ARE THERE ADVANTAGES IN DOUBLE TRANSIT RECONSTRUCTION AFTER TOTAL GASTRECTOMY IN PATIENTS WITH GASTRIC CANCER? A SYSTEMATIC REVIEW.

BACKGROUND: Curative treatment for gastric cancer involves tumor resection, followed by transit reconstruction, with Roux-en-Y being the main technique employed. To permit food transit to the duodenum, which is absent in Roux-en-Y, double transit reconstruction has been used, whose theoretical advantages seem to surpass the previous technique. AIMS: To compare the clinical evolution of gastric cancer patients who underwent total gastrectomy with Roux-en-Y and double tract reconstruction. METHODS: A systematic review was carried out on Web of Science, Scopus, EmbasE, SciELO, Virtual Health Library, PubMed, Cochrane, and Google Scholar databases. Data were collected until June 11, 2022. Observational studies or clinical trials evaluating patients submitted to double tract (DT) and Roux-en-Y (RY) reconstructions were included. There was no temporal or language restriction. Review articles, case reports, case series, and incomplete texts were excluded. The risk of bias was calculated using the Cochrane tool designed for randomized clinical trials. RESULTS: Four studies of good methodological quality were included, encompassing 209 participants. In the RY group, there was a greater reduction in food intake. In the DT group, the decrease in body mass index was less pronounced compared to preoperative values. CONCLUSIONS: The double tract reconstruction had better outcomes concerning body mass index and the time until starting a light diet; however, it did not present any advantages in relation to nutritional deficits, quality of life, and post-surgical complications.

Are there advantages in double transit reconstruction after total gastrectomy in patients with gastric cancer? a systematic review / Há vantagens na reconstrução em duplo trânsito após a gastrectomia total em portadores de câncer gástrico? revisão sistemática

ABSTRACT BACKGROUND: Curative treatment for gastric cancer involves tumor resection, followed by transit reconstruction, with Roux-en-Y being the main technique employed. To permit food transit to the duodenum, which is absent in Roux-en-Y, double transit reconstruction has been used, whose theoretical advantages seem to surpass the previous technique. AIMS: To compare the clinical evolution of gastric cancer patients who underwent total gastrectomy with Roux-en-Y and double tract reconstruction. METHODS: A systematic review was carried out on Web of Science, Scopus, EmbasE, SciELO, Virtual Health Library, PubMed, Cochrane, and Google Scholar databases. Data were collected until June 11, 2022. Observational studies or clinical trials evaluating patients submitted to double tract (DT) and Roux-en-Y (RY) reconstructions were included. There was no temporal or language restriction. Review articles, case reports, case series, and incomplete texts were excluded. The risk of bias was calculated using the Cochrane tool designed for randomized clinical trials. RESULTS: Four studies of good methodological quality were included, encompassing 209 participants. In the RY group, there was a greater reduction in food intake. In the DT group, the decrease in body mass index was less pronounced compared to preoperative values. CONCLUSIONS: The double tract reconstruction had better outcomes concerning body mass index and the time until starting a light diet; however, it did not present any advantages in relation to nutritional deficits, quality of life, and post-surgical complications.

RESUMO RACIONAL: O tratamento curativo do câncer gástrico envolve a ressecção do tumor, seguida de reconstrução do trânsito, sendo o Y-de-Roux a principal técnica empregada. Para permitir o trânsito alimentar para o duodeno, ausente em Y-de-Roux, tem-se utilizado a reconstrução de duplo trânsito, cujas vantagens teóricas parecem superar a técnica anterior. OBJETIVOS: Comparar a evolução clínica de pacientes com câncer gástrico submetidos à gastrectomia total com Y-de-Roux e reconstrução de duplo trânsito. MÉTODOS: Foi realizada uma revisão sistemática nas bases de dados: Web of Science, Scopus, Embase, Scielo, Biblioteca Virtual em Saúde, PubMed e Cochrane. Os dados foram coletados até 11 de junho de 2022. Foram incluídos estudos observacionais ou ensaios clínicos avaliando pacientes que utilizaram reconstruções de duplo trânsito (DT) e Y-de-Roux (RY). Não houve restrição temporal ou de idioma. Foram excluídos artigos de revisão, relatos de casos, séries de casos e aqueles com texto incompleto. O risco de viés foi calculado utilizando a ferramenta Cochrane desenvolvida para ensaios clínicos randomizados. RESULTADOS: Foram incluídos quatro estudos de boa qualidade metodológica, abrangendo 209 participantes. No grupo RY houve maior redução na ingestão alimentar. No grupo DT, a diminuição do índice de massa corporal (IMC) foi menos pronunciada em comparação aos valores pré-operatórios. CONCLUSÕES: A reconstrução de duplo trânsito apresentou melhores resultados em relação ao índice de massa corporal e ao tempo para início de dieta leve, porém não apresentou vantagens em relação aos déficits nutricionais, qualidade de vida e complicações pós-cirúrgicas.

Endoscopic and histopathological assessment of individuals undergoing one anastomosis gastric bypass: a 2-year follow-up.

BACKGROUND: There is much debate over the occurrence of biliary reflux to the gastric pouch after one anastomosis gastric bypass (OAGB) and its potential risks. OBJECTIVE: To assess endoscopic and histopathological findings following a standardized protocol of biopsy collection two years after OAGB. METHODS: A historical cohort study was conducted, based on a prospectively collected database, which involved 39 participants who underwent OAGB. Participants underwent clinical evaluation and esophagogastroduodenoscopy at the time of surgery and 24 months afterward. Post-operatively, biopsy specimens in esophagogastric junction, pouch, and anastomosis were systematically collected. RESULTS: 92.3% of the participants were female and the mean age was 37 ± 8.5 years. The mean body mass index (BMI) significantly decreased from 37.6 ± 5.7 kg/m2 to 27 ± 4.1 kg/m2 after 2 years (p < 0.001). The mean %TWL was 27.2 ± 10.5%. The prevalence of non-erosive gastritis significantly increased from 25.6 to 51.3% (p = 0.02). Erosive gastritis significantly decreased from 28.2 to 10.3% (p = 0.04). Four cases of marginal ulcers were identified (10.3%). The commonest histopathological finding was mild inflammation in 74.3% (esophagogastric junction), 58.9% (pouch), and 71.8% (anastomosis). There was one case of focal intestinal metaplasia in each site of interest and no cases of dysplasia or severe inflammation. CONCLUSIONS: Using a standardized protocol of post-operative biopsy collection, low rates of severe endoscopic and histopathological abnormalities were observed two years after OAGB. Nevertheless, as most patients have histologically proven inflammation, bile in the gastric pouch, and endoscopic gastritis, long-term surveillance is essential because of the uncertain risk of these abnormalities.

EFFICACY ANALYSIS OF ENDOSCOPIC SUBMUCOSAL DISSECTION FOR THE EARLY GASTRIC CANCER AND PRECANCEROUS LESIONS.

BACKGROUND: Endoscopic treatment of precancerous lesions and early gastric cancer has been widely accepted in recent years. Endoscopic submucosal dissection (ESD), following established indication criteria, can lead to cure of the disease in more than 90% of cases. OBJECTIVE: This study aimed to analyze the use of ESD in patients with early gastric cancer and precancerous lesions, as well as the results of the procedure, its complications and effectiveness in controlling the disease. METHODS: This is a retrospective cohort study composed of 41 patients aged from 53 to 87 years (mean age: 65 years; 58.53% male) who were analyzed from 2008 to 2019. The variables collected from the medical records were: comorbidities, classification of the lesion regarding resection criteria, type of resection, histology, degree of invasion, resection margin, complications, disease recurrence. Statistical analysis was performed using the Kruskal-Wallis test, the McNemar's test, and the Mann-Whitney test, with 5% statistical significance (P<0.05). RESULTS: The most frequent site of the lesion was the gastric antrum and the predominant presentation by the Japanese or Paris classification was the one with depressed components in 56.09%. Adenocarcinoma occurred in 75.6% of the biopsies, and the remainder were adenomas without neoplasia. En-bloc resection occurred in 97.57% of cases, and compromise of the safety margin occurred in one patient. The main pre-existing comorbidity was liver cirrhosis in 29.26% of cases. There was a significant increase in post- ESD adenocarcinoma compared to pre-resection diagnosis. The mean follow-up time was 38.4 months, with one recurrence (2.43%) and two metachronous lesions (4.87%). Complications during and after the procedure occurred in three patients (7.31%), being due to bleeding (two cases) and perforation (one case). There was one death due to a cardiac event not directly related to the procedure. CONCLUSION: Endoscopic submucosal dissection proved to be a safe procedure, with a low complication and recurrence rate. Its recommendation must occur within the established criteria; however, it can be indicated for patients outside the criteria, if there is a high risk for surgical treatment.

Análise de eficácia da dissecção endoscópica da submucosa para o câncer gástrico precoce e lesões pré-cancerosas / Efficacy analysis of endoscopic submucosal dissection for the early gastric cancer and precancerous lesions

ABSTRACT Background: Endoscopic treatment of precancerous lesions and early gastric cancer has been widely accepted in recent years. Endoscopic submucosal dissection (ESD), following established indication criteria, can lead to cure of the disease in more than 90% of cases. Objective: This study aimed to analyze the use of ESD in patients with early gastric cancer and precancerous lesions, as well as the results of the procedure, its complications and effectiveness in controlling the disease. Methods: This is a retrospective cohort study composed of 41 patients aged from 53 to 87 years (mean age: 65 years; 58.53% male) who were analyzed from 2008 to 2019. The variables collected from the medical records were: comorbidities, classification of the lesion regarding resection criteria, type of resection, histology, degree of invasion, resection margin, complications, disease recurrence. Statistical analysis was performed using the Kruskal-Wallis test, the McNemar's test, and the Mann-Whitney test, with 5% statistical significance (P<0.05). Results: The most frequent site of the lesion was the gastric antrum and the predominant presentation by the Japanese or Paris classification was the one with depressed components in 56.09%. Adenocarcinoma occurred in 75.6% of the biopsies, and the remainder were adenomas without neoplasia. En-bloc resection occurred in 97.57% of cases, and compromise of the safety margin occurred in one patient. The main pre-existing comorbidity was liver cirrhosis in 29.26% of cases. There was a significant increase in post- ESD adenocarcinoma compared to pre-resection diagnosis. The mean follow-up time was 38.4 months, with one recurrence (2.43%) and two metachronous lesions (4.87%). Complications during and after the procedure occurred in three patients (7.31%), being due to bleeding (two cases) and perforation (one case). There was one death due to a cardiac event not directly related to the procedure. Conclusion: Endoscopic submucosal dissection proved to be a safe procedure, with a low complication and recurrence rate. Its recommendation must occur within the established criteria; however, it can be indicated for patients outside the criteria, if there is a high risk for surgical treatment.

RESUMO Contexto: O tratamento endoscópico das lesões pré-cancerosas e do câncer gástrico precoce tem sido amplamente aceito nos últimos anos. A dissecção endoscópica da submucosa (submucosectomia), obedecendo a critérios estabelecidos de indicação, pode levar a cura da doença em mais de 90% dos casos. Objetivo: Este estudo teve como objetivo analisar o uso da dissecção submucosa endoscópica em pacientes com câncer gástrico precoce e lesões pré-cancerosas, bem como os resultados do procedimento, suas complicações e eficácia no controle da doença. Métodos: Foram analisados 41 pacientes, com idade variando de 53 a 87 anos (média de 65 anos), sendo 58,53% do sexo masculino, no período de 2008 a 2019, sendo este estudo do tipo coorte retrospectivo. As variáveis coletadas dos prontuários foram: comorbidades, classificação da lesão quanto aos critérios de ressecção, tipo de ressecção, histologia, grau de invasão, margem de resseção, complicações, recidiva de doença. A análise estatística foi feita com o uso do teste de Kruskal-Wallis, teste de McNemar e teste de Mann-Whitney, com significância estatística de 5% (P<0,05). Resultados: O local mais frequente da lesão foi o antro gástrico e a apresentação predominante pela classificação japonesa ou de Paris foram os com componentes deprimidos em 56,09%. O adenocarcinoma ocorreu em 75,6% das biópsias e o restante foram adenomas sem neoplasia. A ressecção em bloco ocorreu em 97,57% dos casos, e o comprometimento da margem de segurança ocorreu em um paciente. A principal comorbidade pré-existente foi a cirrose hepática em 29,26% dos casos. Houve um aumento significativo de adenocarcinoma após dissecção endoscópica da submucosa em comparação ao diagnóstico pré ressecção. O tempo médio de seguimento foi de 38,4 meses, sendo registrado uma recidiva (2,43%) e duas lesões metacrônicas (4,87%). As complicações durante e após o procedimento ocorreram em 3 (7,31%) pacientes, sendo por sangramento (dois casos) e perfuração (um caso). Houve um óbito por evento cardiológico, não relacionado diretamente com o procedimento. Conclusão: A ressecção endoscópica da submucosa mostrou ser procedimento seguro, com baixa taxa de complicação e de recidiva. A sua indicação deve ser dentro dos critérios estabelecidos, entretanto, pode ser indicada em pacientes fora de critérios, se há alto risco para o tratamento cirúrgico.

THE ESOPHAGEAL, GASTRIC, AND COLORECTAL TUMORS AND THE ESOPHAGOGASTRODUODENOSCOPIES AND COLONOSCOPIES BY THE BRAZILIAN UNIFIED HEALTH SYSTEM: WHAT IS THE IMPORTANCE?

AIM: Esophagogastroduodenoscopies and colonoscopies are the main diagnostic examinations for esophageal, stomach, and colorectal tumors. This study aimed to evaluate the estimates of the incidence of esophageal, stomach, and colorectal cancer; population growth; and esophagogastroduodenoscopies and colonoscopies performed by the Unified Health System (SUS), from 2010 to 2018, in the five regions of the country, and to analyze the relationship between these values. RESULTS: The colorectal tumor had a significant elevation, while the esophageal and gastric maintained the incidences. In the five regions, there was a significant increase in the number of colonoscopies; however, this increase did not follow the increase in the population in the North and Northeast regions. There was no significant increase in the number of esophagogastroduodenoscopies in the North, Northeast, Midwest, and South regions, and in the North region there was a decrease. In the Northeast region, there was a decreasing number, and in the South and Midwest regions, the number of examinations remained stable in the period. The Southeast region recorded an increase in the number of examinations following the population growth. CONCLUSION: The current number of esophagogastroduodenoscopies and colonoscopies performed by the SUS did not follow the population growth, in order to attend the population and diagnose esophageal, stomach, and colorectal tumors. Therefore, the country needs to have adequate and strategic planning on how it will meet the demand for these tests and serve the population well, incorporating new technologies.

The esophageal, gastric, and colorectal tumors and the esophagogastroduodenoscopies and colonoscopies by the brazilian unified health system: what is the importance? / Os tumores esofágicos, gástricos e colorretais e as esofagogastroduodenoscopias e colonoscopias pelo sistema único de saúde (sus): qual a importância?

ABSTRACT - BACKGROUND: Esophagogastroduodenoscopies and colonoscopies are the main diagnostic examinations for esophageal, stomach, and colorectal tumors. AIM: This study aimed to evaluate the estimates of the incidence of esophageal, stomach, and colorectal cancer; population growth; and esophagogastroduodenoscopies and colonoscopies performed by the Unified Health System (SUS), from 2010 to 2018, in the five regions of the country, and to analyze the relationship between these values. RESULTS: The colorectal tumor had a significant elevation, while the esophageal and gastric maintained the incidences. In the five regions, there was a significant increase in the number of colonoscopies; however, this increase did not follow the increase in the population in the North and Northeast regions. There was no significant increase in the number of esophagogastroduodenoscopies in the North, Northeast, Midwest, and South regions, and in the North region there was a decrease. In the Northeast region, there was a decreasing number, and in the South and Midwest regions, the number of examinations remained stable in the period. The Southeast region recorded an increase in the number of examinations following the population growth. CONCLUSION: The current number of esophagogastroduodenoscopies and colonoscopies performed by the SUS did not follow the population growth, in order to attend the population and diagnose esophageal, stomach, and colorectal tumors. Therefore, the country needs to have adequate and strategic planning on how it will meet the demand for these tests and serve the population well, incorporating new technologies.

RESUMO - RACIONAL: Os tumores de esôfago, estômago e colorretal têm como principal exame diagnóstico as esofagogastroduodenoscopias e colonoscopias. OBJETIVO: Avaliar as estimativas de incidências de câncer de esôfago, estômago e colorretal, o crescimento populacional, e as esofagogastroduodenoscopias e colonoscopias realizadas pelo Sistema Único de Saúde (SUS) de 2010 a 2018, nas cinco regiões do país e a relação entre esses valores. RESULTADOS: O tumor colorretal teve elevação significativa, enquanto os esofágicos e gástricos mantiveram as incidências. Nas cinco regiões foi registrado elevação significativa do número de colonoscopias, entretanto, essa elevação não acompanhou a elevação da população nas regiões Norte e Nordeste. Não ocorreu elevação significativa do número de esofagogastroduodenoscopias nas regiões Norte, Nordeste, Centro-Oeste e Sul e na região Norte ocorreu diminuição. Na região Nordeste ocorreu número decrescente e nas regiões Sul e Centro-Oeste o número de exames manteve-se estável no período. A região Sudeste registrou elevação do número de exames acompanhando o crescimento população. CONCLUSÃO: O número atual de esofagogastroduodenoscopias e colonoscopias realizadas pelo SUS, não acompanhou o crescimento populacional, para atender a população e diagnosticar os tumores de esôfago, estômago e colorretais. Portanto, o país necessita ter um planejamento adequado e estratégico de como irá suprir a demanda desses exames e bem atender a população, incorporando as novas tecnologias.

Surgical treatment of relapsed megaesophagus.

OBJECTIVE: to analyze the surgical treatment of patients with recurrent megaesophagus followed at the esophageal-stomach-duodenal outpatient clinic of the Hospital de Clínicas - UNICAMP. METHODS: a retrospective study, from 2011 to 2017, with 26 patients with Chagas or idiopathic megaesophagus, surgically treated, and who recurred with dysphagia. Clinical, endoscopic and radiographic aspects were assessed and correlated with the performed surgical procedures. RESULTS: 50% had dysphagia for liquids, 69% regurgitation, 65.3% heartburn, 69.2% weight loss and 69.2% had Chagas disease. In addition, 38.4% had megaesophagus stage 1 and 2 and 61.5% stage 3 and 4. Regarding the reoperations, 53% of them underwent Heller-Pinotti surgery by laparoscopy, Serra-Dória in 30.7% and esophageal mucosectomy in 7.9%. In 72% of the reoperations there were no postoperative complications, and 80% of the patients had a good outcome, with reduction or elimination of dysphagia. Among the reoperated patients undergoing the laparoscopic Heller-Pinotti technique, three reported little improvement of dysphagia in the postoperative period and among those who underwent Serra-Dória surgery, 100% had no dysphagia. It was observed that, when the time between the first procedure and the reoperation was longer, the better the surgical result was, with statistical significant decreased dysphagia (p=0.0013, p<0.05). CONCLUSIONS: there was a preference to perform laparoscopic re-miotomy and, as a second option, Serra-Dória surgery, for patients with recurrent megaesophagus. Esophagectomy or esophageal mucosectomy were reserved for more severe patients.

Surgical treatment of relapsed megaesophagus / Tratamento cirúrgico do megaesôfago recidivado

ABSTRACT Objective: to analyze the surgical treatment of patients with recurrent megaesophagus followed at the esophageal-stomach-duodenal outpatient clinic of the Hospital de Clínicas - UNICAMP. Methods: a retrospective study, from 2011 to 2017, with 26 patients with Chagas or idiopathic megaesophagus, surgically treated, and who recurred with dysphagia. Clinical, endoscopic and radiographic aspects were assessed and correlated with the performed surgical procedures. Results: 50% had dysphagia for liquids, 69% regurgitation, 65.3% heartburn, 69.2% weight loss and 69.2% had Chagas disease. In addition, 38.4% had megaesophagus stage 1 and 2 and 61.5% stage 3 and 4. Regarding the reoperations, 53% of them underwent Heller-Pinotti surgery by laparoscopy, Serra-Dória in 30.7% and esophageal mucosectomy in 7.9%. In 72% of the reoperations there were no postoperative complications, and 80% of the patients had a good outcome, with reduction or elimination of dysphagia. Among the reoperated patients undergoing the laparoscopic Heller-Pinotti technique, three reported little improvement of dysphagia in the postoperative period and among those who underwent Serra-Dória surgery, 100% had no dysphagia. It was observed that, when the time between the first procedure and the reoperation was longer, the better the surgical result was, with statistical significant decreased dysphagia (p=0.0013, p<0.05). Conclusions: there was a preference to perform laparoscopic re-miotomy and, as a second option, Serra-Dória surgery, for patients with recurrent megaesophagus. Esophagectomy or esophageal mucosectomy were reserved for more severe patients.

RESUMO Objetivo: analisar o tratamento cirúrgico dos pacientes com megaesôfago recidivado acompanhados no ambulatório de cirurgia de esôfago-estômago-duodeno do Hospital de Clínicas da UNICAMP. Métodos: estudo restrospectivo no período de 2011 a 2017, com 26 pacientes portadores de megaesôfago chagásico ou idiopático, tratados cirurgicamente e que evoluíram com recidiva da disfagia. Foram avaliados aspectos clínicos, endoscópicos e radiográficos, sendo correlacionados com os procedimentos cirúrgicos realizados. Resultados: 50% apresentava disfagia para líquidos, 69% regurgitação, 65,3% pirose, 69,2% perda de peso e 69,2% era chagásico. Além disso, 38,4% apresentavam megaesôfago estágio 1 e 2 e 61,5% estágio 3 e 4. Quanto às reoperações, em 53% foi realizada a cirurgia de Heller-Pinotti videolaparoscópica, seguida de Serra-Dória em 30,7% e mucosectomia esofágica em 7,9%. Em 72% das reoperações não houve complicações pós-operatórias e 80% tiveram evolução satisfatória com redução ou ausência da disfagia. Dentre os pacientes reoperados pela técnica de Heller-Pinotti videolaparoscópica, três referiram pouca melhora da disfagia no pós-operatório. Dentre os pacientes submetidos à cirurgia de Serra-Dória, 100% tiveram evolução satisfatória da disfagia. Foi observado nos pacientes com o tempo entre a primeira cirurgia e a reoperação mais longo, um melhor resultado cirúrgico com diminuição da disfagia, com relevância estatística (p=0,0013, p<0,05). Conclusão: houve preferência nas reoperações de megaesôfago pela realização de re-miotomia por videolaparoscopia e, como segunda opção, a cirurgia de Serra-Dória. A esofagectomia ou mucosectomia esofágica foram reservadas para os casos mais avançados.

Genetic Alterations in Benign Breast Biopsies of Subsequent Breast Cancer Patients.

Background: Fibrocystic changes are associated with an increased risk of breast cancer. Genetic alterations have been found in fibrocystic changes with or without epithelial changes, suggesting that critical oncogenic events are occurring at an early stage. Methods: We investigated a unique collective of 17 breast cancer patients who, prior to the diagnosis of invasive breast cancer, underwent open surgical biopsy showing fibrocystic changes of the breast. The time span between biopsy for fibrocystic changes and invasive carcinoma ranged from 1 to 11 years (average 5.3 years). Ten (58.8%) of the patients had an ipsilateral invasive carcinoma, and 7 (41.2%) of the patients developed an invasive carcinoma of the contralateral breast. Massive parallel sequencing targeting genes frequently mutated in breast cancer was performed on the fibrocystic breast tissue as well as the ensuing cancer tissue. Results: In 9 cases, somatic mutations were found in the tumor tissue, the most prevalent being PIK3CA mutations (n = 4), followed by TP53 mutations (n = 2). None of these mutations were present in the previously removed mastopathy tissue. In one of the cases, an ERBB3 E928G mutation was present in the mastopathy as well as in the tumor tissue, with the variant allele frequency in the mastopathy being <0.1%. In two patients, we found two mutations (MAP3K1 L380fs and PIK3CA I391M, respectively) present in the mastopathy as well as in the subsequent breast cancer. These two mutations, however, could also be due to fixation artifacts. Conclusion: Since no significant somatic mutations in the fibrocystic breast tissue, and only doubtful shared mutations between benign and associated cancer tissue were detected, it remains unclear why women with fibrocystic breast disease have a statistically significant increased risk of breast cancer. Further analyses, maybe on the level of gene expression, could help to clarify the role of these benign alterations in the development of breast cancer and help to identify women at greater risk of developing subsequent invasive cancer.

Analysis of AR/ARV7 Expression in Isolated Circulating Tumor Cells of Patients with Metastatic Castration-Resistant Prostate Cancer (SAKK 08/14 IMPROVE Trial).

Despite several treatment options and an initial high response rate to androgen deprivation therapy, the majority of prostate cancers will eventually become castration-resistant in the metastatic stage (mCRPC). Androgen receptor splice variant 7 (ARV7) is one of the best-characterized androgen receptor (AR) variants whose expression in circulating tumor cells (CTCs) has been associated with enzalutamide resistance. ARV7 expression analysis before and during enzalutamide treatment could identify patients requiring alternative systemic therapies. However, a robust test for the assessment of the ARV7 status in patient samples is still missing. Here, we implemented an RT-qPCR-based assay for detection of AR full length (ARFL)/ARV7 expression in CTCs for clinical use. Additionally, as a proof-of-principle, we validated a cohort of 95 mCRPC patients initiating first line treatment with enzalutamide or enzalutamide/metformin within a clinical trial. A total of 95 mCRPC patients were analyzed at baseline of whom 27.3% (26/95) had ARFL+ARV7+, 23.1% (22/95) had ARFL+ARV7-, 23.1% (22/95) had ARFL-ARV7-, and 1.1% (1/95) had ARFL-ARV7+ CTCs. In 11.6% (11/95), no CTCs could be isolated. A total of 25/95 patients had another CTC analysis at progressive disease, of whom 48% (12/25) were ARV7+. Of those, 50% (6/12) were ARV7- and 50% (6/12) were ARV7+ at baseline. Our results show that mRNA analysis of isolated CTCs in mCRPC is feasible and allows for longitudinal endocrine agent response monitoring and hence could contribute to treatment optimization in mCRPC.

Treatment with long acting muscarinic antagonists stimulates serum levels of irisin in patients with COPD.

Long acting muscarinic antagonists (LAMA) are currently considered the therapeutic mainstay for patients with COPD and have been shown to improve clinical outcomes including symptoms, exercise capacity and airflow limitation. Irisin, is a newly discovered hormone-like myokine generated by skeletal muscle cells in response to exercise and it is suggested to regulate energy expenditure and exercise capacity. The aim of the present study was to investigate if treatment with LAMA alters serum irisin levels in patients with COPD. Irisin was assessed by ELISA in the serum of 506 patients with COPD, GOLD II-IV, with a smoking history >10 PY, who were included in the PROMISE-COPD cohort. The effect of inhaled LAMA on serum irisin levels was evaluated in a proof-of-concept cohort of 40 COPD patients. Univariate linear regression analysis revealed that there was a significant negative association of irisin with age-adjusted Charlson score (p = 0.003) and a positive association of irisin with 6-min walking distance (6MWD) (p = 0.018) and treatment with LAMA (p = 0.004) but not with LABA or ICS. Multivariate analysis revealed that the association of irisin with LAMA treatment remains significant after adjustment for age-adjusted score and 6MWD. In the proof-of-concept cohort a single inhalation of LAMA stimulated serum irisin levels after 4 h. These findings imply that treatment of COPD patients with LAMA increase circulating irisin, thus explaining some of the beneficial extra-pulmonary effects of these drugs when used in the treatment of COPD.

The MNK-1/eIF4E pathway as a new therapeutic pathway to target inflammation and remodelling in asthma.

Therapeutic targets in asthma are reduction of airway inflammation and remodelling, the latter is not affected by available drugs. Here we present data that inhibition of MAPK-activated protein kinase (MNK)-1 reduces inflammation and remodelling. MNK-1 regulates protein expression by controlling mRNA stability, nuclear export and translation through the eukaryotic initiation factor 4E (eIF4E). Airway smooth muscle cells were derived from asthmatic and non-asthmatic donors. Cells were pre-treated with CGP57380 (MNK-1 inhibitor) or MNK-1 siRNA, before TNF-α stimulation. Cytokine and protein expression was analysed by ELISA, real time PCR and immunoblotting. Proliferation was monitored by cell counts. TNF-α activated MNK-1 phosphorylation between 15 and 30min. and subsequently eIF4E between 15 and 60min. EIF4E activity was inhibited by CGP57380 dose-dependently. Inhibition of MNK-1 by CGP57380 or MNK-1 siRNA significantly reduced TNF-α induced CXCL10 and eotaxin mRNA expression and secretion, but had no effect on IL-8. However, CXCL10 mRNA stability or NF-κB activity were not affected by MNK-1 inhibition. Furthermore, eIF4E was detected in the cytosol and the nucleus, but TNF-α did not affected its export from the nucleus. Cytokine array assessment showed that in addition to eotaxin and CXCL10, asthma relevant GRO α and RANTES were down-regulated by MNK-1 inhibition. In addition, MNK-1 inhibition significantly reduced FCS and PDGF-BB induced cell proliferation. We are the first to report that MNK-1 controls chemokine secretion and proliferation in human airway smooth muscle cells. Therefore we suggest that MNK-1 inhibition may present a new target to limit inflammation and remodelling in asthmatic airways.

Mannose-binding lectin protein and its association to clinical outcomes in COPD: a longitudinal study.

BACKGROUND: Functional deficiency of mannose-binding lectin (MBL) may contribute to the pathogenesis of chronic obstructive pulmonary disease. We hypothesized that specific MBL2 gene polymorphisms and circulating MBL protein levels are associated with clinically relevant outcomes in the Predicting Outcome using systemic Markers In Severe Exacerbations of COPD PROMISE-COPD cohort. METHODS: We followed 277 patients with stable COPD GOLD stage II-IV COPD over a median period of 733 days (IQR 641-767) taking survival as the primary outcome parameter. Patients were dichotomized as frequent (≥ 2 AECOPD/year) or infrequent exacerbators. Serum MBL levels and single nucleotide polymorphisms of the MBL2 gene were assessed at baseline. RESULTS: The MBL2-HYPD haplotype was significantly more prevalent in frequent exacerbators (OR: 3.33; 95% CI, 1.24-7.14, p = 0.01). The median serum MBL concentration was similar in frequent (607 ng/ml, [IQR; 363.0-896.0 ng/ml]) and infrequent exacerbators (615 ng/ml, [IQR; 371.0-942.0 ng/ml]). Serum MBL was not associated with lung function characteristics or bacterial colonization in sputum. However, high serum MBL at stable state was associated with better survival compared to low MBL (p = 0.046, log rank test). CONCLUSIONS: In COPD, the HYPD haplotype of MBL2 gene is associated with frequent exacerbations and high serum MBL is linked to increased survival. The PROMISE-COPD study was registered at www.controlled-trials.com under the identifier ISRCTN99586989.

Aclidinium bromide combined with formoterol inhibits remodeling parameters in lung epithelial cells through cAMP.

Combined muscarinic receptor antagonists and long acting ß2-agonists improve symptom control in chronic obstructive pulmonary disease (COPD) significantly. In clinical studies aclidinium bromide achieved better beneficial effects than other bronchodilators; however, the underlying molecular mechanisms are unknown. This study assessed the effect of aclidinium bromide combined with formoterol on COPD lung (n=20) and non-COPD lung (n=10) derived epithelial cells stimulated with TGF-ß1+carbachol on: (i) the generation of mesenchymal cells in relation to epithelial cells, (II) extracellular matrix (ECM) deposition, and (iii) the interaction of ECM on the generation of epithelial and mesenchymal cells. TGF-ß1+carbachol enhanced the generation of mesenchymal cells, which was significantly reduced by aclidinium bromide or formoterol. The effect of combined drugs was additive. Inhibition of p38 MAP kinase and Smad by specific inhibitors or aclidinium bromide reduced the generation of mesenchymal cells. In mesenchymal cells, TGF-ß1+carbachol induced the deposition of collagen-I and fibronectin which was prevented by both drugs dose-dependently. Formoterol alone reduced collagen-I deposition via cAMP, this however, was overruled by TGF-ß1+carbachol and rescued by aclidinium bromide. Inhibition of fibronectin was cAMP independent, but involved p38 MAP kinase and Smad. Seeding epithelial cells on ECM collagen-I and fibronectin induced mesenchymal cell generation, which was reduced by aclidinium bromide and formoterol. Our results suggest that the beneficial effect of aclidinium bromide and formoterol involves cAMP affecting both, the accumulation of mesenchymal cells and ECM remodeling, which may explain the beneficial effect of the drugs on lung function in COPD.

Vasoactive Intestinal Peptide for Diagnosing Exacerbation in Chronic Obstructive Pulmonary Disease.

BACKGROUND: Vasoactive intestinal peptide (VIP) is the most abundant neuropeptide in the lung. VIP has been linked to pulmonary arterial hypertension and hypoxia. OBJECTIVES: We aimed to assess circulating VIP levels at exacerbation and at stable chronic obstructive pulmonary disease (COPD) and to evaluate the diagnostic performance in a well-characterized cohort of COPD patients. METHODS: The nested cohort study included patients with Global Initiative for Chronic Obstructive Lung Disease stage II-IV. Patients were examined at stable state and at acute exacerbation of COPD (AE-COPD), and dedicated serum was collected at both conditions. Serum VIP levels were determined by enzyme-linked immunosorbent assay. Diagnostic accuracy was analyzed by receiver operating characteristic curve and area under the curve (AUC). RESULTS: Patients with acute exacerbation (n = 120) and stable COPD (n = 163) had similar characteristics at baseline. Serum VIP levels did not correlate with oxygen saturation at rest (p = 0.722) or at exercise (p = 0.168). Serum VIP levels were significantly higher at AE-COPD (130.25 pg/ml, 95% CI 112.19-151.83) as compared to stable COPD (40.07 pg/ml, 95% CI 37.13-43.96, p < 0.001). The association of increased serum VIP with AE-COPD remained significant after propensity score matching (p < 0.001). Analysis of the Youden index indicated the optimal serum VIP cutoff value as 56.6 pg/ml. The probability of AE-COPD was very low if serum VIP was ≤35 pg/ml (sensitivity >90%) and very high if serum VIP was ≥88 pg/ml (specificity >90%). Serum VIP levels presented a robust performance to diagnose AE-COPD (AUC 0.849, 95% CI 0.779-0.899). CONCLUSIONS: Increased serum VIP levels are associated with AE-COPD.

Longitudinal Measurement of Serum Vascular Endothelial Growth Factor in Patients with Chronic Obstructive Pulmonary Disease.

BACKGROUND: Impaired vascular endothelial growth factor (VEGF) signaling causes emphysema in animal models. In chronic obstructive pulmonary disease (COPD) patients, alterations in VEGF tissue expression have been observed. We hypothesize that circulating VEGF may be a biomarker to phenotype COPD patients. OBJECTIVE: The aim of this study was to investigate VEGF serum levels in stable and exacerbated COPD. METHODS: VEGF serum levels as well as parameters of short- and long-term outcome were assessed and analyzed in two COPD cohorts [PROMISE, n = 117; ProCOLD (PC), n = 191]. RESULTS: VEGF serum levels at stable COPD were neither related to forced expiratory volume in 1 s nor to the Modified Medical Research Council dyspnea score, 6-min walking distance or BODE index. There was no association between single VEGF levels and COPD exacerbation frequency or mortality at 1 and 2 years of follow-up. In PC an increase in VEGF over time (ΔVEGF) was associated with the exacerbation frequency as well as the 1- and 2-year hospitalization rate (p = 0.046, 0.009 and 0.006, respectively). Furthermore, in PC ΔVEGF was associated with 1- and 2-year survival (p = 0.009 and 0.041, respectively). CONCLUSIONS: Single serum VEGF levels, at stable and exacerbated COPD, were not associated with clinically significant outcomes in COPD. Conversely, the VEGF course seems related to COPD prognosis.

Extracellular matrix composition is modified by ß2-agonists through cAMP in COPD.

Long acting ß2-agonists (LABA) have been reported to modify the extracellular matrix (ECM) composition in the airway wall. Based on our earlier studies we here investigated the mechanism underlying the control of ECM modification by LABA in primary human airway smooth muscle cells. Cells were treated with formoterol or salmeterol (30 min) before TGF-ß1 stimulation (2-3 days) Using RT-PCT, immuno-blotting and ELISA the de novo synthesis and deposition of collagen type-I, -III, -IV and fibronectin were determined. Matrix metalloproteinases (MMP)-2 and -9 were analyzed by zymography. Both LABA activated cAMP and its corresponding transcription factor CREB within 60 min and thus partly reduced TGF-ß1-induced gene transcription of collagen type-I, -III, fibronectin and connective tissue growth factor (CTGF). The inhibitory effect of both LABA on collagen type-I and -III deposition involved a cAMP dependent mechanism, while the inhibitory effect of the two drugs on TGF-ß1-induced fibronectin deposition and on CTGF secretion was independent of cAMP. Interestingly, none of the two LABA reduced CTGF-induced synthesis of collagen type-I or type-III deposition. In addition, none of the two LABA modified collagen type-IV deposition or the expression and activity of MMP-2 or MMP-9. Our results show that LABA can prevent de novo deposition of specific ECM components through cAMP dependent and independent signaling. However, they do not reduce all ECM components by the same mechanism and they do not reduce existing collagen deposits. This might explain some of the controversial reports on the anti-remodeling effect of LABA in chronic inflammatory lung diseases.

Tiotropium sustains the anti-inflammatory action of olodaterol via the cyclic AMP pathway.

Mesenchymal cells (fibroblasts) of the airway wall respond to cholinergic stimulation by releasing pro-inflammatory and chemotactic cytokines and may thus contribute to chronic inflammation of the lung. Here, we studied the anti-inflammatory potential of olodaterol, a long acting ß2-adrenergic receptor agonist, and tiotropium, a long-acting muscarinic receptor antagonist, and whether they interact at the level of the cyclic AMP dependent signaling pathway. Pulmonary fibroblasts of asthmatic (n = 9) and non-asthmatic (n = 8) subjects were stimulated with the muscarinic receptor agonist carbachol and interleukin-1ß (IL-1 beta) in presence or absence of tiotropium or olodaterol alone, or their combination. We also measured cAMP levels and phosphorylation of the cAMP response element binding protein (CREB). As single components, carbachol, olodaterol and tiotropium did not affect IL-6 and IL-8 release. Carbachol concentration-dependently enhanced the production of IL-1ß-induced IL-6 and IL-8, which was blocked by the simultaneous addition of tiotropium. The combination of olodaterol plus tiotropium further reduced IL-6 and IL-8 release. Olodaterol induced cAMP and the phosphorylation of CREB, an effect counteracted by carbachol, but rescued by tiotropium. We conclude that olodaterol plus tiotropium cooperate to decrease the inflammatory response in pulmonary fibroblasts in vitro.

Avaliação nutricional de crianças atendidas em ambulatório de gastroenterologia / Nutritional assessment of children assisted in a ambulatory of gastroenterology

avaliar, nutricionalmente, crianças utilizando os parâmetros do Centro de Controle de Doenças (CDC)-2000 e da Organização Mundial da Saúde (OMS)-2006. MÉTODO: estudo transversal de prevalência das criançasatendidas no Ambulatório de Gastroenterologia Pediátrica da Fundação Santa Casa de Misericórdia do Pará (FSCMPA),entre outubro de 2009 e abril de 2010. RESULTADOS: a maioria (64,7%) das crianças avaliadas eram do sexomasculino; o maior percentual de baixa estatura (BE) foi encontrado pelos critérios do Z-score/OMS-2006 e o menorpelo Percentil/CDC-2000. Com relação ao sobrepeso, foi encontrado 3,9% pelo Percentil e Z-score/OMS-2006 e Z--score/CDC-2000 e 2,6% pelo Percentil/CDC-2000. Obesidade foi encontrada em 2,6%, conforme o Percentil e Z--score/CDC-2000 e Percentil/OMS-2006 e em 1,3% pelo Z-score/OMS-2006. Desnutrição foi mais diagnosticada peloscritérios do Percentil/CDC-2000, seguido pelo Z-score/CDC-2000, Percentil e Z-score da OMS-2006 com 28,9%,22,4%, 17,1% e 11,9% respectivamente; baixo peso (BP) foi encontrado em maior prevalência pelo Percentil/CDC-2000 e em menor prevalência pelo Percentil/OMS-2006. CONCLUSÃO: os resultados são compatíveis com os dadosda literatura, com maior percentual de BE e sobrepeso/obesidade encontrado, respectivamente, pelo Z-score/OMS-2006 e Z-score/Percentil/OMS-2006. Maior prevalência de BP e de desnutrição foram encontrados pelo CDC-2000.

OBJECTIVE: to assesment nutritionally children according to the World Health Organization (WHO) 2006 growthreference and Centers for Disease Control and Prevention (CDC) 2000 growth reference. METHODS: A transversaldescriptive study with children assisted in the ambulatory of Pediatric Gastroenterology of the Santa Casa de Misericórdia do Pará Foundation (FSCMPA), between october of 2006 and april of 2008. RESULTS: there most percentualof short stature was by Z-score/WHO charts and less by percentile/CDC charts. Overweight was 3,9% by Percentile/WHO, Z-score/WHO and z-score/CDC and 2,6% by percentil/CDC. Obesity was in 2,6% according percentile andZ-score/CDC and Percentile/WHO and in 1,3% by Z-score/WHO. Undernutrition was most diagnostic by Percentile/CDC, Z-score/CDC, percentile and Z-score/WHO in 8,9%, 22,4%, 17,1% e 11,9% respectivaly; low weight was accountedin most prevalence by Percentile/CDC and less prevalence by Percentile/WHO. CONCLUSION: the studyshowed similar results to those observed at other studies, whit good agreement between the two curves growth.