Gastroesophageal tube of the Iguana iguana (Iguanidae): histological description, histochemical and immunohistochemical analysis of 5-HT and SS cells

Abstract The work aims were to describe the histological and histochemical structure of the gastroesophageal tube of Iguana iguana and verify the occurrence and distribution of immunoreactive serotonin (5-HT) and somatostatin (SS) cells. Fragments of the gastrointestinal tract (GIT) of five iguanas were which underwent standard histological and immunohistochemistry technique. Immunoreactive cells for 5-HT and SS were quantified using the STEPanizer. The oesophagus has ciliated columnar pseudostratified epithelium with staining Alcian blue (AB) + and goblet cells highly reactive to periodic acid Schiff (PAS). In the cervical oesophagus, the numerical density of 5-HT cells per unit area (QA [5-HT cells]/µm2) was 4.6x10-2 ± 2.0 and celomatic oesophagus presented QA = 4.0x10-2 ± 1.0. The epithelium of the stomach is simple columnar, PAS and AB +. The cranial and middle regions of the stomach presented (QA [5-HT cells]/µm2) = 6.18x10-2 ± 3.2 and the caudal region, QA = 0.6x10-2 ± 0.2. The SS cells were only observed in the caudal stomach, with numerical density (QA [SS cells]/µm2) = 1.4x10-2 ± 0.9 In I. iguana, variation was observed in terms of the distribution of mucus secretions and the pattern of occurrence of serotonin and somatostatin-secreting enteroendocrine cells in the TGI, which possibly will result in an interspecific adaptive response.

Resumo Os objetivos do trabalho foram descrever a estrutura histológica e histoquímica do tubo gastroesofágico da Iguana iguana e verificar a ocorrência e distribuição de células serotonina (5-HT) e somatostatina (SS) imunorreativas. Fragmentos do trato gastrointestinal (TGI) de cinco iguanas foram submetidos à técnica histológica e imunohistoquímica padrão. As células imunorreativas para 5-HT e SS foram quantificadas usando o STEPanizer. O esôfago apresenta epitélio pseudoestratificado colunar ciliado Alcian blue (AB) positivo, com células caliciformes altamente reativas ao ácido periódico de Schiff (PAS). No esôfago cervical, a densidade numérica de células 5-HT por unidade de área (QA [células 5-HT] / µm2) foi de 4.6x10-2 ± 2.0 e o esôfago celomático apresentou QA = 4.0x10-2 ± 1.0. O epitélio do estômago é colunar simples, PAS e AB positivo. As regiões cranial e média do estômago apresentaram (QA [células 5-HT] / µm2) = 6.18x10-2 ± 3.2 e a região caudal, QA = 0.6x10-2 ± 0.2. As células SS foram observadas apenas no estômago caudal, com densidade numérica (QA [células SS] / µm2) = 1.4x10-2 ± 0.9. Em I. iguana, foi observada variações em termos da distribuição das secreções de muco e padrão de ocorrência das células enteroendócrinas secretoras de serotonina e somatostatina no TGI, o que possivelmente reflete uma resposta adaptativa interespecifica.

Gastroesophageal tube of the Iguana iguana (Iguanidae): histological description, histochemical and immunohistochemical analysis of 5-HT and SS cells / Tubo gastroesofágico da Iguana iguana (Iguanidae): descrição histológica, histoquímica e análise imunohistoquímica de células 5-HT e SS

Abstract The work aims were to describe the histological and histochemical structure of the gastroesophageal tube of Iguana iguana and verify the occurrence and distribution of immunoreactive serotonin (5-HT) and somatostatin (SS) cells. Fragments of the gastrointestinal tract (GIT) of five iguanas were which underwent standard histological and immunohistochemistry technique. Immunoreactive cells for 5-HT and SS were quantified using the STEPanizer. The oesophagus has ciliated columnar pseudostratified epithelium with staining Alcian blue (AB) + and goblet cells highly reactive to periodic acid Schiff (PAS). In the cervical oesophagus, the numerical density of 5-HT cells per unit area (QA [5-HT cells]/µm2) was 4.6x10-2 ± 2.0 and celomatic oesophagus presented QA = 4.0x10-2 ± 1.0. The epithelium of the stomach is simple columnar, PAS and AB +. The cranial and middle regions of the stomach presented (QA [5-HT cells]/µm2) = 6.18x10-2 ± 3.2 and the caudal region, QA = 0.6x10-2 ± 0.2. The SS cells were only observed in the caudal stomach, with numerical density (QA [SS cells]/µm2) = 1.4x10-2 ± 0.9 In I. iguana, variation was observed in terms of the distribution of mucus secretions and the pattern of occurrence of serotonin and somatostatin-secreting enteroendocrine cells in the TGI, which possibly will result in an interspecific adaptive response.

Resumo Os objetivos do trabalho foram descrever a estrutura histológica e histoquímica do tubo gastroesofágico da Iguana iguana e verificar a ocorrência e distribuição de células serotonina (5-HT) e somatostatina (SS) imunorreativas. Fragmentos do trato gastrointestinal (TGI) de cinco iguanas foram submetidos à técnica histológica e imunohistoquímica padrão. As células imunorreativas para 5-HT e SS foram quantificadas usando o STEPanizer. O esôfago apresenta epitélio pseudoestratificado colunar ciliado Alcian blue (AB) positivo, com células caliciformes altamente reativas ao ácido periódico de Schiff (PAS). No esôfago cervical, a densidade numérica de células 5-HT por unidade de área (QA [células 5-HT] / µm2) foi de 4.6x10-2 ± 2.0 e o esôfago celomático apresentou QA = 4.0x10-2 ± 1.0. O epitélio do estômago é colunar simples, PAS e AB positivo. As regiões cranial e média do estômago apresentaram (QA [células 5-HT] / µm2) = 6.18x10-2 ± 3.2 e a região caudal, QA = 0.6x10-2 ± 0.2. As células SS foram observadas apenas no estômago caudal, com densidade numérica (QA [células SS] / µm2) = 1.4x10-2 ± 0.9. Em I. iguana, foi observada variações em termos da distribuição das secreções de muco e padrão de ocorrência das células enteroendócrinas secretoras de serotonina e somatostatina no TGI, o que possivelmente reflete uma resposta adaptativa interespecifica.

Gastroesophageal tube of the Iguana iguana (Iguanidae): histological description, histochemical and immunohistochemical analysis of 5-HT and SS cells / Tubo gastroesofágico da Iguana iguana (Iguanidae): descrição histológica, histoquímica e análise imunohistoquímica de células 5-HT e SS

The work aims were to describe the histological and histochemical structure of the gastroesophageal tube of Iguana iguana and verify the occurrence and distribution of immunoreactive serotonin (5-HT) and somatostatin (SS) cells. Fragments of the gastrointestinal tract (GIT) of five iguanas were which underwent standard histological and immunohistochemistry technique. Immunoreactive cells for 5-HT and SS were quantified using the STEPanizer. The oesophagus has ciliated columnar pseudostratified epithelium with staining Alcian blue (AB) + and goblet cells highly reactive to periodic acid Schiff (PAS). In the cervical oesophagus, the numerical density of 5-HT cells per unit area (QA [5-HT cells]/µm2) was 4.6x10-2 ± 2.0 and celomatic oesophagus presented QA = 4.0x10-2 ± 1.0. The epithelium of the stomach is simple columnar, PAS and AB +. The cranial and middle regions of the stomach presented (QA [5-HT cells]/µm2) = 6.18x10-2 ± 3.2 and the caudal region, QA = 0.6x10-2 ± 0.2. The SS cells were only observed in the caudal stomach, with numerical density (QA [SS cells]/µm2) = 1.4x10-2 ± 0.9 In I. iguana, variation was observed in terms of the distribution of mucus secretions and the pattern of occurrence of serotonin and somatostatin-secreting enteroendocrine cells in the TGI, which possibly will result in an interspecific adaptive response.

Os objetivos do trabalho foram descrever a estrutura histológica e histoquímica do tubo gastroesofágico da Iguana iguana e verificar a ocorrência e distribuição de células serotonina (5-HT) e somatostatina (SS) imunorreativas. Fragmentos do trato gastrointestinal (TGI) de cinco iguanas foram submetidos à técnica histológica e imunohistoquímica padrão. As células imunorreativas para 5-HT e SS foram quantificadas usando o STEPanizer. O esôfago apresenta epitélio pseudoestratificado colunar ciliado Alcian blue (AB) positivo, com células caliciformes altamente reativas ao ácido periódico de Schiff (PAS). No esôfago cervical, a densidade numérica de células 5-HT por unidade de área (QA [células 5-HT] / µm2) foi de 4.6x10-2 ± 2.0 e o esôfago celomático apresentou QA = 4.0x10-2 ± 1.0. O epitélio do estômago é colunar simples, PAS e AB positivo. As regiões cranial e média do estômago apresentaram (QA [células 5-HT] / µm2) = 6.18x10-2 ± 3.2 e a região caudal, QA = 0.6x10-2 ± 0.2. As células SS foram observadas apenas no estômago caudal, com densidade numérica (QA [células SS] / µm2) = 1.4x10-2 ± 0.9. Em I. iguana, foi observada variações em termos da distribuição das secreções de muco e padrão de ocorrência das células enteroendócrinas secretoras de serotonina e somatostatina no TGI, o que possivelmente reflete uma resposta adaptativa interespecifica.

Gastroesophageal tube of the Iguana iguana (Iguanidae): histological description, histochemical and immunohistochemical analysis of 5-HT and SS cells / Tubo gastroesofágico da Iguana iguana (Iguanidae): descrição histológica, histoquímica e análise imunohistoquímica de células 5-HT e SS

The work aims were to describe the histological and histochemical structure of the gastroesophageal tube of Iguana iguana and verify the occurrence and distribution of immunoreactive serotonin (5-HT) and somatostatin (SS) cells. Fragments of the gastrointestinal tract (GIT) of five iguanas were which underwent standard histological and immunohistochemistry technique. Immunoreactive cells for 5-HT and SS were quantified using the STEPanizer. The oesophagus has ciliated columnar pseudostratified epithelium with staining Alcian blue (AB) + and goblet cells highly reactive to periodic acid Schiff (PAS). In the cervical oesophagus, the numerical density of 5-HT cells per unit area (QA [5-HT cells]/µm2) was 4.6x10-2 ± 2.0 and celomatic oesophagus presented QA = 4.0x10-2 ± 1.0. The epithelium of the stomach is simple columnar, PAS and AB +. The cranial and middle regions of the stomach presented (QA [5-HT cells]/µm2) = 6.18x10-2 ± 3.2 and the caudal region, QA = 0.6x10-2 ± 0.2. The SS cells were only observed in the caudal stomach, with numerical density (QA [SS cells]/µm2) = 1.4x10-2 ± 0.9 In I. iguana, variation was observed in terms of the distribution of mucus secretions and the pattern of occurrence of serotonin and somatostatin-secreting enteroendocrine cells in the TGI, which possibly will result in an interspecific adaptive response.(AU)

Os objetivos do trabalho foram descrever a estrutura histológica e histoquímica do tubo gastroesofágico da Iguana iguana e verificar a ocorrência e distribuição de células serotonina (5-HT) e somatostatina (SS) imunorreativas. Fragmentos do trato gastrointestinal (TGI) de cinco iguanas foram submetidos à técnica histológica e imunohistoquímica padrão. As células imunorreativas para 5-HT e SS foram quantificadas usando o STEPanizer. O esôfago apresenta epitélio pseudoestratificado colunar ciliado Alcian blue (AB) positivo, com células caliciformes altamente reativas ao ácido periódico de Schiff (PAS). No esôfago cervical, a densidade numérica de células 5-HT por unidade de área (QA [células 5-HT] / µm2) foi de 4.6x10-2 ± 2.0 e o esôfago celomático apresentou QA = 4.0x10-2 ± 1.0. O epitélio do estômago é colunar simples, PAS e AB positivo. As regiões cranial e média do estômago apresentaram (QA [células 5-HT] / µm2) = 6.18x10-2 ± 3.2 e a região caudal, QA = 0.6x10-2 ± 0.2. As células SS foram observadas apenas no estômago caudal, com densidade numérica (QA [células SS] / µm2) = 1.4x10-2 ± 0.9. Em I. iguana, foi observada variações em termos da distribuição das secreções de muco e padrão de ocorrência das células enteroendócrinas secretoras de serotonina e somatostatina no TGI, o que possivelmente reflete uma resposta adaptativa interespecifica.(AU)

Canine mixed tumors as one of the mammary neoplasia varieties

The most significant interest in veterinary oncology is occupied by tumors of a mixed nature, which are represented by the direct interaction of epithelial and mesenchymal components. According to some data, the formation of cell lineage of mesenchymal cells occurs due to the transformation of epidermal basket cells surrounding the glandular epithelium. In this case, the formation of a cartilage anlage with its further transformation into differentiated bone tissue occurs. The article contains information about the structure and features of the location and interaction of mixed tumors' glandular and stromal components in female dogs (n=29). It was revealed that in addition to simple mixed neoplasms, tumors with proplasia of the mesenchymal component into highly differentiated cartilage or bone tissue were also often registered. To confirm subcellular changes in the expression of certain types of proteins, such as a-SMA and vimentin, an IHC study was used. The expression of the studied biomarkers was established in mesenchymal fibroblastic differon cells and myoepithelial cells. The intensity of immunoreactive material expression ranged from moderate (2+) to strong (3+), indicating changes in myoepithelial cells' genotype during the formation of mixed mammary neoplasms.(AU)

Adenosquamous carcinoma in the palate of a dog: case report

Adenosquamous carcinoma is characterized by the simultaneous presence of squamous and glandular components. This article reports a case of adenosquamous carcinoma in the oral mucosa in a dog. Microscopically, the tumour revealed an infiltrative epithelial proliferation with a tubular pattern and areas of squamous differentiation. In the lumina of the tubules, there were eosinophilic material (mucin). The alcian blue staining showed positive intraluminal material and the cytoplasm of neoplastic cells were immunoreactive for cytokeratin, supporting the diagnosis of adenosquamous carcinoma. The cells exhibited reduced or negative immunoreactivity for E-cadherin, which could be associated with more aggressive tumour behaviour and worse prognosis.(AU)

Sarcoma estromal esplênico em canino da raça labrador / Splenic stromal sarcoma in a labrador dog

Background: Splenic stromal neoplasms are a heterogeneous group of tumors that shares the morphology of spindle cells, and are considered a primary mesenchymal neoplasm of the spleen. Among these neoplasms, some are as yet unclassified. Although stromal neoplasms represent 25 a 50% of dog splenic neoplasms in dogs, subtypes rarely occur alone. As these neoplasms are difficult to diagnose in the routine veterinary medical examination, the objective of this article was to report a case of splenic stromal sarcoma in a dog treated at a private veterinary hospital in Recife, Pernambuco, Brazil. Case: An 8-year-old male labrador dog was treated at a veterinary hospital for an initial clinical suspicion of gastroenteritis. Clinical examinations showed anemic ocular mucosa and a distended abdomen. Considering the animal's condition, blood count, serum biochemistry, and abdominal ultrasonography (US) were requested. The laboratory tests revealed anemia, thrombocytopenia, leukocytosis with neutrophilia, and changes in the indices of alanine aminotransferase (ALT) and alkaline phosphatase (ALP), among other findings. Abdominal ultrasound showed hepatomegaly, diffuse splenomegaly, a neoformation in the spleen suggestive of splenic neoplasia, and mild cystic prostatic hyperplasia. The dog underwent exploratory laparotomy with splenectomy. During the procedure, a nodule was found attached to the spleen. It was removed, fixed in a 10% neutral buffered formalin solution, and sent for histopathological and immunohistochemical examinations (vimentin, S100, desmin, MyoD1, HHF35, CD31, and alpha smooth muscle actin). The histopathological diagnosis was poorly differentiated spindle cell splenic sarcoma, compatible with splenic stromal sarcoma. Immunohistochemistry showed neoplastic cells immunoreactive for vimentin and S100, and it was not reactive for other markers. Based on the immunohistochemical and morphological profile, the diagnosis of splenic stromal sarcoma was confirmed. Discussion: Dogs affected by splenic neoplasms are generally aged between three and 17 years, and clinically present with anorexia, emesis, lethargy, weight loss, and abdomen distension, which were observed in the present case. However, a purelyclinical diagnosis is disputable, since other diseases can present the same signs. Thus, complementary exams are essential for a correct diagnosis. Laboratory changes in blood count and serum biochemistry are considered nonspecific for splenic neoplasms, with neutrophilic leukocytosis and anemia being consistent with paraneoplastic syndromes frequently diagnosed in small animals. Microscopically, splenic stromal sarcomas are characterized by focal splenic nodules composed of cohesive polygonal to spindle-shaped cell layers. The cells usually exhibit anisokaryosis and often have large oval to round vesicular nuclei and, occasionally, multinucleated cells. They have large areas of necrosis, diffuse or nodular lymphoid aggregates, and foci of extramedullary hematopoiesis, consistent with the histopathological examination reported in the present case. The immunohistochemical analysis included evaluation of seven markers to elucidate the histogenesis of the neoplasm, with the anti-vimentin and anti-S100 markers demonstrating positive cytoplasmic immunoreactivity, revealing the mesenchymal origin of the neoplasm. Thus, the definitive diagnosis was splenic stromal sarcoma, a rare and aggressive neoplasm.

Pigeon as a model to study peripheral projections from the horizontal semicircular canal vestibular apparatus to a brainstem target immunoreactive for AMPA

ABSTRACT Purpose: To evaluate whether the pigeon (Columba livia) is a good model for evaluating the vestibular system involved with postural maintenance during movement. Methods: This study maps the brainstem targets of the horizontal ampullary inputs from the vestibular periphery of the pigeon. We used biotin dextran amine (BDA) injection in horizontal semicircular canal (HSCC), immunohistochemistry for GluR2/3 and GluR4 AMPA and computerized histomorphology reconstruction. Results: Our results show the same distribution pattern with ipsilateral projections to vestibular nuclear complex (VNC) from the HSCC, with the majority of labeled fibers being, long, thin, with few varicosities and many ramifications. Horizontal semicircular canal projections achieve neurons belonging to all nuclei of the VNC with exception of dorsal portion of lateral vestibular nucleus and this area express GluR2/3 and GluR4 AMPA receptors reinforcing the idea of glutamate participation in these connections. Conclusions: Pigeon is an appropriated experimental model to study of projections of HSCC and reinforcing the information that the vestibular system has strong relation with the fast responses necessary for postural control. Moreover, its phylogenetic organization apparently conservation, also seems to be a fundamental characteristic for vertebrates.

Spinal cord meningioma in a Boxer breed dog

Meningiomas are one of the most common primary tumors of the canine central nervous system. They usually grow within the cranial cavity but occasionally they are found in the spinal cord. A case of a Boxer breed with a spinal cord transitional meningioma is reported. Two years history presenting progressive neurological signs is described. The MRI examination revealed a focal, intradural extramedullary lesion within the right side of the spinal canal, compromising the C1and C2 vertebral segments. Cytoreductive surgery with a dorsolateral laminectomy approach was performed to decompressthe spinal cord and to obtain a definitive diagnosis. Histopathological studies diagnosed a grade I transitional meningioma. Immunohistochemistry and immunofluorescence analysis detected positive cells to vimentin (VIM), pan-cytokeratin (pCk),neuron-specific enolase (NSE), glial fibrillary acidic protein (GFAP) and doublecortin (DXC). Although apparently benign, positivity to pCK and DXC suggest a possible transition into a malignant tumor. Although variations in the behavior, histology and the immunoreactive profile of these tumors are reported, the latter constitutes a good indicator for the diagnosis of the patient.

Spinal cord meningioma in a Boxer breed dog

Meningiomas are one of the most common primary tumors of the canine central nervous system. They usually grow within the cranial cavity but occasionally they are found in the spinal cord. A case of a Boxer breed with a spinal cord transitional meningioma is reported. Two years history presenting progressive neurological signs is described. The MRI examination revealed a focal, intradural extramedullary lesion within the right side of the spinal canal, compromising the C1and C2 vertebral segments. Cytoreductive surgery with a dorsolateral laminectomy approach was performed to decompressthe spinal cord and to obtain a definitive diagnosis. Histopathological studies diagnosed a grade I transitional meningioma. Immunohistochemistry and immunofluorescence analysis detected positive cells to vimentin (VIM), pan-cytokeratin (pCk),neuron-specific enolase (NSE), glial fibrillary acidic protein (GFAP) and doublecortin (DXC). Although apparently benign, positivity to pCK and DXC suggest a possible transition into a malignant tumor. Although variations in the behavior, histology and the immunoreactive profile of these tumors are reported, the latter constitutes a good indicator for the diagnosis of the patient.(AU)

Nephroblastoma in a black-tufted marmoset (Callithrix penicillata) / Nefroblastoma em um sagui-de-tufos-pretos (Callithrix penicillata)

A renal nephroblastoma is described in a free-living black-tufted marmoset (Callithrix penicillata) in Central Brazil. The monkey was found dead and subjected to necropsy. Gross anatomic changes consisted of a ruptured left kidney, which was almost completely effaced by a white to yellow, partially encapsulated friable mass. The left ureter was distended due to obstruction by a red, spherical, 2mm in diameter friable mass. The urinary bladder was also distended. Histologically the renal and ureteral masses consisted of a triphasic embryonal neoplasm composed of embryonic epithelium forming glomeruli and tubules, polygonal blastemal cells, and a mesenchymal stroma. The embryonic epithelium exhibited rare nuclear immunoreactivity for WT-1, whereas blastemal cells exhibited robust cytoplasmic and rare nuclear immunoreactivity for WT-1; blastemal cells were also immunoreactive for vimentin. No immunoreactivity was detected for pan-cytokeratin (AE1/AE3), actin, and desmin. Morphological and immunohistochemical features of the present neoplasm are consistent with those described for renal nephroblastoma.(AU)

Descreve-se um caso de nefroblastoma maligno em um sagui de vida livre no Brasil Central. O macaco foi encontrado morto e encaminhado para necropsia. Na macroscopia, o rim esquerdo apresentava-se rompido e o parênquima estava substituído por um tecido neoplásico friável, parcialmente encapsulado e de superfície natural branca e de corte amarela. O ureter esquerdo apresentava-se distendido devido à obstrução por uma massa friável, vermelha, esférica, de 2mm de diâmetro. Histologicamente, as massas renal e ureteral consistiam de uma neoplasia embrionária composta por três populaçõies de células neoplásicas, composta por epitélio embrionário formando glomérulos e túbulos, células blastemais poligonais e um estroma mesenquimal. O epitélio embrionário exibiu imunorreactividade nuclear rara para WT-1, enquanto que as células blastemais exibiram imunorreactividade nuclear citoplasmática e rara para WT-1; As células blastemais também foram imunorreativas à vimentina. Nenhuma imunorreatividade foi detectada para pan-citoqueratina (AE1/AE3), actina e desmina. As características morfológicas e imuno-histoquímicas da presente neoplasia são consistentes com as descritas para o nefroblastoma renal.(AU)

Nephroblastoma in a black-tufted marmoset (Callithrix penicillata) / Nefroblastoma em um sagui-de-tufos-pretos (Callithrix penicillata)

A renal nephroblastoma is described in a free-living black-tufted marmoset (Callithrix penicillata) in Central Brazil. The monkey was found dead and subjected to necropsy. Gross anatomic changes consisted of a ruptured left kidney, which was almost completely effaced by a white to yellow, partially encapsulated friable mass. The left ureter was distended due to obstruction by a red, spherical, 2mm in diameter friable mass. The urinary bladder was also distended. Histologically the renal and ureteral masses consisted of a triphasic embryonal neoplasm composed of embryonic epithelium forming glomeruli and tubules, polygonal blastemal cells, and a mesenchymal stroma. The embryonic epithelium exhibited rare nuclear immunoreactivity for WT-1, whereas blastemal cells exhibited robust cytoplasmic and rare nuclear immunoreactivity for WT-1; blastemal cells were also immunoreactive for vimentin. No immunoreactivity was detected for pan-cytokeratin (AE1/AE3), actin, and desmin. Morphological and immunohistochemical features of the present neoplasm are consistent with those described for renal nephroblastoma.(AU)

Descreve-se um caso de nefroblastoma maligno em um sagui de vida livre no Brasil Central. O macaco foi encontrado morto e encaminhado para necropsia. Na macroscopia, o rim esquerdo apresentava-se rompido e o parênquima estava substituído por um tecido neoplásico friável, parcialmente encapsulado e de superfície natural branca e de corte amarela. O ureter esquerdo apresentava-se distendido devido à obstrução por uma massa friável, vermelha, esférica, de 2mm de diâmetro. Histologicamente, as massas renal e ureteral consistiam de uma neoplasia embrionária composta por três populaçõies de células neoplásicas, composta por epitélio embrionário formando glomérulos e túbulos, células blastemais poligonais e um estroma mesenquimal. O epitélio embrionário exibiu imunorreactividade nuclear rara para WT-1, enquanto que as células blastemais exibiram imunorreactividade nuclear citoplasmática e rara para WT-1; As células blastemais também foram imunorreativas à vimentina. Nenhuma imunorreatividade foi detectada para pan-citoqueratina (AE1/AE3), actina e desmina. As características morfológicas e imuno-histoquímicas da presente neoplasia são consistentes com as descritas para o nefroblastoma renal.(AU)

Effect of the ethanolic extract of pequi (Caryocar brasiliense) peel on acute cardiotoxicity induced by doxorubicin in Wistar rats (Rattus norvegicus albinus) / Efeito do extrato etanólico da casca do pequi (Caryocar brasiliense) na cardiotoxicidade aguda induzida por doxorrubicina em ratos Wistar (Rattus norvegicus albinus)

This study evaluated the activity of the ethanolic extract of pequi peel (EEPP) and immunostaining with matrix metalloproteinases 2 and 9 (MMP2 and MMP9), and tissue inhibitor of metalloproteinases 1 and 2 (TIMP1 and TIMP2) in the myocardium of rats subjected to acute cardiotoxicity using doxorubicin (DOX). Thirty Wistar rats (six groups of five animals) were used as follows: sham group (SG), water and saline; group G1, 16 mg/kg of DOX and 300 mg/kg of EEPP for 17 days; group G2, 16 mg/kg of DOX and 600 mg/kg of EEPP for 17 days; group G3, 16 mg/kg of DOX and 300 mg/kg of EEPP for 10 days; group G4, 16 mg/kg of DOX and 600 mg/kg of EEPP for 10 days; and control group (CG), 16 mg/kg of DOX. Three days after administering DOX (day 17), euthanasia was performed, and samples were collected for anatomopathological analysis. Myocyte vacuolar degeneration, cardiomyocyte disorganization and myofibrillar fragmentation, necrosis, mononuclear inflammatory infiltrate, Anitschkow cells, fibrosis, congestion, and edema were observed in the hearts of DOX recipients. In G1 and G2, EEPP attenuated myocyte vacuolar degeneration whereas in G4, EEPP attenuated cardiomyocyte disorganization. The percentage of cells immunoreactive for TIMP1 was higher in G1. It was concluded that EEPP minimizes the deleterious effects of DOX on rat myocardium. Doses of 300 and 600 mg/kg for 17 days attenuate the vacuolar degeneration of myocytes. The dose of 600 mg/kg for 10 days reduced cardiomyocyte disorganization, and the dose of 300 mg/kg for 17 days increased TIMP1 expression in the myocardium of DOX-treated rats.(AU)

Avaliou-se a ação do extrato etanólico da casca do pequi (EECP) e a imunomarcação de metaloproteinases de matriz 2 e 9 (MMP2 e MMP9), e inibidores teciduais das metaloproteinases de matriz 1 e 2 (TIMP1 e TIMP2), no miocárdio de ratos submetidos à cardiotoxicidade aguda pela doxorrubicina (DOX). Utilizaram-se 30 ratos Wistar, em seis grupos de cinco animais, sendo Grupo Sham (GS) água e salina; (G1) 16 mg/kg de DOX e 300 mg/kg de EECP por 17 dias; (G2) 16 mg/kg de DOX e 600 mg/kg de EECP por 17 dias; (G3) 16 mg/kg de DOX e 300 mg/kg de EECP por 10 dias; (G4) 16 mg/kg de DOX e 600 mg/kg de EECP por 10 dias; e grupo controle (GC) 16 mg/kg de DOX. Três dias após a aplicação da DOX, no dia 17, realizaram-se a eutanásia e colheita de amostras para análises antomopatológicas. No coração dos ratos que receberam DOX observaram-se degeneração vacuolar miocítica, desorganização dos cardiomiócitos e fragmentação das miofibrilas, necrose, infiltrado inflamatório mononuclear, células de Anitschkow, fibrose, congestão e edema. Nos grupos G1 e G2 o EECP atenuou a degeneração vacuolar miocítica e no G4 atenuou a desorganização dos cardiomiócitos. TIMP1 foi constatada em maior porcentagem de células marcadas no grupo de ratos que recebeu 300 mg/kg do EECP por 17 dias. Conclui-se que o EECP minimiza os efeitos deletérios da DOX no miocárdio de ratos. Nas doses de 300 e 600 mg/kg por 17 dias atenua a degeneração vacuolar miocítica, a 600 mg/kg por 10 dias reduz a desorganização dos cardiomiócitos e a 300 mg/kg por 17 dias aumenta a expressão de TIMP1 no miocárdio de ratos tratados com DOX.(AU)

The expression of endothelial and inducible nitric oxide synthase and apoptosis in intestinal ischemia and reperfusion injury under the action of ischemic preconditioning and pentoxifylline

Purpose:To investigate the expression of nitric oxide synthase (NOS) and apoptosis associated with ischemic preconditioning (IPC) and pentoxifylline (PTX) in intestinal ischemia (I) and reperfusion (R) injury.Methods:Thirty male rats were assigned to 5 groups: (CG), no clamping of the superior mesenteric artery (90 minutes); (IR-SS) saline + ischemia (30 minutes) + reperfusion (60 minutes); (IR-PTX) PTX + ischemia (30 minutes) + reperfusion (60 minutes); (IPC-IR-SS) 5 minutes of ischemia + 5 minutes of reperfusion (IPC) + saline + I(30 minutes)+R(60 minutes); and (IPC-IR-PTX) IPC + PTX + I(30 minutes)+ R(60 minutes).Results:The application of IPC and PTX showed a significantly lower immunohistochemistry reaction for active caspase-3 (P<0.05) compared to IR+SS. The number of cells immunoreactive to BCL-2 was higher in the IR-PTX group (P>0.05). The NOS-2 expression (qRTPCR) in the IR-PTX group (P<0.05) was higher than the values for the IPC+IR-SS and IPC-IR-PTX groups. The NOS-3 expression was significantly upper in the IPC-IR-PTX group than in the CG (P<0.05), the IR-SS (P<0.05) and the IR-PTX (P<0.05) groups.Conclusions:The BCL-2 and active caspase-3 showed beneficial effects on PTX and IPC. The expression of NOS-2 and NOS-3 in the IPC and IPC-PTX groups showed no synergistic effect.(AU)

FOLICULOGÊNESE EM PRIMATAS NEOTROPICAIS (Aotus nancymaae e Sapajus macrocephalus: QUANTIFICAÇÃO FOLICULAR E IMUNOLOCALIZAÇÃO DAS PROTEÍNAS CASPASE-3, BCL-2, PCNA E AMH

Este trabalho objetivou contribuir para o conhecimento da biologia reprodutiva de fêmeas de primatas não humanos (PNH). Foi realizada a morfofisiologia ovariana, identificando a localização in situ de componentes celulares envolvidos no processo de apoptose, antiapoptose, proliferação folicular e da expressão do hormônio anti-Mülleriano (AMH). Foram coletados ovários de seis fêmeas de Aotus nancymaae criadas em ambiente de cativeiro, e de seis fêmeas de Sapajus macrocephalus, de vida livre, nas três fases reprodutivas (luteal, folicular e gestacional), pertencentes à comunidade Nova Esperança localizado em Iquitos, na Amazônia peruana. As amostras foram conservadas em formol, e posteriormente em álcool 70%; submetidas ao processamento histológico, e cortes de 5µm de espessura foram corados em Hematoxilina-Eosina. Em seguida foram realizadas as etapas de aplicações da imunohistoquímica para Caspase-3, Bcl-2, PCNA e AMH. Foi utilizado o programa BioEstat 5.0, para análise de variância da quantificação histológica folicular (Kruskal-Wallis) e da quantificação das células imunorreativas para Caspase-3, Bcl-2, PCNA e AMH (KruskalWallis e Dunn). A associação entre as imunorreações apoptóticas, anti-apoptóticos, prolifarativas e hormonais foi determinada pela Análise das Coordenadas Principais (PCoA) do software R. Em A. nancymaae e S. macrocephalus, houve diferença significativa na quantificação entre os folículos primordiais e antrais pequenos, primordiais e antrais grandes, e primários e antrais grandes (Kruskal-Wallis, p<0,05), nas três fases reprodutivas. Os marcadores proteicos e hormonais variaram significativamente entre os tipos celulares para A. nancymaae (p=0.001) e S. macrocephalus (p=0.002). Em ambas as espécies foi observada imunorreação para Caspase-3-ativada no estroma e células foliculares de folículos primários, antrais pequenos, antrais grandes (células da granulosa, oócitos, células tecais), além do corpo lúteo, nas três fases reprodutivas. Foi observada ainda imunorreação para Bcl-2 em células da granulosa e oócitos de folículos pré-antrais e antrais; e imunorreação para PCNA em núcleos e citoplasmas de oócitos de folículos primordiais e primários, nas gônadas de fêmeas no estágio luteal, em folículos primordiais em fêmeas gestantes e em folículos primordiais normais e atrésicos de fêmeas no estágio folicular; e ainda, imunorreação para o AMH nas células da granulosa e oócitos de folículos antrais em maturação. Assim, as técnicas de imunomarcação para as proteínas Caspase-3, Bcl-2, PCNA e AMH, foram imprescindíveis para determinar a análise da dinâmica ovariana, no sentido de propiciarem dados que sugerem avaliar se folículos estão em estado de atresia ou desenvolvimento e a produção hormonal atuando no processo da foliculogênese dessas espécies.

This work aims to contribute to the knowledge of the reproductive biology of female nonhuman primates (PNH). Therefore, research was carried out on ovarian morphophysiology, identifying the in situ location of cellular components involved in the process of apoptosis, anti-apoptosis and follicular proliferation, in addition to the expression of anti-Müllerian hormone (AMH). For this purpose, ovaries were collected from six females of Aotus nancymaae raised in captivity, and from six females of Sapajus macrocephalus, free-living, in the three reproductive stages (luteal, follicular and pregnant). The animals belonged to the Nova Esperança community located in Iquitos, in the Peruvian Amazon. The samples were preserved in formalin, and later in 70% alcohol; submitted to histological processing, and 5µm-thick sections were stained with Hematoxylin-Eosin. Then, the steps of application of immunohistochemistry for Caspase-3, Bcl-2, PCNA and AMH were carried out. The BioEstat 5.0 program was used for analysis of variance of the follicular histological quantification (Kruskal-Wallis) and the quantification of immunoreactive cells for Caspase-3, Bcl-3, PCNA and AMH (Kruskal-Wallis and Dunn). The association between apoptotic, anti-apoptotic, proliferative and hormonal immunoreactions was determined by the Principal Coordinate Analysis (PCoA) of the R software. In A. nancymaae and S. macrocephalus there was a significant difference in the quantification between primordial and small antral, primordial and large antral, and primary and large antral (Kruskal-Wallis, p<0.05), in the three reproductive stages. Protein and hormonal markers varied significantly between cell types of A. nancymaae (p=0.001) and S. macrocephalus (p=0.002). Immunoreaction to Caspase-3- activated was observed in the stroma and follicular cells of primary follicles, small antral, large antral (granulosa cells, oocytes, thecal cells), in addition to the corpus luteum, in the three reproductive phases. Immunoreaction to Bcl-2 was also observed in granulosa cells and oocytes from preantral and antral follicles; and PCNA immunoreaction in oocyte nuclei and cytoplasm of primordial and primary follicles, in the gonads of females in the luteal stage, in primordial follicles in pregnant females, and in normal and atresic primordial follicles of females in the follicular stage; and also, immunoreaction for AMH in granulosa cells and oocytes from maturing antral follicles. Thus, immunostaining techniques for Caspase-3, Bcl2, PCNA and AMH proteins were essential to determine the analysis of ovarian dynamics, in order to provide data that suggest evaluating whether follicles are in a state of atresia or development and the hormone production acting in the folliculogenesis process of these species.

Oral fibrosarcoma in jararaca (Bothrops pubescens): anatomopathological and immunohistochemical aspects / Fibrossarcoma oral em jararaca (Bothrops pubescens): aspectos anatomopatológicos e imuno-histoquímicos

A 4-year-old female captive-bred snake of the genus Bothrops showed swelling on the left side of the oral cavity, suggesting the development of neoplasia. The mass was removed surgically and sent for pathological examination. Two months later a new increase in volume in the same site was observed, suggesting recurrence. The lesion was completely removed and sent for pathological analysis. Histologically, the two-samples consisted of a mass with highly-cell density composed of spindle-shaped anaplastic cells arranged in interwoven bundles, distributed throughout the tissue extension and, occasionally, polygonal cells arranged in irregular fascicles. The Masson trichrome staining showed modest amount of collagen supporting the neoplastic cells. PAS-positive content was not observed in the cytoplasm of neoplastic cells. Histological and histochemical findings indicated that it was a spindle cell neoplasm, but the classification was not possible. Immunohistochemistry was requested and performed using the streptavidin-biotin-peroxidase method. The markers used were anti-vimentin, anti-PCNA, anti-EMA, anti-melan A and anti-melanosome, anti-desmin, anti-actin, anti-CD68 and anti- S100protein. The neoplastic cells were immunoreactive for vimentin and PCNA and negative for the other antibodies. The morphology characterization, histochemical and immunohistochemical analysis of neoplastic cells allowed the definitive diagnosis of oral fibrosarcoma.(AU)

Uma serpente de cativeiro, fêmea, quatro anos de idade, do gênero Bothrops apresentou aumento de volume no lado esquerdo da cavidade oral, sugerindo tratar-se de neoplasma. A massa foi removida cirurgicamente e enviada para exame anatomopatológico. Dois meses depois foi observado novo aumento de volume no mesmo local, sugerindo recidiva. A lesão foi removida por completo e também enviada para análise. Histologicamente, as duas amostras consistiam de massa altamente celular, composta por células anaplásicas fusiformes organizadas em feixes entrelaçados e distribuídos por toda extensão tecidual e, ocasionalmente, células poligonais arranjadas em fascículos irregulares. A coloração de tricrômico de Masson apresentou quantidade modesta de colágeno sustentando as células neoplásicas. Não foi observado conteúdo PAS-positivo no citoplasma das células neoplásicas. Os achados histológicos e histoquímicos indicavam tratar-se de neoplasma de células fusiformes, porém não era possível sua classificação. A imuno-histoquímica foi requisitada e realizada pelo método streptavidina-biotina-peroxidase, utilizando os anticorpos anti-vimentina, anti-PCNA, Anti-EMA, anti-melan A, anti-HMB45, anti-desmina, anti-actina, anti-CD68 e anti-proteína S-100. As células neoplásicas foram imunorreativas para vimentina e PCNA e, negativas para os demais anticorpos. A caracterização morfológica, histoquímica e imuno-histoquímica das células neoplásicas permitiu o diagnóstico definitivo de fibrossarcoma oral.(AU)

Evaluation of immunohistochemical expression of ghrelin-producing rectal cells in Wistar rats underwent to the cafeteria diet

PURPOSE:To investigate the impact of cafeteria diet on ghrelin expression in rectal tissue and identify the morphologic cell type. METHODS:Twenty-four male Wistar rats were divided into four subgroups of six animals each: RC1 (rat chow 1) and CAF1 (cafeteria diet 1) for a period of 30 days; RC2 (rat chow 2) and CAF2 (cafeteria diet 2) for a period of 60 days. The animal and rectal weight, the number and the type of immunoreactive ghrelin cells were recorded and compared between the subgroups. The statistical study was established by ANOVA and Student's t test. RESULTS:There was no difference in the total of immunoreactive cells (p=0.685) between the subgroups nor between weight and presence or absence of ghrelin expression (p=0.993). All the immunoreactive cells identified were closed-type. CONCLUSION:The cafeteria diet did not have influence on the amount of immunoreactive rectal cells of ghrelin and only one type (closed-type) of immunoreactive cells was expressed in the rectum.(AU)

The influence of selected pathological states on the somatostatin-like immunoreactive (SOM-LI) endocrine cells in the mucosal layer of the porcine descending colon

This study reports on changes in the number of somatostatin-like immunoreactive (SOM-LI) endocrine cells in the porcine descending colon, caused by chemically driven inflammation, axotomy and proliferative enteropathy (PE). The distribution pattern of SOM-LI endocrine cells has been studied using the routine single-labelling immunofluorescence technique. Semi-quantitative evaluation of the number of the SOM-immunostained endocrine cells within the mucosal layer of the porcine descending colon has been based on counting of all endocrine cells immunoreactive to SOM per unit area (0,1 mm²). Under physiological conditions the number of SOM-LI endocrine cells has been shown to constitute 3,30±0,22. All applied pathological processes resulted in changes in the SOM-like immunoreactivity, which varied in particular processes studied. The number of SOM-LI endocrine cells increased to 6,28±0,31 and 4,43±0,35 during chemically driven inflammation and proliferative enteropathy, respectively, and decreased to 1,17%±0,16 after axotomy. The obtained results suggest that SOM-LI endocrine cells may participate in various pathological states within porcine descending colon and their functions probably depend on the type of pathological factor.(AU)

The influence of selected pathological states on the somatostatin-like immunoreactive (SOM-LI) endocrine cells in the mucosal layer of the porcine descending colon

This study reports on changes in the number of somatostatin-like immunoreactive (SOM-LI) endocrine cells in the porcine descending colon, caused by chemically driven inflammation, axotomy and proliferative enteropathy (PE). The distribution pattern of SOM-LI endocrine cells has been studied using the routine single-labelling immunofluorescence technique. Semi-quantitative evaluation of the number of the SOM-immunostained endocrine cells within the mucosal layer of the porcine descending colon has been based on counting of all endocrine cells immunoreactive to SOM per unit area (0,1 mm²). Under physiological conditions the number of SOM-LI endocrine cells has been shown to constitute 3,30±0,22. All applied pathological processes resulted in changes in the SOM-like immunoreactivity, which varied in particular processes studied. The number of SOM-LI endocrine cells increased to 6,28±0,31 and 4,43±0,35 during chemically driven inflammation and proliferative enteropathy, respectively, and decreased to 1,17%±0,16 after axotomy. The obtained results suggest that SOM-LI endocrine cells may participate in various pathological states within porcine descending colon and their functions probably depend on the type of pathological factor.(AU)

DESENVOLVIMENTO DOS PRÉ-ESTÔMAGOS DE BEZERROS MESTIÇOS ALEITADOS COM LEITE EM PÓ ASSOCIADO AO SORO DE QUEIJO EM PÓ

Objetivou-se avaliar o desenvolvimento dos pré-estômagos, o crescimento e a taxa de proliferação celular das papilas ruminais, em bezerros aleitados com sucedâneos (leite em pó e soro de queijo em pó) durante os 60 primeiros dias de vida. Verificar se a dieta pode influenciar as células produtoras de quimosina e pepsinogênio. Para isso, 24 bezerros mestiços com uma semana de vida foram distribuídos em um delineamento inteiramente casualizado com 4 (quatro) tratamentos e 6 (seis) repetições: LI: leite integral (Controle); LP: leite em pó; LPS1: 80% leite em pó + 20% de soro de queijo em pó; LPS2: 60% leite em pó + 40% de soro de queijo em pó. Para os cálculos de consumo de matéria seca (MS) foram feitas medições diárias de consumo de feno e concentrado pelo método de oferta/sobra. Os animais foram abatidos aos 60 dias de vida. Os órgãos cavitários foram pesados com conteúdo, e logo em seguida, esvaziados e novamente pesados, registrando-se, os pesos cheios e vazios, respectivamente. Foi determinado o peso dos estômagos e de seus compartimentos com valores absolutos (kg) e relativos (%) em função do peso de corpo vazio (% PCVZ) e em função do peso do estômago completo (% PE). Foram retirados fragmentos de aproximadamente um centímetro quadrado (1cm2) de tecidos de 4 regiões do rúmen: átrio ruminal, saco dorsal, saco cego caudo-dorsal e saco ventral de onde foram medidas as alturas das papilas ruminais. A atividade proliferativa foi avaliada em amostras de tecidos retiradas do rúmen por meio da contagem de Regiões Organizadoras de Nucléolos Argirofílicos (AgNOR). Foram coletadas amostras teciduais de 1cm2 do abomaso para a realização da imunohistoquímica. Não houve diferença significativa para o consumo de matéria seca (P>0,05). As médias obtidas para pesos absolutos e relativos em função do peso do corpo vazio dos compartimentos gástricos não apresentaram diferenças significativas (P>0,05) entre os tratamentos. Quanto ao desenvolvimento de papilas no rúmen houve diferenças significativas (P<0,05) para a variável altura no saco ventral, onde os animais que receberam leite integral possuíram maiores médias. Não houve diferenças significativas (P>0,05) para a taxa de proliferação celular nas papilas ruminais. Os percentuais de ácinos contendo células imunorreativas aos anticorpos quimosina e pepsinogênio apresentaram diferença estatística (P<0,05), onde os bezerros que receberam a dieta 100% leite integral, apresentaram média percentual de marcação para quimosina superior (95,11%), aqueles animais que consumiram a dieta 60% leite em pó + 40% de soro de queijo em pó (89,59%).

The aim of this study was to evaluate the use of whey powder associated with powdered milk as a substitute for whole milk without breastfeeding until 60 days of age on the development of pre-stomachs, growth and proliferation rate Cellular of the ruminal papillae, as well as an influence of the diets on as chymosin and pepsinogen producing cells of the calf abomasum through the immunohistochemical technique. To that end, 24 crossbred calves with one week of life were distributed in a completely randomized design with four treatments and six replicates: LI: whole milk (Control); LP: milk powder; LPS1: 80% milk powder + 20% whey powder; LPS2: 60% milk powder + 40% whey powder cheese. For the methods of dry matter (DM) consumption, measurements of hay consumption and concentrate are made by the supply / surplus method. The animals were slaughtered at 60 days of age. The cavity organs were weighed with contents, and soon thereafter emptied and newer, registering, full and empty weights, respectively. The weight of the stomach and its compartments were determined with absolute values (kg) and relative (%) as a function of the empty body weight (%PCVZ) and as a function of the weight of the whole stomach (%PE). Fragments of approximately one square centimeter (1cm2) of tissues were collected from 4 regions of the world: rumen atrium, dorsal sac, caudate-dorsal blind sac and ventral sac from where they are measured as heights of ruminal papillae. Proliferative activity was evaluated in tissue samples collected by counting Argirophilic Nucléole Organisms (AgNOR). Tissue samples of 1 cm2 of the abomasum were collected for immunohistochemistry. There was no difference for dry matter intake (P> 0.05). As averages obtained for absolute and relative weights as a function of the empty body weight of the digital compartments did not present significant residues (P> 0.05) between the treatments. Regarding the development of non-rumen papillae There were significant (P <0.05) for a height variable in the ventral sac, where animals receiving whole milk had higher averages. There were no significant differences (P> 0.05) for the rate of cell proliferation in ruminal papillae. The percentage of acini with immunoreactive cells to the chymosin and pepsinogen antibodies presented a statistical difference (P <0.05), where the calves that received a 100% whole milk diet presented a mean percentage of marking for upper chymosin (95.11%), Animals that consumed 60% milk powder + 40% whey powder (89.59%).