Inhibitory Effects of a Reengineered Anthrax Toxin on Canine Oral Mucosal Melanomas.

Canine oral mucosal melanomas (OMM) are the most common oral malignancy in dogs and few treatments are available. Thus, new treatment modalities are needed for this disease. Bacillus anthracis (anthrax) toxin has been reengineered to target tumor cells that express urokinase plasminogen activator (uPA) and metalloproteinases (MMP-2), and has shown antineoplastic effects both, in vitro and in vivo. This study aimed to evaluate the effects of a reengineered anthrax toxin on canine OMM. Five dogs bearing OMM without lung metastasis were included in the clinical study. Tumor tissue was analyzed by immunohistochemistry for expression of uPA, uPA receptor, MMP-2, MT1-MMP and TIMP-2. Animals received either three or six intratumoral injections of the reengineered anthrax toxin prior to surgical tumor excision. OMM samples from the five dogs were positive for all antibodies. After intratumoral treatment, all dogs showed stable disease according to the canine Response Evaluation Criteria in Solid Tumors (cRECIST), and tumors had decreased bleeding. Histopathology has shown necrosis of tumor cells and blood vessel walls after treatment. No significant systemic side effects were noted. In conclusion, the reengineered anthrax toxin exerted inhibitory effects when administered intratumorally, and systemic administration of this toxin is a promising therapy for canine OMM.

Slc11a1 (Nramp-1) gene modulates immune-inflammation genes in macrophages during pristane-induced arthritis in mice

OBJECTIVE AND DESIGN: Pristane-induced arthritis (PIA) in AIRmax mice homozygous for Slc11a1 R and S alleles was used to characterize the influence of Slc11a1 gene polymorphism on immune responses during disease manifestation. Previous reports demonstrated that the presence of the Slc11a1 S allele increased the incidence and severity of PIA in AIRmax SS , suggesting that this gene could interact with inflammatory loci to modulate PIA. We investigated the effects of Slc11a1 alleles on the activation of phagocytes during PIA. TREATMENT: Mice were injected intraperitoneally with two doses of 0.5 mL of mineral oil pristane at 60-day intervals. Arthritis development was accompanied for 180 days. RESULTS: AIRmax SS mice showed differential peritoneal macrophage gene expression profiles during PIA, with higher expression and production of H2O2, NO, IL-1ß, IL-6, TNF-α, and several chemokines. The presence of the Slc11a1 R allele, on the other hand, diminished the intensity of macrophage activation, restricting arthritis development. CONCLUSION: Our data demonstrated the fine-tuning roles of Slc11a1 alleles modulating macrophage activation, and consequent PIA susceptibility, in those mouse lines. (AU)

Slc11a1 (Nramp-1) gene modulates immune-inflammation genes in macrophages during pristane-induced arthritis in mice.

OBJECTIVE AND DESIGN: Pristane-induced arthritis (PIA) in AIRmax mice homozygous for Slc11a1 R and S alleles was used to characterize the influence of Slc11a1 gene polymorphism on immune responses during disease manifestation. Previous reports demonstrated that the presence of the Slc11a1 S allele increased the incidence and severity of PIA in AIRmax SS , suggesting that this gene could interact with inflammatory loci to modulate PIA. We investigated the effects of Slc11a1 alleles on the activation of phagocytes during PIA. TREATMENT: Mice were injected intraperitoneally with two doses of 0.5 mL of mineral oil pristane at 60-day intervals. Arthritis development was accompanied for 180 days. RESULTS: AIRmax SS mice showed differential peritoneal macrophage gene expression profiles during PIA, with higher expression and production of H2O2, NO, IL-1ß, IL-6, TNF-α, and several chemokines. The presence of the Slc11a1 R allele, on the other hand, diminished the intensity of macrophage activation, restricting arthritis development. CONCLUSION: Our data demonstrated the fine-tuning roles of Slc11a1 alleles modulating macrophage activation, and consequent PIA susceptibility, in those mouse lines.

Adenoma hipofisário em pônei: relato de caso / Pituitary Adenoma on pony: report case / Adenoma Hipofisario en poni: reporte de un caso

A disfunção adeno hipofisária (DAH) é a mais comum endocrinopatia dos equídeos idosos. A presença de adenomas em região correspondente a hipófise pode ser associada a esta disfunção. Apesar de várias descrições desta afecção como doença clínica bem definida, suas características fisiopatológicas impõem a ela inúmeras possíveis interferências ao diagnóstico presuntivo. Relata-se neste artigo um caso de DAH por adenoma hipofisário em pônei idoso que apresentava diversos sinais associados a infecções secundárias, comumente predispostas pela endocrinopatia, definindo condição clínica diferenciada. Em função de sua senilidade e de seu estado geral debilitado não houve tempo hábil para diagnóstico in vivo sendo sua condição esclarecida somente em exame post-mortem.

The Adenohypophyseal dysfunction (AHD) is the most common endocrine disease of older horses.The presence of adenomas in pituitary corresponding region can be commonly associated with this disorder. Despite several descriptions of this condition as well-defined clinical disease, its pathophysiological characteristics impose numerous possible interference to the presumptive diagnosis. It's reported in this paper a case of AHD for pituitary adenomas in older pony that showed several signs associated with secondary infections, commonly . predisposed by endocrinopathy, defining distinct clinical condition. Because of their senility and its weakenedgeneral condition there wasn't enough time for in vivo diagnosis and their condition was only clarified in post mortem exam.

La disfunción adenohipofisarias (DAH) es Ia enfermedad endocrina más común de los caballos másviejos. La presencia de adenomas en Ia región correspondiente de hipófisis pueden ser comúnmente asociadoscon este trastorno. A pesar de varias descripciones de esta condición como uma enfermedad clínica bien definida,su características fisiopatológicas imponen numerosas posibles interferencias en diagnóstico presuntivo. Se informaen este trabajo un caso de DAH debido a adenomas hipofisarios en poni ancianos que mostraban varios signos asociados a infecciones secundarias, comúnmente predispuestos por endo crinopatía, que definen una condición clínica distinta. Debido a su senilidad y su estado general de debilidad no hubo tiempo suficiente para el diagnóstico in vivo y su condición aclaró sólo en examen post- mortem.

Adenoma hipofisário em pônei: relato de caso / Pituitary Adenoma on pony: report case / Adenoma Hipofisario en poni: reporte de un caso

A disfunção adeno hipofisária (DAH) é a mais comum endocrinopatia dos equídeos idosos. A presença de adenomas em região correspondente a hipófise pode ser associada a esta disfunção. Apesar de várias descrições desta afecção como doença clínica bem definida, suas características fisiopatológicas impõem a ela inúmeras possíveis interferências ao diagnóstico presuntivo. Relata-se neste artigo um caso de DAH por adenoma hipofisário em pônei idoso que apresentava diversos sinais associados a infecções secundárias, comumente predispostas pela endocrinopatia, definindo condição clínica diferenciada. Em função de sua senilidade e de seu estado geral debilitado não houve tempo hábil para diagnóstico in vivo sendo sua condição esclarecida somente em exame post-mortem.(AU)

The Adenohypophyseal dysfunction (AHD) is the most common endocrine disease of older horses.The presence of adenomas in pituitary corresponding region can be commonly associated with this disorder. Despite several descriptions of this condition as well-defined clinical disease, its pathophysiological characteristics impose numerous possible interference to the presumptive diagnosis. It's reported in this paper a case of AHD for pituitary adenomas in older pony that showed several signs associated with secondary infections, commonly . predisposed by endocrinopathy, defining distinct clinical condition. Because of their senility and its weakenedgeneral condition there wasn't enough time for in vivo diagnosis and their condition was only clarified in post mortem exam.(AU)

La disfunción adenohipofisarias (DAH) es Ia enfermedad endocrina más común de los caballos másviejos. La presencia de adenomas en Ia región correspondiente de hipófisis pueden ser comúnmente asociadoscon este trastorno. A pesar de varias descripciones de esta condición como uma enfermedad clínica bien definida,su características fisiopatológicas imponen numerosas posibles interferencias en diagnóstico presuntivo. Se informaen este trabajo un caso de DAH debido a adenomas hipofisarios en poni ancianos que mostraban varios signos asociados a infecciones secundarias, comúnmente predispuestos por endo crinopatía, que definen una condición clínica distinta. Debido a su senilidad y su estado general de debilidad no hubo tiempo suficiente para el diagnóstico in vivo y su condición aclaró sólo en examen post- mortem. (AU)

Ceratoplastia lamelar em cães usando membrana amniótica equina. Estudo clínico e morfológico / Lamelar keratoplasty in dogs using equine amniotic membrane. Clinical and morphological study

A membrana amniótica (MA) consolidou-se no tratamento de afecções na superfície ocular. O objetivo deste estudo foi avaliar a viabilidade do implante de MA equina em ceratoplastia lamelar de cães. As membranas amnióticas foram preservadas em glicerina (98%). A eficácia do implante foi acompanhada por avaliação clínica, tempo de cicatrização, resposta inflamatória e reconstrução da arquitetura da córnea. Foram selecionados 12 cães, que foram divididos em quatro grupos de três animais. Em cada animal, foi realizada ceratotomia lamelar com 5 mm de diâmetro, seguida de aplicação do implante de MA. Após cirurgia, os animais foram avaliados em diferentes tempos: 2, 7, 21 e 40 dias. Duran- te o período de observação, os exames oftalmológicos foram realizados com intervalo de 48 h e, após a última avaliação, os animais foram submetidos à eutanásia. Os olhos foram enucleados, fixados e corados com hematoxilina-eosina (HE), ácido periódico de Schiff (PAS) e picrossirius. Os implantes foram completamente epitelizados em cerca de 10 dias após a cirurgia. Os neovasos apresentaram involução progressiva a partir de 21 dias e não foram detectados ao final de 40 dias pós-cirurgia, restando apenas uma nébula no local da lesão. À microscopia óptica, observou-se resposta inflama- tória moderada, presença de epitélio pavimentoso estratificado aos sete dias e epitelização completa aos 21 dias. Aos 40 dias, a membrana basal do epitélio apresentou-se reconstituída. Assim, concluímos que a membrana amniótica equina é viável como implante em córnea de cão, sendo incorporada ao estroma e resultando em restabelecimento parcial da transparência.(AU)

Amniotic membranes have been successfully used in the treatment of superficial corneal diseases. The purpose of this study was to assess the feasibility of use of equine amniotic membrane (AM) in lamellar keratoplasty of dogs. Amniotic membranes were preserved in glycerol (98%). Implant efficacy was assessed by clinical evaluation, time of healing, in-flammatory response, and the corneal architectural configuration. Twelve dogs were divided into four groups of three animals. In each animal, a 5-mm diameter lamellar keratectomy was performed, and followed by implantation of the AM. The animals were evaluated at the 2th, 7th, 21th, and 40th days after surgery. During this period, ophthalmological exams were performed at 48-h intervals, and the animals were euthanized after the last evaluation. The eyes were enu-cleated, included, fixed, and stained with hematoxylin-eosin (HE), periodic acid-Schiff (PAS), and picrosirius. Epitheli-zation of the implants was completed in about ten days after surgery. New vessels progressively diminished from 21 days on, were not detected on day 40 after surgery, and only a nebula was observed in the implant area. Optical microscopy revealed a mild inflammatory response, presence of squamous epithelium on day seven, and total epithelization on day 21 after surgery. On day 40, the basal epithelial membrane was shown to be reformed. Therefore, we conclude that use of equine amniotic membranes is feasible as implant for dog cornea since it is incorporated to the corneal stroma, and a relative transparency can be obtained.(AU)

Ceratoplastia lamelar em cães usando membrana amniótica equina: Estudo clínico e morfológico / Lamelar keratoplasty in dogs using equine amniotic membrane: Clinical and morphological study

A membrana amniótica (MA) consolidou-se no tratamento de afecções na superfície ocular. O objetivo deste estudo foi avaliar a viabilidade do implante de MA equina em ceratoplastia lamelar de cães. As membranas amnióticas foram preservadas em glicerina (98%). A eficácia do implante foi acompanhada por avaliação clínica, tempo de cicatrização, resposta inflamatória e reconstrução da arquitetura da córnea. Foram selecionados 12 cães, que foram divididos em quatro grupos de três animais. Em cada animal, foi realizada ceratotomia lamelar com 5 mm de diâmetro, seguida de aplicação do implante de MA. Após cirurgia, os animais foram avaliados em diferentes tempos: 2, 7, 21 e 40 dias. Duran- te o período de observação, os exames oftalmológicos foram realizados com intervalo de 48 h e, após a última avaliação, os animais foram submetidos à eutanásia. Os olhos foram enucleados, fixados e corados com hematoxilina-eosina (HE), ácido periódico de Schiff (PAS) e picrossirius. Os implantes foram completamente epitelizados em cerca de 10 dias após a cirurgia. Os neovasos apresentaram involução progressiva a partir de 21 dias e não foram detectados ao final de 40 dias pós-cirurgia, restando apenas uma nébula no local da lesão. À microscopia óptica, observou-se resposta inflama- tória moderada, presença de epitélio pavimentoso estratificado aos sete dias e epitelização completa aos 21 dias. Aos 40 dias, a membrana basal do epitélio apresentou-se reconstituída. Assim, concluímos que a membrana amniótica equina é viável como implante em córnea de cão, sendo incorporada ao estroma e resultando em restabelecimento parcial da transparência.

Amniotic membranes have been successfully used in the treatment of superficial corneal diseases. The purpose of this study was to assess the feasibility of use of equine amniotic membrane (AM) in lamellar keratoplasty of dogs. Amniotic membranes were preserved in glycerol (98%). Implant efficacy was assessed by clinical evaluation, time of healing, in-flammatory response, and the corneal architectural configuration. Twelve dogs were divided into four groups of three animals. In each animal, a 5-mm diameter lamellar keratectomy was performed, and followed by implantation of the AM. The animals were evaluated at the 2th, 7th, 21th, and 40th days after surgery. During this period, ophthalmological exams were performed at 48-h intervals, and the animals were euthanized after the last evaluation. The eyes were enu-cleated, included, fixed, and stained with hematoxylin-eosin (HE), periodic acid-Schiff (PAS), and picrosirius. Epitheli-zation of the implants was completed in about ten days after surgery. New vessels progressively diminished from 21 days on, were not detected on day 40 after surgery, and only a nebula was observed in the implant area. Optical microscopy revealed a mild inflammatory response, presence of squamous epithelium on day seven, and total epithelization on day 21 after surgery. On day 40, the basal epithelial membrane was shown to be reformed. Therefore, we conclude that use of equine amniotic membranes is feasible as implant for dog cornea since it is incorporated to the corneal stroma, and a relative transparency can be obtained.

Estudo casuístico retrospectivo de neoformações primárias esplênicas / Retrospective case study of primary splenic neoformations

O baço é alvo de inúmeras afecções não neoplásicas e neoplásicas. O hemangiossarcoma correspondea um das proliferações neoplásicas esplênicas mais frequentes. Esse possui origem vascular endoteliale alto potencial metastático. Todas as afecções do baço provocam uma esplenomegalia perceptível aoexame físico (por palpação) ou por ultrassonografia abdominal. O objetivo do presente trabalho foi verificara incidência, na espécie canina, do hemangiossarcoma dentre todas as lesões esplênicas da rotinaclínico-cirúrgica de um hospital veterinário universitário. Foram colhidos, durante os anos de 2005 a2011, dados de 45 caninos portadores de neoformações esplênicas, a partir de prontuários oriundosdo Hospital Veterinário Anhembi Morumbi (São Paulo, SP). Considerou-se o tipo histológico de cadaproliferação. Para os animais acometidos por hemangiossarcoma, também se obtiveram informaçõesrelacionadas à raça, idade e sexo. O estudo observou que as lesões não neoplásicas compreenderam64,4% dos casos, com maior frequência para as hiperplasias nodulares (31,1%) e alterações circulatórias(24,4%), como congestão, hematoma e hemorragia. As neoplasias esplênicas demonstraram uma frequênciade 35,6%, sendo 69,23% dessas equivalentes a hemangiossarcomas. O perfil epidemiológico usualdo paciente acometido por tal afecção correspondeu a animais de raça de grande porte, idade de noveanos, porém sem predisposição sexual. Com o presente trabalho pode-se concluir que os hemangiossarcomassão as afecções neoplásicas esplênicas mais comuns na espécie canina, precedida, somente,pelas hiperplasias nodulares demonstrando a importância no diagnóstico diferencial em relação a tratamentoe prognóstico.(AU)

The spleen is the subject of numerous non-neoplastic and neoplastic diseases. The hemangiosarcomacorresponds to a proliferation of neoplastic splenic more frequent. This has vascular endothelial and highmetastatic potential. All the conditions cause a spleen noticeable splenomegaly on physical examination(by palpation) or abdominal ultrasonography. The objective of this study was determine the incidence,in dogs, hemangiosarcoma among the all splenic injuries from routine clinical and surgical hospital veterinarycollege. Were collected during the years 2005 to 2011, data from 45 patients with canine splenicneoformation, the from records from the Veterinary Hospital Anhembi Morumbi (Sao Paulo, SP). Weconsidered the histological type of each spread. For those animals afflicted with hemangiosarcoma, alsoobtained information related to race, age and sex. The study noted that non-neoplastic lesions comprised64,4% of cases, more often for nodular hyperplasia (31,1%) and circulatory disorders (24,4%), as congestion, bruising and bleeding. Splenic neoplasms showed a frequency of 35,6% and 69,23% of theequivalent hemangiosarcomas. The usual epidemiological profile of the affected patient on such pathologycorresponded to animals of large breed, age nine years, but no sexual predisposition. The presentwork it can be concluded that are neoplastic hemangiosarcomas splenic more common in dogs, precededonly by the nodular hyperplasia demonstrating the importance in the diagnosis differential in relation totreatment and prognosis.(AU)

Estudo casuístico retrospectivo de neoformações primárias esplênicas / Retrospective case study of primary splenic neoformations

O baço é alvo de inúmeras afecções não neoplásicas e neoplásicas. O hemangiossarcoma correspondea um das proliferações neoplásicas esplênicas mais frequentes. Esse possui origem vascular endoteliale alto potencial metastático. Todas as afecções do baço provocam uma esplenomegalia perceptível aoexame físico (por palpação) ou por ultrassonografia abdominal. O objetivo do presente trabalho foi verificara incidência, na espécie canina, do hemangiossarcoma dentre todas as lesões esplênicas da rotinaclínico-cirúrgica de um hospital veterinário universitário. Foram colhidos, durante os anos de 2005 a2011, dados de 45 caninos portadores de neoformações esplênicas, a partir de prontuários oriundosdo Hospital Veterinário Anhembi Morumbi (São Paulo, SP). Considerou-se o tipo histológico de cadaproliferação. Para os animais acometidos por hemangiossarcoma, também se obtiveram informaçõesrelacionadas à raça, idade e sexo. O estudo observou que as lesões não neoplásicas compreenderam64,4% dos casos, com maior frequência para as hiperplasias nodulares (31,1%) e alterações circulatórias(24,4%), como congestão, hematoma e hemorragia. As neoplasias esplênicas demonstraram uma frequênciade 35,6%, sendo 69,23% dessas equivalentes a hemangiossarcomas. O perfil epidemiológico usualdo paciente acometido por tal afecção correspondeu a animais de raça de grande porte, idade de noveanos, porém sem predisposição sexual. Com o presente trabalho pode-se concluir que os hemangiossarcomassão as afecções neoplásicas esplênicas mais comuns na espécie canina, precedida, somente,pelas hiperplasias nodulares demonstrando a importância no diagnóstico diferencial em relação a tratamentoe prognóstico.

The spleen is the subject of numerous non-neoplastic and neoplastic diseases. The hemangiosarcomacorresponds to a proliferation of neoplastic splenic more frequent. This has vascular endothelial and highmetastatic potential. All the conditions cause a spleen noticeable splenomegaly on physical examination(by palpation) or abdominal ultrasonography. The objective of this study was determine the incidence,in dogs, hemangiosarcoma among the all splenic injuries from routine clinical and surgical hospital veterinarycollege. Were collected during the years 2005 to 2011, data from 45 patients with canine splenicneoformation, the from records from the Veterinary Hospital Anhembi Morumbi (Sao Paulo, SP). Weconsidered the histological type of each spread. For those animals afflicted with hemangiosarcoma, alsoobtained information related to race, age and sex. The study noted that non-neoplastic lesions comprised64,4% of cases, more often for nodular hyperplasia (31,1%) and circulatory disorders (24,4%), as congestion, bruising and bleeding. Splenic neoplasms showed a frequency of 35,6% and 69,23% of theequivalent hemangiosarcomas. The usual epidemiological profile of the affected patient on such pathologycorresponded to animals of large breed, age nine years, but no sexual predisposition. The presentwork it can be concluded that are neoplastic hemangiosarcomas splenic more common in dogs, precededonly by the nodular hyperplasia demonstrating the importance in the diagnosis differential in relation totreatment and prognosis.

Estudo quantitativo e morfológico das regiões organizadoras de nucléolo coradas pela prata (AgNORs) em neoplasias benignas e malignas da glândula mamária da espécie canina / Quantitative and morphological study of silver staining nucleolar organizer regions (AgNORs) in benign and malignant neoplasms of the canine mammary gland

O alvo deste estudo foi apresentar um novo método de avaliação das regiões organizadoras de nucléolos (NORs), objetivando verificar a correlação entre malignidade das células tumorais com proliferação celular. Vinte e nove tumores foram analisados quanto aos aspectos morfológicos e quantitativos de regiões organizadoras de nucléolos coradas pela prata (AgNORs) e comparados à quatro glândulas mamárias normais utilizadas como controle. Os dados obtidos foram tratados pelos testes não-paramétrico de Kruskal-Wallis e ôUõ de Mann-Whitney. Os números médios de NORs dos grupos neoplásicos malignos incluindo carcinomas tubulares (2,14 ± 0,54), carcinomas papilíferos (1,79 ± 0,24) e carcinomas sólidos (1,95 ± 0,30) mostraram diferença significativa quando comparados ao grupo controle (1,16 ± 0,08). A análise morfológica das AgNORs mostrou que o padrão de apresentação da NORs do tipo I prevaleceu nos grupos neoplásicos benignos (75% dos casos de adenoma tubular e 50% dos casos de carcinoma tubular), enquanto o tipo II prevaleceu nos grupos neoplásicos de comportamento maligno (54,5 % dos carcinomas papilíferos e 100% dos carcinomas sólidos). O grupo controle mostrou na sua totalidade (100% dos casos) predominância de NORs do tipo IV. O padrão do Tipo III não predominou em nenhum dos grupos envolvidos neste estudo. A análise da morfologia das AgNORs é uma ferramenta útil na determinação do prognóstica de alterações neoplásicas da glândula mamária canina.(AU)

The aim of this study was try to present a new method of evaluation of the nucleolar organizer regions (NORs) being related its morphology and distribution, objectifying to verify the correlation between malignancy of tumoral cells with cellular proliferation. Twenty-nine tumors were analyzed with regard to quantitative and morphological aspects of the silver staining nucleolar organizer regions (AgNORs) and compared to four normal mammary glands used as control. The data obtained were submitted to non-parametric Kruskal- Wallis and Mann-Whitney ôUõ tests. The mean NORs counts of the malignant neoplastic groups tubular carcinomas (2.14 ± 0.54), papillary carcinomas (1.79 ± 0.24) and solid carcinomas (1.95 ± 0.30) showed significant difference when compared with the control group (1.16 ± 0.08). Morphological analysis of AgNORs revealed that the expression pattern of type I NORs predominate in the benign histological groups (75% of the tubular adenoma cases and 50% of the tubular carcinoma cases), whereas type II NORs pattern prevailed in the malignant behavior neoplastic groups (54,5% of the papillary carcinoma and 100% of the solid carcinoma cases). Control group showed total predominance (100% of the cases) of type IV NORs. Expression pattern of the type III NORs did not prevail in neither histological group encompassed by this study. Evaluation of AgNORs morphology reveals an useful tool for prognosis assessment of neoplastic alterations of the canine mammary gland.(AU)

Aspectos macroscópicos da anatomia do sistema reprodutor de escargots: Achatina fulica e Achatina monochromatica / Macroscopic aspects of the anatomy of the reproductive system of escargots: Achatina fulica and Achatina monochromatica

Foi estudada a macro-anatomia do sistema reprodutor do escargot das espécies Achatina fulica e Achatina monochromatica e estabelecidos os parâmetros morfológicos comparativos de ambas espécies. Foram utilizados 15 exemplares de cada espécie com quase 12 meses de idade pertencentes à mesma família. E estes foram sacrificados por congelamento (-2ºC). A coleta do sistema reprodutor foi feita após a retirada da concha e dissecação do animal. Verificou-se que, apesar dos animais terem sido mantidos nas mesmas condições ambientais e alimentares bem como submetidos ao mesmo tipo de seleção, o desenvolvimento dos órgãos reprodutores apresentou certa variação. Os aspectos morfológicos de alguns segmentos do sistema reprodutor, nas duas espécies, diferem macroscopicamente, porém, a disposição e a localização destes segmentos são idênticas.(AU)

The macro-anatomy of the reproductive system of both Achatina fulica and Achatina monochromatica was studied, by establishing the morphologic parameters of both species. Fifteen samples of each species with nearly twelve years old from the same family were used. The specimens were sacrificed by freezing (-2ºC). The collection of the reproductive system was made after the retreat from the shell and dissection of the animal. It was observed that even if the specimens have been maintained in the same environmental and alimentary conditions, as well as submitted to the same type of selection, the development of their reproductive organs show some variation. In both species, some segments of the reproductive system morphologic aspects differ macroscopically, although the disposition and the location of these segments are identical.(AU)

Descrição histológica da via genital masculina e hermafrodita de escargots das espécies Achatina fulica e Achatina monochromatica / Histology description of the male and hermafrodite genital vias of the escargots Achatina fulica and Achatina monochromatica

O objetivo deste trabalho foi descrever a histologia da via genital masculina e hermafrodita dos escargots das espécies Achatina fulica e Achatina monochromatica. Foram usados 10 exemplares adultos de cada espécie para análise histológica. Após o processo seletivo, pesagem e tomada das medidas da concha, os espécimes foram sacrificados por congelamento em freezer, a -2ºC por aproximadamente 10min. Para a exposição dos órgãos reprodutores foi efetuada uma incisão longitudinal iniciada na extremidade cranial e, acompanhando a torção do corpo, até a extremidade caudal. Cada animal foi dissecado, sobre uma placa de Petri, sob imersão em formol a 10% para melhor preservação dos tecidos. Foram coletados fragmentos representativos de cada segmento do sistema reprodutor e observou-se que a estrutura histológica dos mesmos não apresentou diferenças notórias, com exceção do ovotestis do Achatina fulica, no qual se constatou um número maior de células pigmentadas no epitélio germinativo do que, nesse mesmo epitélio, no Achatina monochromatica.(AU)

The aim of this work was to make histological description of the male and hermaphrodite genital system of escargots of the species Achatina fulica and Achatina monochromatica. Ten adult copies of each species were used for such analysis. After a selective process, weighting and measuring the shell, the specimens were sacrificed by freezing in freezer to -2ºC for approximately 10min. Reproductive organs were exposed by longitudinal incision made in the cranial extremity and by accompanying the sprain of the body, until the tail extremity. Each animal was dissected over a Petri plate, under immersion in 10% phormol for better preservation of the tissues. Representative fragments of each segment of the reproductive system were collected and it was observed that their histological structure did not present evident differences, except for the Achatina fulica ovotestis in which a larger number of pigmented cells was observed in the germinative epithelium than in the same epithelium in the Achatina monochromatica.(AU)

Estudo quantitativo e morfológico das regiões organizadoras de nucléolo coradas pela prata (AgNORs) em neoplasias benignas e malignas da glândula mamária da espécie canina / Quantitative and morphological study of silver staining nucleolar organizer regions (AgNORs) in benign and malignant neoplasms of the canine mammary gland

O alvo deste estudo foi apresentar um novo método de avaliação das regiões organizadoras de nucléolos (NORs), objetivando verificar a correlação entre malignidade das células tumorais com proliferação celular. Vinte e nove tumores foram analisados quanto aos aspectos morfológicos e quantitativos de regiões organizadoras de nucléolos coradas pela prata (AgNORs) e comparados à quatro glândulas mamárias normais utilizadas como controle. Os dados obtidos foram tratados pelos testes não-paramétrico de Kruskal-Wallis e ôUõ de Mann-Whitney. Os números médios de NORs dos grupos neoplásicos malignos incluindo carcinomas tubulares (2,14 ± 0,54), carcinomas papilíferos (1,79 ± 0,24) e carcinomas sólidos (1,95 ± 0,30) mostraram diferença significativa quando comparados ao grupo controle (1,16 ± 0,08). A análise morfológica das AgNORs mostrou que o padrão de apresentação da NORs do tipo I prevaleceu nos grupos neoplásicos benignos (75% dos casos de adenoma tubular e 50% dos casos de carcinoma tubular), enquanto o tipo II prevaleceu nos grupos neoplásicos de comportamento maligno (54,5 % dos carcinomas papilíferos e 100% dos carcinomas sólidos). O grupo controle mostrou na sua totalidade (100% dos casos) predominância de NORs do tipo IV. O padrão do Tipo III não predominou em nenhum dos grupos envolvidos neste estudo. A análise da morfologia das AgNORs é uma ferramenta útil na determinação do prognóstica de alterações neoplásicas da glândula mamária canina.

The aim of this study was try to present a new method of evaluation of the nucleolar organizer regions (NORs) being related its morphology and distribution, objectifying to verify the correlation between malignancy of tumoral cells with cellular proliferation. Twenty-nine tumors were analyzed with regard to quantitative and morphological aspects of the silver staining nucleolar organizer regions (AgNORs) and compared to four normal mammary glands used as control. The data obtained were submitted to non-parametric Kruskal- Wallis and Mann-Whitney ôUõ tests. The mean NORs counts of the malignant neoplastic groups tubular carcinomas (2.14 ± 0.54), papillary carcinomas (1.79 ± 0.24) and solid carcinomas (1.95 ± 0.30) showed significant difference when compared with the control group (1.16 ± 0.08). Morphological analysis of AgNORs revealed that the expression pattern of type I NORs predominate in the benign histological groups (75% of the tubular adenoma cases and 50% of the tubular carcinoma cases), whereas type II NORs pattern prevailed in the malignant behavior neoplastic groups (54,5% of the papillary carcinoma and 100% of the solid carcinoma cases). Control group showed total predominance (100% of the cases) of type IV NORs. Expression pattern of the type III NORs did not prevail in neither histological group encompassed by this study. Evaluation of AgNORs morphology reveals an useful tool for prognosis assessment of neoplastic alterations of the canine mammary gland.

Aspectos macroscópicos da anatomia do sistema reprodutor de escargots: Achatina fulica e Achatina monochromatica / Macroscopic aspects of the anatomy of the reproductive system of escargots: Achatina fulica and Achatina monochromatica

Foi estudada a macro-anatomia do sistema reprodutor do escargot das espécies Achatina fulica e Achatina monochromatica e estabelecidos os parâmetros morfológicos comparativos de ambas espécies. Foram utilizados 15 exemplares de cada espécie com quase 12 meses de idade pertencentes à mesma família. E estes foram sacrificados por congelamento (-2ºC). A coleta do sistema reprodutor foi feita após a retirada da concha e dissecação do animal. Verificou-se que, apesar dos animais terem sido mantidos nas mesmas condições ambientais e alimentares bem como submetidos ao mesmo tipo de seleção, o desenvolvimento dos órgãos reprodutores apresentou certa variação. Os aspectos morfológicos de alguns segmentos do sistema reprodutor, nas duas espécies, diferem macroscopicamente, porém, a disposição e a localização destes segmentos são idênticas.

The macro-anatomy of the reproductive system of both Achatina fulica and Achatina monochromatica was studied, by establishing the morphologic parameters of both species. Fifteen samples of each species with nearly twelve years old from the same family were used. The specimens were sacrificed by freezing (-2ºC). The collection of the reproductive system was made after the retreat from the shell and dissection of the animal. It was observed that even if the specimens have been maintained in the same environmental and alimentary conditions, as well as submitted to the same type of selection, the development of their reproductive organs show some variation. In both species, some segments of the reproductive system morphologic aspects differ macroscopically, although the disposition and the location of these segments are identical.

Estudo quantitativo e morfológico das regiões organizadoras de nucléolo coradas pela prata (AgNORs) em neoplasias benignas e malignas da glândula mamária da espécie canina / Quantitative and morphological study of silver staining nucleolar organizer regions (AgNORs) in benign and malignant neoplasms of the canine mammary gland

O alvo deste estudo foi apresentar um novo método de avaliação das regiões organizadoras de nucléolos (NORs), objetivando verificar a correlação entre malignidade das células tumorais com proliferação celular. Vinte e nove tumores foram analisados quanto aos aspectos morfológicos e quantitativos de regiões organizadoras de nucléolos coradas pela prata (AgNORs) e comparados à quatro glândulas mamárias normais utilizadas como controle. Os dados obtidos foram tratados pelos testes não-paramétrico de Kruskal-Wallis e ôUõ de Mann-Whitney. Os números médios de NORs dos grupos neoplásicos malignos incluindo carcinomas tubulares (2,14 ± 0,54), carcinomas papilíferos (1,79 ± 0,24) e carcinomas sólidos (1,95 ± 0,30) mostraram diferença significativa quando comparados ao grupo controle (1,16 ± 0,08). A análise morfológica das AgNORs mostrou que o padrão de apresentação da NORs do tipo I prevaleceu nos grupos neoplásicos benignos (75% dos casos de adenoma tubular e 50% dos casos de carcinoma tubular), enquanto o tipo II prevaleceu nos grupos neoplásicos de comportamento maligno (54,5 % dos carcinomas papilíferos e 100% dos carcinomas sólidos). O grupo controle mostrou na sua totalidade (100% dos casos) predominância de NORs do tipo IV. O padrão do Tipo III não predominou em nenhum dos grupos envolvidos neste estudo. A análise da morfologia das AgNORs é uma ferramenta útil na determinação do prognóstica de alterações neoplásicas da glândula mamária canina.

The aim of this study was try to present a new method of evaluation of the nucleolar organizer regions (NORs) being related its morphology and distribution, objectifying to verify the correlation between malignancy of tumoral cells with cellular proliferation. Twenty-nine tumors were analyzed with regard to quantitative and morphological aspects of the silver staining nucleolar organizer regions (AgNORs) and compared to four normal mammary glands used as control. The data obtained were submitted to non-parametric Kruskal- Wallis and Mann-Whitney ôUõ tests. The mean NORs counts of the malignant neoplastic groups tubular carcinomas (2.14 ± 0.54), papillary carcinomas (1.79 ± 0.24) and solid carcinomas (1.95 ± 0.30) showed significant difference when compared with the control group (1.16 ± 0.08). Morphological analysis of AgNORs revealed that the expression pattern of type I NORs predominate in the benign histological groups (75% of the tubular adenoma cases and 50% of the tubular carcinoma cases), whereas type II NORs pattern prevailed in the malignant behavior neoplastic groups (54,5% of the papillary carcinoma and 100% of the solid carcinoma cases). Control group showed total predominance (100% of the cases) of type IV NORs. Expression pattern of the type III NORs did not prevail in neither histological group encompassed by this study. Evaluation of AgNORs morphology reveals an useful tool for prognosis assessment of neoplastic alterations of the canine mammary gland.

Descrição histológica da via genital masculina e hermafrodita de escargots das espécies Achatina fulica E Achatina monochromatica / Histology description of the male and hermafrodite genital vias of the escargots Achatina fulica AND Achatina monochromatica

O objetivo deste trabalho foi descrever a histologia da via genital masculina e hermafrodita dos escargots das espécies Achatina fulica e Achatina monochromatica. Foram usados 10 exemplares adultos de cada espécie para análise histológica. Após o processo seletivo, pesagem e tomada das medidas da concha, os espécimes foram sacrificados por congelamento em freezer, a -2ºC por aproximadamente 10min. Para a exposição dos órgãos reprodutores foi efetuada uma incisão longitudinal iniciada na extremidade cranial e, acompanhando a torção do corpo, até a extremidade caudal. Cada animal foi dissecado, sobre uma placa de Petri, sob imersão em formol a 10% para melhor preservação dos tecidos. Foram coletados fragmentos representativos de cada segmento do sistema reprodutor e observou-se que a estrutura histológica dos mesmos não apresentou diferenças notórias, com exceção do ovotestis do Achatina fulica, no qual se constatou um número maior de células pigmentadas no epitélio germinativo do que, nesse mesmo epitélio, no Achatina monochromatica.

The aim of this work was to make histological description of the male and hermaphrodite genital system of escargots of the species Achatina fulica and Achatina monochromatica. Ten adult copies of each species were used for such analysis. After a selective process, weighting and measuring the shell, the specimens were sacrificed by freezing in freezer to -2ºC for approximately 10min. Reproductive organs were exposed by longitudinal incision made in the cranial extremity and by accompanying the sprain of the body, until the tail extremity. Each animal was dissected over a Petri plate, under immersion in 10% phormol for better preservation of the tissues. Representative fragments of each segment of the reproductive system were collected and it was observed that their histological structure did not present evident differences, except for the Achatina fulica ovotestis in which a larger number of pigmented cells was observed in the germinative epithelium than in the same epithelium in the Achatina monochromatica.

Histological alterations in the livers of Cx43-deficient mice submitted to a cholestasis model.

Gap junction intercellular communication capacity and connexin expression are reportedly involved in cell proliferation. To understand the participation of connexins in biliary duct hyperplasia, a cholestasis model was applied to mice with heterologous deletion of Gja 1, the connexin 43 (Cx43) gene. Heterozygous (Cx43+/-) knockout (KO) and wild-type mice (Cx43+/+) (WT) were submitted to bile duct ligation and euthanized at different time points (48 h, 7 days, and 14 days) after surgery. After euthanasia, the macroscopic and microscopic liver alterations were examined. A histomorphometric study of the livers was performed. For this purpose, a grid containing 100 points was applied to each liver section. The volumetric fraction of bile ducts, hepatocytes, arterioles, and terminal hepatic vein were quantified. Cell proliferation was also quantified by western blot PCNA. High mortality was observed in both genotypes. The heterologous deletion of Cx43 did not affect the biliary duct hyperplasia or most of the other parameters analyzed; however, the Cx43-deficient mice showed decrease in hepatic vein angiogenesis in comparison with the wild-type mice 48 h after surgery. In conclusion, our results indicate that the Cx43 gene heterologous deletion does not affect the biliary duct hyperplasia; on the other hand, connexin 43 deficiencies do affect the hepatic vein angiogenesis, although other studies to understand the details of this influence will be necessary.

Walker 256 tumor-bearing rats as a model to study cancer pain.

UNLABELLED: An animal model of cancer pain induced by injection of Walker 256 carcinoma cells into the plantar surface of rat hind paw is described. Tumor growth and the occurrence of metastasis were investigated by histopathological analysis. Tumor cell growth was also analyzed plethysmographically by the increase in paw volume. For characterization of pain symptoms, hyperalgesia, allodynia, and spontaneous pain were evaluated 5 to 8 days after cell injection. The volume of the inoculated paw started to increase on day 2 after inoculation, being 40% higher on day 5 after injection. At this time, there was a marked proliferation of tumor cells, with the presence of anaplastic and pleomorphic cells, nucleoli, and atypical mitotic features. On days 7 and 8 after injection, histopathological analysis of popliteal lymph nodes showed the presence of tumor cells. The intraplantar injection of Walker 256 cells caused hyperalgesia at day 5 after cell inoculation. Low-threshold mechanical allodynia was significant 2 days after cell injection, being increased on day 5. In addition, inoculation of tumor cells induced gross behavior, characterized by a significant increase in licking and lifting of the injected paw 5 days after injection. The pain-enhancing effect caused by cell inoculation was partially inhibited by indomethacin on day 2 after cell injection, whereas morphine blocked allodynia on days 2 and 5. These results indicate that intraplantar injection of Walker 256 cells cause pain symptoms characteristic of cancer pain. This experimental model can then be used to investigate new analgesic or anti-tumor drugs. PERSPECTIVE: This article presents a new animal model for studying cancer pain and metastasis. This model could help in understanding the mechanisms involved in cancer pain symptoms and may be used for the investigation of new analgesic or anti-tumor drugs.

Hepatic granulomas induced by Schistosoma mansoni in mice deficient for connexin 43 present lower cell proliferation and higher collagen content.

Granuloma formation involves a coordinated interaction between monocytes and macrophages, epithelioid cells, lymphocytes, eosinophils, neutrophils and fibroblasts. It has been established that extracellular communication via cytokines is important for the assembly of granulomas. However, the importance of gap junctions and intercellular communication to granuloma formation and development had never been assessed. Connexins are proteins that form gap junctions, and connexin 43 (Cx43) is present in macrophages, lymphoid cells, myelogenous cells, fibroblasts and others. We analyzed the effect of heterologous deletion of Gja1 (Cx43 gene) on the formation and development of hepatic granulomas induced by Schistosoma mansoni eggs. Heterozygous (Cx43(+/-)) and wild-type (Cx43(+/+)) mice were infected subcutaneously with S. mansoni cercarie and evaluated after 6, 8 and 12 weeks. Granuloma cells express Cx43, as revealed by real-time PCR in isolated granulomas, and by immunohistochemistry. Cx43 expression was reduced in Cx43(+/-) mice, as expected. No differences in the average area of granulomas or number of cells per granuloma were observed between mice of different genotypes. However, granuloma cells from Cx43(+/-) mice displayed a reduced index of the proliferating cell nuclear antigen (PCNA) labeling at 8 and 12 weeks post-infection. Moreover, Cx43(+/-) granulomas unexpectedly presented a higher degree of fibrosis, quantified by morphometric analysis in Sirius Red-stained slides. Our results indicate that the deletion of one allele of the Cx43 gene, and possibly the reduced gap junction intercellular communication capacity (GJIC), may impair the interactions between granuloma cells, reducing their proliferation and increasing their collagen content, thereby modifying the characteristics of S. mansoni granuloma in mice.

Walker 256 Tumor-Bearing Rats as a Model to Study Cancer Pain

An animal model of cancer pain induced by injection of Walker 256 carcinoma cells into the plantar surface of rat hind paw is described. Tumor growth and the occurrence of metastasis were investigated by histopathological analysis. Tumor cell growth was also analyzed plethysmographically by the increase in paw volume. For characterization of pain symptoms, hyperalgesia, allodynia, and spontaneous pain were evaluated 5 to 8 days after cell injection. The volume of the inoculated paw started to increase on day 2 after inoculation, being 40% higher on day 5 after injection. At this time, there was a marked proliferation of tumor cells, with the presence of anaplastic and pleomorphic cells, nucleoli, and atypical mitotic features. On days 7 and 8 after injection, histopathological analysis of popliteal lymph nodes showed the presence of tumor cells. The intraplantar injection of Walker 256 cells caused hyperalgesia at day 5 after cell inoculation. Low-threshold mechanical allodynia was significant 2 days after cell injection, being increased on day 5. In addition, inoculation of tumor cells induced gross behavior, characterized by a significant increase in licking and lifting of the injected paw 5 days after injection. The pain-enhancing effect caused by cell inoculation was partially inhibited by indomethacin on day 2 after cell injection, whereas morphine blocked allodynia on days 2 and 5. These results indicate that intraplantar injection of Walker 256 cells cause pain symptoms characteristic of cancer pain. This experimental model can then be used to investigate new analgesic or anti-tumor drugs. Perspective: This article presents a new animal model for studying cancer pain and metastasis. This model could help in understanding the mechanisms involved in cancer pain symptoms and may be used for the investigation of new analgesic or anti-tumor drugs.