Veridiana Maria Brianezi Dignani de Moura

Angiogenic activity of Calendula officinalis flowers L. in rats

PURPOSE In this work, angiogenic activity of Calendula officinalis L. (Asteraceae) ethanolic extract and dichloromethane and hexanic fractions were evaluated, considering medicinal properties, especially healing activity, are attributed to this plant. METHODS Models using 36 rats and 90 embryonated eggs were used to evaluate healing and angiogenic activities of extracts and fractions of the plant, through the induction of skin wounds and the chorioallantoic membrane, respectively. The effect of vascular proliferation was also tested from the study to verify the intensity of expression of vascular endothelial growth factor (VEGF) in cutaneous wounds in rats. RESULTS The angiogenic activity of the extract and the fractions was evidenced in both experimental models. It was verified that this effect is not directly related to the expression of VEGF and it could be associated to other pro-angiogenic factors. CONCLUSION The healing activity referred to C. officinalis is related, among other factors, to its positive effect on angiogenesis, characterized by the induction of neovascularization.

Alterations of C-MYC, NKX3.1, and E-Cadherin Expression in Canine Prostate Carcinogenesis

The dog (canis lupus familiaris) is the only other species besides humans that develop spontaneous prostatic carcinomas (PCa) at a high frequency. The canine model is primarily utilized for the study of the PCa molecular mechanisms and provides a natural animal model for the study of potential therapies. In humans, the PCa frequently exhibits mutations in the C‐MYC and a reduced expression of the E‐cadherin and NKX3.1 proteins. This studys objective was to evaluate the NKX3.1, C‐MYC, and E‐cadherin expression in the canine normal prostate, benign prostatic hyperplasia (BPH), proliferative inflammatory atrophy (PIA) and PCa and to verify differences in expression and subcellular localization of these proteins in the prostatic carcinogenesis. A tissue microarray (TMA) slide was constructed, and immunohistochemistry with antibodies raised against C‐MYC, NKX3.1, E‐cadherin and p63 was performed using the peroxidase and DAB methods. The C‐MYC protein expression was elevated in the cytoplasm and nuclei of the canine PCa and PIA compared with the normal prostate (P = 0.004. The NKX3.1 protein expression was reduced in 94.75% of the PCa and 100% of the PIA compared with the normal prostate (P = 0.0022). In fact, the expression of E‐cadherin trended towards a decrease in carcinomas when compared to normal prostate and PIA. By immunohistochemistry, more p63‐positive basal cells were observed in the PCa and PIA when compared with the normal prostate (P = 0.0002). This study has demonstrated that the carcinogenesis of canine prostatic tissue may be related to basal cell proliferation, the gain of C‐MYC function and the loss of NKX3.1 protein expression. Microsc. Res. Tech. 76:1250–1256, 2013.

Caracterização histomorfológica da atrofia inflamatória proliferativa na próstata canina

Proliferative inflammatory atrophy (PIA) is a premalignant prostatic change with high occurrence in human prostate and it has been recently observed in dogs. This research aimed to verify the morphologic aspects of the PIA in canine prostate. It was studied 43 glands of adult dogs of several breeds and sizes and with or without a history of prostatic disease. PIA focus was characterized by dysplastic epithelium, atrophic acini formed by several layers of cells with variable degrees of anisocytosis, anisokaryosis, reduced amount of cytoplasm and nuclei with prominent nucleoli. The epithelial changes were always accompanied by periacinar interstitial infiltrate composed of mononuclear cells, especially lymphocytes. The histomorphological evaluation showed occurrence of 65% of PIA. Out of these, 39% were PIA with discrete inflammatory infiltration, 42% PIA with moderate infiltrate and 19% PIA with marked infiltration. Thus, it was possible to characterize the PIA in the canine prostate which had high occurrence. The histomorphological characterization of canine PIA is considered of great importance because the premalignant characteristic of it have been studied in human prostate. The canine prostate can be used as an experimental model of human PIA, because the lesion is similar in both species.

Paracoccidioides brasiliensis presents metabolic reprogramming and secretes a serine proteinase during murine infection

ABSTRACT Paracoccidoides brasiliensis and Paracoccidioides lutzii, the etiologic agents of paracoccidioidomycosis, cause disease in healthy and immunocompromised persons in Latin America. We developed a method for harvesting P. brasiliensis yeast cells from infected murine lung to facilitate in vivo transcriptional and proteomic profiling. P. brasiliensis harvested at 6 h post-infection were analyzed using RNAseq and LC-MSE. In vivo yeast cells had 594 differentially expressed transcripts and 350 differentially expressed proteins. Integration of transcriptional and proteomic data indicated that early in infection (6 h), P. brasiliensis yeast cells underwent a shift in metabolism from glycolysis to β-oxidation, upregulated detoxifying enzymes to defend against oxidative stress, and repressed cell wall biosynthesis. Bioinformatics and functional analyses also demonstrated that a serine proteinase was upregulated and secreted in vivo. To our knowledge this is the first study depicting transcriptional and proteomic data of P. brasiliensis yeast cells upon 6 h post-infection of mouse lung.

Matrix metalloproteinases 2 and 9 expression in canine normal prostate and with proliferative disorders

In this study the expression of metalloproteinases 2 (MMP-2) and 9 (MMP-9) in canine normal prostates and with proliferative disorders was evaluated to verify the role of these enzymes in extracellular matrix remodeling (ECM) and in the tissue invasion process. A total of 355 prostatic samples were obtained, from which 36 (10.1%) were normal prostates, 46 (13.0%) with benign prostatic hyperplasia (BPH), 128 (36.1%) with proliferative inflammatory atrophy (PIA), 74 (20.8%) with prostatic intraepithelial neoplasia (PIN), and 71 (20.0%) with prostatic carcinoma (PC). Difference in cytoplasmic immunohistochemical staining of MMP-2 and MMP-9 between acinar epithelium and periacinar stroma was found regarding the different diagnosis. The correlation between MMP-2 and MMP-9 expression in relation to the number of labeled cells in acinar epithelium and periacinar stroma, as well as to the staining intensity in the periacinar stromal cells was evidenced in canine prostates with PIA. In conclusion, MMP-2 and MMP-9 expression has a variation in canine prostate according to the lesion, with lower expression in normal tissue and with BPH, and higher expression in those with PIA, PIN and PC. Moreover, the inflammatory microenvironment of the PIA has influence in the activity of both enzymes.

Efeito EFEITO DO EXTRATO ETANÓLICO DA CASCA DO PEQUI SOBRE A ENZIMA GLUTATIONA REDUTASE EM RATOS SUBMETIDOS A CARDIOTOXICIDADE POR DOXORRUBICINA

Léa Resende Moura1, Ângela Alves Viegas2, Loanny Maria de Almeida3, Raquel Emanuelle de Jesus Neves3, Claudinei Sousa Lima4* & Veridiana Maria Brianezi Dignani de Moura5 1Veterinary, Dr. Escola de Medicina, Centro Universitário UniEvangélica, Anápolis, GO, Brasil 2Biomedical, MSc. Escola de Medicina, Centro Universitário Unievangélica, Anápolis, GO, Brasil 3Graduate students in Biological Science. Faculdade Metropolitana de Anápolis, Anápolis, GO, Brasil 4Pharmaceutical, Dr. Unidade Universitária de Ciências Exatas e Tecnológicas, Universidade Estadual de Goiás, Anápolis, GO, Brasil 5Veterinary, Dr. Escola de Veterinária e Zootecnia, Universidade Federal de Goiás, Federal University of Goiás, Goiânia, GO, Brasil

Insertion of the LINE-1 element in the C-MYC gene and immunoreactivity of C-MYC, p53, p21 and p27 proteins in different morphological patterns of the canine TVT

The canine transmissible venereal tumor (TVT) affects the external genitalia of dogs by the natural transplant of viable tumor cells. Thus, this research aimed to diagnose and characterize TVT morphological patterns, identify the insertion of the LINE-1 element in C-MYC gene, by means of the polymerase chain reaction (PCR), and evaluate the immunohistochemical expression of C-MYC, p53, p21 and p27 proteins. The relationship between C-MYC and p53 proteins and their interference on the expression of p21 and p27 were also studied. For that, 20 samples of naturally occurring TVT were used, subjected to cytopathological, histopathological and immunohistochemical analysis, and to molecular diagnosis of neoplasia. The increased tissue expression and the correlation among C-MYC, p53, p21 and p27 proteins indicate reduction and/or loss of their functionality in the TVT microenvironment, with consequent apoptotic suppression, maintenance of cell growth and progression of neoplasia.The canine transmissible venereal tumor (TVT) affects the external genitalia of dogs by the natural transplant of viable tumor cells. Thus, this research aimed to diagnose and characterize TVT morphological patterns, identify the insertion of the LINE-1 element in C-MYC gene, by means of the polymerase chain reaction (PCR), and evaluate the immunohistochemical expression of C-MYC, p53, p21 and p27 proteins. The relationship between C-MYC and p53 proteins and their interference on the expression of p21 and p27 were also studied. For that, 20 samples of naturally occurring TVT were used, subjected to cytopathological, histopathological and immunohistochemical analysis, and to molecular diagnosis of neoplasia. The increased tissue expression and the correlation among C-MYC, p53, p21 and p27 proteins indicate reduction and/or loss of their functionality in the TVT microenvironment, with consequent apoptotic suppression, maintenance of cell growth and progression of neoplasia.

Topical and intraluminal Carolina Rinse Solution in p44/42 and p38 MAP Kinase activation profile in rabbit jejunum after ischemia and reperfusion

The effects of topical and intraluminal Carolina Rinse Solution (CRS) in p44/42 (ERK 1/2) and p38 MAPK activation profile, in rabbit jejunum after ischemia and reperfusion (I/R), were investigated in this study. Fifteen New Zealand rabbits were allocated in three groups: Sham-operated (A), Ischemia and reperfusion (B) and CRS (C). Groups B and C were subjected to one hour of ischemia and two hours of reperfusion. In group C, ten minutes prior to reperfusion, the bowel lumen was filled with CRS, and the segment was immersed in CRS until reperfusion onset. Ischemia and reperfusion stimulated the phosphorylation of the p44/42 MAPK and p38 MAPK pathways in some layers of jejunum. Progressive activation of p44/42 MAPK was chiefly localized in the crypts of Lieberkuhn, circular and longitudinal muscle layers, whereas p38 MAPK was prominently activated in myenteric plexus and both muscle layers. The results of this research indicate that the chosen model of topical and intraluminal CRS does not interfere in p44/42 and p38 MAPK activation profile in rabbit jejunum subjected to I/R.

Expressão de Cyr61 em glândulas mamárias normais e neoplásicas de cadelas

Mammary glands are the most usual sites of tumors in bitches, representing 25 to 30% of all types of cancer. During cancer genesis, several genetic changes alter the expression of different proteins like Cyr61, a peptide involved in cell proliferation and angiogenesis. This work aimed to determine through immunohistochemistry the expression profile of Cyr61 in normal and neoplastic mammary glands from bitches. So, 10 cases of each diagnose were selected: simple adenoma, complex carcinoma, simple solid carcinoma and fragments of normal mammary glands, in a total of 40 cases. Sections were subjected to immunohistochemistry, employing primary antibodies directed Cyr61. Cyr61 labeling of normal mammary epithelial cells is probably linked to its role in apoptosis and cellular proliferation. The labeling was more intense in normal and adenoma fragments and was discrete in the labeling malignant tumor sections, a different feature of expression from previously reports in human breast cancer. Immunohistochemical analysis with polyclonal antibody Cyr61 demonstrated the constitutive expression of the protein and contributed to neoplastic research alternative for tumors in bitches.

Caracterização imunofenotípica e avaliação quantitativa do infiltrado linfocítário junto às áreas de displasia epitelial da próstata de cães adultos sexualmente intactos

Canine prostatic studies have been common due to high incidence of prostatic diseases in these animals and similarities with alterations in human prostatic gland. Due the high frequency of dysplasia associated with interstitial lymphocitary infiltrate and acinar atrophy in canine prostate, the aims in this study were the immunophenotypic characterization and the quantitative evaluation of the same infiltrated using anti-CD3 and anti-CD79a to T and B lymphocytes, respectively. Forty two epithelial dysplasic lesions were graduated in discrete (48%), moderate (38%) and accentuated (14%). Lymphocitary periacinar infiltrate in dysplasic areas was T type and interaction between dysplasia grade and marker was observed, with oscillation of T and B cells in according with epithelial dysplasia grade.