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CagA EPIYA polymorphisms in Colombian Helicobacter pylori strains and their influence on disease-associated cellular responses

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World Journal of Gastrointestinal Oncology 2013年第3期5卷 50-59页

作者： Carlos Alberto Fajardo Andrés Javier Quiroga Andrea Coronado Karen Labrador Nicole Acosta Pilar Delgado Carlos Jaramillo maría mercedes bravo Cancer and Infectious Agents Research Group National Cancer Institute Molecular Diagnostics and Bioinformatics Laboratory Biological Sciences Department Los Andes University

AIM: To investigate the influence of the CagA diversity in Helicobacter pylori (H. pylori ) strains from Colombia on the host cell biology. METHODS: Eighty-four H. pylori-cagA positive strains with different Glu-Pro-Ile-Tyr-Ala (EPIYA) motifs patterns, isolated from patients with gastritis (n=17), atrophic gastritis (n=17), duodenal ulcer (n=16), intestinal metaplasia (n=16) and gastric cancer (n=18), were included. To determine the integrity of the cag pathogenicity island (cag PAI) we evaluated the presence of cagA, cagT, cagE, and cag10 genes by polymerase chain reaction. AGS gastric epithelial cellswere infected with each strain and assayed for translo-cation and tyrosine phosphorylation of CagA by western blot, secretion of interleukin-8 (IL-8) by enzyme-linked immuno sorbent assay after taking supernatants from cocultures and cell elongation induction. For cell elongation quantification, coculture photographs were taken and the proportion of "hummingbird" cells (>15 μm) was determined. RESULTS: Overall 72% (60/84) of the strains were found to harbor a functional cag PAI. Levels of phos-phorylated CagA were significantly higher for isolates from duodenal ulcer than the ones in strains from gas-tritis, atrophic gastritis, intestinal metaplasia and gastric cancer (49.1% ± 23.1% vs 21.1% ± 19.5%, P < 0.02; 49.1% ± 23.1% vs 26.2%±14.8%, P<0.045; 49.1% ± 23.1% vs 21.5% ± 19.5%, P<0.043 and 49.1% ± 23.1% vs 29.5% ± 27.1%, P < 0.047 respectively). We observed variable IL-8 expression levels ranging from 0 to 810 pg/mL and from 8.8 to 1442 pg/mL at 6 h and 30 h post-infection, respectively. cagPAI-defective strains did not induce detectable levels of IL-8 at 6 h post-infection. At 30 h post-infection all strains induced IL-8 expression in AGS cells, although cagPAI-defective strains induced significantly lower levels of IL-8 than strains with a functional cagPAI (57.1 ± 56.6 pg/mL vs 513.6 ± 338.6 pg/mL,P < 0.0001). We did not observe differences in the extent of cell

关键词： Helicobacter pylori cagA 3’region CagA protein Interleukin 8 Cell elongation Glu-Pro-Ile-Tyr-Ala

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Helicobacter pylori CagA protein polymorphisms and their lack of association with pathogenesis

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World Journal of Gastroenterology 2010年第31期16卷 3936-3943页

作者： Nicole Acosta Andrés Quiroga Pilar Delgado maría mercedes bravo Carlos Jaramillo Molecular Diagnostics and Bioinformatics Laboratory Biological Sciences DepartmentLos Andes University Infectious Agents and Cancer Research Group National Cancer Institute(Colombia)

AIM: To investigate Helicobacter pylori (H. pylori) CagA diversity and to evaluate the association between protein polymorphisms and the occurrence of gastric pathologies. METHODS: One hundred and twenty-two clinical isolates of H. pylori cultured from gastric biopsies obtained from Colombian patients with dyspepsia were included as study material. DNA extracted from isolates was used to determine cagA status, amplifying the C-terminal cagA gene region by polymerase chain reaction. One hundred and six strains with a single amplicon were sequenced and results were used to characterize the 3' variable region of the cagA gene. To establish the number and type of tyrosine phosphorylation motifs Glutamine acid-Proline-Isoleucine-Tyrosine-Alanine (EPI-YA) bioinformatic analysis using Amino Acid Sequence Analyzer-Amino Acid Sequence Analyzer software was conducted. Analysis of the association between the number of EPIYA motifs and the gastric pathology was performed using χ2 test and analysis of the presence of EPIYA-C motifs in relation to the pathology was made by logistic regression odds ratios. Comparisons among EPIYA types found and those reported in GenBank were performed using a proportion test in Statistix Analytical Software version 8.0. RESULTS: After amplification of the 3' of the cagA gene, 106 from 122 isolates presented a single amplicon and 16 showed multiple amplicons. As expected, diversity in the size of the cagA unique fragments among isolates was observed. The 106 strains that presented a single amplicon after 3' cagA amplification came from patients with gastritis (19 patients), atrophic gastritis (21), intestinal metaplasia (26), duodenal ulcer (22) and gastric cancer. DNA sequence analysis showed that the differences in size of 3' cagA unique fragments was attributable to the number of EPIYA motifs: 1.9% had two EPIYA motifs, 62.3% had three, 33.0% had four and 2.8% had five motifs. The majority of tested clinical strains (62.3%) were found to harbor

关键词： Helicobacter pylori CagA-protein polymorphisms Molecular characterization Bioinformatic analysis Pathogenesis Cancer

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Translational pancreatic cancer research:a comparative study on patient-derived xenograft models

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World Journal of Gastroenterology 2018年第7期24卷 794-809页

作者： mercedes Rubio-Manzanares Dorado Luis Miguel marín Gómez Daniel Aparicio Sánchez Sheila Pereira Arenas Juan Manuel Praena-Fernández Juan Jose Borrero martín Francisco Farfán López Miguel ángel Gómez bravo Jordi Muntané Relat Javier Padillo Ruiz Department of Hepatobiliary and Pancreatic Surgery Virgen del Rocio University Hospital Oncology Surgery Cell Therapy and Organ Transplantation Group Institute of Biomedicine of Seville (IBiS) Virgen del Rocio University Hospital University of Seville Statistics Methodology and Evaluation of Research Unit Virgen del Rocio University Hospital Pathology Department Virgen del Rocio University Hospital

AIM To assess the viability of orthotopic and heterotopic patient-derived pancreatic cancer xenografts implanted into nude *** This study presents a prospective experimental analytical follow-up of the development of tumours in mice upon implantation of human pancreatic adenocarcinoma samples. Specimens were obtained surgically from patients with a pathological diagnosis of pancreatic adenocarcinoma. Tumour samples from pancreatic cancer patients were transplanted into nude mice in three different locations(intraperitoneal, subcutaneous and pancreatic). Histological analysis(haematoxylin-eosin and Masson's trichrome staining) and immunohistochemical assessment of apoptosis(TUNEL), proliferation(Ki-67), angiogenesis(CD31) and fibrogenesis(α-SMA) were performed. When a tumour xenograft reached the target size, it was reimplanted in a new nude mouse. Three sequential tumour xenograft generations were generated(F1, F2 and F3).RESULTS The overall tumour engraftment rate was 61.1%. The subcutaneous model was most effective in terms of tissue growth(69.9%), followed by intraperitoneal(57.6%) and pancreatic(55%) models. Tumour development was faster in the subcutaneous model(17.7 ± 2.6 wk) compared with the pancreatic(23.1 ± 2.3 wk) and intraperitoneal(25.0 ± 2.7 wk) models(P = 0.064). There was a progressive increase in the tumour engraftment rate over successive generations for all three models(F1 28.1% vs F2 71.4% vs F3 80.9%, P < 0.001). There were no significant differences in tumour xenograft differentiation and cell proliferation between human samples and the three experimental models among the sequential generations of tumour xenografts. However, a progressive decrease in fibrosis, fibrogenesis, tumour vascularisation and apoptosis was observed in the three experimental models compared with the human samples. All three pancreatic patient-derived xenograft models presented similar histological and immunohistochemical *** In our experie

关键词： Immunohistological analysis Pancreatic cancer Patient-derived xenograft Animal model Nude mice

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Characterization Of Peripherical Blood And Tumor Infiltrating Lymphocytes In Cervical Carcinoma Patients By Flow Cytometry

Characterization Of Peripherical Blood And Tumor Infiltratin...

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24th International Papillomavirus Conference and Clinical Workshop

作者： maria Angelica Cespedes Ardila maria mercedes bravo Hernandez Susana Fiorentino Alba Lucia Combita Rojas Instituto Nacional de Cancerologia Instituto Nacional de Cancerologia Pontificia Universidad Javeriana Instituto Nacional de Cancerologia

＜正＞Decreases of host immune function,alterations in frequencies of lymphocyte subsets and increase of regulatory T cells（T reg ）have been observed in patients with different types of *** cervical cancer,the changes of immune responses in tumor milieu,especially the composition of TILs,may have important implications in terms of the progression of *** characterize the tumor infiltrating lymphocytes and peripheral blood lymphocytes in patients with cervical cancer,we analyzed 39 peripheral

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Survival after curative pancreaticoduodenectomy for ampullary adenocarcinoma in a South American population:A retrospective cohort study

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World Journal of Gastrointestinal Surgery 2022年第1期14卷 24-35页

作者： Ramiro Manuel Fernandez-Placencia Paola Montenegro Melvy Guerrero mariana Serrano Emperatriz Ortega mercedes bravo Lourdes Huanca Stéphane Bertani Juan Manuel Trejo Patricia Webb Jenny Malca-Vasquez Luis Taxa Alberto Lachos-Davila Juan Celis-Zapata Carlos Luque-Vasquez Eduardo Payet Eloy Ruiz Francisco Berrospi Hepato-Pancreato-Biliary Section Department of Abdominal SurgeryInstituto Nacional de Enfermedades NeoplasicasLima 15038Peru Department of Medical Oncology Instituto Nacional de Enfermedades NeoplasicasLima 15038Peru Department of Pathology Instituto Nacional de Enfermedades NeoplasicasLima 15038Peru International Joint Laboratory of Molecular Anthopological Oncology Instituto Nacional de Enfermedades NeoplasicasLima 15038Peru Unite Pharmacochim&Pharmacol Dev UMR152F-31062 ToulouseFrance Department of Radiation Oncology Instituto Nacional de Enfermedades NeoplasicasLima 15038Peru Department of Abdominal Surgery Instituto Nacional de Enfermedades NeoplasicasLima 15038Peru

BACKGROUND Ampullary adenocarcinoma(AAC)is a rare neoplasm that accounts for only 0.2%of all gastrointestinal *** incidence rate is lower than 6 cases per million *** prognostic factors have been described for AAC and are associated with a wide range of survival ***,these studies have been exclusively conducted in patients originating from Asian,European,and North American *** To evaluate the histopathologic predictors of overall survival(OS)in South American patients with AAC treated with curative pancreaticoduodenectomy(PD).METHODS We analyzed retrospective data from 83 AAC patients who underwent curative(R0)PD at the National Cancer Institute of Peru between January 2010 and October 2020 to identify histopathologic predictors of *** Sixty-nine percent of patients had developed intestinal-type AAC(69%),23%had pancreatobiliary-type AAC,and 8%had other ***-one percent of patients were classified as Stage I,according to the AJCC 8 th *** occurred primarily in the liver(n=8),peritoneum(n=4),and lung(n=4).Statistical analyses indicated that T3 tumour stage[hazard ratio(HR)of 6.4,95%confidence interval(CI)of 2.5-16.3,P<0.001],lymph node metastasis(HR:4.5,95%CI:1.8-11.3,P=0.001),and pancreatobiliary type(HR:2.7,95%CI:1.2-6.2,P=0.025)were independent predictors of *** Extended tumour stage(T3),pancreatobiliary type,and positive lymph node metastasis represent independent predictors of a lower OS rate in South American AAC patients who underwent curative PD.

关键词： Gastrointestinal neoplasms Adenocarcinoma Ampulla Pancreaticoduodenectomy Survival South America

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Loss of heterozygosity at chromosomes 6p and 15q;HLA-I,HLA-G and IL-10 expression in cervical cancer biopsies.

Loss of heterozygosity at chromosomes 6p and 15q;HLA-I,HLA-G...

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24th International Papillomavirus Conference and Clinical Workshop

作者： Josefa Antonia Rodriguez Garcia martha Lucia Serrano Lopez Liliana Galeano Petro Constanza Gomez Diana maria Palacios Ortiz maria mercedes bravo Hernandez Alba Lucia Combita Rojas Instituto Nacional de Cancerologia Instituto Nacional de Cancerologia Instituto Nacional de Cancerologia Instituto Nacional de Cancerologia Fundacion Santa Fe Instituto Nacional de Cancerologia Instituto Nacional de Cancerologia

＜正＞Immune evasion in cancer can result from structural and functional alterations in HLA-1 molecules and from the local release of immunosuppressive *** of heterozygosity（LOH）in HLA（6p21.3）and B2-m（15q21）regions is associated with HLA-1 down regulation in tumours HLA-G,a no classical class I molecule,and IL-10 are implicated in immune evasion in cancer

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