Diagnóstico, estadiamento e tratamento cirúrgico do adenocarcinoma de pâncreas

O adenocarcinoma pancreático é um dos tumores sólidos de pior prognóstico, sendo o tratamento cirúrgico o único potencialmente curativo. Na grande maioria dos pacientes o tumor é diagnosticado em fase avançada, comumente na presença de doença metastática. A introdução de modernos métodos diagnósticos associados ao aperfeiçoamento dos já existentes tem gerado controvérsia quanto à melhor maneira de se estabelecer o diagnóstico e estadiamento do tumor. Da mesma forma, o papel da cirurgia na paliação e aspectos técnicos da ressecção de lesões localizadas estão longe de alcançarem consenso na prática. Método - Revisão da literatura sobre os aspectos controversos relacionados ao tema e um algoritmo para a abordagem dos pacientes com suspeita de tumor de pâncreas são apresentados. Foram utilizados os descritores: “adenocarcinoma” e “pâncreas” para pesquisa no PubMed (www.pubmed.com) e na Bireme (www.bireme.br) e a seguir selecionadas as publicações pertinentes a cada tópico escolhido com atenção especial para metanálises, estudos clínicos controlados, revisões sitemáticas e ainda publicações de grandes centros especializados em doenças pancreáticas. Conclusões - Na suspeita de adenocarcinoma de pâncreas é possível realizar estadiamento muito próximo do real sem a necessidade da exploração cirúrgica sistemática em virtude da disponibilidade na prática de exames modernos e eficientes. Isso permite que paliação menos invasiva seja praticada na maioria dos pacientes com lesões avançadas e incuráveis. Nos em que a cura é possível, a operação deve ser realizada objetivando-se, essencialmente, a remoção da lesão com margens livres e com aceitáveis índices de morbi-mortalidade

Purificação, caracterização e análise da atividade bioinseticida, de um inibidor de tripsina em sementes de catanduva (Piptadenia moniliformis)

One Kunitz-type trypsin inhibitors (PmTI) was purified from Piptadenia moniliformis seeds, a tree of the sub-family Mimosoideae, by TCA precipitation, affinity chromatography on immobilized trypsin-Sepharose, DEAE cellulose (ion exchange) and Superose 12 (molecular exclusion) column FPLC/AKTA. The inhibitor has Mr of 25 kDa by SDS-PAGE and chromatography molecular exclusion. The N-terminal sequence of this inhibitor showed high homology with other family Kunitz inhibitors. This also stable variations in temperature and pH and showed a small decrease in its activity when incubated with DDT in the concentration of 100mM for 120 minutes. The inhibition of trypsin by PmTI was competitive, with Ki of 1.57 x10-11 M. The activity of trypsin was effectively inhibited by percentage of inhibition of 100%, among enzymes tested, was not detected inhibition for the bromelain, was weak inhibitor of pancreatic elastase (3.17% of inhibition) and inhibited by 76.42% elastase of neutrophils, and inhibited in a moderate, chymotrypsin and papain with percentage of inhibition of 42.96% and 23.10% respectively. In vitro assays against digestive proteinases from Lepidoptera, Diptera and Coleoptera pests were carried out. Several degrees of inhibition were found. For Anthonomus grandis and Ceratitis capitata the inhibition was 89.93% and 70.52%, respectively, and the enzymes of Zabrotes subfasciatus and Callosobruchus maculatus were inhibited by 5.96% and 9.41%, respectively, and the enzymes of Plodia. interpunctella and Castnia licus were inhibited by 59.94% and 23.67, respectively. In vivo assays, was observed reduction in the development of larvae in 4rd instar of C. capitata, when PmTI was added to the artificial diet, getting WD50 and LD50 of 0.30% and 0.33%, respectively. These results suggest that this inhibitor could be a strong candidate to plant management programs cross transgenic

Purificação, caracterização e avaliação da atividade antiproliferativa de um inibidor de quimotripsina tipo kunitz de semente de Eryhrina velutina

A chymotrypsin inhibitor was purified from Erythrina velutina seeds by ammonium sulphate fractionation, affinities chromatographies on Trypsin-Sepharose, Quimotrypsin-Sepharose and reversed phase C-18 FPLC/AKTA system. The inhibitor, named EvCI, shown molecular mass of 17 kDa, as determined by SDSPAGE. 2D-PAGE showed four isoinhibitors with pI values of 4,42, 4,63, 4,83 and 5,06, with molecular mass of 17 kDa each. The aminoacid sequence of EvCI was determined by MALDI-TOF-MS and showed a high similarity with other Kunitz-type inhibitor of Erythrina variegata. EvCI competitively inhibited chymotrypsin, with Ki of 4 x10-8 M, but did not inhibited trypsin, pancreatic elastase, bromelain and papain. The inhibitory activity of EvCI was stable over wide pH and temperature ranges. In the presence of DTT 100 mM for 120 min, EvCI lost 50 % of activity. Cytotoxicity was studied in HeLa, MDA, HepG2, K562 and PC3 cells after 72-h incubation period. EvCl inhibited HeLa cells growth with an IC50 value of 50 μg/ml. Subsequent studies in HeLa cells analysis of cell death by annexin V/PI double-staining and cell cycle, using flow cytometry. The results provide evidence for a cytostatic activity of EvCl and support further studies on potential application of this inhibitors as an antiproliferative agent in combined therapy against cervical cancer

Structure-function relationship of new crotamine isoform from the Crotalus durissus cascavella

In this work we isolated a novel crotamine like protein from the Crotalus durissus cascavella venom by combination of molecular exclusion and analytical reverse phase HPLC. Its primary structure was:YKRCHKKGGHCFPKEKICLPPSSDLGKMDCRWKRK-CCKKGS GK. This protein showed a molecular mass of 4892.89 da that was determined by Matrix Assisted Laser Desorption Ionization Time-of-flight (MALDI-TOF) mass spectrometry. The approximately pI value of this protein was determined in 9.9 by two-dimensional electrophoresis. This crotamine-like protein isolated here and that named as Cro 2 produced skeletal muscle spasm and spastic paralysis in mice similarly to other crotamines like proteins. Cro 2 did not modify the insulin secretion at low glucose concentration (2.8 and 5.6 mM), but at high glucose concentration (16.7 mM) we observed an insulin secretion increasing of 2.7-3.0-fold than to control. The Na+ channel antagonist tetrodoxin (6 mM) decreased glucose and Cro 2-induced insulin secretion. These results suggested that Na+ channel are involved in the insulin secretion. In this article, we also purified some peptide fragment from the treatment of reduced and carboxymethylated Cro 2 (RC-Cro 2) with cyanogen bromide and protease V8 from Staphylococcus aureus. The isolated pancreatic beta-cells were then treated with peptides only at high glucose concentration (16.7 mM), in this condition only two peptides induced insulin secretion. The amino acid sequence homology analysis of the whole crotamine as well as the biologically-active peptide allowed determining the consensus region of the biologically-active crotamine responsible for insulin secretion was KGGHCFPKE and DCRWKWKCCKKGSG.

Abdominal hyperalgesia in secretory phospholipase A(2)-induced rat pancreatitis: Distinct roles of NK1 receptors

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

Efeito do uso de probiótico sobre o desempenho e atividade de enzimas digestivas de frangos de corte

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

Effect of acute administration of Clenbuterol on athletic performance in horses

The aim of the study was to determine the effect of clenbuterol on the anaerobic-threshold of horses on a tread-mill with increasing physical stress, measuring heart rate (HR) and blood levels of lactate, glucose, and insulin. Twelve Arabian horses. were submitted to two physical tests separated by a 10-day interval. Clenbuterol (CL) at 0.8 mu g/kg or saline (control-C) was administered intravenously 30 minutes, before the test. The treadmill exercise test consisted of an initial warmup followed by a gradually increasing effort. There was no statistical difference in either V-2 or V-4 (velocity at which plasma lactate concentration reached 4 and 2 mmol/L, respectively) between the two-experimental groups. For the CL group, V-200, V-180, V-160, and V-140 (velocity at which the rate heart is 140, 160, 180, and 200 beats/minute, respectively) decreased significantly. At rest as well as times 4, 6, and 10 minutes, insulin levels were higher in the group that recieved clenbuterol (P < .05). Contrary to what was expected, apparently, there was no improvement in aerobic metabolism in animals when given a therapeutic dose of the bronchodilator. The elevated heart rate observed could have been attributable to the stimulation of cardiac beta(1) adrenoceptors and the increased insulin levels to the stimulation of pancreatic beta(2) receptors.

Intestinal and pancreas enzyme activity of broilers exposed to thermal stress

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Estudo do sistema portal hepático no pato doméstico (Cairina moshata)

Estudou-se o comportamento do sistema portal hepático em 30 patos domésticos, adultos, machos e fêmeas. O sistema apresenta-se constituído por duas veias portais hepáticas: direita e esquerda. A veia portal hepática esquerda é formada por veias gástricas esquerdas (em número de 1 a 2), veias da margem ventral do ventrículo, veia pilórica e veia proventricular caudal. A veia portal hepática direita é formada pela veia mesentérica caudal, veia mesentérica cranial, veia proventrículo-esplênica e veia gastropancreaticoduodenal. A veia mesentérica caudal recebe tributárias do mesorreto, cloaca e junção ileocecocólica. A veia mesentérica cranial recebe tributárias jejunais (em número de 12 a 21) e se anastomosa com a veia mesentérica caudal, formando a veia mesentérica comum. A veia pancreaticoduodenal recebe duas veias gástricas direitas, constituindo assim a veia gastropancreaticoduodenal. A veia proventrículo-esplênica é formada pelas veias proventriculares dorsal e direita e pelas veias esplênicas.

Atividade enzimática pancreática de frangos de corte alimentados com dietas contendo enzima ou probiótico

Este trabalho foi realizado para avaliar o efeito da adição de enzima e/ou probiótico, bem como do estresse calórico sobre a atividade de enzimas digestivas de frangos de corte aos 7, 14, 28 e 42 dias de idade. O delineamento experimental utilizado foi o inteiramente casualizado em parcelas subdivididas com 6 tratamentos primários, constituindo um esquema fatorial 6x2 (rações x temperatura) e 4 tratamentos secundários (idades de amostragem), com 2 repetições. Não foi verificado efeito significativo dos tratamentos sobre a atividade específica das enzimas, exceto para as atividades de amilase aos 14 dias e tripsina aos 28 dias de idade, nas quais a adição de enzima proporcionou maiores atividades. Já a idade de criação afetou significativamente todas as enzimas quantificadas, sendo que a atividade específica de lipase diminuiu e as atividades específicas de amilase, tripsina e quimotripsina aumentaram com a idade das aves. A temperatura ambiente (calor) também afetou a produção enzimática de acordo com a idade dos frangos de corte, com um aumento na atividade de lipase e redução na tripsina e amilase.

Efeitos de fontes protéicas na dieta sobre a morfologia intestinal e o desenvolvimento pancreático de leitões recém-desmamados

Objetivou-se neste trabalho analisar os efeitos do uso de hidrolisado protéico do conteúdo celular de levedura (HPCCL), da proteína isolada de soja (PIS), do hidrolisado protéico de mucosa intestinal de suínos (HPMIS) e do leite em pó integral em substituição parcial ao farelo de soja sobre a morfologia do intestino delgado e o desenvolvimento pancreático de leitões aos 7 e 14 dias pós-desmame. Foram utilizados 44 leitões desmamados aos 21 dias de idade, com peso de 5,5 ± 0,6 kg, alimentados desde o desmame com as seguintes dietas isonutritivas: FS - ração à base de milho e farelo de soja; LPI - ração FS + leite em pó integral; LPI+HPMIS - ração LPI mais HPMIS; LPI+PIS - ração LPI mais PIS; LPI+HPCCL- ração LPI mais HPCCL. Os tratamentos não influenciaram a morfologia intestinal dos leitões, evidenciando que nenhuma das fontes protéicas utilizadas foi capaz de minimizar os efeitos deletérios da mudança da alimentação sobre a vilosidade intestinal. Os animais alimentados com LPI+PIS e LPI+HPMIS apresentaram, aos sete dias pós-desmame, o maior desenvolvimento pancreático. Concluiu-se, portanto, que todas as fontes protéicas estudadas foram igualmente adequadas para a formulação de dietas de desmame.

Effects and toxicity of Eugenia punicifolia extracts in streptozotocin-diabetic rats

Extracts and decoctions of Eugenia jambolana Lam., Eugenia uniflora L., and Eugenia punicifolia (Humb., Bonpl. & Kunt) DC. are used in traditional medicine to treat diabetes mellitus. Although there have been reports that Eugenia jambolana and Eugenia uniflora have antidiabetic effects, no study has yet been made on Eugenia punicifolia . We investigated the effects of aqueous, butanol, and methanol extracts of Eugenia punicifolia leaves administered by gavage to streptozotocin-diabetic rats for 26 to 29 days. Body weight, food and fluid intake, urine volume, and urinary glucose and urea were evaluated every 7 days. At the end of the experiment, we measured serum cholesterol, high-density lipoprotein (HDL)-cholesterol, triglycerides and bilirubin, hepatic glycogen and serum marker-enzymes (alanine and aspartate aminotransferases, alkaline phosphatase, gamma-glutamyltransferase, L-lactate dehydrogenase, creatine kinase, alpha-amylase, and angiotensin I converting enzyme). We found that in rats treated with the aqueous extracts, food and liquid intake, urinary volume, and body weight were all reduced, while for rats treated with the methanol extract, not only were liquid intake, urinary volume and body weight reduced, but urinary glucose and urea also decreased. Rats treated with the butanol extract showed no significant alterations in any of the parameters measured. Chronic treatment with extracts had no effect on the marker enzymes nor on serum bilirubin levels. The results indicate that aqueous extracts of Eugenia punicifolia leaves produced an anorexic effect and that methanol extracts had a beneficial effect on the diabetic state by improving carbohydrate and protein metabolism without provoking hepatobiliary, microvascular, muscular, or pancreatic toxic effects.

Signal transducer and activator of transcription 3-regulated sarcoendoplasmic reticulurn Ca2+-ATPase 2 expression by prolactin and glucocorticoids is involved in the adaptation of insulin secretory response during the peripartum period

During pregnancy, the maternal endocrine pancreas undergoes, as a consequence of placental lactogens and prolactin (PR,L) action, functional changes that are characterized by increased glucose-induced insulin secretion. After delivery, the maternal endocrine pancreas rapidly returns to nonpregnant state, which is mainly attributed to the increased serum levels of glucocorticoids (GCs). Although GCs are known to decrease insulin secretion and counteract PRL action, the mechanisms for these effects are poorly understood. We have previously demonstrated that signal transducer and activator of transcription 3 (STAT3) is increased in islets treated with PRL. In the present study, we show that STAT3 expression and serine phosphorylation are increased in pancreatic islets at the end of pregnancy (P19). STAT3 serine phosphorylation rapidly returned to basal levels 3 days after delivery (U). The expression of the sarcoendoplasmic reticulum Ca2+-ATPase 2 (SERCA2), a crucial protein involved in the regulation of calcium handling in P-cells, was also increased in P19, returning to basal levels at L3. PRL increased SERCA2 and STAT3 expressions and STAT3 serine phosphorylation in RINm5F cells. The upregulation of SERCA2 by PRL was abolished after STAT3 knockdown. Moreover, PRL-induced STAT3 serine phosphorylation and SERCA2 expression were inhibited by dexamethasone (DEX). Insulin secretion from islets of PI 9 rats pre-incubated with thapsigargin and L3 rats showed a dramatic suppression of first phase of insulin release. The present results indicate that PRL regulates SERCA2 expression by a STAT3-dependent mechanism. PRL effect is counteracted by DEX and might contribute to the adaptation of maternal endocrine pancreas during the peripartum period.

Light microscopy and ultrastructure of the liver of Astyanax altiparanae Garutti and Britski, 2000 (Teleostei, Characidae).

Livers of thirty specimens of Astyanax altiparanae obtained from a commercial fish farm were subjected to light and transmission electron microscopy, in order to describe the hepatic parenchyma and the intrahepatic exocrine pancreatic tissue. Anatomically, the liver showed only three hepatic lobes. Histological analysis demonstrated that the hepatocytes were spread out as anastomotic cords, arranged in two cellular layers and surrounded by sinusoids. The intrahepatic exocrine pancreatic tissue exhibited an acinar arrangement and was diffused in the hepatic parenchyma. Ultrastructural analysis showed that the hepatocytes had a rounded nucleus and a rough endoplasmatic reticulum, with a parallel disposition to the nuclear membrane. The exocrine pancreatic cells showed secretion granules at the apical portion, and the rough endosplasmatic reticulum was concentrically distributed.

Protection of insulin-producing cells against toxicity of dexamethasone by catalase overexpression

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