Activity of selenium on cell proliferation, cytotoxicity, and apoptosis and on the expression of CASP9, BCL-XL and APC in intestinal adenocarcinoma cells

Intestinal cancers are correlated with diet. Thus, determining and understanding nutrient-genome interactions is important. The present work assessed the action of the oligoelement selenium on cell proliferation, cytotoxicity, and in situ apoptosis induction and on the expression CASP9, BCL-XL and APC genes in intestinal adenocarcinoma cells (HT29). HT29 cells were cultured and treated with selenium at concentrations of 5, 50 and 500 ng/mL with or without the damage-inducing agent doxorubicin. These cells were then evaluated for cytotoxicity (MTT), cell proliferation and in situ apoptosis induction. To evaluate gene expression, only the cells treated with 500 ng/mL of selenium were used. RNA was extracted from these cells, and the expressions of CASP9, BCL-XL and APC were analyzed by the RT-PCR method. The GAPDH gene was used as a reference gene. The MTT assay showed that selenium was not cytotoxic at any of the concentrations tested. The cell proliferation assay showed that selenium did not interfere with cell proliferation at the three concentrations tested. In contrast, when the three concentrations were combined with doxorubicin, a significant decrease in the proliferation rate was observed. The apoptosis rate was significantly increased in the selenium (500 ng/mL) and doxorubicin group. CASP9 expression was increased and BCL-XL expression decreased in the selenium (500 ng/mL) and doxorubicin group. APC was significantly increased in the selenium group alone. These results show that selenium increases apoptosis, especially when it is associated with a damage-inducing agent. Also, selenium has an important role in the expression of the APC gene, which is related to cell cycle regulation. (C) 2011 Elsevier B.V. All rights reserved.

Deletion (1)(q12) and double minutes in a metastatic adenocarcinoma of the prostate

Cytogenetic analysis after short-term cultures were performed on an invasive and moderately-differentiated prostatic adenocarcinoma. The results showed a normal male chromosomal complement in most metaphases examined. Furthermore, several abnormalities were found, including del(1)(q12), double minute and ring chromosomes, acentric fragments, triradial figures, and near-tetraploid cells. (C) Elsevier B.V., 1999. All rights reserved.

Comparison of histological and molecular diagnosis of Helicobacter pylori in benign lesions and gastric adenocarcinoma

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Experimental ischemia and reperfusion in equine small colon

Sob anestesia geral, com constante controle sobre a pressão arterial e a saturação de oxigênio da hemoglobina arterial, realizou-se celiotomia em 12 eqüinos. No cólon menor exposto foram demarcados três segmentos de 25cm, separados entre si por igual distância. Dois desses segmentos foram submetidos à isquemia arteriovenosa completa por 90 (grupo A) ou 180 minutos (grupo B). O terceiro segmento foi o grupo-controle. Amostras para histopatologia foram colhidas ao final dos períodos de isquemia e após 90 e 180 minutos de reperfusão no grupo A e após 90 minutos de reperfusão no grupo B. No controle, colheram-se amostras no início e final do procedimento. Avaliaram-se as lesões produzidas na mucosa e na submucosa pelos métodos semiquantitativos-escores para desprendimento de epitélio, edema, hemorragia e infiltrado de neutrófilos, e pelos quantitativos-porcentagem de perda de mucosa (PM) e razão cripta:interstício (C:I). As lesões isquêmicas foram mais intensas no grupo B do que no A para PM, C:I, desprendimento de epitélio e edema de mucosa. As amostras obtidas após a reperfusão revelaram que houve agravamento na PM, C:I, desprendimento de epitélio e edema de submucosa em ambos os grupos. Concluiu-se que a reperfusão agravou as lesões isquêmicas no cólon menor e que o modelo proposto é viável para produção dessas lesões.

Rolling technique for treatment of left displacement of the large colon in horses: 11 cases (2004-2009)

Durante o período de 2004 a 2009, 11 animais foram tratados para o deslocamento à esquerda do cólon maior, por meio da técnica de rolamento sob anestesia geral, com uma técnica distinta das demais previamente descritas. A seleção dos casos foi baseada nos achados da palpação retal e confirmada ultrassonograficamente. Nove animais foram tratados com sucesso e dois foram submetidos ao tratamento cirúrgico após três tentativas de rolamento. Não foram observadas complicações decorrentes do procedimento a curto e a longo prazos. Somente um animal apresentou recidiva do quadro clínico 10 meses após o tratamento e foi novamente submetido ao rolamento. Apesar da diferença com relação às outras técnicas, esta também se mostrou eficaz. O rolamento, mesmo quando realizado mais de uma vez, mostrou ser um procedimento seguro, porém enfatiza-se a necessidade de cuidados especiais aos animais após a sua realização, pois medidas emergenciais podem ser necessárias caso complicações decorrentes desta técnica ocorram.

Wound healing and colon carcinogenesis. Enhancing effects of skin wounding on development of colon tumors induced by 1,2 Dimethylhydrazine in the rat

Este estudo foi desenvolvido com a finalidade de investigar a possível influência do processo de cicatrização sobre o desenvolvimento neoplásico à distância, em um modelo experimental de carcinogênese do colon induzido pela 1,2 dimetil-hidrazina ( DMH). Ratos Wistar machos receberam injeções semanais de DMH ( 20mg/Kg, via subcutânea) ou solução salina, durante oito semanas. Na nona semana, um grupo tratado com DMH e um controle, foram submetidos à intervenção cirúrgica para retirada de um retalho cutâneo de 4cm no flanco direito, que cicatrizou por segunda intenção. Na 12ª semana, logo após o fechamento da ferida cutânea, os animais foram submetidos à eutanásia. O colon foi dividido em segmentos proximal e distal e examinado a nível macroscópico e histológico. Foram analisadas a incidência, distribuição e morfologia das lesões. O número total de tumores na mucosa do colon e o número de tumores por animal foi significantimente maior no grupo submetido à ferida cutânea do que no grupo tratado somente com DMH. O número de carcinomas pouco diferenciados foi significantimente maior no grupo com ferida cutânea do que em seu respectivo controle. Estes resultados sugerem que o processo de reparação de uma ferida cutânea favorece o desenvolvimento neoplásico em um órgão à distância, tal como o colon e que este efeito parece estar relacionado ao tipo histológico da neoplasia.

TOXIC DILATATION OF THE COLON IN CHAGAS-DISEASE

Toxic megacolon occurs in colitis of differing aetiology. This report describes 15 patients with chagasic megacolon with this complication. The clinical signs and symptoms in all patients were pain and progressive abdominal distension accompanied by fever, severe toxaemia and shock. Seven patients developed this clinical pattern after manual removal of faeces. The remaining patients had pain and abdominal distension followed by signs of severe toxaemia when first examined. Nine patients underwent total colectomy with ileostomy (one death), four partial colectomy (all died) and two received medical treatment (both died). At autopsy, three of the four patients undergoing partial colectomy had residual colitis and enteritis. The surgical procedure of choice for this complication of chagasic megacolon is total colectomy.

Characterization of the cellular component of polymorphous low-grade adenocarcinoma by immunohistochemistry and electron microscopy

In order to characterize the cellular component of the polymorphous low-grade adenocarcinoma (PLGA) of the salivary gland, a morphological and immunohistochemical study was carried out. Thirty cases of PLGA were studied by light microscopy and immunohistochemistry and five cases by transmission electron microscopy (TEM). The expression of cytokeratins (CKs) 7,8,10,13,14,18,19, vimentin and muscle-specific actin (MSA) was investigated through the streptavidin-biotin method. The majority of tumor cells stained for vimentin, CKs 8,18 and 7. CK 14 was positive in most cells of the papillary and trabecular sub-types. Although the expression of CKs 8,18 and 14 varied among the tumors sub-types, a straight relationship between each histologic pattern and the CK expression could not be delineated. MSA was reactive in only three tumors while CKs 10 and 13 were not detected in any tumor studied. The absence of MSA and the expression of CKs 8,18 and 7, in most of the tumor cells, lead to the hypothesis that myoepithelial cells are not the major cellular component of the PLGA. TEM revealed cells exhibiting microvilli and variable amounts of secretory granules, some of them suggesting an excretory activity. The presence of CKs 8, 18 and 7, added to the secretory granules, indicates that PLGA originates from cells located at the acinar-intercalated duct junction. (C) 1999 Elsevier B.V. Ltd. All rights reserved.

Effect of intraluminal distention on microvascular perfusion in the equine small colon

Objective-To determine the effect of experimental intraluminal distention on microvascular perfusion of the small colon in horses.Animals-6 mixed-breed healthy horses (mean age [+/- SD], 9.1 +/- 2 years).Procedure-Under general anesthesia, the small colon was exposed by celiotomy and 3 segments were demarcated. In 1 of these segments, intraluminal obstruction was created by placement of a latex balloon inflated to a pressure of 40 mm Hg (obstructed segment). The other segments were the sham-operated segment and the control segment. Microvascular perfusion was evaluated in the mucosal, submucosal, muscular, and serosal layers by injection of 15-mum-diameter colored microspheres into branches of the caudal mesenteric artery. Recovery of microspheres was performed by tissue digestion, washing, and centrifugation. Distribution of microspheres in the intestinal layers was evaluated by direct observation of stained frozen sections by light microscopy.Results-A significant reduction was observed in total microvascular perfusion of obstructed segments, which was 26.4% of that of control segments. This reduction was not evident in the mucosal layer.Conclusions and Clinical Relevance-Intraluminal distention of the equine small colon wall can promote ischemia by a reduction in microvascular perfusion in the intestinal wall. Intestinal layers do not seem to be affected to the same extent, because the absolute value for mucosal perfusion did not decrease in the obstructed segment.

INVITRO ACTION OF VASOPRESSIN ON WATER-ABSORPTION BY RAT COLON

The action of vasopressin on water absorption was studied in vitro in the colon of rats, weighing approximately 250 g. Everted sacs of intestinal segments (ascending and descending colons) prepared with mucosa only were used. Vasopressin (10 mM) significantly stimulated water absorption in both the proximal (4.85 +/- 3.78 vs 1.51 +/- 1.16 ml/mg) and distal (10.39 +/- 3.52 vs 7.22 +/- 3.58 ml/mg) colon, which corresponds to an increase of 220% and 50%,respectively. The results indicate the need for a study of the possible physiological function of vasopressin in enhancing intestinal water absorption, when it is released in response to plasma hyperosmolarity.

Gastric adenocarcinoma and Helicobacter pylori: Correlation with p53 mutation and p27 immunoexpression

Introduction: Helicobacter pylori infection is an established risk factor for gastric cancer development, but the exact underlying mechanism still remains obscure. The inactivation of tumor suppressor genes such as p53 and p27(KIP1) is a hypothesized mechanism, although there is no consensus regarding the influence of H. pylori cagA(+) in the development of these genetic alterations. Goals: To verify the relationship among H. pylori infection, p53 mutations and p27(Kip1) Protein (p27) expression in gastric adenocarcinomas (GA) seventy-four tissues were assayed by PCR for H. pylori and cagA presence. Mutational analysis of p53 gene was performed by single-strand conformation polymorphism (SSCP). Seventy tissues were analyzed by an immunohistochemical method for p27 expression. Results: From the samples examined, 95% (70/74) were H. pylori positive, 63% cagA(+). Altered p53 electrophoretic mobility was found in 72% of cases and significantly more frequent in the presence of cagA. Considerable reduction in p27 expression (19%) was found with a tendency for association between cagA(+) and p27(-), although the results were not statistically significant. Concomitant alterations of both suppressor genes were detected in 60% of cases. In the cases cagA(+), 66.7% of them had these concomitant alterations. Conclusions: The data suggest that H. pylori cagA(+) contributes to p53 alteration and indicate that concomitant gene inactivation, with reduced p27 expression, may be a mechanism in which H. pylori can promote the development and progression of gastric cancer. (C) 2010 Elsevier Ltd. All rights reserved.

Local and remote lesions in horses subjected to small colon distension and decompression

The purpose of this study was to observe and characterize colonic and lung lesions in horses subjected to experimental distension and decompression of the small colon. Sixteen healthy adult horses were divided into 2 groups: 9 horses that were subjected to distension of the small colon by means of a latex balloon surgically implanted in the lumen and inflated to a pressure of 40 mm Hg for 4 h, and 7 horses in which the balloon was implanted but not inflated. Colonic biopsy specimens were collected before balloon implantation, at the end of the period of obstruction, and 1.5 and 12 h after decompression and were examined for hemorrhage, edema, and neutrophil infiltration; myeloperoxidase (MPO) activity and hemoglobin concentration were measured as well. At the end of the experiment, lung samples were also collected and examined for neutrophil accumulation and MPO activity. The mucosa was not affected by luminal distension; lesions were restricted to the seromuscular layer. Neutrophil accumulation and edema were observed in the samples from both groups of horses but were greater in those from the distension group, in which there was also hemorrhage, fibrin deposition, and increased MPO activity in the seromuscular layer. Similarly, there was greater accumulation of neutrophils in the lung samples from the distension group than in those from the sham-operated group, as determined by histologic evaluation and MPO assay. These findings provide new evidence of reperfusion injury and a systemic inflammatory response, followed by remote lesions, in horses with intestinal obstruction.