Cloning and functional analysis of RADIALIS-like 1 gene from Antirrhinum majus.

MYB is one of the largest transcription factor families in plants. Among them, the R3-MYB transcription factor RADIALIS (RAD) plays a very important role in the flowers development in Antirrhinum majus. In this study, a R3-MYB gene similar to RAD was found by analyzing the genome of A. majus, which was named AmRADIALIS-like 1 (AmRADL1). The gene function was predicted through bioinformatics. The relative expression levels in different tissues and organs of wild-type A. majus were analyzed by qRT-PCR. AmRADL1 was overexpressed in A. majus, and the transgenic plants were analyzed by morphological observation and histological staining. The results showed that the open reading frame (ORF) of AmRADL1 gene was 306 bp in length, encoding 101 amino acids. It has typical SANT domain, and the C-terminal contains a CREB motif, which was highly homologous to tomato SlFSM1. The results of qRT-PCR showed that AmRADL1 was expressed in roots, stems, leaves and flowers, and the expression level was higher in flowers. Further analysis of its expression in different floral organs showed that AmRADL1 had the highest expression in carpel. The results of histological staining analysis of the transgenic plants showed that compared with the wild type, although the size of the carpel cells of the transgenic plants did not change significantly, the placenta area in the carpel became smaller and the number of cell decreased. In summary, AmRADL1 may be involved in the regulation of carpel development, but the specific mechanism of action in carpel remains to be further studied.

Tissue fibrosis and its correlation with malignancy in canine mammary tumors / Fibrosis tisular y su correlación con malignidad en neoplasias mamarias caninas / Fibrose tecidual e sua correlação com a malignidade em neoplasias mamárias em cadelas

Abstract Background: Fibrosis is present in several pathologies associated with mammary carcinogenesis. Objective: To evaluate and quantify the fibrosis present in malignant and benign mammary neoplasms in bitches. Methods: Eighty-three samples were divided according to histopathological diagnosis into benign (n= 21) and malignant (n= 62) neoplasms. Haematoxylin-eosin and Masson's trichrome were used to locate the connective tissue, and the extent of fibrosis was assessed with image software. Results: Benign neoplasms were classified into adenomas (cystic, complex, and tubular), benign mixed tumor, and ductal and lobular hyperplasia. Malignant neoplasms were classified as carcinomas (complex, mixed tumor, in situ tubular, tubulopapillary, and solid). Grade I was the most prevalent histopathological class, followed by grade II and III. Fibrosis was classified as severe, moderate, or discrete. No significant (p >0.05) difference was observed for the percentage of fibrosis between malignant and benign group neoplasms. However, difference (p=0.028) was found for fibrosis percentage between histopathological subtypes of tumors. The benign subtype of lobular hyperplasia presented differences between cystic adenoma and benign mixed tumor. The in situ malignant tubular carcinoma subtype presented differences between solid and tubulopapillary carcinoma. Conclusions: Fibrosis in canine mammary tumors can be estimated with Massons's trichrome staining.

Resumen Antecedentes: La fibrosis está presente en diversas patologías asociadas con carcinogénesis mamaria. Objetivo: Analizar si existe una correlación entre fibrosis tisular y malignidad en tumores mamarios caninos. Métodos: 83 muestras de tejido mamario fueron divididas en masas benignas (n=21) y malignas (n=62), de acuerdo con sus características histopatológicas. En estas muestras se utilizaron las coloraciones de hematoxilina-eosina y tricromo de Masson para localizar el tejido conectivo y se analizó la proporción y cuantificación de fibrosis en los mismos con un software de imagen especializado. Resultados: Las masas mamarias benignas se diagnosticaron como adenomas (quístico, complejo y tubular), tumor mixto benigno, e hiperplasia ductal y lobular; las masas malignas, como carcinomas (complejo, tumor mixto, tubular in situ, túbulopapilar y solido). Los tumores grado I fueron los más prevalentes, seguidos por los grados II y III. La fibrosis se clasifico como discreta, moderada o severa. No se observó diferencia (p>0,05) en el porcentaje de fibrosis entre neoplasias benignas y malignas. No obstante, el porcentaje de fibrosis mostro diferencias (p=0,028) entre subtipos tumorales. La hiperplasia lobular fue diferente en relación a los adenomas quísticos y tumor mixto mamario. El subtipo maligno carcinoma tubular in situ fue diferente respecto a los carcinomas complejo, mixto maligno, sólido, y tubulopapilar. Conclusiones: La fibrosis en los tumores mamarios caninos se puede estimar por la tinción con tricromo de Masson.

Resumo Antecedentes: A fibrose participa em diversas patologias e ainda possui função adicional associada à carcinogênese mamária. Objetivo: Objetivou-se avaliar a fibrose e correlacionar com a malignidade nas neoplasias mamárias em cadelas. Métodos: 83 amostras foram divididas em dois grupos baseado no diagnóstico histopatológico: neoplasias benignas (n= 21) e neoplasias malignas (n= 62). Hematoxilina e eosina e Tricômico de Masson foram usadas para vizibilização de tecido conjuntivo e avaliação de fibrose através de programa de imagens. Resultados: As neoplasias benignas foram: adenoma (cístico, complexo e tubular), tumor misto benigno, hiperplasia ductal e lobular. As malignas foram: carcinoma complexo, em tumor misto maligno, tubular in situ, tubulopapilar e sólido. O grau histopatológico prevalente foi grau I, seguido do grau II e III. A fibrose nas neoplasias mamárias malignas foi classificada como severa, moderada e discreta. Não houve diferença significativa (p>0,05) na porcentagem de fibrose entre neoplasias malignas e benignas. A estatística revelou diferença significativa (p=0,028) na porcentagem de fibrose em relação ao diagnóstico histopatológico. O subtipo benigno hiperplasia lobular apresentou diferença entre o adenoma cístico e o tumor misto benigno. O subtipo maligno carcinoma tubular in situ apresentou diferença entre carcinoma complexo, misto maligno, sólido e tubulopapilar. Conclusões: A fibrose nos tumores mamários caninos pode ser estimada através da coloração de tricrômio de Masson.

Selective A2A receptors blockade reduces degeneration of substantia nigra dopamine neurons in a rotenone-induced rat model of Parkinson's disease: A histological study.

Parkinson's disease (PD) is characterized by a loss of dopaminergic neurons in the substantia nigra pars compacta (SNpc). Adenosine is a neuromodulator that inhibit the release of dopamine via a disinhibitory mechanism. In this study, we investigated the neuroprotective effect of 8-cyclopentyl-1,3-dipropylxanthine and ZM241385 (respectively, A1 and A2A receptors antagonists), on nigrostriatal dopamine neurons degradation reduction in a rotenone-induced PD model using histopathological and immunohistochemical methods. 32 male rats were randomly divided into 4 groups, 8 in each one: vehicle control (1ml/kg/48h), rotenone (1.5mg/kg/48h,s.c.), ZM241385 (3.3mg/kg/day, i.p) and 8-cyclopentyl-1,3-dipropylxanthine (5mg/kg/day, i.p). 24h after the last rotenone injection, animals were sacrificed and their brains were sectioned and prepared for histopathological staining with hematoxylin and eosin, cresyl violet for Nissl-staining, Mallory's phosphotungestic acid haematoxylin staining as well as for immunohistochemical staining for tyrosine hydroxylase. Our study showed that A2A-receptor blockade by ZM241385, but not A1 receptor blocking by 8-cyclopentyl-1,3-dipropylxanthine, decreased histopathological degeneration in SNpc neurons and hindered the reduction in dopamine levels caused by rotenone application. These results indicate that the selective A2A, but not A1 receptor blocking, has a neuroprotective effect, and may provide a more selective pharmacological strategy for the treatment of PD symptoms.