Mature tertiary lymphoid structures are key niches of tumour-specific immune responses in pancreatic ductal adenocarcinomas.

OBJECTIVE: To better understand the immune microenvironment of pancreatic ductal adenocarcinomas (PDACs), here we explored the relevance of T and B cell compartmentalisation into tertiary lymphoid structures (TLSs) for the generation of local antitumour immunity. DESIGN: We characterised the functional states and spatial organisation of PDAC-infiltrating T and B cells using single-cell RNA sequencing (scRNA-seq), flow cytometry, multicolour immunofluorescence, gene expression profiling of microdissected TLSs, as well as in vitro assays. In addition, we performed a pan-cancer analysis of tumour-infiltrating T cells using scRNA-seq and sc T cell receptor sequencing datasets from eight cancer types. To evaluate the clinical relevance of our findings, we used PDAC bulk RNA-seq data from The Cancer Genome Atlas and the PRINCE chemoimmunotherapy trial. RESULTS: We found that a subset of PDACs harbours fully developed TLSs where B cells proliferate and differentiate into plasma cells. These mature TLSs also support T cell activity and are enriched with tumour-reactive T cells. Importantly, we showed that chronically activated, tumour-reactive T cells exposed to fibroblast-derived TGF-ß may act as TLS organisers by producing the B cell chemoattractant CXCL13. Identification of highly similar subsets of clonally expanded CXCL13 + tumour-infiltrating T cells across multiple cancer types further indicated a conserved link between tumour-antigen recognition and the allocation of B cells within sheltered hubs in the tumour microenvironment. Finally, we showed that the expression of a gene signature reflecting mature TLSs was enriched in pretreatment biopsies from PDAC patients with longer survival after receiving different chemoimmunotherapy regimens. CONCLUSION: We provided a framework for understanding the biological role of PDAC-associated TLSs and revealed their potential to guide the selection of patients for future immunotherapy trials.

Angiolymphatic invasion as a prognostic fator in resected N0 pancreatic adenocarcinoma.

Background: Pancreatic adenocarcinoma remains one of the worst digestive cancers. Surgical resection is the main target when treating a patient with curative intent. Aim: To assess angiolymphatic invasion as a prognostic factor in resected pN0 pancreatic cancer. Methods: Thirty-eight patients were submitted to pancreatoduodenectomy due to head pancreatic cancer. Tumor size, margins, lymph nodes, pTNM staging, angiolymphatic and perineural invasion were described in the pathologists' reports. Results: Most patients were female. Overall median survival was 13 months. Gemcitabine was the regimen of choice for chemotherapy in selected patients; however, it did not improve overall survival. pR0 resection had better survival compared with pR1. Within the pN0 group, survival was significantly better in patients without angiolymphatic invasion. Conclusion: Angiolymphatic invasion in N0 pancreatoduodenectomy can be demonstrated by the Hematoxylin-Eosin stain and may predict a poor prognosis factor for those patients.

Racional: Adenocarcinoma pancreático continua sendo um dos piores cânceres do aparelho digestivo. A ressecção cirúrgica é o principal objetivo quando se trata de intenção curativa. Objetivo: Avaliar a invasão angiolinfática como um fator prognóstico no câncer da cabeça do pâncreas ressecado pN0. Método: Trinta e oito pacientes foram submetidos a duodenopancreatectomia por câncer da cabeça do pâncreas. Tamanho do tumor, margens, linfonodos, estadiamento pTNM, invasão angiolinfática e perineural foram descritos nos laudos anatomopatológicos. Resultados: A maioria foi de mulheres. A sobrevida mediana global foi de 13 meses. Gencitabina foi a droga de escolha para quimioterapia nos pacientes selecionados, entretanto não aumentou a sobrevida global. Pacientes com ressecção pR0 tiveram sobrevida global superior quando comparados com ressecção pR1. Dentro do grupo de pacientes com pN0, a sobrevida foi significativamente melhor no grupo de pacientes que não apresentavam invasão angiolinfática. Conclusão: A invasão angiolinfática da duodenopancreatectomia N0 pode ser demonstrada utilizando apenas a hematoxilina-eosina e pode predizer prognóstico ruim para estes pacientes.

Systematic training model for teaching, development and training of instructors in inguinal hernia treatment using the Lichtenstein technique. Hernia campaign 2014 & 2015.

OBJECTIVE:: to evaluate the method of training and continuing education of 18 surgeons in 2014, and 28 surgeons in 2015, in the Holy Homes of Ribeirao Preto, Araraquara, Franca and San Carlos of São Paulo, in the performance of Lichtenstein inguinal herniorrhaphy, tutored by the Faculty of Medical Sciences of the São Paulo Holy Home and the organization HERNIA HELP - "Hernia Repair for the Underserved". METHODS:: the training was tutored and systematized through an active methodology of teaching and learning, aiming to offer competence, skills and attitudes, measured by a previously validated Qualification Form, qualifying leaders in trainees' improvement. RESULTS:: in 2014 the outcomes were: the difficulty of the case, direction, incision, dissection, mesh preparation, mesh cutting, mesh setting, closing, instruments, respect to tissues, flow, time and motion, and performance, all presented change in the general rating (p=0.000002); there was greater confidence in the execution of the procedure in 80% of trainees, considered "very valuable" in 93.3% of the interventions. In 2015, 28 surgeons were trained by ten surgeons previously qualified in 2014. The nerve identification rate, a relevant time the Lichtenstein technique, was 95.5% for the Iliohypogastric, 98.5% for the ilioinguinal and 89.4% for the genital branch of the genitofemoral nerve. CONCLUSION:: the applied teaching method is possible, reproducible, reliable and valid. The joint efforts offer enormous opportunity of directed education, reaching underserved populations, revealing the great teacher-student social responsibility. OBJETIVO:: avaliar resultados do método de treinamento e educação continuada de 18 cirurgiões, em 2014, e 28 cirurgiões, em 2015, nas Santas Casas de Ribeirão Preto, Araraquara, Franca e São Carlos do Estado de São Paulo, na realização da Herniorrafia Inguinal à Lichtenstein, tutorados pela Faculdade de Ciências Médicas da Santa Casa de São Paulo e pela organização HERNIA HELP - "Hernia Repair for the Underserved". MÉTODOS:: treinamento tutorado e sistematizado, através de metodologia ativa de ensino e aprendizagem, visando a oferecer competência, habilidade e atitudes, auferidas por um Formulário de Qualificação previamente validado, qualificando líderes no aperfeiçoamento de treinandos. RESULTADOS:: em 2014, os desfechos foram: dificuldade do caso, direção, incisão, dissecção, preparo da tela, corte da tela, fixação da tela, fechamento, instrumentos, respeito aos tecidos, fluxo, tempo e movimento e desempenho, apresentaram mudança na Classificação Geral (p=0,000002); houve maior confiança na execução do procedimento em 80% dos treinandos, sendo considerado "Muito Valioso" em 93,3% das participações. Em 2015, os 28 cirurgiões foram treinados por dez cirurgiões previamente qualificados em 2014. A taxa de identificação dos nervos, tempo relevante da técnica de Lichtenstein, foi 95,5 % para o ílio-hipogástrico, 98,5% para o ilioinguinal e 89,4% para o ramo genital do nervo genitofemoral. CONCLUSÃO:: o método de ensino aplicado é possível, reprodutível, confiável e válido. Os mutirões oferecem a enorme oportunidade do ensino, dirigido, atingindo populações carentes, revelando a grande responsabilidade social docente-discente.

Mirizzi syndrome grades III and IV: surgical treatment.

OBJECTIVE: : to evaluate the epidemiology and outcomes of surgical treatment of patients with Mirizzi Syndrome (MS) grades III and IV, the most advanced according to Csendes classification. METHODS: : we conducted a retrospective, cross-sectional study by reviewing records of thirteen patients with grades III and IV MS operated from December 2001 to September 2013, among the 3,691 cholecystectomies performed in the period. RESULTS: : the incidence of MS was 0.6% (23 cases) and grades III and IV amounted to 0.35% of this number. There was a predominance of type IV (12 cases). The preoperative diagnosis was possible in 53.8% of cases. The preferred approach was biliary-digestive derivation (10 cases), and "T" tube drainage with suture of the bile duct was the choice in three special occasions. Three patients had biliary fistula resolved with clinical management, and one coliperitoneum case required reoperation. In the outpatient follow-up of patients who underwent biliodigestive anastomosis (eight), 50% are asymptomatic, 25% had anastomotic stricture and 25% lost follow-up. The mean follow-up was 41.8 months. CONCLUSION: : MS in advanced degrees has low incidence, preoperative diagnosis in only half of cases, and has the biliodigestive anastomosis as the best conduct, but not without morbidity. OBJETIVO: avaliar a epidemiologia e os resultados do tratamento cirúrgico de doentes portadores de graus III e IV, mais avançados, da Síndrome de Mirizzi (SM) de acordo com a classificação de Csendes. MÉTODOS: estudo retrospectivo, de corte transversal através da revisão de prontuários de 13 pacientes portadores de graus III e IV da SM operados de dezembro de 2001 a setembro de 2013, entre 3691 colecistectomias realizadas neste período. RESULTADOS: a incidência da SM foi 0,6% (23 casos) e os graus III e IV perfizeram 0,35% deste número. Houve um predomínio de tipo IV (12 casos). O diagnóstico pré-operatório foi possível em 53,8% dos casos. A conduta preferencial foi derivação biliodigestiva (10 casos) e foi optado por drenagem com tubo "T" e sutura da via biliar em três ocasiões especiais. Três pacientes apresentaram fístula biliar resolvida com conduta expectante e um caso de coleperitônio necessitou reoperação. No seguimento ambulatorial dos pacientes que realizaram a anastomose biliodigestiva (oito), 50% estão assintomáticos, 25% apresentaram estenose da anastomose e 25% perderam seguimento. O tempo médio de acompanhamento foi 41,8 meses. CONCLUSÃO: de incidência baixa e de diagnóstico pré-operatório em apenas metade dos casos, a SM em graus avançados tem na anastomose biliodigestiva sua melhor conduta, porém não isenta de morbimortalidade.

QUALIS OF TECHNICAL PRODUCTION: METRIC DEVELOPMENT FOR MANUALS.

There are few authors focused on the study of preparing manuals criteria, often criticized for methodological rigor of the information. However, it is interesting to understand what is going wrong; to understand prognosis in self-handling; in prevention of complications; in identifying better services and information; in encouraging the pursuit of excellence in the healthcare. In its preparation should be valued the basic requirements of a manual, which are: 1) wrote in simple words, being concise, efficient, clear with index or table of contents; 2) using the existing institutional rules; 3) being flexible; and 4) having ongoing process of review, update and distribution. In the process of evaluation of Medicine III manuals may have the following stratification: M4 - 100 points: when there is ISBN registration and disclosure in other languages; M3 - 75 points: registration with the ISBN only in our country; M2 - 50 points: registered in regional libraries; M1 - 25 points: manual use for inter-departmental or intra-institutional groups; M0 - 0 point: use of manual in intra-departmental teaching. In conclusion, it is known that the use of manuals is focused to the ones who do not know the subject; however, it is also an important tool that can minimize errors, avoid professional misconduct, optimize resources, and at the end allowing assessment of the subject at hand. Poucos são os autores que se debruçam no estudo dos critérios de elaboração de manuais, sendo muitas vezes criticados pelo rigor metodológico das informações. Contudo, eles são interessantes no entender o que é errado, compreender prognóstico, na autoassistência, na prevenção de complicações, para identificar melhores serviços e informações, motivando a busca pela excelência no atendimento assistencial. Em sua elaboração devem ser valorizados os requisitos básicos de um manual, que são: 1) conter redação simples, concisa, eficiente e clara, contendo índice ou sumário; 2) utilizar as normas institucionais vigentes; 3) serem flexíveis; e 4) ter processo contínuo de revisão, atualização e distribuição. No processo de avaliação da Medicina III poderá ter os seguintes estratos: M4 - 100 pontos: registro no ISBN e divulgação em outros idiomas; M3 - 75 pontos: registro no ISBN apenas em nível nacional; M2 - 50 pontos: registrado em bibliotecas regionais; M1 - 25 pontos: manual de uso intrainstitucional por grupos interdepartamentais; M0 - 0 ponto: manual de uso intradepartamental ou de ensino. Em conclusão, sabe-se que o uso de manuais está destinado a quem não sabe do assunto; no entanto, trata-se de ferramenta importante que pode minimizar erros, consolidar condutas, otimizar recursos, permitindo avaliação criteriosa e aprofundada do tema em pauta.

Modified "Roux en Y" hepaticojejunostomy to permit transgastric endoscopic access.

PURPOSE: To present an option technique during the accomplishment of a "Roux-en-Y" hepaticojejunostomy that, allows future transgastric endoscopic access to the biloenteric anastomosis. METHODS: The side-to-side "Roux-en-Y" hepaticojejunostomy is carried through transmesocolic, retrogastric short tunnel. A distal jejunal loop of approximately 10,0 to 15,0 cm must be kept far from the anastomosis that allows its setting, without tension, in the anterior gastric wall bellow the small bending, between body and of the stomach antrum. Once the jejunal segment is fixed on the anterior gastric wall, this will allow future endoscopic access to the bilioenteric anastomosis, carried through transgastric punction guided by endoscopic-ultrasound (EUS). CONCLUSION: The modified "Roux en Y" hepaticojejunostomy is a simple and new technique to permit an alternative transgastric endoscopic access to bilioenteric anastomosis.

Correlation analysis between post-pancreatoduodenectomy pancreatic fistula and pancreatic histology.

OBJECTIVE: To evaluate the relationship between the occurrence of pancreatic fistula after pancreatojejunal anastomosis in patients undergoing pancreaticoduodenectomy for periampullary malignancy and the histological fibrosis and inflammation found in pancreatic tissue and the caliber of the main pancreatic duct. METHODS: We conducted a retrospective study with patients that were treated with surgical resection. The rate of pancreatic fistulae was recorded. Histology classification was performed according to fibrosis and pancreatic inflammation. RESULTS: We identified 77 patients, mean age was 57.6 years; 62.4% were male. As for the type of operation performed, 66.3% were gastroduodenopancreatectomies and 33.7% pancreatoduodenectomies with pylorus preservation. Regarding the number of fistulas diagnosed, it was found that 23.4% patients displayed this postoperative complication and in 66.7% the cause was cancer of the papilla. As for intraoperative macroscopic findings, we identified the classification of pancreatic texture, seen as normal in 85.8%, and the caliber of the main pancreatic duct, finding an average of 4.9 mm. There was a significant relationship between the hardened pancreatic stump and the absence of fistula. In patients with normal or soft tissue, the rate of fistula was 25.4%. Regarding ductal diameter, we identified a higher number of pancreatic anastomotic dehiscences in the absence of ductal dilation (p <0.05). We noticed that patients with an average ductal diameter of 5.4 mm (76.7%) did not show this complication. CONCLUSION: The presence of fibrosis and ductal dilation usually coexist and is related to a lower percentage of pancreatic fistulae.

Prevalence of Helicobacter spp in chronic cholecystitis and correlation with changes on the histological pattern of the gallbladder.

PURPOSE: Establish the prevalence of Helicobacter spp in chronic cholecystitis and its correlation with the gallbladder's histological findings. METHODS: 100 patients were operated for chronic cholecystitis with cholecystolithiasis. In pathological examination of the gallbladder, were evaluated the presence of metaplasia, dysplasia, lymphoid follicles, anaplasia and tumors that might be related to the presence of Helicobacter plus the presence of the bacilli Giemsa(+) by optical microscopy. From the DNA extracted from the gallbladder's bile, PCR was performed by using specific primers for the identification of Helicobacter spp with amplification of the 400bp segment of rRNA gene16S, with positive control DNA from Helicobacter pylori. All the cases negative for isolation of genetic material were excluded. The cases of PCRTheta and GiemsaTheta were used as negative control group. The histological findings were compared to the presence of bacilli and PCR data using a chi-square and Fisher's Exact test (CI = 95.0%, p <0.05). RESULTS: Of 68 patients, 42 (61.8%) were PCR(+) for Helicobacter spp and 19 (27.9%) had Giemsa(+). There was no correlation between the two findings. The PCR(+) for Helicobacter spp was not correlated to the histological findings. The presence of lymphoid follicles and metaplasia was related to the Giemsa(+) (p = 0.025 and p= 0.039). CONCLUSION: There is high prevalence of Helicobacter spp in patients with chronic cholecystitis and cholecystolithiasis without be correlated with the histological patterns studied.

Analysis of the bactericidal effect of ozone pneumoperitoneum.

PURPOSE: To assess the bactericidal action of ozone pneumoperitonium, and to compare the results with CO2. METHODS: It was used 36 Wistar rats. The animals, under anesthesia, were inoculated with 2ml of E. coli ATCC at a concentration of 10(10)UFC, and 1ml of BaSO4, into the peritoneal cavity. They were divided into three groups: Group 1, CO2 pneumoperitoneum was performed for 15 minutes; Group 2, ozone pneumoperitoneum was performed for 5 minutes at a concentration of 42microg/ml, and Group 3, ozone pneumoperitoneum was performed for 5 minutes at a concentration of 62microg/ml. Six animals from each group were sacrificed after the experiment, and the remaining 6 observed for 24 hours. Material was collected from the cavity of all animals for microbiological study. RESULTS: Ozone presented a greater bactericidal effect than CO2 in those animals sacrificed immediately after pneumoperitoneum. In the animals studied 24 hours after pneumoperitoneum evidenced no difference in bactericidal effect between the two gases. Moreover, no difference in mortality was observed. CONCLUSION: Ozone has a more potent bactericidal effect than carbon dioxide gas, although this did not influence survival of the animals.

Modelo experimental de icterícia obstrutiva: avaliação por meio da bioimpedância / Experimental model obstructive jaundice: assessment by bioimpedance

OBJETIVO: A bioimpedância é um método utilizado, como medida de massa corporal, volume líquido e volume de gordura corporal. Os autores analisam a utilização e desempenho da impedância em modelo de pesquisa experimental de icterícia obstrutiva. MÉTODO: Utilizaram-se oito ratos da linhagem Wistar, que foram divididos em dois grupos de quatro animais: grupo 1(experimento) e grupo 2 (controle). No grupo 1 foi realizado ligadura do colédoco e no grupo 2 laparotomia. Realizou-se a mensuração da resistência e da reatância capacitiva com aparelho de análise de bioimpedância no pré e pós-operatórios. RESULTADOS: O estudo revelou valores médios de resistência para o grupo controle maiores do que para o experimento, mostrando uma diferença significante. Também os valores de reatância capacitiva apresentaram médias maiores no grupo controle com p<0,0001. CONCLUSÕES: A análise dos resultados permite concluir que no rato: há diferenças nos valores de impedância entre os animais do grupo controle e experimental; os valores médios de resistência e reatância capacitiva são menores no grupo experimento e que a aplicabilidade da impedância no presente modelo pode ser sugerida.

[Predictors of choledocholithiasis in patients sustaining acute biliary pancreatitis]. / Fatores preditivos de coledocolitíase em doentes com pancreatite aguda biliar.

BACKGROUND: To assess the role of alkaline phosphatase (AP), gamil-glutamyltransferase (gammaGT) and abdominal ultrasound (US) as predictors of choledocholithiasis in patients sustaining acute biliary pancreatitis. METHODS: Data was prospectively collected during a period of 31 months. Forty patients were included, 30 were female and the mean age was 49 +/- 16. All patients sustaining acute biliary pancreatitis were enrolled. Patients with clinical jaundice and severe pancreatitis were excluded. Serum content of AP and gGT as well as US were assessed at admission and 48 hours before cholecistectomy. All patients underwent intra-operative cholangiography (IOC) or pre-operative endoscopic retrograde cholangiography (ERCP), which was indicated based on the odds of choledocholithiasis. In order to identify the predictors of choledocholithiasis, variables were compared between patients sustaining or not such alteration in cholangiography. Student t, Fisher and chi square tests were used for statistical analysis, considering p<0.05 as significant. Positive (PPV) and negative predictor values (NPV) were calculated for each variable. RESULTS: Upon admission, 15 (37%) patients sustained biliary tract dilatation and 5 (12%) choledocholithiasis at the US. Forty eight hours before the operation, 34 (85%) patients had altered levels of gGT and 16 (40%) of AP. Pre-operative US showed biliary tract dilatation in nine patients and choledocholithiasis in three. ERCP was performed in 15 (37%) cases. Higher PPV (55%) was attributed to pre-operative US, which had also a NPV of 96%. CONCLUSION: The best predictor of choledocholithiasis in patients sustaining mild acute pancreatitis was the biliary tract dilatation in pre-operative US.