in healthy participants. A systematic review with meta-analysis.

Tensiomyography™ (TMG) is a non-invasive method to monitor skeletal muscle mechanical characteristics. This systematic review and meta-analysis reports on diagnostic accuracy, validity, and reliability of TMG (maximal radial displacement [Dm], contraction time [Tc], delay time [Td], and velocity of contraction [Vc]) to assess exercise-induced fatigue in healthy volunteers, with the specific aim to determine the current level of supporting evidence. Systematic literature searches within Medline, Embase and Sportdiscus databases were conducted from January 1990 through November 2018. Methodological quality was evaluated by the Assessment of Diagnostic Accuracy Studies (QUADAS) tool or the Validity and Reliability Critical Appraisal Tool (CAT) or the Quality Appraisal of Diagnostic Reliability checklist (QAREL). Meta-analytical methods were utilised to summarize relative reliabilities of Dm, Tc, Td (95%, CI). The methodological quality of the 19 included studies (n = 373; female = 13.0%) ranged from low to high quality. The analysis revealed insufficient diagnostic accuracy and validity, mixed results regarding absolute reliability, and high to excellent relative reliability for the assessed measures. To conclude, robust evidence for diagnostic accuracy/validity of TMG has yet to be determined, whereas there is substantial evidence for its reliability. Higher methodological standards need to be established, including the avoidance of gender bias.

Myonecrosis in three horses with colic: evidence for endotoxic injury.

Three horses with colic, clinical evidence of endotoxaemia and high serum activities of creatine kinase and aspartate aminotransferase were examined postmortem. The horses were diagnosed with severe ulcerative colitis, pyloric ulceration and stenosis with colonic sand impaction, and colonic obstruction due to faecaliths. There was no gross or histological evidence of muscle trauma. Their semimembranosus muscles had scattered acute to subacute segmental necrosis of the myofibres, suggestive of endotoxin-induced muscle injury.

Cardiac-gated computed tomography angiography in three alpacas with complex congenital heart disease.

BACKGROUND: The prevalence of congenital heart disease is higher in camelids than in other domestic species and complex defects, often involving the great vessels, are more frequently encountered in llamas and alpacas than in other species. Some of these complex defects can be difficult to accurately characterize via echocardiography, the most commonly used diagnostic imaging technique to evaluate the heart in veterinary patients. Contrast-enhanced, electrocardiogram (ECG)-gated computed tomography (CT) has proven utility for the evaluation of human patients with certain congenital heart defects, including those with conotruncal septation defects and other abnormalities involving the formation of the great vessels. METHODS: Three alpaca crias, 4 days, 5 weeks and 14 months of age were clinically evaluated and subjected to a complete color-flow Doppler echocardiogram and a contrast-enhanced ECG-gated CT. RESULTS: These alpacas exhibited a variety of clinical findings including lethargy, failure to thrive, exercise intolerance, heart murmur, and/or respiratory difficulty. All three crias were subsequently diagnosed with complex cardiac defects including pulmonary atresia with a ventricular septal defect (VSD), a truncus arteriosus with a large VSD, and a double outlet right ventricle with a large VSD and aortic hypoplasia. In each case, the diagnosis was confirmed by postmortem examination. CONCLUSION: Color flow echocardiographic evaluation identified all of the intra-cardiac lesions and associated flow anomalies but contrast-enhanced ECG-gated CT permitted more accurate assessment of the morphology of the extracardiac structures and permitted a more precise determination of the exact nature and anatomy of the great vessels.

Calcium signalling in glial cells.

Calcium signals are the universal way of glial responses to the various types of stimulation. Glial cells express numerous receptors and ion channels linked to the generation of complex cytoplasmic calcium responses. The glial calcium signals are able to propagate within glial cells and to create a spreading intercellular Ca2+ wave which allow information exchange within the glial networks. These propagating Ca2+ waves are primarily mediated by intracellular excitable media formed by intracellular calcium storage organelles. The glial calcium signals could be evoked by neuronal activity and vice versa they may initiate electrical and Ca2+ responses in adjacent neurones. Thus glial calcium signals could integrate glial and neuronal compartments being therefore involved in the information processing in the brain.

Calcium influx into dendrites of the leech Retzius neuron evoked by 5-hydroxytryptamine.

5-Hydroxytryptamine (5-HT) is a ubiquitous neurotransmitter and neuromodulator that affects neural circuits and behaviours in vertebrates and invertebrates. In the present study, we have investigated 5-HT-induced Ca(2+) transients in subcellular compartments of Retzius neurons in the leech central nervous system using confocal laser scanning microscopy, and studied the effect of 5-HT on the electrical coupling between the Retzius neurons. Bath application of 5-HT (50mM) induced a Ca(2+) transient in axon, dendrites and cell body of the Retzius neuron. This Ca(2+) transient was significantly faster and larger in dendrites than in axon and cell body, and was half-maximal at a 5-HT concentration of 5-12mM. The Ca(2+) transient was suppressed in the absence of extracellular Ca(2+) and by methysergide (100mM), a non-specific antagonist of metabotropic 5-HT receptors, and was strongly reduced by bath application of the Ca(2+) channel blocker Co(2+) (2mM). Injection of the non-hydrolysable GTP analogue GTPgammaS increased and prolonged the dendritic 5-HT-induced Ca(2+) transient. The non-selective protein kinase inhibitor H7 (100mM) and the adenylate cyclase inhibitor SQ22536 (500 mM) did not affect the Ca(2+) transient, and the membrane-permeable cAMP analogue dibutyryl-cAMP (500 mM) did not mimic the effect of 5-HT application. 5-HT reduced the apparent electrical coupling between the two Retzius neurons, whereas suppression of the Ca(2+) influx by removal of external Ca(2+) improved the transmission of action potentials at the electrical synapses which are located between the dendrites of the adjacent Retzius neurons. The results indicate that 5-HT induces a Ca(2+) influx through calcium channels located primarily in the dendrites, and presumably activated by a G protein-coupled 5-HT receptor. The dendritic Ca(2+) increase appears to modulate the excitability of, and the synchronization between, the two Retzius neurons.

Activity-induced Ca2+ transients in nerve and glial cells in the leech CNS.

We have measured activity-induced Ca2+ transients in Retzius neurones, neuropile glial cells, and extracellular spaces of isolated ganglia of the leech Hirudo medicinalis using the fluorescent dye fura-2 and Ca(2+)-sensitive microelectrodes. Neuronal activity, induced by electrical side nerve stimulation (20 Hz/1 min), elicited transient rises of intracellular Ca2+ in both neurones and glial cells, which amounted to 24 +/- nM (n = 15) and 17 +/- 14 nM (n = 7), respectively. The extracellular Ca2+ declined by 160 +/- 73 microM (n = 6) during stimulation. Intra- and extracellular Ca2+ transients were reduced by the glutamate/kainate receptor blocker CNQX (6-cyano-7-dinitroquinozaline-2,3-dione; 50 microM). Our results show that neuronal activity evokes Ca2+ signals not only in neurones, but also in glial cells and suggest that these Ca2+ transients are partly mediated via activation of glutamate/kainate receptors.

Activity-dependent accumulation of Ca2+ in axon and dendrites of the leech Leydig neuron.

We have investigated Ca2+ changes evoked by single action potentials (APs) in axon and dendrites of leech Leydig neurons. Dendritic Ca2+ transients induced by an AP were twice as large as in the axon, and Ca2+ recovery was significantly faster in the dendrites as compared to the axon. The AP-induced Ca2+ transients were blocked by Co2+ and suppressed in Ca2+-free saline, indicating Ca2+ influx through voltage-activated channels. During a train of APs, Ca2+ accumulated significantly more in the axon than in the dendrites. Suppression of the Ca2+ influx changed the shape of the action potential and increased the firing frequency. The results suggest a functional role of Ca2+ influx and Ca2+ accumulation during electrical activity in different neuronal subcompartments.

Extracellular Ca2+ changes during transmitter application in the leech central nervous system.

Changes in extracellular Ca2+ concentration ([Ca2+]e) evoked by transmitters and transmitter agonists, respectively, and by elevation of bath K+ concentration were recorded in isolated segmental ganglia of the leech Hirudo medicinalis using Ca(2+)-selective microelectrodes. A 1-min bath application of kainate (10 microM), glutamate (1 mM), aspartate (1 mM), or carbachol (200 microM) decreased [Ca2+]e by up to 1 mM, whereas the inhibitory transmitters gamma-amino butyric acid (GABA, 100 microM) and serotonin (5-HT, 100 microM) did not change [Ca2+]e. The amplitude of the kainate-induced changes in [Ca2+]e increased with repetitive applications, and changes were blocked by 6-cyano-7-dinitroquinozaline-2,3-dione (CNQX). Elevation of bath K+ concentration from 4 to 40 mM led to a Ni(2+)-sensitive decrease in [Ca2+]e by 0.9 mM. Our results suggest that excitatory transmission in the leech central nervous system might be accompanied by substantial decreases in [Ca2+]e.

Detection of canine oral papillomavirus-DNA in canine oral squamous cell carcinomas and p53 overexpressing skin papillomas of the dog using the polymerase chain reaction and non-radioactive in situ hybridization.

Nineteen cutaneous and mucocutaneous papillomas, as well as 29 oral and 25 non-oral squamous cell carcinomas of dogs were analyzed immunohistologically for the presence of papillomavirus (PV)-antigens. Canine oral papillomavirus (COPV)-DNA was detected in formalin-fixed, paraffin-embedded tissues by polymerase chain reaction (PCR) and non-radioactive in situ hybridization (ISH). Furthermore, the expression of the tumor suppressor protein p53 was investigated. PV-antigens were detectable in more than 50% of the oral and cutaneous papillomas, while no PV-antigens could be demonstrated in venereal papillomas. One squamous cell carcinoma was PV-antigen positive. Only two cutaneous papillomas of the head showed a strong p53-specific immunostaining, while overexpressed p53 was detectable in approximately 35% of all squamous cell carcinomas. It was possible to amplify fragments of the E6, E7 and L1 gene by polymerase chain reaction (PCR) from five of eight oral and from five of eight cutaneous papillomas as well as from three oral squamous cell carcinomas. Nine of 10 papillomas showed a strong nucleus-associated hybridization signal typical for COPV-DNA. In three squamous cell carcinomas COPV-DNA was located in nests of the epithelial tumor cells surrounding 'horn pearls' or disseminated in the carcinoma tissue. These observations support the view that COPV may also induce non-oral papillomas in the dog and confirm the opinion that a progression of viral papillomas into carcinomas in dogs may occur.

Laparoscopic management of Chilaiditi's syndrome.

Chilaiditi's syndrome refers to the symptoms of abdominal pain, distention, vomiting, anorexia, and constipation caused by hepatodiaphragmatic interposition of the intestine. Although patients with this radiographic finding are commonly asymptomatic, presentation with symptoms is rare and accurately refers to this syndrome. There is an increased incidence of Chilaiditi's syndrome among mentally ill adults. Traditionally, Chilaiditi's syndrome is managed medically by discontinuing causative medicines. However, among the mentally ill population whose psychotropic medications precipitate the interposition of the colon, ceasing these psychotropic medications is not an appropriate option. The case presented involves a mentally ill patient with Chilaiditi's syndrome who was successfully managed with laparoscopic colopexy. At follow-up, the patient reported marked improvement of abdominal symptoms.

Diagnosis of feline herpesvirus infection by immunohistochemistry, polymerase chain reaction, and in situ hybridization.

An adult domestic shorthair cat had severe chemosis due to purulent and necrotizing blepharitis and conjunctivitis. Purulent rhinitis, necrotizing glossitis, and dermatitis were also diagnosed. The cat was positive for feline immunodeficiency virus and feline leukemia virus. Histologically, intranuclear Cowdry type A inclusions were found within numerous epithelial cells adjacent to the lesions in skin, conjunctiva, and tongue. Electron microscopic examination revealed herpesviral particles within the lesions. Paraffin-embedded skin and tongue tissues were processed in a polymerase chain reaction, using primers to amplify a 306-bp region of the thymidine kinase gene of feline herpesvirus type 1, resulting in a distinct amplification product of the predicted size. The distribution of feline herpesvirus was demonstrated by immunohistochemistry and nonradioactive in situ hybridization. Positive immunostaining was found in nuclei and cytoplasm of numerous epithelial cells within and next to the lesions, whereas in situ hybridization, performed with a digoxigenin-labeled double-stranded DNA probe, revealed hybridization signal only in nuclei of intact epithelial cells. Neither immunohistochemistry nor in situ hybridization showed feline herpesvirus type 1 in tissues of lungs, liver, spleen, intestine, or brain.

Immunohistochemical detection of P53 overexpression in paraffin wax-embedded squamous cell carcinomas of cattle, horses, cats and dogs.

One hundred and six squamous cell carcinomas (SCCs) of cattle, horses, cats and dogs were analysed immunohistochemically for overexpression of p53 protein. The monoclonal antibody pAb 240, which recognizes only mutant p53, was used. Of 41 bovine ocular SCCs, 26 (63.4%) showed p53 nuclear reactivity. All of six (100%) equine ocular SCCs and seven of nine (77.7%) SCCs of the equine penis or vulva gave positive reactions. In nine of 11 (81.8%) feline SCCs of the ear and in seven of 14 (50%) feline SCCs of other locations, p53 immunoreactivity was detected. Only seven of 25 (29.5%) canine cutaneous SCCs gave a positive reaction. Thus p53 antigen could be detected immunohistochemically in formalin-fixed and paraffin wax-embedded tissues of SCCs of domestic animals. The results support the view that, as in man, p53 overexpression plays an important role in the development of most SCCs of the animal species studied. This was in particular true for feline SCCs of the ear and for bovine and equine ocular SCCs, which are assumed to be related to ultraviolet radiation.