Autophagy and proinflammatory cytokine expression in the intestinal mucosa and mesenteric fat tissue of patients with Crohn's disease / Autofagia e expressão de citocinas pro-inflamatórias na mucosa intestinal e no tecido mesenterial de pacientes com doença de Crohn

BACKGROUND: Recently, mesenteric fat has been proposed to play a role in the pathophysiology of Crohn's disease (CD), as fat hypertrophy is detected close to the affected intestinal area; however, there are few studies regarding autophagy and creeping fat tissue in CD. OBJECTIVE: Evaluate autophagy-related proteins and proinflammatory cytokines in intestinal mucosa and mesenteric fat in patients with CD and controls. PATIENTS AND METHODS: Ten patients with CD, eight with non-inflammatory disease who underwent surgery, and eight with normal ileocolonoscopy were studied. The expression of LC3-II, TNF-alpha and IL-23 was determined by immunoblot of protein extracts. In addition, total RNA of LC3 and Atg16-L1 were determined using RT-PCR. RESULTS: The expression of LC3-II was significantly lower in the mesenteric tissue of CD when compared to controls (p < 0.05). In contrast, the intestinal mucosa of the CD group had higher levels of LC3-II (p < 0.05). However, mRNA expression of autophagy-related proteins was similar when compared to mesenteric fat groups. TNF-alpha and IL-23 expressions were higher in intestinal mucosa of CD than in control (p < 0.05). CONCLUSION: These findings suggest a defect in the autophagic activity of the creeping fat tissue in CD, which could be involved with the maintenance of the inflammatory process in the intestinal mucosa. (AU)

INTRODUÇÃO: Recentemente, tem se proposto que o tecido mesenterial possa participar da fisiopatologia da DC, uma vez que é notória a hipertrofia da gordura mesenterial próxima ao segmento intestinal afetado pela doença. Entretanto, há poucos estudos relacionando autofagia e tecido mesenterial na DC. OBJETIVO: Avaliar autofagia e citocinas na mucosa intestinal e no mesentério de pacientes com DC. PACIENTES E MÉTODOS: Dez pacientes com DC, oito sem doença inflamatória intestinal que foram submetidos à cirurgia, e oito com ileocolonoscopia normal, foram estudados. As expressões de LC3-II, TNF-alfa e IL-23 foram determinadas por imunoblot de extrato protéico total. Além disso, expressão gênica de LC3 e de Atg16-L1 foi realizada por RT-PCR. RESULTADOS: A expressão de LC3-II foi significativamente menor no tecido mesenterial de pacientes com DC quando comparada à dos controles (p < 0,05); as amostras de tecido intestinal do grupo DC apresentaram maior expressão de LC3-II (p < 0,05). Entretanto, as expressões gênicas relacionadas à autofagia foram similares nos grupos de tecido mesenterial. Os níveis de TNF-alfa e de IL-23 foram maiores na mucosa intestinal do grupo CD (p < 0,05). CONCLUSÃO: Estes achados sugerem alteração da autofagia no mesentério na DC, o que pode estar envolvido com a manutenção da inflamação na mucosa intestinal. (AU)

Histological description of the midgut and the pyloric valve of Tropidacris collaris (Stoll, 1813) (Orthopetera: Romaleidae)

The present research describes the histology of the midgut, gastric caeca, and pyloric valve of Tropidacris collaris (Stoll, 1813), (Orthopetera: Romaleidae). We used light microscopy, staining (Gomori's trichrome and periodic acid-Schiff (PAS)), and a routine histological analysis method (hematoxilin-eosin). The insects were obtained from, and also bred in, the Laboratory of Entomology, Department of Biology, of the Rural Federal University of Pernambuco (UFRPE). The collected material was fixed in alcoholic Boüin and embedded in paraplast. The results demonstrated that the midgut wall is composed of an inner epithelial layer and two outer layers of striate muscles: one internal (circular) and the other external (longitudinal), with connective tissue between the muscle fibers. The epithelium is single-layered, with two cell types: regenerative and elongated columnar. The gastric caeca presents muscle layers similar to those of the midgut. Simple columnar epithelium lines the gastric caeca, which presents villi and projects towards the lumen. The pyloric valve is of striate muscle tissue, covered by a single epithelial-cell layer.

A presente pesquisa teve como objetivo descrever a histologia do mesêntero, cecos gástricos e válvula pilórica de Tropidacris collaris (Stoll, 1813) (Orthoptera: Romaleidae), por meio da microscopia de luz, utilizando-se colorações especiais (Tricrômico de Gomori e P.A.S. - Acido periódico de Schiff) e de rotina (Hematoxilina-Eosina). Os insetos foram obtidos da criação existente no Laboratório de Entomologia, do Departamento de Biologia, da UFRPE. O material coletado foi fixado em Boüin alcoólico e processado para inclusão em "paralast". Os resultados mostraram que a parede do mesêntero é constituída por duas camadas de músculo estriado: uma interna (circular) e outra externa (longitudinal), com a presença de tecido conjuntivo entre as fibras musculares. O epitélio é do tipo simples, constituído por dois tipos de células: células regenerativas e células colunares altas. Os cecos gástricos também apresentam as camadas de músculo semelhantes às observadas no mesêntero. O tecido epitelial é do tipo simples colunar projetando-se para a luz do órgão, formando vilosidades. A válvula pilórica é constituída por tecido muscular estriado, recoberto por tecido epitelial.

A modified thionin acridine-orange stain for mast cells.

For in vitro studies in mesentery, toluidine blue and thionin dyes are generally used. We have tried thionin and acridine orange stains and found it to be good as it showed good colour contrast between mast cells and the background tissue. The two dyes were selected to give good colour contrast.

Heterogeneous response of adipose tissue to cancer cachexia

Cancer cachexia causes disruption of lipid metabolism. Since it has been well established that the various adipose tissue depots demonstrate different responses to stimuli, we assessed the effect of cachexia on some biochemical and morphological parameters of adipocytes obtained from the mesenteric (MES), retroperitoneal (RPAT), and epididymal (EAT) adipose tissues of rats bearing Walker 256 carcinosarcoma, compared with controls. Relative weight and total fat content of tissues did not differ between tumor-bearing rats and controls, but fatty acid composition was modified by cachexia. Adipocyte dimensions were increased in MES and RPAT from tumor-bearing rats, but not in EAT, in relation to control. Ultrastructural alterations were observed in the adipocytes of tumor-bearing rat RPAT (membrane projections) and EAT (nuclear bodies)

On the Degranulation of Rat's Mesenteric Mast Cells Caused by Morphine and Meperidine in Vitro

Histological studies were carried out on the degranulation of mesenteric mast cells of albino rats in which excised pieces of rat mesentery were incubated in media containing morphine and meperidine hydrochloride. The following conclusions were obtained. 1. The experimental dose of 0.04mg./ml. of morphine hydrochloride in Tyrode solution for the incubated mesenteric pieces brought about the degranulation of mast cells. 2. The experimental dose of 0.04mg./ml. of meperidine hydrochloride in Tyrode solution for the incubation of the mesenteric pieces did not effect the cytological changes of the mast cells. 3. By the addition of metabolic inhibitor such as iodoacetic acid to the incubating medium the degranulation of the mast cells was remarkably inhibited for the group in which the incubation was carried out for 20 minutes. However, the inhibition of the degranulation of the mast cells due to the metabolic inhibitor was abolished after 30 minutes of incubation. Consequently the authors have demonstrated the effect of morphine hydrochloride in its ability to induce a degranulation of mesenteric mast cells in vitro.

Mast Cell Degranulation with Special Reference to the Effect of Lipid Administration upon the Mesenteric Mast Cell of Albino Rats

Morphological effects of degranulation upon me-senteric mast cells of albino rats (SPrague-Dawley strain) by means of lipid administration were studied. An evident degranulation of metachromatic granules from mesenteric tissue mast cells was observed in more than half of experimental rats which were intraperitoneally given 10cc of stearic monoglyceride suspension in warm Tyrode solution (5Omg. of stearic monoglyceride in 10cc of Tyrode solution). A fairly light degranulation of metachro-matic granules from mesenteric mast cells was also displayed by the rats fed ad libitum with butter for 6 hours after being deprived of food for 24 hours.