Linfonodo axilar como sentinela de neoplasia mamária em cadelas / Axillary lymph node as sentinel for mammary neoplasia in bitches

Linfonodo axilar como sentinela de neoplasia mamária em cadelas. O estudo dos tumores de mama em cadelas é de grande importância devido à alta frequência com que surgem na clínica de pequenos animais. O presente estudo teve como objetivo avaliar a importância do linfonodo axilar como sentinela em neoplasias mamárias de cadelas. Foram avaliadas 49 fêmeas com neoplasia mamária, submetidas à mastectomia unilateral total, utilizando o corante azul patente para a identificação do linfonodo axilar, o qual foi submetido à análise histopatológica com a coloração de hematoxilina-eosina e imuno-histoquímica (IHQ) com anticorpo citoqueratina (AE1/AE3) para procura de metástase. Oito cadelas apresentaram metástases em linfonodo axilar, sendo sete detectadas por histopatologia e por IHQ e uma somente pela IHQ (micrometástase). Uma paciente que apresentava tumor em mamas abdominal caudal e inguinal tinha metástase no linfonodo axilar e inguinal. Assim, observa-se que o tumor pode causar alteração na drenagem linfática provocando metástase em linfonodos que normalmente não drenam determinadas mamas, por isso a retirada do linfonodo axilar deve ser incluída como técnica de rotina para permitir melhor estadiamento das neoplasias mamárias de cadelas.(AU)

Mammary tumors research in bitches is important due to their high incidence. The aim of this study was to evaluate the importance of the axillary lymph node as a sentinel lymph node for mammary neoplasms in female dogs. Forty-nine bitches with mammary neoplasia were submitted to total unilateral mastectomy, and the axillary lymph node was identified using the patent blue dye. This lymph node was processed routinely for histopathological analysis and stained with hematoxylin-eosin and by immunohistochemistry (IHC) with cytokeratin antibody (AE1/AE3) to search for metastasis. Eight dogs had axillary lymph node metastases, seven of which were detected by histopathology and by IHC and only one by IHC (micrometastasis). One dog who presented tumor in caudal and inguinal abdominal mammary glands had metastases in the axillary and inguinal lymph nodes. It is concluded that the mammary tumor can cause alteration in lymphatic drainage leading to metastases in lymph nodes which normally do not drain certain glands; so the removal of the axillary lymph node should be included as a routine technique to allow better staging of mammary neoplasms of bitches.(AU)

NM23 protein expression in colorectal carcinoma using TMA (tissue microarray): association with metastases and survival / Expressão da proteína NM23 no carcinoma colorretal preparado em TMA (tissue microarray): associação com metástases e sobrevivência

CONTEXT: NM23, a metastasis suppressor gene, may be associated with prognosis in patients with colorectal carcinoma. OBJECTIVE: To analyze NM23 expression and its association with the presence of lymph node and liver metastases and survival in patients operated on for colorectal carcinoma. METHODS: One hundred thirty patients operated on for colorectal carcinoma were investigated. Tissue microarray blocks containing neoplastic tissue and tumor-adjacent non-neoplastic mucosa were obtained and analyzed by immunohistochemical staining using a monoclonal anti-NM23 antibody. Immunohistochemical expression was assessed using a semiquantitative scoring method, counting the percentage of stained cells. The results were compared regarding morphological and histological characteristics of the colorectal carcinoma, presence of lymph node and liver metastases, tumor staging, and patient survival. Statistical analysis was performed using the Mann-Whitney test, the Kruskal-Wallis test and Fisher's exact test. Survival analysis was performed using the Kaplan-Meier method and the log-rank test. RESULTS: NM23 expression was higher in colorectal carcinoma tissue than in adjacent non-neoplastic mucosa (P<0.0001). NM23 protein expression did not correlate with degree of cell differentiation (P = 0.57), vascular invasion (P = 0.85), lymphatic invasion (P = 0.41), perineural infiltration (P = 0.46), staging (P = 0.19), lymph node metastases (P = 0.08), or liver metastases (P = 0.59). Disease-free survival showed significant association (P = 0.01) with the intensity of NM23 protein immunohistochemical expression in colorectal carcinoma tissue, whereas overall survival showed no association with NM23 protein expression (P = 0.13). CONCLUSIONS: NM23 protein expression was higher in neoplastic colorectal carcinoma tissue than in adjacent non-neoplastic mucosa, showing no correlation with morphological aspects, presence of lymph node or liver metastases, colorectal carcinoma staging, or overall survival. Disease-free survival was higher in patients with increased NM23 expression.

CONTEXTO: O NM23, denominado de gene supressor de metástases, pode estar relacionado com o prognóstico em doentes com carcinoma colorretal. OBJETIVOS: Analisar a expressão do marcador tumoral NM23 relacionando-a com a presença de metástases linfonodais e hepáticas e com a sobrevivência dos doentes operados por carcinoma colorretal. MÉTODO: Cento e trinta doentes operados por carcinoma colorretal foram analisados. Blocos de "tissue microarray" foram obtidos com tecido neoplásico e com mucosa não neoplásica adjacente ao tumor e submetidos ao estudo imunoistoquímico com o anticorpo monoclonal NM23. A imunoexpressão foi avaliada por método semiquantitativo, com contagem do percentual de células coradas. Os resultados encontrados foram relacionados com as características morfológicas e histopatológicas do carcinoma colorretal, presença de metástases linfonodais e hepáticas, estádio e sobrevivência dos doentes. O estudo estatístico foi realizado com os testes de Mann-Whitney, Kruskal-Wallis e exato de Fisher. A análise da sobrevivência foi calculada pelo método de Kaplan-Meier e pelo teste de long-rank. RESULTADOS: A expressão do marcador NM23 foi maior no tecido do carcinoma colorretal do que na mucosa não-neoplásica adjacente (P<0,0001). A expressão da proteína NM23 não apresentou relação com o grau de diferenciação celular (P = 0,57), invasão vascular (P = 0,85), invasão linfática (P = 0,41), infiltração perineural (P = 0,46), estádio (P = 0,19), metástases linfonodais (P = 0,08) ou metástases hepáticas (P = 0,59). A sobrevivência livre de doença mostrou relação significante (P = 0,01) com a intensidade de imunoexpressão da proteína NM23 no tecido do carcinoma colorretal, e a sobrevivência global não mostrou relação com a expressão da proteína NM23 (P = 0,13). CONCLUSÕES: A expressão da proteína NM23 foi mais intensa no tecido neoplásico do carcinoma colorretal do que na mucosa não-neoplásica adjacente. A expressão da proteína NM23 não se relacionou com os aspectos morfológicos, presença de metástases linfonodais ou hepáticas, estádio do carcinoma colorretal ou com a sobrevivência global. A sobrevivência livre de doença foi maior nos doentes com expressão aumentada do gene supressor de metástases NM23.

Effects of soluble antigen-induced immune cell activation on steroidogenesis in murine lymphoid organ.

The effect of soluble antigenic (bovine serum albumin, BSA) stimulation to induce steroidogenesis in murine lymphoid organs with concomitant changes in proinflammatory or inflammatory cytokine levels and its implication in the alteration of T-cell response was studied in the mice. Male Swiss albino mice (6-8 weeks old) with average body weight (20 +/- 4 g) were randomly assigned to 3 groups and injected with BSA in presence and absence of Freund's complete or incomplete adjuvant, whereas the control group received only saline. After 3 weeks, animals were sacrificed, and serums as well as lymphoid organs were collected. From the lymphoid tissue homogenate, the activities of steroidogenic enzymes and corticosterone and cytokine levels of the serum were estimated. Steroidogenic enzyme activities in murine lymphoid organs, as well as the pro-inflammatory and inflammatory cytokines levels in serum increased after Freund's complete adjuvant-emulsified BSA administration, as compared to control. The serum corticosterone and serum cytokine profile were also elevated. Results suggested that soluble protein antigen (BSA) administration stimulated steroidogenesis in murine lymphoid tissues and rise in the pro-inflammatory or inflammatory cytokine levels might indicate monocyte recruitment as well as TH1 activation.

The association of co-expression of CD44v4/MMP-9 with different nodal status in high-grade breast carcinoma patients.

BACKGROUND: Breast carcinoma is one of the most common tumors in female patients, and its metastasis is a major cause of death. An experimental model has recently found the association of CD44 with MMP-9 that facilitates tumor cell invasion and metastasis. MATERIAL AND METHOD: The CD44v4 and MMP-9 were performed on tissue in paraffin blocks of 50 cases of high-grade breast carcinoma with node positive and 50 cases with node negative. RESULTS: Increased expression of MMP-9(60%) significantly observed in high-grade breast carcinoma patients with node positive (p = 0. 004), whereas CD44v4 displays no significant difference between the two groups (p-value = 0.81). Significant co-expression of CD44v4+ / MMP-9+ (46%) was observed and correlated with node-positive patients whereas the CD44v4+ / MMP-9- (54%) express in node-negative patient (p-value = 0.01). CONCLUSION: The solely expression of CD44v4 does not associate with node status. MMP-9 plays an important role to enhance breast carcinoma cell invasion and associates with lymph node metastasis. The combined expression of CD44v4 (overexpression) and derangement of MMP-9 expression was significantly associated with nodal status.

Effect of protein malnutrition on the glycolytic and glutaminolytic enzyme activity of rat thymus and mesenteric lymph nodes

The activity of important glycolytic enzymes (hexokinase, phosphofructokinase, aldolase, phosphohexoseisomerase, pyruvate kinase and lactate dehydrogenase) and glutaminolytic enzymes (phosphate-dependent glutaminase) was determined in the thymus and mesenteric lymph nodes of wistar rats submited to protein malnutrition (6 percent protein in the diet rather than 20 percent) from conception to 12 weeks after birth. The wet weight (g) of the thymus and mesenteric lymph nodes decreased due to protein malnutrition by 87 percent (from 0.30 + 0.05 to 0.04 + 0.01) and 75 percent (0.40 + 0.04 to 0.10 + 0.02), respectively. The protein content was reduced only in the thymus from 102.3 + 4.4 (control rats) to 72.6 + 6.6 (malnourished rats). The glycolytic enzymes were not affected by protein malnutrition, but the glutaminase activity of the thymus and lymph nodes was reduced by halfing in protein-malnourished rats as compared to controls. This fact may lead to a decrease in the cellularity of the organ and thus in its size, weight and protein content.

Antioxidant enzyme activities in the lymphoid organs and macrophages of rats trained to perform moderate exercise

Strenous exercise and high levels of athletic competition may suppress immune function, increasing susceptibility to infections. Infections are often associated with a reduction in athletic performance and can have permanent or lethal consequences. Recent research, however; suggests that regular paraticipation in moderate exercise has an immunoenhancing effect but the mechanism involved remains unknown. This study examined the effect of moderate exercise (70 per cent of maximal oxygen consumption - swimming for 1 hour daily at 32 degrees Celsius with 5 per cent body weight extra load attached to the tail) training on antioxidant enzyme activities and lipid peroxidation in the lymphoid organs (mesenteric lymph nodes, thymus and spleen) and macrophages of rats. This modality of physical effort reduced the content of lipide peroxides in the lymphoid organs. The authors assumed that this effect of exercise training resulted in increased activity of antioxidant enzymes: Glutathione peroxidase in the mesenteric lymph nodes (2.1 fold) and spleen (3-fold), catalase in the spleen (5-fold) and Mn-superoxide dismutase (SOD) in the thymus (28 per cent). The exercise training increased the hydrogen peroxide production and phagocytic capacity in macrophages which was accompanied by a higher Mn-SOD activity. Therefore, a moderate exercise may be the able to improve immune function due to changes in the oxidative metabolism of the lymphoid organs and macrophages.

Effect of aging on the glutaminase activity of neoplastic and immune tissues

1. The effect of age and Walker 256 tumor on maximal phosphate-dependent glutaminase activity of rat immune tissue was determined. Glutaminase is a key enzyme in the metabolism of glutamine, an important fuel for normal and neoplastic cells. 2. Maximal activity of phosphate-dependent glutaminase was measured in immune tissues and tumors of Walker 256 tumor-bearing young (28 days old), mature (3 months old) and aged (15 months old) Wistar rats. The following tissues were examined: thymus, spleen, mesenteric lymph nodes and tumor. 3. Tumor implantation for 14 days reduced glutaminase activity in the thymus and mesenteric lymph nodes. Tumor glutaminase activity was lowest in aged rats and highest in the mature group. 4. Comparison of glutaminase activity in immune and tumor tissues suggested the flux of glutamine between these tissues in the 3 groups. Glutaminase activity was 2.8-fold higher in immune tissues in aged rats (2.58 +/- 0.35 vs 0.93 +/- 0.16 mumol min-1 g tissue wet weight-1, mean +/- SEM, 5 rats), and 1.9- (4.14 +/- 0.47 vs 8.36 +/- 1.29 mumol min-1 g tissue wet weight-1, mean +/- SEM, 5 rats) and 2.5-fold increased (2.41 +/- 0.20 vs 5.92 +/- 0.22 mumol min-1 g tissue wet weight-1, mean +/- SEM, 5 rats) in tumor tissue in the mature and young groups, respectively. These results suggest the deviation of glutamine flux from defense cells to the neoplastic tissue in tumor-bearing young and mature rats and may partially explain the slow cancer growth in elderly patients