The ß2-adrenergic receptor agonist terbutaline upregulates T helper-17 cells in a protein kinase A-dependent manner.

BACKGROUND: T helper 17 (Th17) cells produce IL-17A cytokine and can exacerbate autoimmune diseases and asthma. The ß2 adrenergic receptor is a g protein-coupled receptor that induces cAMP second messenger pathways. We tested the hypothesis that terbutaline, a ß2-adrenergic receptor-specific agonist, promotes IL-17 secretion by memory Th17 cells in a cAMP and PKA-dependent manner. METHODS: Venous peripheral blood mononuclear cells (PBMC) from healthy human participants were activated with anti-CD3 and anti-CD28 antibodies. Secreted IL-17A was measured by enzyme linked immunosorbent assay, intracellular IL-17A, and RORγ were measured using flow cytometry, and RORC by qPCR. Memory CD3+CD4+CD45RA-CD45RO+ T cells were obtained by immunomagnetic negative selection and activated with tri-antibody complex CD3/CD28/CD2. Secreted IL-17A, intracellular IL-17A, RORC were measured, and phosphorylated-serine133-CREB was measured by western blotting memory Th cells. RESULTS: Terbutaline increased IL-17A (p < 0.001), IL-17A+ cells (p < 0.05), and RORC in activated PBMC and memory Th cells. The PKA inhibitors H89 (p < 0.001) and Rp-cAMP (p < 0.01) abrogated the effects of terbutaline on IL-17A secretion in PBMC and memory T cells. Rolipram increased IL-17A (p < 0.01) to a similar extent as terbutaline. P-Ser133-CREB was increased by terbutaline (p < 0.05) in memory T cells. CONCLUSION: Terbutaline augments memory Th17 cells in lymphocytes from healthy participants. This could exacerbate autoimmune diseases or asthma, in cases where Th17 cells are considered to be pro-inflammatory.

Detection of vector-borne pathogens in owned dogs with cranial cruciate ligament rupture living in the Mediterranean area.

BACKGROUND: Cranial cruciate ligament rupture (CCLR) results from a multifactorial degenerative process that leads to rupture of the ligament. Vector-borne pathogens (VBP) in dogs can induce joint disease but their role in CCLR has not been previously investigated. The aim of the present work is to evaluate the prevalence of VBP in dogs with CCLR. METHODS: This was a prospective study that included 46 dogs presented for CCLR surgical treatment and 16 control dogs euthanized for diseases unrelated to the joints. Specimens collected included blood, synovial fluid, and synovial membrane biopsy. Pathogen testing consisted of serology for Leishmania infantum (quantitative ELISA), Ehrlichia canis/ewingii, Borrelia burgdorferi, Anaplasma phagocytophilum/platys, and Dirofilaria immitis (4DX IDEXX test), and PCR for L. infantum, Ehrlichia/Anaplasma spp., Bartonella spp., piroplasms (Babesia spp. and Theileria spp.), and filariae (D. immitis, Dirofilaria repens, Acanthocheilonema dracunculoides, Acanthocheilonema reconditum, and Cercopithifilaria spp.) on both EDTA-whole blood (EB) and synovial fluid (SF) samples. SF cytology and histopathological evaluation of synovial membrane were also performed. RESULTS: The prevalence of VBP was 19.6% in the CCLR group and 18.8% in the control group, with no statistical difference among them. The presence of synovitis was not more frequent in CCLR dogs (45.6%) than in control dogs (43.7%). Lymphoplasmacytic infiltration was the most common inflammatory pattern detected in the joints of both groups of dogs. CONCLUSIONS: This study failed to demonstrate a role of canine VBP in CCLR or the presence or different pattern of joint inflammation in pathogen-positive dogs.

Leishmaniosis in a cat with chronic diarrhea as the only clinical manifestation.

A 10-year-old male domestic shorthaired cat was presented with chronic diarrhea unresponsive to treatment. Laboratory testing identified hyperglobulinemia and mild nonregenerative anemia, and nongastrointestinal causes of diarrhea were ruled out. Gastrointestinal endoscopy and biopsy were performed and disclosed diffuse generalized granulomatous and lymphoplasmocytic inflammatory reaction in all segments of gastrointestinal tract evaluated, with numerous Leishmania spp. amastigotes within the cytoplasm of macrophages. The organism also was detected in spleen and bone marrow and Leishmania spp. serology was positive (immunofluorescence assay 1 : 160). A diagnosis of granulomatous enteritis secondary to leishmaniosis was made. Gastrointestinal signs resolved after treatment with allopurinol and a dietary supplement of nucleotides and active hexose-correlated compounds (N-AHCC), but seropositivity and gammopathy persisted 8 months later. The cat died of unrelated causes after an additional 3 months and permission for necropsy was not granted. Leishmaniosis as a cause of chronic diarrhea has not been reported previously in cats and should be considered in endemic areas in cats with chronic gastrointestinal signs.

Stereocilia Bundle Imaging with Nanoscale Resolution in Live Mammalian Auditory Hair Cells.

Inner ear hair cells detect sound-induced displacements and transduce these stimuli into electrical signals in a hair bundle that consists of stereocilia that are arranged in rows of increasing height. When stereocilia are deflected, they tug on tiny (~5 nm in diameter) extracellular tip links interconnecting stereocilia, which convey forces to the mechanosensitive transduction channels. Although mechanotransduction has been studied in live hair cells for decades, the functionally important ultrastructural details of the mechanotransduction machinery at the tips of stereocilia (such as tip link dynamics or transduction-dependent stereocilia remodeling) can still be studied only in dead cells with electron microscopy. Theoretically, scanning probe techniques, such as atomic force microscopy, have enough resolution to visualize the surface of stereocilia. However, independent of imaging mode, even the slightest contact of the atomic force microscopy probe with the stereocilia bundle usually damages the bundle. Here we present a detailed protocol for the hopping probe ion conductance microscopy (HPICM) imaging of live rodent auditory hair cells. This non-contact scanning probe technique allows time lapse imaging of the surface of live cells with a complex topography, like hair cells, with single nanometers resolution and without making physical contact with the sample. The HPICM uses an electrical current passing through the glass nanopipette to detect the cell surface in close vicinity to the pipette, while a 3D-positioning piezoelectric system scans the surface and generates its image. With HPICM, we were able to image stereocilia bundles and the links interconnecting stereocilia in live auditory hair cells for several hours without noticeable damage. We anticipate that the use of HPICM will allow direct exploration of ultrastructural changes in the stereocilia of live hair cells for better understanding of their function.

Retinal detachment secondary to vitreoretinopathy in two closely related warmblood horses.

BACKGROUND: To describe the clinical, diagnostic imaging, and histopathological findings of two visually impaired closely related horses and to identify a possible cause. CASE DESCRIPTION: Two warmblood horses, with a common dam and sire, were presented to the ophthalmology department of Liège for investigation of impaired vision. Information collected included physical and ophthalmic examination findings, results of ocular ultrasound, electroretinogram, magnetic resonance imaging (MRI), and histopathology. Ophthalmic examination, ocular ultrasound and MRI revealed a complete retinal detachment (RD) in the left eye and vitreous synaeresis in both eyes of both horses. Electroretinograms showed a normal response in both right eyes but a total loss of the retinal response in their left eyes. Histopathologic examination revealed multifocal retinal dysplasia in both left eyes. CONCLUSION: In these two horses, RD has likely been caused by the congenital posterior segment abnormalities of the vitreous and the retina. A vitreoretinopathy is highly suspected and is possibly hereditary in these closely related siblings.

Cryopreservation (-20°C) of equine corneoscleral tissue: Microbiological, histological, and ultrastructural study.

OBJECTIVE: To evaluate microbiological, histological, and ultrastructural characteristics of short-term cryopreserved (STC) equine corneoscleral tissue (<1 year), and to compare it with long-term cryopreserved (LTC) tissue (>7 years). ANIMALS STUDIED: Thirty-four healthy equine globes. PROCEDURE: After a decontamination protocol, globes were enucleated and stored at -20°C in broad-spectrum antibiotics. Corneoscleral tissue was evaluated at different storage periods: 1 month-1 year (20 eyes) and 7-9 years (12 eyes). Two eyes were used as controls. Microbiologic study included direct (blood, McConkey, and Sabouraud agars) and enrichment (brain-heart infusion broth) cultures. Cryopreservation artifacts were evaluated by hematoxylin-eosin. Corneoscleral collagen organization and number of normal and dead keratocytes were established by transmission electron microscopy. RESULTS: All microbiologic direct cultures were negative. Enrichment cultures were positive in 12.5% of corneal and 59.4% of scleral tissues (pcornea  = 0.136; psclera  = 1.000). Cryopreservation artifacts were most commonly observed in LTC tissues (P = 0.002). Normal keratocytes were predominant in STC corneas (STC 60% and LTC 0%) and apoptotic ones in LTC (STC 40% and LTC 90%), whereas necrotic keratocytes were only seen in LTC (LTC 10%) (P = 0.001). No structural differences were detected in collagen organization between STC and LTC (pcornea  = 1.000; psclera  = 0.703). CONCLUSIONS: Cryopreservation of equine corneoscleral tissue did not yield direct bacterial contamination. Apoptosis is the main cause of death of cryopreserved equine keratocytes. Based on the lack of significant structural differences between STC and LTC samples, these cryopreserved tissues could potentially be used for tectonic support for at least 9 years without structural or microbiological impediment.

Problemas relacionados con medicamentos que causan ingresos por urgencias en un hospital de alta complejidad / Drug-related problems causing hospital admissions in the emergency rooms at of high complexity hospital

Objetivo: Determinar la prevalencia de ingresos hospitalarios asociados a problemas relacionados con medicamentos en los servicios de urgencias de un hospital de alta complejidad, realizando las intervenciones farmacéuticas pertinentes. Método: Estudio observacional descriptivo de corte transversal. Se revisaron las historias clínicas de pacientes que ingresaron en los servicios de urgencias, seleccionando aquellas que reportaban ingresos por problemas relacionados con medicamentos. Estas se clasificaron según la adaptación del Tercer Consenso de Granada. Se realizó un perfil farmacoterapéutico a los pacientes hospitalizados y se establecieron las intervenciones farmacéuticas necesarias para evitar futuros problemas relacionados con medicamentos. Resultados: El 3,8% de los pacientes fueron incluidos en el estudio. Los problemas relacionados con medicamentos tuvieron una evitabilidad del 87,7% y los más frecuentes fueron los de necesidad, con un 42,2%. Se realizó un perfil farmacoterapéutico a los pacientes hospitalizados (137) y se llevaron a cabo 150 intervenciones farmacéuticas, las cuales fueron aceptadas en un 95,3%. El riesgo más intervenido fue administrar un medicamento innecesario (62,7%). Conclusiones: La falta de supervisión y análisis de problemas relacionados con los medicamentos podría ocasionar el fracaso del abordaje terapéutico y la no consecución de la mejoría de la salud y la calidad de vida de los pacientes. El químico farmacéutico cumple un papel fundamental en el cuidado de la salud de los pacientes, ayudando desde la prevención y el uso adecuado de los medicamentos. El programa de Atención Farmacéutica demuestra que brinda un aporte inestimable al servicio de la salud pública al mejorar la farmacoseguridad de los tratamientos, disminuyendo costes y problemas de salud pública

Objective: Determining hospital admissions prevalence associated with problems related to medicines in the emergency services of high complexity hospital; performing the pertinent pharmaceutical interventions. Method: Descriptive observational cross-sectional study. The medical records of the patients admitted to the emergency services were reviewed. Those that reported hospitalization due to problems related to medication were selected. These were classified according to the Third Consensus of Granada adaptation. A pharma-therapeutic profile was made to hospitalized patients; performing the necessary pharmaceutical interventions to avoid future medication related problems. Results: 3.8% of patients were included in the study. The problems related to medications had a preventability of 87.7% and the most frequent were those of need with 42.2%. A pharma-therapeutic profile was done to hospitalized patients (137). 150 pharmaceutical interventions were done, which had an acceptance of 95.3%. The most intervened risk was administering an unnecessary medication 62.7%. Conclusions: Lack of supervision and analysis of problems related to medication could cause therapeutic approach failure, therefore, health and life quality improvement of the patients is not achieve. The pharmaceutical chemist plays a fundamental role in the health care of patients, helping to the prevention and proper use of medicines. The Pharmaceutical Care program proves that it provides an invaluable contribution to public health service by improving the pharmacological safety of treatments, reducing costs and public health problems

Problemas relacionados con medicamentos que causan ingresos por urgencias en un hospital de alta complejidad.

OBJECTIVE: Determining hospital admissions prevalence associated with problems related to medicines in the emergency services of high complexity hospital; performing the pertinent pharmaceutical interventions. METHOD: Descriptive observational cross-sectional study. The medical records of the patients admitted to the emergency services were reviewed. Those that  reported hospitalization due to problems related to medication were selected.  These were classified according to the Third Consensus of Granada adaptation. A pharma-therapeutic profile was made to hospitalized patients; performing the  necessary pharmaceutical interventions to avoid future medication related  problems. RESULTS: 3.8% of patients were included in the study. The problems related to  medications had a preventability of 87.7% and the most frequent were those of  need with 42.2%. A pharma-therapeutic profile was done to hospitalized  patients (137). 150 pharmaceutical interventions were done, which had an  acceptance of 95.3%. The most intervened risk was administering an  unnecessary medication 62.7%. CONCLUSIONS: Lack of supervision and analysis of problems related to medication could cause therapeutic approach failure, therefore, health and life quality improvement of the patients is not achieve. The pharmaceutical chemist  plays a fundamental role in the health care of patients, helping to the prevention  and proper use of medicines. The Pharmaceutical Care program  proves that it provides an invaluable contribution to public health service by  improving the pharmacological safety of treatments, reducing costs and public  health problems.

Objetivo: Determinar la prevalencia de ingresos hospitalarios asociados a  problemas relacionados con medicamentos en los servicios de urgencias de un  hospital de alta complejidad, realizando las intervenciones farmacéuticas  pertinentes.Método: Estudio observacional descriptivo de corte transversal. Se revisaron las historias clínicas de pacientes que ingresaron en los servicios de urgencias,  seleccionando aquellas que reportaban ingresos por problemas relacionados con  medicamentos. Estas se clasificaron según la adaptación del Tercer Consenso de  Granada. Se realizó un perfil farmacoterapéutico a los pacientes hospitalizados y se establecieron las intervenciones farmacéuticas necesarias para evitar futuros  problemas relacionados con medicamentos.Resultados: El 3,8% de los pacientes fueron incluidos en el estudio. Los problemas relacionados con medicamentos tuvieron una evitabilidad del 87,7% y los más frecuentes fueron los de necesidad, con un 42,2%. Se realizó un perfil farmacoterapéutico a los pacientes hospitalizados (137) y se  llevaron a cabo 150 intervenciones farmacéuticas, las cuales fueron aceptadas  en un 95,3%. El riesgo más intervenido fue administrar un medicamento  innecesario (62,7%).Conclusiones: La falta de supervisión y análisis de problemas relacionados con  los medicamentos podría ocasionar el fracaso del abordaje terapéutico y la no  consecución de la mejoría de la salud y la calidad de vida de los pacientes. El  químico farmacéutico cumple un papel fundamental en el cuidado de la salud de  los pacientes, ayudando desde la prevención y el uso adecuado de los  medicamentos. El programa de Atención Farmacéutica demuestra que brinda un  aporte inestimable al servicio de la salud pública al mejorar la farmacoseguridad  de los tratamientos, disminuyendo costes y problemas de salud pública.

Cryopreservation (-20 °C) of canine corneoscleral tissue: histological, microbiological, and ultrastructural study.

OBJECTIVE: To evaluate microbiological, histological, and ultrastructural characteristics of short-term cryopreserved (STC) canine corneoscleral tissue (<1 year) and to compare it with long-term cryopreserved (LTC) tissue (>6 years). ANIMALS STUDIED: Thirty-six healthy canine globes. PROCEDURE: After a decontamination protocol, globes were enucleated and stored at -20 °C. Corneoscleral tissue was evaluated at different periods: <1 year (20 eyes) and >6 years (12 eyes). Four eyes were used as controls. Microbiologic study included direct (blood, McConkey and Sabouraud agars) and enrichment (brain-heart infusion broth) cultures. Cryopreservation artifacts were evaluated by hematoxylin-eosin. Corneoscleral collagen organization and number of normal and dead keratocytes were established by transmission electron microscopy (TEM). Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) was also used for keratocyte characterization. RESULTS: Corneal microbial growth was observed in 25% of the direct STC cultures, and in 47.4% and 16.7% of the enriched STC and LTC cultures, respectively. Scleral STC direct cultures were 30% positive, while enrichment cultures were positive in 66.7% and 16.7% of the STC and LTC, respectively (P = 0.011). Cryopreservation artifacts were higher in LTC tissues (P < 0.001). Apoptotic keratocytes were predominant by TEM and TUNEL, in both STC and LTC. Minimal structural differences were detected in collagen organization between STC and LTC. CONCLUSIONS: Cryopreservation of canine corneoscleral tissue seems to reduce bacterial contamination over time. Apoptosis is the main way of death of cryopreserved canine keratocytes. Based on the lack of significant structural differences between STC and LTC samples, these cryopreserved tissues could potentially be used for tectonic support for at least 8 years without structural or microbiological impediment.

Immediate effects of diamond burr debridement in patients with spontaneous chronic corneal epithelial defects, light and electron microscopic evaluation.

PURPOSE: To evaluate immediate effects of diamond burr debridement (DBD) on the cornea of canine patients diagnosed with spontaneous chronic corneal epithelial defects (SCCEDs). ANIMALS STUDIED: Eight client owned dogs with SCCEDs. METHODS: Nine eyes from eight dogs with SCCEDs underwent superficial keratectomy (SK). The ulcerated area was divided into quadrants with a 300-micron restricted depth knife. Two of four quadrants underwent DBD for 40-60 s. A SK followed immediately. One burred section and one nonburred section were fixed with formaldehyde 10% and underwent light microscopy (LM). The remaining quadrants from five eyes were fixed with glutaraldehyde 2.5% and underwent transmission electron microscopy (TEM). Masked pathologists evaluated the samples. A student's paired t-test was used to analyze the data. RESULTS: With LM all nonburred samples had a superficial stromal hyaline acellular zone (HAZ), seven of the burred samples had an intermittent HAZ and in two burred samples this zone was absent. The HAZ thickness of burred samples (1.062 ± 0.664 µm) was significantly thinner than that of the nonburred samples (4.309 ± 1.348 µm) (P < 0.0001). Transmission electron microscopy showed an absence of basement membrane and the presence of an amorphous, fine fibrillar material in the superficial stroma in nonburred samples. This material was intermittent or absent in burred samples. CONCLUSION: DBD significantly reduces the superficial stromal HAZ in SCCEDs. A reduction of its thickness may be responsible for the healing rates reported with DBD.

Cryopreservation (-20 °C) of feline corneoscleral tissue: histologic, microbiologic, and ultrastructural study.

OBJECTIVE: To evaluate microbiological, histologic, and ultrastructural characteristics of short-term cryopreserved (STC) feline corneoscleral tissue (<1 year) and to compare it with long-term cryopreserved (LTC) tissue (>7 years). ANIMALS STUDIED: Twenty healthy feline globes were obtained from 2003 to 2013. PROCEDURE: After a decontamination protocol, globes were enucleated and stored at -20 °C in broad-spectrum antibiotics. Corneoscleral tissue was evaluated at different storage periods: <1 year (10 eyes) and >7 years (8 eyes). Two eyes were used as controls. Microbiologic study included direct (blood, McConkey, and Sabouraud agars) and enrichment (brain-heart infusion broth) cultures. Cryopreservation artifacts were evaluated by hematoxylin-eosin. Corneoscleral collagen organization and number of normal and dead keratocytes were established by transmission electron microscopy. RESULTS: Although microbiologic cultures were positive only in STC [direct (20.8%); enrichment (37.5%)], significant differences between periods were only found in enrichment cultures (P = 0.006). Cryopreservation artifacts were most commonly observed in LTC tissues (P < 0.001). Normal keratocytes were predominant in STC corneas (STC 58.3%, LTC 12.5%) and apoptotic ones in LTC (STC 41.7%, LTC 75%), whereas necrotic keratocytes were only seen in LTC (LTC 12.5%) (P = 0.046). No structural differences were detected in collagen organization between STC and LTC (Pcornea = 0.147; Psclera = 0.362). CONCLUSIONS: Cryopreservation of feline corneoscleral tissue seems to reduce bacterial contamination over time. Apoptosis is the main cause of death of cryopreserved feline keratocytes. Based on the lack of significant structural differences between STC and LTC samples, these cryopreserved tissues could potentially be used for tectonic support for at least 10 years without structural or microbiological impediment.

Prácticas de prescripción y conocimientos médicos acerca de los anticoagulantes orales directos en un hospital de referencia / Prescription practices and medical knowledge on direct oral anticoagulants in a reference hospital / Práticas de prescrição e conhecimentos médicos a respeito dos anticoagulantes orais diretos num hospital de referência

Introducción y objetivo: se desconocen la experiencia y el conocimiento con los que cuenta el personal médico del medio en el manejo de los anticoagulantes orales directos. Se pretende describir el uso de estos medicamentos en los pacientes atendidos en el Hospital Pablo Tobón Uribe y evaluar el nivel de conocimientos al respecto en los profesionales de la misma institución. Materiales y métodos: estudio de corte transversal en el período comprendido entre enero del 2012 y enero del 2013. Para recolectar la información se utilizaron dos estrategias: el análisis de las historias clínicas pertinentes y la evaluación de conocimientos sobre la prescripción y el uso de anticoagulantes orales directos en el grupo de especialistas facultados. Resultados: en el análisis se incluyeron 114 historias; el rivaroxaban fue el medicamento prescrito con mayor frecuencia (87%) seguido del dabigatrán (13%). La principal indicación fue la profilaxis en cirugía ortopédica (69%). Entre los especialistas evaluados, el promedio de respuestas acertadas fue del 67% sin diferencias aparentes entre los mismos. Conclusión: el rivaroxaban se prescribió con mayor frecuencia que el dabigatrán; sin embargo, este hallazgo no parece asociarse con un conocimiento médico claro y suficiente acerca de estos medicamentos. No se encontraron reportes de eventos adversos asociados al uso de los mismos.

Introduction and objective: The experience and knowledge concerning the use of direct oral anticoagulants among specialists in Medellin, Colombia, are not known. Our goal was to describe the use of these drugs in patients treated at Hospital Pablo Tobón Uribe and to assess the level of knowledge regarding this issue in professionals from this institution. Materials and methods: Cross sectional study conducted between January 2012 and January 2013. Two strategies to collect information were used, namely: Analysis of relevant medical records and evaluation of knowledge about the appropriate use and prescription of direct oral anticoagulants in the group of medical specialists. Results: 114 records were included in the analysis; rivaroxaban was the most frequently prescribed drug (87%) followed by dabigatran (13%). The main indication was prophylaxis in orthopedic surgery (69%). Average of correct answers among the different specialists evaluated was 67% with no apparent difference between them. Conclusion: rivaroxaban was prescribed more often than dabigatran; however, this fact does not appear to be associated with a clear and sufficient medical knowledge about these drugs. No reports of adverse events associated with this therapy were found.

Introdução e objetivo: desconhecem-se a experiência e o conhecimento com os que conta o pessoal médico do meio no manejo dos anticoagulantes orais diretos. Pretende-se descrever o uso destes medicamentos nos pacientes atendidos no Hospital Pablo Tobón Uribe e avaliar o nível de conhecimentos com respeito aos profissionais da mesma instituição. Materiais e métodos: estudo de corte transversal no período compreendido entre janeiro do 2012 e janeiro do 2013. Para coletar a informação se utilizaram duas estratégias: a análise das histórias clínicas pertinentes e a avaliação de conhecimentos sobre a prescrição e o uso de anticoagulantes orais diretos no grupo de especialistas facultados. Resultados: na análise se incluíram 114 histórias; o Rivaroxabana foi o medicamento prescrito com maior frequência (87%) seguido do Dabigatran (13%). A principal indicação foi a profilaxia em cirurgia ortopédica (69%). Entre os especialistas avaliados, a média de respostas acertadas foi de 67% sem diferenças aparentes entre os mesmos. Conclusão: o Rivaroxabana se prescreveu com maior frequência do que o Dabigatran; no entanto, este achado não parece associar-se com um conhecimento médico claro e suficiente a respeito destes medicamentos. Não se encontraram reportes de eventos adversos sócios ao uso dos mesmos.

Clinical and histological characteristics of canine ocular gliovascular syndrome.

OBJECTIVE: To characterize the clinical, diagnostic, and histopathologic findings in dogs with canine ocular gliovascular syndrome (COGS). PROCEDURES: The archives at the Comparative Ocular Pathology Laboratory of Wisconsin (COPLOW) were used to identify eyes with COGS. Histopathological inclusion criteria included: a neovascular membrane extending from the optic nerve head or retina, clusters of spindle cells lacking vascularization within the vitreous, and histological signs of glaucoma. Special and immunohistochemical (IHC) staining techniques were performed. Clinical data, treatments, and outcomes were obtained from case records and information provided by submitting veterinarians. RESULTS: Thirty-seven eyes of 36 dogs were identified with COGS. The average age at diagnosis was 8.8 years (±2.2). The relative risk for a Labrador retriever affected by COGS was significantly greater (9.3 times) (P < 0.0001) when compared to all other dog breeds within the COPLOW database. Most dogs presented with hyphema and secondary glaucoma; average intraocular pressure was 39 mmHg (±19). Average time to enucleation or evisceration was 27 days. Vitreal cells stained positive with IHC for glial fibrillary acidic protein in 14 of 17 globes, and vascular endothelial growth factor was expressed in the vitreal cells in five of five globes. CONCLUSIONS: We have defined a syndrome associated with vitreal glial cell aggregates and neovascular proliferation from the optic nerve or retina, which leads to neovascular glaucoma. The inflammation and secondary glaucoma resulting from this syndrome appear poorly responsive to conventional medical therapies. The exact etiology of COGS remains undetermined, but a systemic etiology is unlikely.

Histopathological study of the causes for failure of intrascleral prostheses in dogs and cats.

OBJECTIVE: To characterize the histopathological causes for failure of intrascleral prosthesis placement in dogs and cats. PROCEDURES: The Comparative Ocular Pathology Laboratory of Wisconsin database was searched to find canine and feline evisceration samples that were diagnosed with neoplasia. A second population included canine and feline scleral shells that were removed after an evisceration surgery had been performed. The causes for removal were divided into: neoplasia, corneal abnormalities, and other causes. RESULTS: In dogs, 163 of 1985 evisceration samples (8.21%) contained a neoplasm, whereas 17 of 88 (19.31%) evisceration samples in cats contained a neoplasm. In dogs, severe corneal disease was diagnosed in 38 of 80 scleral shells (46.25%) and neoplasia was diagnosed in 31 of 80 scleral shells (38.75%). Malignant melanoma was the most frequently diagnosed tumor, in 14 of 31 scleral shells. In cats, eight of 12 scleral shells contained a tumor (66.7%), with feline diffuse iris melanoma being diagnosed most commonly (six of eight shells). Two of 12 feline scleral shells had severe corneal disease (16.7%). Epithelial downgrowth, lining the inner aspect of the fibrous tunic, was seen in 14 of 38 canine scleral shells and in two of two feline scleral shells with severe corneal disease. CONCLUSIONS: Severe corneal disease and neoplasia are the most common causes for intrascleral prosthesis failure in dogs, whereas neoplasia is the single most common cause for intrascleral prosthesis failure in cats.

Evaluation of the presence of Leishmania spp. by real-time PCR in the lacrimal glands of dogs with leishmaniosis.

Leishmania infantum infection is highly prevalent in endemic areas. Dogs with leishmaniosis may develop keratoconjunctivitis sicca (KCS). The goals of this study were (1) to quantify Leishmania amastigotes in the Meibomian glands (MG), main lacrimal gland (MLG) and nictitating membrane gland (NMG) from dogs with leishmaniosis; (2) to compare these results to immunohistochemistry (IHC), and (3) to explore the association between the Leishmania parasite load and the presence of ocular clinical signs. Twenty-five dogs diagnosed with leishmaniosis were included. MG, MLG and NMG from both eyes were collected. Histopathology, IHC and real-time PCR were performed. All specimens yielded positive real-time PCR results. For all three glands, samples from dogs with ocular clinical signs had mean ΔCt (cycle threshold) values significantly lower (higher parasite loads) than those from dogs without signs. Cut-off values of ΔCt<0, ΔCt<4 and ΔCt<4.9 for MG, MLG and NMG, resulted in a likelihood ratio of positives of 5.9, 6.38 and 6.38, respectively. Samples with ΔCt values below the reported cut-off were significantly more likely to display clinical signs related to KCS than those with results above the cut-off, for all three glands. Similarly, ΔCt values below the cut-off were significantly associated with positive IHC. In this study real-time PCR has been standardised for use in MG, MLG and NMG. A cut-off value established for each of these tissues may aid the clinician in the discrimination between ocular signs related to Leishmania from those associated with other causes of KCS.

Allelic variants of melanocortin 3 receptor gene (MC3R) and weight loss in obesity: a randomised trial of hypo-energetic high- versus low-fat diets.

INTRODUCTION: The melanocortin system plays an important role in energy homeostasis. Mice genetically deficient in the melanocortin-3 receptor gene have a normal body weight with increased body fat, mild hypophagia compared to wild-type mice. In humans, Thr6Lys and Val81Ile variants of the melanocortin-3 receptor gene (MC3R) have been associated with childhood obesity, higher BMI Z-score and elevated body fat percentage compared to non-carriers. The aim of this study is to assess the association in adults between allelic variants of MC3R with weight loss induced by energy-restricted diets. SUBJECTS AND METHODS: This research is based on the NUGENOB study, a trial conducted to assess weight loss during a 10-week dietary intervention involving two different hypo-energetic (high-fat and low-fat) diets. A total of 760 obese patients were genotyped for 10 single nucleotide polymorphisms covering the single exon of MC3R gene and its flanking regions, including the missense variants Thr6Lys and Val81Ile. Linear mixed models and haplotype-based analysis were carried out to assess the potential association between genetic polymorphisms and differential weight loss, fat mass loss, waist change and resting energy expenditure changes. RESULTS: No differences in drop-out rate were found by MC3R genotypes. The rs6014646 polymorphism was significantly associated with weight loss using co-dominant (p = 0.04) and dominant models (p = 0.03). These p-values were not statistically significant after strict control for multiple testing. Haplotype-based multivariate analysis using permutations showed that rs3827103-rs1543873 (p = 0.06), rs6014646-rs6024730 (p = 0.05) and rs3746619-rs3827103 (p = 0.10) displayed near-statistical significant results in relation to weight loss. No other significant associations or gene*diet interactions were detected for weight loss, fat mass loss, waist change and resting energy expenditure changes. CONCLUSION: The study provided overall sufficient evidence to support that there is no major effect of genetic variants of MC3R and differential weight loss after a 10-week dietary intervention with hypo-energetic diets in obese Europeans.

Detection of Leishmania spp. and associated inflammation in ocular-associated smooth and striated muscles in dogs with patent leishmaniosis.

OBJECTIVE: Canine leishmaniosis is a disease characterized by the wide distribution of the parasite throughout the tissues of the host. The purpose of this study was to describe the presence of Leishmania spp. and associated inflammation in ocular-associated muscles of dogs with patent leishmaniosis. PROCEDURES: Smooth muscles (iris dilator muscle, iris sphincter muscle, ciliary muscle, Müller muscle, smooth muscle of the periorbita and smooth muscle of the nictitating membrane) and striated muscles (orbicularis oculi muscle, obliquus dorsalis muscle and dorsal rectus muscle) were evaluated. Routine staining with hematoxylin and eosin and immunohistochemistry to detect Leishmania spp. were performed on tissue sections. RESULTS: Granulomatous inflammation was seen surrounding muscular fibers and was composed mainly of macrophages with scattered lymphocytes and plasma cells. This infiltrate could be seen in 52/473 (10.99%) samples of smooth muscle and 36/142 (25.35%) samples of striated muscle. Parasites were detected in 43/473 (9.09%) samples of smooth muscle and in 28/142 (19.71%) samples of striated muscle. CONCLUSIONS: To the authors' knowledge, this is the first report assessing the presence of Leishmania spp. and associated infiltrate in intraocular, extraocular and adnexal smooth and striated muscles. The inflammation present in those muscles could contribute to clinical signs already described, such as blepharitis, uveitis, and orbital cellulitis.

Ligneous conjunctivitis in a plasminogen-deficient dog: clinical management and 2-year follow-up.

A 1-year-old-female Yorkshire Terrier was referred to the Veterinary Teaching Hospital of the Autonomous University of Barcelona (VTH-UAB) (Spain) with a 6-month history of unilateral chronic proliferative conjunctivitis and intermittent vomiting and cough. Several medical and surgical treatment efforts to manage conjunctival lesions had resulted in no improvement of the clinical signs. Complete general and ophthalmic examinations revealed several proliferative 'wood-like' masses in the conjunctiva, oral cavity and an interscapular subcutaneous nodule. Conjunctival and buccal biopsies were performed as diagnostic procedures. A diagnosis of ligneous conjunctivitis was made on the basis of histopathology findings and clinical presentation. The only biochemical abnormalities found were severe proteinuria and low plasminogen activity in plasma. No other analytical abnormalities were observed. Topical treatment with heparin and anti-inflammatory and immunosuppressive drugs have controlled the ophthalmological clinical signs. To our knowledge, this is the first case report of a dog with plasminogen deficiency and ligneous conjunctivitis with a long survival period and 2-year follow-up.

Canine normal corneal epithelium bears a large population of CD45-positive cells.

This investigation sought to identify the presence of immune cells in normal canine corneal epithelium. A whole-mount immunofluorescence study of normal canine epithelium using monoclonal antibodies against CD45, CD11c, CD1c and MHC class II was performed. CD45-positive cells were located in all epithelial layers throughout the cornea, occurring in greater numbers (51.98+/-4.1/mm(2)) at the periphery and decreasing towards the central region (11.8+/-3.1/mm(2)). CD11c-positive cells were also observed, but were fewer in number. The findings show that the normal canine cornea carries a significant number of cells of immune origin; these cells seem to be of an inactive phenotype as they do not express MHC class II. Further studies are needed to determine whether these cells can express co-stimulatory molecules and act as antigen presenting cells if stimulated.