Bilateral canine seminoma with ocular metastasis: histochemical and immunohistochemical characterization

A 10-year old male dog was examined due to a buphthalmia in the left eye and a nodule in the two testicles. Due to the limited resources of the owner and loss of visual acuity of the patient, the enucleation and castration were chosen as treatment. Microscopic analysis of the testicular tissue revealed neoplastic germ cells. Morphologically, neoplastic cells were characterized by distinct cell borders, scarce and eosinophilic cytoplasm, large round nucleus, with thick chromatin and a prominent nucleolus. Binucleated and multinucleated neoplastic cells were also frequently observed. In 10 high powerfields (400x), 62 typical and atypical mitosis were counted. Similar neoplastic cells were identified within the vessels of theretina, sclera and in the sub-epithelial conjunctive tissue of the eyelid. The neoplastic cells observed in the testicle and in the eye were positive for PAS. By immunochemistry technique was identified an intense immunostaining of the neoplastic cells for Vimentin and Ki-67 in both testicular and ocular tissue. While, discrete immunoreactivity was identified to c-KIT from the neoplastic cells in both organs. Based on morphological, histochemical and immunohistochemical analysis, it was possible to characterize the ocular lesion as seminoma metastasis.

Bilateral canine seminoma with ocular metastasis: histochemical and immunohistochemical characterization

A 10-year old male dog was examined due to a buphthalmia in the left eye and a nodule in the two testicles. Due to the limited resources of the owner and loss of visual acuity of the patient, the enucleation and castration were chosen as treatment. Microscopic analysis of the testicular tissue revealed neoplastic germ cells. Morphologically, neoplastic cells were characterized by distinct cell borders, scarce and eosinophilic cytoplasm, large round nucleus, with thick chromatin and a prominent nucleolus. Binucleated and multinucleated neoplastic cells were also frequently observed. In 10 high powerfields (400x), 62 typical and atypical mitosis were counted. Similar neoplastic cells were identified within the vessels of theretina, sclera and in the sub-epithelial conjunctive tissue of the eyelid. The neoplastic cells observed in the testicle and in the eye were positive for PAS. By immunochemistry technique was identified an intense immunostaining of the neoplastic cells for Vimentin and Ki-67 in both testicular and ocular tissue. While, discrete immunoreactivity was identified to c-KIT from the neoplastic cells in both organs. Based on morphological, histochemical and immunohistochemical analysis, it was possible to characterize the ocular lesion as seminoma metastasis.(AU)

Molecular characterization and potential sources of aqueous humor bacterial contamination during phacoemulsification with intraocular lens implantation in dogs.

Bacterial contamination of the anterior chamber during cataract surgery is one of the main responsible for endophthalmitis postoperative. Phacoemulsification is a less invasive technique for cataract treatment, although it does not exclude the possibility of contamination. In this study, bacterial contaminants of aqueous humor collected pre- and post-phacoemulsification with intraocular lens implantation (IOL) of twenty dogs were identified. As the conjunctival microbiota constitute a significant source of anterior chamber contamination, bacterial isolates from aqueous humor were genetically compared with those present in the conjunctival surface of the patients. Three dogs presented bacterial growth in both aqueous humor and conjunctival surface samples. Bacterial isolates from these samples were grouped according to their genetic profiles by repetitive-element PCR (rep-PCR) and their representatives were identified by 16S rRNA sequencing. Isolates from conjunctival surface were identified as Enterobacter spp., Staphylococcus spp. and S. aureus; and from aqueous humor samples as Enterobacter spp., Pantoea spp., Streptococcus spp. and Staphylococcus spp., respectively in decreasing order of prevalence. According to the rep-PCR analysis, 16.6% of Enterobacter spp. isolates from conjunctival surface were genetically similar to those from aqueous humor. The rest of isolates encountered in aqueous humor were genetically distinct from those of conjunctival surface. The significant genetic diversity of bacterial isolates found in the aqueous humor samples after surgery denoted the possibility of anterior chamber contamination during phacoemulsification by bacteria not only from conjunctival surface but also from different sources related to surgical environment.

Histological, morphometric, protein and gene expression analyses of rat retinas with ischaemia-reperfusion injury model treated with sildenafil citrate.

The aim of this study was to better understand the role of apoptosis in a retinal ischaemia-reperfusion injury model and to determine whether sildenafil citrate treatment can prevent retinal cell apoptosis. Thirty-six rats were divided into a control group (n = 6) and two experimentally induced ischaemia-reperfusion groups (7 and 21 days; n = 15 per group). The induced ischaemia-reperfusion groups were treated with sildenafil for 7 and 21 days (n = 10 per group), and 10 animals were treated with a placebo for the same period (n = 5 per group). Paracentesis of the anterior chamber was performed with a 30-G needle attached to a saline solution (0.9%) bag positioned at a height of 150 cm above the eye for 60 min. Intraocular pressure was measured by rebound tonometer (TonoVet® ). The eyes were analysed by histology and morphometry, and by immunohistochemistry and qRT-PCR for expression of Caspase-7, Caspase-6, Caspase-9, Tnf-r2, Fas-l, Bcl-2 and Bax. Sildenafil-treated animals showed lower levels of histopathological changes (inflammatory, cellular and tissue) than their placebo-treated counterparts at both 7 and 21 days. The retinal ganglion cell layer (RGC) was preserved in the sildenafil groups (SG), with a cell count closer to control than in the placebo groups (PG). Caspase-7 expression was significantly higher in both treated groups at 7 days compared to controls. Gene expression levels in both treatment groups differed from the controls, but in SG Bax and Caspase-6 expression levels were similar to control animals. These results suggest that the main mechanism of retinal cell death in this model is apoptosis, as there is an increase in pro-apoptotic factors and decrease in the anti-apoptotic ones. Also, sildenafil seems to protect the retinal ganglion cell layer from apoptosis. Cell survival was evident in the histological and morphometric analyses, and sildenafil treatment had a protective effect in the apoptosis pathways, with gene expression levels in SG similar to the controls.

Immunohistochemistry characterization of a bronchioloalveolar carcinoma mixed subtype in cat

A 12 year-old mixed breed female cat was present with signs of dyspnea, cyanosis and drooling, followed by sudden death. At necropsy we observed nodules measuring 0.5 x 0.5 to 2.0 x 2.0 cm in the pulmonary parenchyma showed multifocal atelectasis associated with discrete emphysema. The histopathological examination of lung masses revealed neoplastic epithelial cells compatible with bronchioloalveolar carcinoma (BAC). The immunohistochemical analysis of tumor cells showed positive labeling for pan-Cytokeratin, CK 7, CK 20 and TTF-1. According to macroscopic features, as well as histological and immunohistochemical findings, this tumor was diagnostic as BAC mixed subtype.(AU)

Immunohistochemistry characterization of a bronchioloalveolar carcinoma mixed subtype in cat

A 12 year-old mixed breed female cat was present with signs of dyspnea, cyanosis and drooling, followed by sudden death. At necropsy we observed nodules measuring 0.5 x 0.5 to 2.0 x 2.0 cm in the pulmonary parenchyma showed multifocal atelectasis associated with discrete emphysema. The histopathological examination of lung masses revealed neoplastic epithelial cells compatible with bronchioloalveolar carcinoma (BAC). The immunohistochemical analysis of tumor cells showed positive labeling for pan-Cytokeratin, CK 7, CK 20 and TTF-1. According to macroscopic features, as well as histological and immunohistochemical findings, this tumor was diagnostic as BAC mixed subtype.

Effects of morphine on the expression of cytokines and inflammatory mediators in a rabbit model of endotoxin-induced experimental uveitis.

PURPOSE: To evaluate the effects of 1% morphine instillation on clinical parameters, aqueous humor turbidity, and expression levels of tumor necrosis factor alpha (TNF-α), interleukin-1 beta (IL-1beta), prostaglandin E2 (PGE2), and myeloperoxidase (MPO) in rabbits with endotoxin-induced experimental uveitis. METHODS: Twenty four New Zealand white rabbits were divided into four groups (n=6 each): control (CG), morphine (MG), naloxone (NG), and morphine-naloxone (MNG) groups. Under dissociative anesthesia, 0.1 mL of solution containing 0.2 µg of lipopolysaccharide (LPS) endotoxin from the Salmonella typhimurium cell wall was injected in the vitreous chamber. Clinical evaluations (conjunctical hyperemia, chemosis blepharospasm, and ocular discharge) and laser flaremetry were performed before (baseline), and 10 and 20 hours after induction of uveitis. Rabbits were subsequently euthanized and eyes were enucleated to quantify expression levels of TNF-α, IL-1 beta, PGE2, and MPO. RESULTS: No significant differences in clinical parameters and flare values were observed between the study groups. TNF-α and IL-1 beta levels increased significantly in the CG, MG, NG, and MNG groups compared to baseline (P<0.05). Significant differences in PGE2 levels were observed between the MG and NMG groups (P<0.05). A trend toward increased MPO activity was observed in response to uveitis induction; however, this trend did not reach statistical significance (P>0.05). CONCLUSIONS: Morphine has no effect on clinical parameters, flare, or expression levels of inflammatory mediators in a rabbit model of uveitis induced by intravitreal injection of LPS.

Effects of morphine on the expression of cytokines and inflammatory mediators in a rabbit model of endotoxin-induced experimental uveitis / Efeitos da morfina na expressão de citocinas e de mediadores inflamatórios em coelhos com uveítes experimental induzida por endotoxina

ABSTRACT Purpose: To evaluate the effects of 1% morphine instillation on clinical parameters, aqueous humor turbidity, and expression levels of tumor necrosis factor alpha (TNF-α), interleukin-1 beta (IL-1beta), prostaglandin E2 (PGE2), and myeloperoxidase (MPO) in rabbits with endotoxin-induced experimental uveitis. Methods: Twenty four New Zealand white rabbits were divided into four groups (n=6 each): control (CG), morphine (MG), naloxone (NG), and morphine-naloxone (MNG) groups. Under dissociative anesthesia, 0.1 mL of solution containing 0.2 µg of lipopolysaccharide (LPS) endotoxin from the Salmonella typhimurium cell wall was injected in the vitreous chamber. Clinical evaluations (conjunctical hyperemia, chemosis blepharospasm, and ocular discharge) and laser flaremetry were performed before (baseline), and 10 and 20 hours after induction of uveitis. Rabbits were subsequently euthanized and eyes were enucleated to quantify expression levels of TNF-α, IL-1 beta, PGE2, and MPO. Results: No significant differences in clinical parameters and flare values were observed between the study groups. TNF-α and IL-1 beta levels increased significantly in the CG, MG, NG, and MNG groups compared to baseline (P<0.05). Significant differences in PGE2 levels were observed between the MG and NMG groups (P<0.05). A trend toward increased MPO activity was observed in response to uveitis induction; however, this trend did not reach statistical significance (P>0.05). Conclusions: Morphine has no effect on clinical parameters, flare, or expression levels of inflammatory mediators in a rabbit model of uveitis induced by intravitreal injection of LPS.

RESUMO Objetivo: Estudaram-se os efeitos da instilação de morfina 1% sobre parâmetros clínicos, turbidez do humor aquoso e expressão de fator de necrose tumoral alfa (TNF-alfa), de interleucina-1 beta (IL-1beta), de prostaglandina E2 (PGE2) e de mieloperoxidase (MPO), em olhos de coelhos com uveíte induzida por endotoxina. Material e Métodos: Vinte e quatro coelhos da raça Nova Zelândia Branco foram distribuídos em quatro grupos (n=6, em cada): grupo controle (GC), morfina (GM), naloxona (GN) e morfina-naloxona (GMN). Sob anestesia dissociativa, injetou-se 0,1 mL de solução contendo 0,2 µg de lipossacarídeo (LPS) endotóxico da parede celular de Salmonella typhimurium na câmara vítrea. Realizou-se avaliação clínica (hiperemia conjuntival, quemose, blefaroespasmo e secreção ocular) e a flaremetria a “laser” antes (basal) e após 10 e 20 horas da indução da uveíte. No final, os coelhos foram submetidos à eutanásia e os olhos com uveíte foram enucleados para a quantificação dos níveis de TNF-alfa, IL-1 beta, PGE2 e MPO. Diferenças foram consideradas significativas quando p<0,05. Resultados: Os grupos da pesquisa não diferiram quanto aos parâmetros clínicos e os valores de “flare”. Observou-se elevação significativa nos níveis de TNF-alfa e de IL-1 beta, comparativamente ao basal, nos grupos GC, GM, GN e GMN (p<0,05). Valores de PGE2 variaram entre os grupos GM e GNM (p<0,05). A atividade de MPO aumentou após a indução da uveíte, porém, sem significância estatística (p>0,05). Conclusões: A morfina não atuou sobre parâmetros clínicos, “flare” e expressão dos mediadores inflamatórios estudados, quando instilada em olhos de coelhos com uveíte induzida por injeção intravítrea de LPS.

Clinical presentation and renal evaluation of human visceral leishmaniasis (kala-azar): a retrospective study of 57 patients in Brazil.

Visceral leishmaniasis is an endemic disease caused by various species of Leishmania. We made a retrospective study of 57 consecutive patients with visceral leishmaniasis in Brazil. Patients with visceral leishmaniasis were identified using the registries of the São José Infectious Diseases Hospital. The sample was divided into two groups: patients with serum creatinine (Scr) <1.3mg/dL and Scr > or = 1.3mg/dL. We compared these two groups for differences in clinical manifestations and laboratory features. Patients' mean age was 28 +/- 18 years old; 74% were male. The main clinical symptoms and signs presented in the initial evaluation were: fever (97%), splenomegaly (96.4%), weight loss (95.5%), pallor (93.6%), cough (89.7%), hepatomegaly (87.2%), asthenia (83.3%), anorexia (82.9%) and vomiting (73.9%). Acute renal failure was found in 15 patients (26.3%) and eight of these patients had ARF before amphotericin B administration. The mean age was higher in the group with Scr > or =1.3mg/dL. Death occurred in three cases; all deaths occurred with Scr > or =1.3mg/dL. There were no significant differences in the frequencies of the clinical symptoms and signs between the two groups. The laboratory data and demographic characteristics were significantly worse in the Scr > or =1.3mg/dL group. Renal dysfunction is an important feature of this disease; it is associated with important morbidity and can increase mortality.

Clinical presentation and renal evaluation of human visceral leishmaniasis (kala-azar): a retrospective study of 57 patients in Brazil

Visceral leishmaniasis is an endemic disease caused by various species of Leishmania. We made a retrospective study of 57 consecutive patients with visceral leishmaniasis in Brazil. Patients with visceral leishmaniasis were identified using the registries of the São José Infectious Diseases Hospital. The sample was divided into two groups: patients with serum creatinine (Scr) <1.3mg/dL and Scr > 1.3mg/dL. We compared these two groups for differences in clinical manifestations and laboratory features. Patients' mean age was 28 ± 18 years old; 74 percent were male. The main clinical symptoms and signs presented in the initial evaluation were: fever (97 percent), splenomegaly (96.4 percent), weight loss (95.5 percent), pallor (93.6 percent), cough (89.7 percent), hepatomegaly (87.2 percent), asthenia (83.3 percent), anorexia (82.9 percent) and vomiting (73.9 percent). Acute renal failure was found in 15 patients (26.3 percent) and eight of these patients had ARF before amphotericin B administration. The mean age was higher in the group with Scr > 1.3mg/dL. Death occurred in three cases; all deaths occurred with Scr > 1.3mg/dL. There were no significant differences in the frequencies of the clinical symptoms and signs between the two groups. The laboratory data and demographic characteristics were significantly worse in the Scr > 1.3mg/dL group. Renal dysfunction is an important feature of this disease; it is associated with important morbidity and can increase mortality.