IgA production, coliforms analysis and intestinal mucosa morphology of piglets that received probiotics with viable or inactivated cells

Two types of probiotics were used in piglets. One product is a mixed culture of viable Lactobacillus acidophilus, Enterococcus faecium e Bifidobacterium bifidum. The second product is composed of inactivated Lactobacillus acidophilus cells. The piglets received two weekly oral doses for 30 days while a control group did not receive probiotics. All piglets were euthanized at the 30th day of life and the mesenteric lymph nodes, the small intestine, and blood samples were collected. The tissue samples were studied by light microscopy and the blood serum was analyzed by ELISA method. The treatment with the probiotic with viable cells produced higher serum levels of IgA (P<0.05) and more IgA expressing cells were found in the mesenteric lymph nodes than observed in the inactivated cells treatment or control groups (P<0.05). Also, intestinal villi were longer, crypts were deeper (P<0.05) and fecal coliform count was lower than found in the inactivated product (P<0.05). These results suggest that viable probiotics are more efficient than inactivated probiotics to induce immunostimulation and intestinal modifications in piglets, thus improving their health and development.

Avaliação histomorfométrica e ultra-estrutural da mucosa do cólon menor eqüino submetido a distensão

Estudos recentes demonstraram que a distensão luminal do cólon menor, apesar de ter promovido redução da irrigação da camada seromuscular resultando em inflamação intensa, não acarretou em lesões histopatológicas aparentes na mucosa. O objetivo do presente estudo foi avaliar, por meio de histomorfometria e microscopia eletrônica de varredura (MEV), os efeitos da distensão intraluminal sobre a mucosa do cólon menor. Utilizaram-se 16 eqüinos. No Grupo distendido (GD), nove deles foram submetidos a quatro horas de distensão intraluminal do cólon menor por um balão inflado com pres são de 40mm de Hg. No Grupo instrumentado (GI), o balão foi introduzido no lume, mas sem provocar distensão. Colheram-se amostras intestinais antes e ao fim da obstrução e após 1,5 e 12 horas de reperfusão. À MEV observou-se em GD que a superfície da mucosa adquiriu um aspecto liso, aplainado, após a distensão, retornando ao aspecto rugoso após descompressão. Ao fim do experimento, a superfície da mucosa se apresentava com aspecto um pouco mais irregular, com pontos de fragmentação. À histomorfometria observou-se em GD uma redução da espessura da mucosa em relação ao controle após a distensão, retornando a valores semelhantes aos basais após descompressão. Concluiu-se que a mucosa, apesar de ter seu aspecto alterado e sua espessura reduzida, conseguiu suportar a distensão promovida pelo aumento de pressão intraluminal, retornando às suas características originais sem apresentar lesões significativas.

Mecanoreceptores da mucosa palatina de avestruz (Struthio camelus): estudo ao microscópio de luz

Foram estudados corpúsculos de Herbst da mucosa palatina de avestruz em nível de microscopia de luz. Os corpúsculos compõem-se de uma cápsula externa, cápsula interna e axônio central. A cápsula externa apresentou numerosas lamelas, enquanto que a cápsula interna mostrou estrutura de folhas compactas. Os corpúsculos apresentaram formato ovalado ou circular e circundado por espessos feixes de fibras colágenas. Cada lamela estava composta de uma densa rede de fibras espessas. Os axônios terminais estavam situados ao longo do eixo, terminando em um bulbo terminal. As fibras da cápsula externa, coradas por Picrosirius e examinadas no microscópio óptico sob luz polarizada, revelou a presença de fibras colágenas do tipo I em verde e na região periférica observou-se grande quantidade de fibras colágenas do tipo III. Os corpúsculos apresentaram-se envoltos por células planas e envoltos por fibras colágenas.

Nerve endings of filliform, fungiform and vallate papillae of dorsal tongue mucosa of White-lipped peccary (Tayassu pecari): Neurohistological observations

The neurohistologic observations were performed using the specimens prepared by Winkelmann and Schmitt silver impregnation method. The tissues were fixed in 10% formalin solution and sections of 40µm thickness were obtained by Leica Cryostat at -30ºC. The sections of dorsal mucosa of White-lipped peccary tongue showed numerous filliform and fungiform papillae, and two vallate papillae on the caudal part. The epithelial layer revealed queratinized epithelial cells and the connective tissue papillae of different sizes and shapes. Thick nerve fiber bundles are noted into the subepithelial connective tissue of the papillae. The connective tissue of fungiform and vallate papillae contained numerous sensitive nerves fibers bundles forming a complex nerve plexus.

Queilite angular traumática em eqüinos associada à ingestão de Panicum maximum

Uma condição com aumento da fenda bucal de eqüinos por lesão na comissura labial foi estudada. Este aumento tinha extensão variável e era uni ou bilateral. Na mucosa da bochecha da comissura labial exposta havia pequenas erosões. Durante a mastigação havia perda de pequena quantidade de capim e saliva pela fenda bucal aumentada. Os animais apresentavam bom estado nutricional. O exame histopatológico de tecido retirado da comissura labial revelou epidermite superficial. Nas quatro propriedades onde se verificou o problema, constatou-se que os eqüinos eram mantidos em sistema extensivo de criação em pastagem de Panicum maximum (variedades Tanzânia, Mombaça, Tobiatã e Colonião), com folhas maduras, altas, lignificadas e de bordos cortantes. De acordo com os dados epidemiológicos, com os achados clínicos e histopatológicos, concluí-se que essas lesões foram causadas pela ação cortante das folhas de Panicum maximum, associada à forma de apreensão da pastagem alta e mastigação pelos eqüinos.

Utilização de biópsias da terceira pálpebra e mucosa retal em ovinos para diagnóstico de scrapie em uma propriedade da região sul do Brasil

Scrapie é uma encefalopatia espongiforme transmissível (EET) que causa lesões cerebrais degenerativas em ovinos e caprinos. Caracteriza-se pelo acúmulo, no tecido encefálico e linforreticular, da forma anormal da proteína priônica (PrP Sc) que provoca a morte maciça de neurônios e células gliais, além de vacuolização intensa no tecido afetado. Esse trabalho descreve a utilização da técnica de imuno-histoquímica (IHQ) para proteína priônica em tecido linforreticular de biópsias de terceira pálpebra e mucosa retal, como método diagnóstico de scrapie em ovinos. Realizaram-se exames de IHQ para scrapie em amostras de uma propriedade de origem de um ovino com diagnóstico dessa enfermidade. Utilizaram-se anticorpos monoclonais antipríon para diagnóstico ante mortem pela técnica de IHQ. Nas 318 amostras de biópsias analisadas, encontrou-se 19 resultados positivos para PrP Sc nos folículos de terceira pálpebra e não foi obtida marcação no tecido linfático de mucosa retal em nenhuma das amostras coletadas. Realizaram-se 18 necropsias dos animais positivos anteriormente por biópsia e 21 necropsias de ovinos parentes dos positivos de scrapie. Confirmou-se o resultado de scrapie pela IHQ após a necropsia dos animais positivos nas biópsias de terceira pálpebra. Nesses animais, os órgãos com maior número de cortes positivos foram a terceira pálpebra (18/18) e a tonsila (8/18). Nos ovinos com parentesco com os positivos, nenhum resultado de scrapie ocorreu. A utilização de tecidos linfoides no diagnóstico de scrapie por IHQ através de biópsias mostrou-se um método viável e eficaz para o diagnóstico pré-clínico.

Patogenicidade e virulência de Toxoplasma gondii isolado de suínos de criação artesanal no sul do Brasil

Estudos com Toxoplasma gondii em suínos são relevantes porque seus produtos e subprodutos fazem parte da cadeia alimentar do ser humano. As principais vias de transmissão deste agente são o carnivorismo, fecal-oral e congênita. Seis isolados de Toxoplasma gondii de suínos de criação artesanal foram avaliados quanto à patogenicidade e virulência em camundongos suíços albinos. A suspensão de taquizoítos utilizada nos testes foi obtida através da punção ou lavagem da cavidade peritoneal de camundongos que apresentaram ascite. Cada amostra foi inoculada em grupos de cinco camundongos, com inóculo de 10¹, 10², 10³, 10(4), 10(5) e 10(6) taquizoítos vivos, via intraperitoneal. Dos isolados, 50% (3/6) foram letais e causaram sinais clínicos nos camundongos. A dose mínima letal foi de 10³ taquizoítos. A morte dos animais que apresentaram infecção aguda ocorreu entre 12 e 26 dias após a inoculação. Todos os isolados da região estudada apresentam alta capacidade de formar cistos, o que pode aumentar o risco de infecção pela ingestão de tecidos dos animais infectados pelos mesmos.

Morphological changes of the jejunal mucosa in protracted diarrhea and their correlation with disease duration, weight loss and serum albumin levels

The pathogenesis of protracted diarrhea is multifactorial. In developing countries, intestinal infectious processes seem to play an important role in triggering the syndrome. Thirty-four children aged 1 to 14 months, mean 6.5 months, with protracted diarrhea were studied clinically and in terms of small intestinal mucosal morphology. Mild, moderate or severe hypotrophy of the jejunal mucosa was detected in 82% of cases, and mucosal atrophy was observed in 12%. The intensity of the morphological changes of the jejunal mucosa correlated negatively with serum albumin levels. No correlation was detected between mucosal grading and duration of diarrhea or between mucosal grading and weight reported as percentile. After nutritional support was instituted, serial jejunal biopsies were obtained from 12 patients: five patients submitted to parenteral nutrition for 7 to 38 days, mean 17 days, and 7 patients receiving a hypoallergenic oral diet (semi-elemental formula, 3; chicken formula, 3; human milk, 1). In seven cases (58%) a progressive increase in villus height and a decrease in the number of inflammatory cells were noted. Recovery of the morphologic pattern was accompanied by clinical improvement in all patients

Immunoexpression of CD95 in chronic gastritis and gastric mucosa-associated lymphomas

CD95 (Fas/APO-1)-mediated apoptosis plays an important role in immunological regulation and is related to the pathogenesis of autoimmune diseases. Immunoexpression of CD95 has been reported to frequently occur in low grade non-Hodgkin lymphomas, especially of post-germinal center histogenesis, among which those originating in mucosa-associated lymphoid tissue (MALT lymphomas). However, there is no report comparing in situ immunoexpression of this marker in lymphomas and the hyperplastic lymphoid reaction (chronic gastritis) related to Helicobacter pylori infection. The purpose of the present research was to compare the intensity of lymphoid CD95 immunoexpression in 15 cases of H. pylori-related chronic gastritis and 15 gastric MALT lymphomas. CD95 (anti-CD95) was detected by an immunoperoxidase technique in paraffin sections using the catalyzed amplification system. Graduation of reaction intensity (percentage of CD95-positive cells) was semiquantitative, from 1+ to 4+. Nine cases of chronic gastritis were 4+, five 2+ and one 1+. Three lymphomas were 4+, three 3+, four 2+, four 1+, and one was negative. Although 14 of 15 lymphomas were positive for CD95, the intensity of the reaction was significantly weaker compared to that obtained with gastric tissue for patients with gastritis (P = 0.03). The difference in CD95 immunoexpression does not seem to be useful as an isolated criterion in the differential diagnosis between chronic gastritis and MALT lymphomas since there was overlapping of immunostaining patterns. However, it suggests the possibility of a pathogenetic role of this apoptosis-regulating protein in MALT lymphomas.

Immunopathology of the duodenal mucosa of HIV-positive patients during combined antiretroviral therapy

The objective of the present study was to evaluate the duodenal mucosa of HIV-infected patients during antiretroviral therapy. This was an observational study conducted on HIV-positive patients and a control group. Group 1 comprised 22 HIV-negative individuals while 38 HIV-positive individuals were classified according to the CDC 1993 classification into group 2 (A1 or A2) or group 3 (B2, A3, B3, C2, C3). All subjects were submitted to upper gastrointestinal endoscopy with duodenal biopsies. Qualitative, semi-quantitative and quantitative histological analyses were performed. Results were considered significant when P < 0.05. A higher prevalence of inflammatory infiltrate and eosinophilia was observed in the HIV group, together with a reduction in mucosal CD4+ lymphocyte (L) counts [median (lower-upper quartiles), 12.82 (8.30-20.33), 6.36 (1.75-11.66) and 1.75 (0.87-3.14) in groups 1, 2 and 3, respectively] which was not correlated with disease stage. The extent of CD4+L count reduction was similar in blood and duodenal mucosa. Normal CD8+L and CD45RO+L counts, and normal numbers of macrophages and antigen-presenting cells were also found in the HIV patients. The cytokine pattern did not differ among groups. Tissue HIV, assessed by p24 antigen, correlated with a higher CD45RO+L count (77.0 (61-79.8) and 43.6 (31.7-62.8) in p24+ and p24-, respectively, P = 0.003), and IL-4 positivity (100 and 48.2% in p24+ and p24-, respectively, P = 0.005). The duodenal mucosa of HIV+ patients showed a relatively preserved histological architecture. This finding may be characteristic of a population without opportunistic infections and treated with potent antiretroviral therapy, with a better preservation of the immune status.

The role of IFN-gamma and IL-4 in gastric mucosa inflammation associated with Helicobacter heilmannii type 1 infection

Although Helicobacter heilmannii infection is less common than H. pylori infection in humans, it is considered to be of medical importance because of its association with gastritis, gastric ulcer, carcinoma, and mucosa-associated lymphoid tissue lymphoma of the stomach. However, there have been no studies evaluating the role of the Th cell response in H. heilmannii gastric infection. We evaluated the participation of pro-inflammatory and anti-inflammatory cytokines, IFN-gamma and IL-4, in H. heilmannii gastric infection in genetically IFN-gamma- or IL-4-deficient mice. The serum IFN-gamma and IL-4 concentrations were determined by ELISA. The gastric polymorphonuclear infiltrate was higher (P = 0.007) in H. heilmannii-positive than in H. heilmannii-negative wild-type (WT) C57BL/6 mice, whereas no significant inflammation was demonstrable in the stomach of H. heilmannii-positive IFN-gamma-/- C57BL/6 mice. The degree of gastric inflammatory cells, especially in oxyntic mucosa, was also higher (P = 0.007) in infected IL-4-/- than in WT BALB/c mice. Serum IFN-gamma levels were significantly higher in IL-4-/- than in WT BALB/c mice, independently of H. heilmannii-positive or -negative status. Although no difference in serum IFN-gamma levels was seen between H. heilmannii-positive (11.3 ± 3.07 pg/mL, mean ± SD) and -negative (11.07 ± 3.5 pg/mL) WT BALB/c mice, in the group of IL-4-/- animals, the serum concentration of IFN-g was significantly higher in the infected ones (38.16 ± 10.5 pg/mL, P = 0.04). In contrast, serum IL-4 levels were significantly decreased in H. heilmannii-positive (N = 10) WT BALB/c animals compared to the negative (N = 10) animals. In conclusion, H. heilmannii infection induces a predominantly Th1 immune response, with IFN-gamma playing a central role in gastric inflammation.

Beneficial effect of recombinant human growth hormone on the intestinal mucosa barrier of septic rats

The objective of the present study was to investigate the effects of recombinant human growth hormone (rhGH) on the intestinal mucosa barrier of septic rats and explore its possible mechanism. Female Sprague-Dawley rats were randomized into three groups: control, Escherichia coli-induced sepsis (S) and treatment (T) groups. Groups S and T were subdivided into subgroups 1d and 3d, respectively. Expression of liver insulin-like growth factor-1 (IGF-1) mRNA, Bcl-2 and Bax protein levels and the intestinal Bax/Bcl-2 ratio, and plasma GH and IGF-1 levels were determined. Histological examination of the intestine was performed and bacterial translocation was determined. rhGH significantly attenuated intestinal mucosal injuries and bacterial translocation in septic rats, markedly decreased Bax protein levels, inhibited the decrease of Bcl-2 protein expression and maintained the Bax/Bcl-2 ratio in the intestine. rhGH given after sepsis significantly improved levels of plasma GH (T1d: 1.28 ± 0.24; T3d: 2.14 ± 0.48 µg/L vs S1d: 0.74 ± 0.12; S3d: 0.60 ± 0.18 µg/L; P < 0.05) and IGF-1 (T1d: 168.94 ± 65.67; T3d: 201.56 ± 64.98 µg/L vs S1d: 116.72 ± 13.96; S3d: 107.50 ± 23.53 µg/L; P < 0.05) and expression of liver IGF-1 mRNA (T1d: 0.98 ± 0.20; T3d: 1.76 ± 0.17 vs S1d: 0.38 ± 0.09; S3d: 0.46 ± 0.10; P < 0.05). These findings indicate that treatment with rhGH had beneficial effects on the maintenance of the integrity of the intestinal mucosa barrier in septic rats.

Over-expression of cyclooxygenase-2 in endoscopic biopsies of ectopic gastric mucosa

Ectopic gastric mucosa (EGM) is considered to be a congenital condition. Rare cases of adenocarcinoma have been described. There are no data justifying regular biopsies or follow-up. Cyclooxygenase-2 (COX-2) is a protein involved in gastrointestinal tumor development by inhibiting apoptosis and regulating angiogenesis. The aim of this prospective study was to evaluate COX-2 expression in EGM and compare it with normal tissue and Barrett's esophagus. We evaluated 1327 patients. Biopsies were taken from the inlet patch for histological evaluation and from the gastric antrum to assess Helicobacter pylori infection. Biopsies taken from normal esophageal, gastric antrum and body mucosa and Barrett's esophagus were retrieved from a tissue bank. EGM biopsies were evaluated with respect to type of epithelium, presence of H. pylori, and inflammation. COX-2 was detected by immunohistochemistry using the avidin-biotin complex. EGM islets were found in 14 patients (1.1%). Histological examination revealed fundic type epithelium in 58.3% of cases, H. pylori was present in 50% and chronic inflammation in 66.7%. Expression of COX-2 was negative in normal distal esophagus, normal gastric antrum and normal gastric body specimens (10 each). In contrast, EGM presented over-expression of COX-2 in 41.7% of cases and Barrett's esophagus in 90% of cases (P = 0.04 and 0.03, respectively). COX-2 immunoexpression in EGM was not related to gender, age, epithelium type, presence of inflammation or intestinal metaplasia, H. pylori infection, or any endoscopic finding. Our results demonstrate up-regulation of COX-2 in EGM, suggesting a possible malignant potential of this so-called harmless mucosa.

Argyrophil cell density in the oxyntic mucosa is higher in female than in male morbidly obese patients

Obesity is a multifactorial disorder often associated with many important diseases such as diabetes, hypertension and other metabolic syndrome conditions. Argyrophil cells represent almost the total population of endocrine cells of the human gastric mucosa and some reports have described changes of specific types of these cells in patients with obesity and metabolic syndrome. The present study was designed to evaluate the global population of argyrophil cells of the gastric mucosa of morbidly obese and dyspeptic non-obese patients. Gastric biopsies of antropyloric and oxyntic mucosa were obtained from 50 morbidly obese patients (BMI >40) and 50 non-obese patients (17 dyspeptic overweight and 33 lean individuals) and processed for histology and Grimelius staining for argyrophil cell demonstration. Argyrophil cell density in the oxyntic mucosa of morbidly obese patients was higher in female (238.68 ± 83.71 cells/mm2) than in male patients (179.31 ± 85.96 cells/mm2) and also higher in female (214.20 ± 50.38 cells/mm2) than in male (141.90 ± 61.22 cells/mm2) morbidly obese patients with metabolic syndrome (P = 0.01 and P = 0.02, respectively). In antropyloric mucosa, the main difference in argyrophil cell density was observed between female morbidly obese patients with (167.00 ± 69.30 cells/mm2) and without (234.00 ± 69.54 cells/mm2) metabolic syndrome (P = 0.001). In conclusion, the present results show that the number of gastric argyrophil cells could be under gender influence in patients with morbid obesity. In addition, gastric argyrophil cells seem to behave differently among female morbidly obese patients with and without metabolic syndrome.

Estudo da atividade antimicrobiana dos ácidos anacárdicos do óleo da casca da castanha de caju (CNSL) dos clones de cajueiro-anão-precoce CCP-76 e CCP-09 em cinco estágios de maturação sobre microrganismos da cavidade bucal

A atividade antimicrobiana dos ácidos anacárdicos do óleo da casca da castanha de caju (CNSL) Anacardium occidentale (Anacardiaceae) foi estudada sobre os microrganismos da cavidade bucal Streptococcus mutans ATCC 25175, Staphylococcus aureus ATCC 12598, Candida albicans ATCC 10231 e Candida utilis. Os ácidos anacárdicos obtidos dos extratos etílicos do CNSL apresentaram atividade antibacteriana contra os microganismos citados, porém a maior atividade inibitória ocorreu sobre a bactéria Gram positiva Streptococcus mutans, considerada predominante na cárie dentária. As cáries dentárias são uma das mais freqüentes doenças infecciosas nos países em desenvolvimento. Os elementos que influenciam na cárie dentária incluem o estado nutricional, a ingestão de açúcar e a presença da microbiota cariogênica.