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2.
Front Immunol ; 10: 1394, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31281317

RESUMO

In colorectal cancer (CRC), cancer-associated fibroblasts (CAFs) are the most abundant component from the tumor microenvironment (TM). CAFs facilitate tumor progression by inducing angiogenesis, immune suppression and invasion, thus altering the organization/composition of the extracellular matrix (i.e., desmoplasia) and/or activating epithelial-mesenchymal transition (EMT). Soluble factors from the TM can also contribute to cell invasion through secretion of cytokines and recently, IL-33/ST2 pathway has gained huge interest as a protumor alarmin, promoting progression to metastasis by inducing changes in TM. Hence, we analyzed IL-33 and ST2 content in tumor and healthy tissue lysates and plasma from CRC patients. Tissue localization and distribution of these molecules was evaluated by immunohistochemistry (using localization reference markers α-smooth muscle actin or α-SMA and E-cadherin), and clinical/histopathological information was obtained from CRC patients. In vitro experiments were conducted in primary cultures of CAFs and normal fibroblasts (NFs) isolated from tumor and healthy tissue taken from CRC patients. Additionally, migration and proliferation analysis were performed in HT29 and HCT116 cell lines. It was found that IL-33 content increases in left-sided CRC patients with lymphatic metastasis, with localization in tumor epithelia associated with abundant desmoplasia. Although ST2 content showed similarities between tumor and healthy tissue, a decreased immunoreactivity was observed in left-sided tumor stroma, associated to metastasis related factors (advanced stages, abundant desmoplasia, and presence of tumor budding). A principal component analysis (including stromal and epithelial IL-33/ST2 and α-SMA immunoreactivity with extent of desmoplasia) allowed us to distinguish clusters of low, intermediate and abundant desmoplasia, with potential to develop a diagnostic signature with benefits for further therapeutic targets. IL-33 transcript levels from CAFs directly correlated with CRC cell line migration induced by CAFs conditioned media, with rhIL-33 inducing a mesenchymal phenotype in HT29 cells. These results indicate a role of IL-33/ST2 in tumor microenvironment, specifically in the interaction between CAFs and epithelial tumor cells, thus contributing to invasion and metastasis in left-sided CRC, most likely by activating desmoplasia.

3.
Medwave ; 19(11): e7750, 2019 Dec 23.
Artigo em Espanhol, Inglês | MEDLINE | ID: mdl-31999675

RESUMO

Background: Laparoscopy has become the standard of care in the surgical management of deep infiltrating endometriosis (DIE). However, it is a challenging procedure with a high complication rate. Despite the benefits of the minimally invasive approach, DIE resection is often performed by surgeons without adequate training, especially in developing countries like Chile. Objective: To asses our experience in the diagnosis and laparoscopic management of DIE during seven years. Methods: A retrospective cohort study of data including 137 patients with pathology-proven DIE. Surgical and fertility outcomes were evaluated. Results: All procedures were performed laparoscopically without conversion. Dysmenorrhea and dyspareunia were the most common symptoms in 85.4% and 56.9%, respectively. Uterosacral ligaments were the most common DIE location. Endometrioma was present in 48.9% of cases. Median operative time was 140 minutes; however, it was longer in cases requiring bowel surgery (p < 0.0001). The complication rate was 10.9%. Median follow-up was 24.5 months. The pregnancy rate was 58.1% and 90% of patients reported significant symptom relief after surgery. Conclusion: Laparoscopic surgical management of DIE is effective and safe but it must be performed in tertiary centers with the availability of multidisciplinary teams.

4.
Tumour Biol ; 40(11): 1010428318810059, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30419802

RESUMO

A complex network of chemokines can influence cancer progression with the recruitment and activation of hematopoietic cells, including macrophages to the supporting tumor stroma promoting carcinogenesis and metastasis. The aim of this study was to investigate the relation between tissue and plasma chemokine levels involved in macrophage recruitment with tumor-associated macrophage profile markers and clinicopathological features such as tumor-node-metastases stage, desmoplasia, tumor necrosis factor-α, and vascular endothelial growth factor plasma content. Plasma and tumor/healthy mucosa were obtained from Chilean patients undergoing colon cancer surgery. Chemokines were evaluated from tissue lysates (CCL2, CCL3, CCL4, CCL5, and CX3CL1) by Luminex. Statistical analysis was performed using Wilcoxon match-paired test ( p < 0.05). Macrophage markers (CD68, CD163, and iNOS) were evaluated by immunohistochemistry samples derived from colorectal cancer patients. Correlation analysis between chemokines and macrophage markers and clinicopathological features were performed using Spearman's test. Plasmatic levels of chemokines and inflammatory mediators' vascular endothelial growth factor and tumor necrosis factor-α were evaluated by Luminex. Tumor levels of CCL2 (mean ± standard deviation = 530.1 ± 613.9 pg/mg), CCL3 (102.7 ± 106.0 pg/mg), and CCL4 (64.98 ± 48.09 pg/mg) were higher than those found in healthy tissue (182.1 ± 116.5, 26.79 ± 22.40, and 27.06 ± 23.69 pg/mg, respectively p < 0.05). The tumor characterization allowed us to identify a positive correlation between CCL4 and the pro-tumor macrophages marker CD163 ( p = 0.0443), and a negative correlation of iNOS with desmoplastic reaction ( p = 0.0467). Moreover, we identified that tumors with immature desmoplasia have a higher CD163 density compared to those with a mature/intermediated stromal tissue ( p = 0.0288). Plasmatic CCL4 has shown a positive correlation with inflammatory mediators (tumor necrosis factor-α and vascular endothelial growth factor) that have previously been associated with poor prognosis in patients. In conclusion High expression of CCL4 in colon cancer could induce the infiltration of tumor-associated macrophages and specifically a pro-tumor macrophage profile (CD163+ cells). Moreover, plasmatic chemokines could be considered inflammatory mediators associated to CRC progression as well as tumor necrosis factor-α and vascular endothelial growth factor. These data reinforce the idea of chemokines as potential therapeutic targets or biomarker in CRC.


Assuntos
Biomarcadores Tumorais/metabolismo , Quimiocina CCL2/metabolismo , Quimiocina CCL3/metabolismo , Quimiocina CCL4/metabolismo , Neoplasias Colorretais/patologia , Macrófagos/patologia , Microambiente Tumoral , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Neoplasias Colorretais/metabolismo , Feminino , Seguimentos , Humanos , Macrófagos/metabolismo , Masculino , Pessoa de Meia-Idade , Prognóstico
5.
Rev. bras. anestesiol ; 68(2): 135-141, Mar.-Apr. 2018. tab, graf
Artigo em Inglês | LILACS-Express | ID: biblio-897816

RESUMO

Abstract Background: Post-operative delirium is a serious complication in patients undergoing major abdominal surgery. It remains unclear whether peri-operative hemodynamic and perfusion variables affect the risk for postoperative delirium. The objective of this pilot study was to evaluate the association between perfusion and hemodynamics peri-operative with the appearance of post-operative delirium. Methods: Prospective cohort study of adults 60 years or older undergoing elective open colon surgery. Multimodal hemodynamic and perfusion variables were monitored, including central venous oxygenation (ScvO2), lactate levels, and non-invasive cerebral oxygenation (rSO2), according to a standard anesthesia protocol. Fisher's exact test or Student's t-test were used to compare patients who developed post-operative delirium with those who did not (p < 0.05). Results: We studied 28 patients, age 73 ± 7 years, 60.7% female. Two patients developed post-operative delirium (7.1%). These two patients had fewer years of education than those without delirium (p = 0.031). None of the peri-operative blood pressure variables were associated with incidence of post-operative delirium. In terms of perfusion parameters, postoperative ScvO2 was lower in the delirium than the non-delirium group, without reaching statistical significance (65 ± 10% vs. 74 ± 5%; p = 0.08), but the delta-ScvO2 (the difference between means post-operative and intra-operative) was associated with post-operative delirium (p = 0.043). Post-operative lactate and rSO2 variables were not associated with delirium. Conclusions: Our pilot study suggests an association between delta ScvO2 and post-operative delirium, and a tendency to lower post-operative ScvO2 in patients who developed delirium. Further studies are necessary to elucidate this association.


Resumo Justificativa: O delírio pós-operatório é uma complicação séria em pacientes submetidos à cirurgia abdominal de grande porte. Ainda não está claro se as variáveis hemodinâmicas e de perfusão no período perioperatório afetam o risco de delírio pós-operatório. O objetivo deste estudo piloto foi avaliar a associação entre perfusão e hemodinâmica no perioperatório com o surgimento de delírio pós-operatório. Métodos: Estudo prospectivo de coorte de adultos com 60 anos ou mais, submetidos à cirurgia eletiva aberta do cólon. As variáveis multimodais de hemodinâmica e perfusão foram monitoradas, inclusive oxigenação venosa central (ScvO2), níveis de lactato e oxigenação cerebral não invasiva (rSO2), de acordo com um protocolo-padrão de anestesia. O teste exato de Fisher ou o teste t de Student foram usados para comparar os pacientes que desenvolveram delírio pós-operatório com aqueles que não desenvolveram p < 0,05. Resultados: Avaliamos 28 pacientes, 73 ± 7 anos, 60,7% do sexo feminino. Dois pacientes desenvolveram delírio pós-operatório (7,1%). Esses dois pacientes tinham menos anos de escolaridade do que aqueles sem delírio pós-operatório (p = 0,031). Nenhuma das variáveis de pressão arterial no perioperatório foi associada à incidência de delírio. Quanto aos parâmetros de perfusão, ScvO2 foi menor no grupo que apresentou delírio pós-operatório do que no grupo que não apresentou delírio, sem atingir significância estatística (65 ± 10% vs. 74 ± 5%; p = 0,08), mas o delta-ScvO2 (a diferença entre as médias no pós-operatório e intraoperatório) foi associado ao delírio (p = 0,043). As variáveis de lactato e rSO2 no pós-operatório não foram associadas ao delírio. Conclusões: Nosso estudo piloto sugere uma associação entre delta-ScvO2 e delírio e uma tendência à diminuição da ScvO2 no pós-operatório de pacientes com delírio. Estudos adicionais são necessários para elucidar essa associação.

6.
Rev Bras Anestesiol ; 68(2): 135-141, 2018.
Artigo em Português | MEDLINE | ID: mdl-29287672

RESUMO

BACKGROUND: Post-operative delirium is a serious complication in patients undergoing major abdominal surgery. It remains unclear whether peri-operative hemodynamic and perfusion variables affect the risk for postoperative delirium. The objective of this pilot study was to evaluate the association between perfusion and hemodynamics peri-operative with the appearance of post-operative delirium. METHODS: Prospective cohort study of adults 60 years or older undergoing elective open colon surgery. Multimodal hemodynamic and perfusion variables were monitored, including central venous oxygenation (ScvO2), lactate levels, and non-invasive cerebral oxygenation (rSO2), according to a standard anesthesia protocol. Fisher's exact test or Student's t-test were used to compare patients who developed post-operative delirium with those who did not (p<0.05). RESULTS: We studied 28 patients, age 73±7 years, 60.7% female. Two patients developed post-operative delirium (7.1%). These two patients had fewer years of education than those without delirium (p=0.031). None of the peri-operative blood pressure variables were associated with incidence of post-operative delirium. In terms of perfusion parameters, postoperative ScvO2 was lower in the delirium than the non-delirium group, without reaching statistical significance (65±10% vs. 74±5%; p=0.08), but the delta-ScvO2 (the difference between means post-operative and intra-operative) was associated with post-operative delirium (p=0.043). Post-operative lactate and rSO2 variables were not associated with delirium. CONCLUSIONS: Our pilot study suggests an association between delta ScvO2 and post-operative delirium, and a tendency to lower post-operative ScvO2 in patients who developed delirium. Further studies are necessary to elucidate this association.


Assuntos
Doenças do Colo/cirurgia , Delírio/epidemiologia , Complicações Pós-Operatórias/epidemiologia , Idoso , Doenças do Colo/complicações , Doenças do Colo/fisiopatologia , Delírio/etiologia , Procedimentos Cirúrgicos do Sistema Digestório , Feminino , Humanos , Hipotensão/complicações , Masculino , Projetos Piloto , Complicações Pós-Operatórias/etiologia , Estudos Prospectivos , Fluxo Sanguíneo Regional
7.
Rev. chil. cir ; 70(1): 13-18, 2018. tab, graf
Artigo em Espanhol | LILACS | ID: biblio-899650

RESUMO

Resumen Introducción La resistencia a antibióticos es un problema mundial. En los pacientes que requieren cirugía de urgencia, los antibióticos son un apoyo importante concomitante al acto quirúrgico. Objetivo Analizar los cultivos de líquido peritoneal obtenidos de pacientes operados por patología quirúrgica abdominal de urgencia. Materiales y Métodos Se realiza una cohorte prospectiva de los pacientes operados de urgencia. Se tomó cultivo de líquido peritoneal y se procesó según técnica estandarizada. Resultados Se encontró un 39,4% de cultivos positivos. E. coli fue el germen más frecuente. Destacan 5 cultivos positivos para P. aeruginosa. Existe un 25% de resistencia a ampicilina/sulbactam y 19% a quinolonas para E. coli. Conclusión La resistencia encontrada fue menor a lo reportado en la literatura, pero aún destacable. El conocimiento del perfil de bacterias y sus resistencias a antimicrobianos son importantes para las políticas hospitalarias locales de uso racional de antibióticos.


Background Antimicrobial resistance is a worldwide problem. In patients requiring emergency surgery, antibiotics are an important assistance additional to surgical intervention. Objective Analize peritoneal fluid cultures obtaines from patients who underwent emergency surgery. Methods A prospective cohort of emergency abdominal surgical patients were enrolled. Peritoneal fluid cultures were taken and processed according to standarized technique. Results A 39.4% of positive cultures was found. E. coli was the most common bacteria identified. Five positive cultures for P. aeruginosa stand out. E. coli had 25% resistance to ampicillin/sulbactam and 19% for quinolones. Conclusion Resistance found was lower than international reports, but still noteworthy. Knowledge of local bacteria profile and antimicrobial resistance is important for local antibiotic hospital policy.


Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Idoso , Bactérias/efeitos dos fármacos , Líquido Ascítico/microbiologia , Abdome/cirurgia , Antibacterianos/farmacologia , Pseudomonas aeruginosa/efeitos dos fármacos , Bactérias/isolamento & purificação , Testes de Sensibilidade Microbiana , Farmacorresistência Bacteriana , Emergências , Escherichia coli/efeitos dos fármacos , Klebsiella pneumoniae/efeitos dos fármacos
8.
Rev. chil. cir ; 69(1): 44-48, feb. 2017. ilus
Artigo em Espanhol | LILACS | ID: biblio-844323

RESUMO

Introducción: El tratamiento en el cáncer de recto ha progresado en la última década. Hoy es factible ofrecer una cirugía con preservación de esfínteres, realizando anastomosis colorrectales bajas o anastomosis coloanales. Esto ha determinado que muchos pacientes desarrollen disfunción intestinal que puede llegar a ser severa, agrupando una serie de alteraciones que se conocen como síndrome de resección anterior baja. Objetivo: Efectuar una adaptación cultural de la versión 1.0 en español neutro del cuestionario acerca de la función intestinal o Low Anterior Resection Syndrome Score (LARS Score), efectuando traducción, comparación de traducciones, traducción inversa y prueba piloto. Resultados: Los resultados obtenidos de la prueba piloto revelan que la población encuestada logró comprender el instrumento, por lo que no se realizaron modificaciones posteriores. Conclusión: Se cuenta con una versión adaptada del cuestionario LARS para ser usada en Chile, la cual puede someterse a procesos de validación y establecer las características psicométricas para ser usada en pacientes con cáncer de recto operados.


Introduction: The treatment of rectal cancer has progressed in the past decade. Nowadays, it's feasible to provide sphincter sparing surgery with low colorectal anastomosis or coloanal anastomosis. This has determined that many patients develop intestinal dysfunctions that can become severe, grouping a number of disorders known as low anterior resection syndrome. Objective: To perform a cultural adaptation of the version 1.0 questionnaire about bowel function or Low Resection Syndrome Score (LARS Score) in neutral Spanish, making a translation, comparing translations, back translation and pilot test. Results: The results of the pilot test showed that the population surveyed understood the instrument, so that no further modifications were made. Conclusion: We now have an adapted version of the LARS questionnaire for use in Chile, which can undergo validation processes to establish the psychometric characteristics for use in patients with rectal cancer surgery.


Assuntos
Humanos , Complicações Pós-Operatórias/diagnóstico , Neoplasias Retais/cirurgia , Inquéritos e Questionários , Chile , Comparação Transcultural , Defecação , Procedimentos Cirúrgicos do Sistema Digestório/efeitos adversos , Flatulência , Complicações Pós-Operatórias/psicologia , Psicometria , Neoplasias Retais/psicologia , Reto/fisiopatologia , Reprodutibilidade dos Testes , Índice de Gravidade de Doença , Síndrome , Traduções
9.
Dis Colon Rectum ; 57(11): 1324-8, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25285701

RESUMO

BACKGROUND: The aim of the current study was to demonstrate the use of a modified stapling technique, called the apex technique, to treat rectal intussusception and full rectal mucosal prolapse. It was conducted as a retrospective study at 3 centers (2 in Brazil and 1 in Chile). TECHNIQUE: The apex technique is performed by using a HEM/EEA-33 stapler. A pursestring suture is placed at the apex of the prolapse, on the 4 quadrants, independent of the distance to the dentate line. A second pursestring is then placed to define the band of rectal mucosa to be symmetrically resected. MAIN OUTCOME MEASURES: Outcome measures included width of the resected full-thickness rectal wall; the intensity of postoperative pain on a visual analog scale from 1 to 10; full mucosal prolapse and rectal intussusception assessed by physical examination, cinedefecography, or echodefecography; and change in the constipation scale. RESULTS: Forty-five patients (30 women/15 men; mean age, 59.5 years) with rectal intussusception and full mucosal prolapse were included. The median operative time was 17 (range, 15-30) minutes. Bleeding after stapler fire requiring manual suture occurred in 3 patients (6.7%); 25 (55.6%) patients reported having no postoperative pain. Hospital stay was 24 hours. The mean width of the resected rectal wall was 5.9 (range, 5.0-7.5) cm. Stricture at the staple line was seen in 4 patients, of whom 1 required dilation under anesthesia. The median follow-up time was 120 (range, 90-120) days. A small residual prolapse was identified in 6 (13.3%) patients. Imaging demonstrated complete disappearance of rectal intussusception in all patients, and the mean postoperative constipation score decreased from 13 (range, 8-15) to 5 (range, 3-7). CONCLUSIONS: The apex technique appears to be a safe, quickly performed, and low-cost method for the treatment of rectal intussusception. In this series, imaging examinations showed the disappearance of rectal intussusception, and a significant decrease in constipation score suggested improvement in functional outcomes.


Assuntos
Constipação Intestinal/cirurgia , Intussuscepção/cirurgia , Prolapso Retal/cirurgia , Grampeamento Cirúrgico/métodos , Adulto , Idoso , Brasil , Constipação Intestinal/etiologia , Constipação Intestinal/patologia , Feminino , Humanos , Intussuscepção/complicações , Intussuscepção/patologia , Masculino , Pessoa de Meia-Idade , Prolapso Retal/complicações , Prolapso Retal/patologia , Estudos Retrospectivos , Síndrome , Resultado do Tratamento
10.
Immunobiology ; 217(6): 634-42, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22101184

RESUMO

Toll-like receptor 2 (TLR2) is a type I pattern recognition receptor that has been shown to participate in intestinal homeostasis. Its increased expression in the lamina propria has been associated with the pathogenesis in inflammatory bowel disease (IBD), such as ulcerative colitis (UC) and Crohn's disease (CD). Recently, soluble TLR2 (sTLR2) variants have been shown to counteract inflammatory responses driven by the cognate receptor. Despite the evident roles of TLR2 in intestinal immunity, no study has elucidated the production and cellular source of sTLR2 in IBD. Furthermore, an increase in the population of activated macrophages expressing TLR2 that infiltrates the intestine in IBD has been reported. We aimed first to assess the production of the sTLR2 by UC and CD organ culture biopsies and lamina propria mononuclear cells (LPMCs) as well as the levels of sTLR2 in serum, and then characterize the cell population from lamina propria producing the soluble protein. Mucosa explants, LPMCs and serum were obtained from UC, CD patients and control subjects. The level of sTLR2 was higher in conditioned media from organ culture biopsies and LPMCs from UC patients in comparison to CD and controls. Moreover, an inverse correlation between the content of intestinal and serum sTLR2 levels was observed in UC patients. Additionally, when characterizing the cellular source of the increased sTLR2 by LPMCs from UC patients, an increase in TLR2(+)/CD33(+) cell population was found. Also, these cells expressed CX3CR1, which was related to the increased levels of intestinal FKN in UC patients, suggesting that a higher proportion of TLR2(+) mononuclear cells infiltrate the lamina propria. The increased production of sTLR2 suggests that a differential regulating factor of the innate immune system is present in the intestinal mucosa of UC patients.


Assuntos
Colite Ulcerativa/metabolismo , Doença de Crohn/metabolismo , Leucócitos Mononucleares/metabolismo , Membrana Mucosa/metabolismo , Receptor 2 Toll-Like/biossíntese , Adulto , Idoso , Idoso de 80 Anos ou mais , Biópsia , Receptor 1 de Quimiocina CX3C , Movimento Celular/imunologia , Quimiocina CX3CL1/genética , Quimiocina CX3CL1/imunologia , Colite Ulcerativa/imunologia , Colite Ulcerativa/patologia , Doença de Crohn/imunologia , Doença de Crohn/patologia , Feminino , Expressão Gênica , Humanos , Imunidade Inata , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/patologia , Masculino , Pessoa de Meia-Idade , Membrana Mucosa/imunologia , Membrana Mucosa/patologia , Receptores de Quimiocinas/genética , Receptores de Quimiocinas/imunologia , Lectina 3 Semelhante a Ig de Ligação ao Ácido Siálico/genética , Lectina 3 Semelhante a Ig de Ligação ao Ácido Siálico/imunologia , Solubilidade , Técnicas de Cultura de Tecidos
12.
DNA Cell Biol ; 25(12): 684-95, 2006 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-17233117

RESUMO

High levels of the pro-inflammatory cytokines, interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha), are present in the gut mucosa of patients suffering form various diseases, most notably inflammatory bowel diseases (IBD). Since the inflammatory milieu can cause important alterations in epithelial cell function, we examined the cytokine effects on the expression of the enterocyte differentiation marker, intestinal alkaline phosphatase (IAP), a protein that detoxifies bacterial lipopolysaccharides (LPS) and limits fat absorption. Sodium butyrate (NaBu), a short-chain fatty acid and histone deacetylase (HDAC) inhibitor, was used to induce IAP expression in HT-29 cells and the cells were also treated +/- the cytokines. Northern blots confirmed IAP induction by NaBu, however, pretreatment (6 h) with either cytokine showed a dose-dependent inhibition of IAP expression. IAP Western analyses and alkaline phosphatase enzyme assays corroborated the Northern data and confirmed that the cytokines inhibit IAP induction. Transient transfections with a reporter plasmid carrying the human IAP promoter showed significant inhibition of NaBu-induced IAP gene activation by the cytokines (100 and 60% inhibition with IL-1beta and TNF-alpha, respectively). Western analyses showed that NaBu induced H4 and H3 histone acetylation, and pretreatment with IL-1beta or TNF-alpha did not change this global acetylation pattern. In contrast, chromatin immunoprecipitation showed that local histone acetylation of the IAP promoter region was specifically inhibited by either cytokine. We conclude that IL-1beta and TNF-alpha inhibit NaBu-induced IAP gene expression, likely by blocking the histone acetylation within its promoter. Cytokine-mediated IAP gene silencing may have important implications for gut epithelial function in the setting of intestinal inflammatory conditions.


Assuntos
Antígenos de Neoplasias/metabolismo , Interleucina-1beta/farmacologia , Fator de Necrose Tumoral alfa/farmacologia , Acetilação/efeitos dos fármacos , Acetiltransferases/efeitos dos fármacos , Fosfatase Alcalina , Butiratos/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Proteínas Ligadas por GPI , Regulação da Expressão Gênica/efeitos dos fármacos , Inativação Gênica/efeitos dos fármacos , Genes Reporter/efeitos dos fármacos , Células HCT116 , Células HT29 , Histona Acetiltransferases/efeitos dos fármacos , Humanos , Mediadores da Inflamação/farmacologia , Luciferases/metabolismo , Regiões Promotoras Genéticas/efeitos dos fármacos , Ativação Transcricional
13.
Dis Colon Rectum ; 48(2): 335-40; discussion 340-3, 2005 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-15812585

RESUMO

PURPOSE: The mayor symptoms of chronic anal fissure are permanent pain, intense pain during defecation that lasts for hours, blood in the stools, and sphincter cramps. It is subsequent to formation of fibrosis infiltrate that leads to an increased anal tone with poor healing tendency. This vicious circle leads to fissure recurrence and chronicity. This study was designed to show the efficacy of gonyautoxin infiltration in healing patients with anal fissures. METHODS: Gonyautoxin is a paralyzing phytotoxin produced by dinoflagellates. Fifty recruited patients received clinical examination, including proctoscopy and questionnaire to evaluate the symptoms. Anorectal manometries were performed before and after toxin injection. Doses of 100 units of gonyautoxin in a volume of 1 ml were infiltrated into both sides of the anal fissure in the internal anal sphincter. RESULTS: Total remission of acute and chronic anal fissures were achieved within 15 and 28 days respectively. Ninety-eight percent of the patients healed before 28 days with a mean time healing of 17.6 +/- 9 days. Only one relapsed during 14 months of follow-up. Neither fecal incontinence nor other side effects were observed. All patients showed immediate sphincter relaxation. The maximum anal resting pressures recorded after two minutes decreased to 56.2 +/- 12.5 percent of baseline. CONCLUSIONS: Gonyautoxin breaks the vicious circle of pain and spasm that leads to anal fissure. This study proposes gonyautoxin anal sphincter infiltration as safe and effective alternative therapeutic approach to conservative, surgical, and botulinum toxin therapies for anal fissures.


Assuntos
Fissura Anal/tratamento farmacológico , Toxinas Marinhas/uso terapêutico , Saxitoxina/análogos & derivados , Saxitoxina/uso terapêutico , Doença Aguda , Adolescente , Adulto , Idoso , Animais , Doença Crônica , Dinoflagelados , Método Duplo-Cego , Feminino , Humanos , Masculino , Manometria , Pessoa de Meia-Idade , Medição da Dor , Resultado do Tratamento
14.
Mol Endocrinol ; 18(8): 1941-62, 2004 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-15143152

RESUMO

Thyroid hormone (T3) is a critical regulator of intestinal epithelial development and homeostasis, but its mechanism of action within the gut is not well understood. We have examined the molecular mechanisms underlying the T3 activation of the enterocyte differentiation marker intestinal alkaline phosphatase (IAP) gene. RT-PCR and Western blotting showed that thyroid hormone receptors TRalpha1 and TRbeta1 were expressed in human colorectal adenocarcinoma Caco-2 cells. Northern blotting detected expression of two IAP transcripts, which were increased approximately 3-fold in response to T3. Transient transfection studies with luciferase reporter plasmids carrying various internal and 5' deletion mutations of the IAP promoter localized a putative thyroid hormone response element (TRE) to a region approximately 620 nucleotides upstream (-620) of the ATG start codon. EMSAs using TRalpha1-retinoid X receptor alpha (RXRalpha) on sequential 5' and 3' single nucleotide deletions defined the TRE between -632 and -612 (5'-TTGAACTCAgccTGAGGTTAC-3'). Compared with the consensus TRE, the IAP-TRE is novel in that it contains an everted repeat of two nonamers (not hexamers) separated by three nucleotides. Neither TRalpha1 nor RXRalpha binds to the IAP-TRE; however, TRbeta1 binds to this TRE with minimal affinity. In the presence of TR and RXRalpha, only the TR-RXRalpha heterodimer binds to the IAP-TRE. Mutagenesis of either nonamer abolishes the biological activity of IAP promoter. We have thus identified a novel response element that appears to mediate the T3-induced activation of the enterocyte differentiation marker, intestinal alkaline phosphatase.


Assuntos
Fosfatase Alcalina/genética , Antígenos de Neoplasias/genética , Diferenciação Celular/efeitos dos fármacos , Enterócitos/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Elementos de Resposta/genética , Hormônios Tireóideos/farmacologia , Fosfatase Alcalina/metabolismo , Anticorpos/imunologia , Antígenos de Neoplasias/metabolismo , Biomarcadores , Células CACO-2 , Núcleo Celular/metabolismo , Enterócitos/citologia , Enterócitos/enzimologia , Enterócitos/metabolismo , Proteínas Ligadas por GPI , Genes Reporter/genética , Humanos , Ligantes , Luciferases/genética , Luciferases/metabolismo , Mutação/genética , Nucleotídeos/genética , Ligação Proteica , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Receptores X Retinoide/metabolismo , Especificidade por Substrato , Receptores alfa dos Hormônios Tireóideos/genética , Receptores alfa dos Hormônios Tireóideos/imunologia , Receptores alfa dos Hormônios Tireóideos/metabolismo , Receptores beta dos Hormônios Tireóideos/genética , Receptores beta dos Hormônios Tireóideos/metabolismo
15.
Am J Physiol Gastrointest Liver Physiol ; 286(1): G23-30, 2004 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-12919939

RESUMO

We have examined the role that the transcription factor gut-enriched Krüppel-like factor (KLF4 or GKLF) plays in activating the enterocyte differentiation marker gene intestinal alkaline phosphatase (IAP). A yeast one-hybrid screen was used to identify proteins interacting with a previously identified cis-element (IF-III) located within the human IAP gene promoter. DNA-protein interactions were determined by using EMSA. Northern blot analysis was used to study RNA expression in human colon cancer RKO cells engineered to overexpress KLF4. Transient transfections with IAP-luciferase reporter constructs were used to characterize the mechanisms by which KLF4 activates IAP transcription. The yeast one-hybrid screen and EMSA identified KLF4 as binding to IF-III. RKO cells induced to overexpress KLF4 demonstrated a corresponding dose-dependent increase in IAP expression, and EMSA with nuclear extract from these cells confirmed that KLF4 binds to the IF-III element. Transient transfections revealed that KLF4 transactivated the IAP gene largely via a critical segment in the IAP promoter that includes the IF-III cis-element. Mutant KLF4 constructs failed to fully activate IAP. We have identified the enterocyte differentiation marker IAP as a KLF4 target gene. IAP transactivation by KLF4 is likely mediated through a critical region located within the proximal IAP promoter region.


Assuntos
Fosfatase Alcalina/genética , Fosfatase Alcalina/metabolismo , Proteínas de Ligação a DNA/fisiologia , Enterócitos/fisiologia , Intestinos/enzimologia , Fatores de Transcrição/fisiologia , Animais , Autorradiografia , Biomarcadores , Northern Blotting , Células COS , Diferenciação Celular/genética , Diferenciação Celular/fisiologia , Linhagem Celular , Ensaio de Desvio de Mobilidade Eletroforética , Humanos , Fatores de Transcrição Kruppel-Like , Luciferases/genética , Regiões Promotoras Genéticas/genética , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Saccharomyces cerevisiae/enzimologia , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Ativação Transcricional/genética , Transfecção
17.
J Gastrointest Surg ; 7(8): 1053-61; discussion 1061, 2003 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-14675715

RESUMO

The gut-enriched Krüppel-like factor (KLF4) and the ligand-bound thyroid hormone receptor (TR) have each been shown to play a critical role in mammalian gut development and differentiation. We investigated an interrelationship between these two presumably independent pathways using the differentiation marker gene, intestinal alkaline phosphatase (IAP). Transient transfections were performed in Cos-7 cells using luciferase reporter plasmids containing a 2.5 kb segment of the proximal human IAP 5' regulatory region, as well as multiple deletions. Cells were cotransfected with TR and/or KLF4 expression vectors and treated+/-100 nmol/L thyroid hormone (T3). IAP reporter gene transactivation was increased independently by KLF4 (ninefold) and ligand-bound TR beta 1 (sevenfold). Cells cotransfected with KLF4 and TR beta 1 in the presence of T3 showed synergistic activation (70-fold). A similar pattern was seen with the other T3 receptor isoform, TR alpha 1. The synergistic effect was lost with deletions of the T3 and KLF4 response elements in the IAP promoter and was completely or partially abolished in the case of mutant KLF4 expression vectors. The thyroid hormone receptor complex and KLF4 synergistically activate the enterocyte differentiation marker gene IAP, suggesting a previously unrecognized interrelationship between these two transcription factor pathways.


Assuntos
Fosfatase Alcalina/genética , Antígenos de Neoplasias/genética , Biomarcadores Tumorais/genética , Proteínas de Ligação a DNA/genética , Fatores de Transcrição/genética , Tri-Iodotironina/genética , Animais , Células COS , Diferenciação Celular/genética , Enterócitos/fisiologia , Proteínas Ligadas por GPI , Humanos , Fatores de Transcrição Kruppel-Like , Transdução de Sinais/genética
18.
J Gastrointest Surg ; 7(2): 237-44; discussion 244-5, 2003 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-12600448

RESUMO

Enterocyte differentiation is thought to occur through the transcriptional regulation of a small subset of specific genes. A recent growing body of evidence indicates that post-translational modifications of chromatin proteins (histones) play an important role in the control of gene transcription. Previous work has demonstrated that one such modification, histone acetylation, occurs in an in vitro model of enterocyte differentiation, butyrate-treated HT-29 cells. In the present work, we sought to determine if the epigenetic signal of histone acetylation occurs in an identifiable pattern in association with the transcriptional activation of the enterocyte differentiation marker gene intestinal alkaline phosphatase (IAP). HT-29 cells were maintained under standard culture conditions and differentiated with sodium butyrate. The chromatin immunoprecipitation (ChIP) assay was used to compare the acetylation state of histones associated with specific regions of the IAP promoter in the two cell populations (undifferentiated vs. differentiated). Chromatin was extracted from cells and cleaved by sonication or enzymatic digestion to obtain fragments of approximately 200 to 600 base-pairs, as confirmed by polymerase chain reaction using primers designed to amplify the IAP segments of interest. The ChIP assay selects DNA sequences that are associated with acetylated histones by immunoprecipitation. Unbound segments represent DNA sequences whose histones are not acetylated. After immunoprecipitation, sequences were detected by radiolabeled polymerase chain reaction, and the relative intensity of the bands was quantified by densitometry. The relative acetylation state of histones at specific sites was determined by comparing the ratios of bound/unbound segments. We determined that in a segment of the IAP promoter between -378 and -303 base-pairs upstream from the transcriptional start site, the acetylation state of histone H3 increased twofold in the differentiated, IAP expressing cells, whereas that of histone H4 remained essentially constant. Additionally, at a distant site, between -1378 and -1303 base-pairs, the acetylation state of H3 and H4 did not change appreciably between the undifferentiated and differentiated cells. We conclude that butyrate-induced differentiation is associated with specific and localized changes in the histone acetylation state within the IAP promoter. These changes within the endogenous IAP gene may underlie its transcriptional activation in the context of the enterocyte differentiation program.


Assuntos
Fosfatase Alcalina/metabolismo , Antígenos de Neoplasias/metabolismo , Cromatina/metabolismo , Enterócitos/fisiologia , Histonas/genética , Regiões Promotoras Genéticas , Ativação Transcricional , Sequência de Bases , Divisão Celular/genética , Proteínas Ligadas por GPI , Marcadores Genéticos/genética , Células HT29 , Histonas/metabolismo , Humanos , Técnicas In Vitro , Região de Controle de Locus Gênico , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Testes de Precipitina , Sensibilidade e Especificidade
20.
Rev. chil. cir ; 50(6): 637-41, dic. 1998. graf
Artigo em Espanhol | LILACS | ID: lil-243817

RESUMO

Se presenta una serie de 34 pacientes operados por cáncer del recto con operación de Miles, sin evidencias de recidiva y con seguimiento mínimo de 18 meses. En ellos se analizó la función urinaria, sexual y el impacto que esta operación significó en la relación de los pacientes con el medio ambiente familiar y laboral. Se puede concluir que el grupo de pacientes sufre consecuencias permanentes en todas las funciones investigadas, las que afectan a un porcentaje mayoritario de ellos


Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Procedimentos Cirúrgicos Urológicos/métodos , Neoplasias Retais/cirurgia , Colostomia , Ereção Peniana/fisiologia , Fenômenos Fisiológicos do Sistema Urinário , Libido/fisiologia , Estomia , Complicações Pós-Operatórias , Comportamento Sexual/fisiologia , Incontinência Urinária/reabilitação
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