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Detection of KRAS G12D in colorectal cancer stool by droplet digital PCR

World Journal of Gastroenterology 2017年第39期23卷 7087-7097页

作者： Susana Olmedillas-López Dennis César Lévano-Linares carmen laura aúz alexandre Luz Vega-Clemente Edurne León Sánchez Alejandro Villagrasa Jaime Ruíz-Tovar Mariano García-Arranz Damián García-Olmo Foundation Health Research Institute-Fundación Jiménez Díaz University Hospital Department of Surgery School of Medicine Universidad Autónoma de Madrid Department of Surgery Rey Juan Carlos University Hospital Department of Pathology Fundación Jiménez Díaz University Hospital Department of Biomedicine and Biotechnology Universidad de Alcalá Department of Surgery Fundación Jiménez Díaz University Hospital

AIMTo assess KRAS G12D mutation detection by droplet digital PCR (ddPCR) in stool-derived DNA from colorectal cancer (CRC) *** this study, tumor tissue and stool samples were collected from 70 patients with stage I-IV CRC diagnosed by preoperative biopsy. KRAS mutational status was determined by pyrosequencing analysis of DNA obtained from formalin-fixed paraffin-embedded (FFPE) tumor tissues. The KRAS G12D mutation was then analyzed by ddPCR in FFPE tumors and stool-derived DNA from patients with this point mutation. Wild-type (WT) tumors, as determined by pyrosequencing, were included as controls; analysis of FFPE tissue and stool-derived DNA by ddPCR was performed for these patients as *** the total 70 patients included, KRAS mutations were detected by pyrosequencing in 32 (45.71%), whereas 38 (54.29%) had WT tumors. The frequency of KRAS mutations was higher in left-sided tumors (11 located in the right colon, 15 in the left, and 6 in the rectum). The predominant point mutation was KRAS G12D (14.29%, n = 10), which was more frequent in early-stage tumors (I-IIA, n = 7). In agreement with pyrosequencing results, the KRAS G12D mutation was detected by ddPCR in FFPE tumor-derived DNA, and only a residual number of mutated copies was found in WT controls. The KRAS G12D mutation was also detected in stool-derived DNA in 80% of all fecal samples from CRC patients with this point *** is a reliable and sensitive method to analyze KRAS G12D mutation in stool-derived DNA from CRC patients, especially at early stages. This non-invasive approach is potentially applicable to other relevant biomarkers for CRC management.

关键词： Droplet digital PCR KRAS Stool Formalin-fixed paraffin-embedded Pyrosequencing Colorectal cancer

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Effect of dephytinization on bioavailability of iron,calcium and zinc from infant cereals assessed in the Caco-2 cell model

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World Journal of Gastroenterology 2009年第16期15卷 1977-1984页

作者： carmen Frontela Maria laura Scarino Simonetta Ferruzza Gaspar Ros carmen Martínez Department of Food Science and Nutrition Faculty of Veterinary Science and Food Science and TechnologyMurcia University INRAN National Research Institute for Food and Nutrition

AIM： To test the effect of the dephytinization of three different commercial infant cereals on iron, calcium, and zinc bioavailability by estimating the uptake, retention, and transport by Caco-2 cells. METHODS： Both dephytinized （by adding an exogenous phytase） and non-dephytinized infant cereals were digested using an in vitro digestion protocol adapted to the gastrointestinal conditions of infants younger than 6 too. Mineral cell retention, transport, and uptake from infant cereals were measured using the soluble fraction of the simulated digestion and the Caco-2 cells. RESULTS： Dephytinization of infant cereals significantly increased （P 〈 0.05） the cell uptake efficiency （from 0.66%-6.05% to 3.93%-13%）, retention （from 6.04%-16.68% to 14.75%-20.14%） and transport efficiency （from 0.14%-2.21% to 1.47%-6.02%）, of iron, and the uptake efficiency （from 5.0%-35.4% to 7.3%-41.6%） and retention （from 4.05%-20.53% to 14.45%-61.3%） of zinc, whereas calcium only cell uptake showed a significant increase （P 〈 0.05） after removing phytate from most of the samples analyzed. A positive relationship （P 〈 0.05） between mineral solubility and the cell uptake and transport efficiencies was observed. CONCLUSION： Removing phytate from infant cereals had a beneficial effect on iron and zinc bioavailability when infant cereals were reconstituted with water. Since in developing countries cereal-based complementary foods for infants are usually consumed mixed with water, exogenous phytase additions could improve the nutritional value of this weaning food.

关键词： Infant cereals Phytate Iron Calcium zinc Caco-2 cells BioavaUability

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Noninvasive investigations for non alcoholic fatty liver disease and liver fi brosis

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World Journal of Gastroenterology 2010年第38期16卷 4784-4791页

作者： carmen Fierbinteanu-Braticevici Ion Dina Ana Petrisor laura Tribus Lucian Negreanu Catalin Carstoiu carmen Fierbinteanu-Braticevici Ana PetrisorLaura TribusLucian NegreanuCatalin CarstoiuMedical Clinic Ⅱ and Gastroenterology University Hospital Bucharest Department of Gastroenterology St John’s Hospital 7001 Bucharest carmen Fierbinteanu-Braticevici Ana PetrisorLaura TribusLucian NegreanuCatalin CarstoiuMedical Clinic Ⅱ and GastroenterologyUniversity Hospital Bucharest

Non-alcoholic fatty liver disease (NAFLD) includes a spectrum of diseases that have insulin resistance in common and are associated with metabolic conditions such as obesity, type 2 diabetes mellitus, and dyslipidemia. NAFLD ranges from simple liver steatosis, which follows a benign course, to nonalcoholic steatohepatitis (NASH), a more severe entity, with necroinflmmation and f ibrosis, which can progress to cryptogenic cirrhosis and end-stage liver disease. Liver biopsy remains the gold standard for evaluating the degree of hepatic necroinflammation and f ibrosis; however, several noninvasive investigations, such as serum biomarkers, have been developed to establish the diagnosis and also to evaluate treatment response. These markers are currently neither available in all centers nor validated in extensive studies. Examples include high-sensitivity C reactive protein and plasma pentraxin 3, which are associated with extensive liver f ibrosis in NASH. Interleukin-6 correlates with inflammation, and cytokeratin-18 represents a marker of hepatocyte apoptosis (prominent in NASH and absent in simple steatosis). Tissue polypep-tide specif ic antigen seems to have a clinical utility in the follow-up of obese patients with NASH.

关键词： Non-alcoholic fatty liver disease Biomarkers Necroinflammation Liver fibrosis

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Murine models based on acute myeloid leukemia-initiating stem cells xenografting

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World Journal of Stem Cells 2018年第6期10卷 57-65页

作者： Cristina Mambet Mihaela Chivu-Economescu Lilia Matei laura Georgiana Necula Denisa laura Dragu Coralia Bleotu carmen Cristina Diaconu Cellular and Molecular Pathology Department Stefan S. Nicolau Institute of Virology Nicolae Cajal Institute Titu Maiorescu University

Acute myeloid leukemia(AML) is an aggressive malignant disease defined by abnormal expansion of myeloid blasts. Despite recent advances in understanding AML pathogenesis and identifying their molecular subtypes based on somatic mutations, AML is still characterized by poor outcomes, with a 5-year survival rate of only 30%-40%, the majority of the patients dying due to AML relapse. Leukemia stem cells(LSC) are considered to be at the root of chemotherapeutic resistance and AML relapse. Although numerous studies have tried to better characterize LSCs in terms of surface and molecular markers, a specific marker of LSC has not been found, and still the most universally accepted phenotypic signature remains the surface antigens CD34+CD38- that is shared with normal hematopoietic stem cells. Animal models provides the means to investigate the factors responsible for leukemic transformation, the intrinsic differences between secondary post-myeloproliferative neoplasm AML and de novo AML, especially the signaling pathways involved in inflammation and hematopoiesis. However, AML proved to be one of the hematological malignancies that is difficult to engraft even in the most immunodeficient mice strains, and numerous ongoing attempts are focused to develop "humanized mice" that can support the engraftment of LSC. This present review is aiming to in-troduce the field of AML pathogenesis and the concept of LSC, to present the current knowledge on leukemic blasts surface markers and recent attempts to develop best AML animal models.

关键词： Acute myeloid leukemia Leukemia-initiating stem cells Antigen markers Murine models Xenografts

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Gastrointestinal cancer stem cells as targets for innovative immunotherapy

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World Journal of Gastroenterology 2020年第14期26卷 1580-1593页

作者： Mihaela Chivu-Economescu laura G Necula Lilia Matei Denisa laura Dragu Ana I Neagu Irina Alexiu Coralia Bleotu carmen Cristina Diaconu Department of Cellular and Molecular Pathology Stefan S.Nicolau Institute of VirologyBucharest 030304Romania Nicolae Cajal Institute Titu Maiorescu UniversityBucharest 040441Romania

The role of cancer stem cells in gastrointestinal cancer-associated death has been widely *** cancer stem cells(GCSCs)are considered to be responsible for tumor initiation,growth,resistance to cytotoxic therapies,recurrence and metastasis due to their unique *** properties make the current therapeutic trials against GCSCs ***,recent studies have shown that targeting stem cell surface markers or stemness associated pathways might have an additional off-target effect on the immune *** advances in oncology and precision medicine have opened alternative therapeutic strategies in the form of cancer *** approach differs from classical anti-cancer therapy through its mechanism of action involving the activation and use of a functional immune system against tumor cells,instead of aiming physically destruction of cancer cells through radio-or *** immunological approaches for GCSCs targeting involve the use of different immune cells and various immune mechanisms like targeting specific surface antigens,using innate immune cells like the natural killer and T cells,T-cell chimeric antigen receptor technology,dendritic cell vaccine,or immune checkpoint *** this respect,better understandings of immune regulatory mechanisms that govern anti-tumor response bring new hope in obtaining long-term remission for cancer therapy.

关键词： Immunotherapy Gastrointestinal cancer Cancer stem cells CAR-T Dendritic cells vaccines Immune checkpoints inhibitors

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New therapeutic options opened by the molecular classification of gastric cancer

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World Journal of Gastroenterology 2018年第18期24卷 1942-1961页

作者： Mihaela Chivu-Economescu Lilia Matei laura G Necula Denisa L Dragu Coralia Bleotu carmen C Diaconu Department of Cellular and Molecular Pathology Stefan S. Nicolau Institute of Virology Nicolae Cajal Institute Titu Maiorescu University

Gastric cancer(GC) is one of the most lethal and aggressive cancers, being the third cause of cancer related death worldwide. Even with radical gastrectomy and the latest generation of molecular chemotherapeutics, the numbers of recurrence and mortality remains high. This is due to its biological heterogeneity based on the interaction between multiple factors, from genomic to environmental factors, diet or infections with various pathogens. Therefore, understanding the molecular characteristics at a genomic level is critical to develop new treatment strategies. Recent advances in GC molecular classification provide the unique opportunity to improve GC therapy by exploiting the biomarkers and developing novel targeted therapy specific to each subtype. This article highlights the molecular characteristics of each subtype of gastric cancer that could be considered in shaping a therapeutic decision, and also presents the completed and ongoing clinical trials addressed to those targets. The implementation of the novel molecular classification system will allow a preliminary patient selection for clinical trials, a mandatory issue if it is desired to test the efficacy of a certain inhibitor to the given target. This will represent a substantial advance as well as a powerful tool for targeted therapy. Nevertheless, translating the scientific results into new personalized treatment opportunities is needed in order to improve clinical care, the survival and quality of life of patients with GC.

关键词： Gastric cancer Molecular classification Immunotherapy Targeted therapy Clinical trials

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Earth Map:A Novel Tool for Fast Performance of Advanced Land Monitoring and Citation:Morales C,DiazAS,Climate Assessment Climate Assessment

Journal of Remote Sensing

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Journal of Remote Sensing 2023年第1期3卷 1-18页

作者： carmen Morales Alfonso Sanchez-Paus Diaz Daniel Dionisio laura Guarnieri Giulio Marchi Danae Maniatis Danilo Mollicone Food and Agriculture Organization of the United Nations 00153 RomeItaly

Earth Map(https://***/)is an innovative and free application developed by the Food and Agriculture Organization of the United Nations that was designed in the framework of the Food and Agriculture Organization of the United Nations-Google partnership and facilitates the visualization,processing,and analysis ofland and climate *** Map makes petabytes of multitemporal,multiscale,multiparametric,and quasi-real-time satellite imagery and geospatial datasets available to any user thanks to the power of Google Earth Engine(https://***/)and a point-and-click graphical user *** are further complemented with more planetary-scale analytical capabilities so that global and local changes and trends on Earth's surface can be easily detected,quantified,and *** does not require users to master coding techniques,thereby avoiding bottlenecks in terms of technical capacities of nonexpert *** ultimately paves the way for countries,research institutes,farmers,and members of the general public to access critical knowledge to develop science-based policy interventions,leverage investments,and sustain *** provide a full overview of Earth Map's software architecture,design,features,and *** illustrate the possible applications of the tool,different examples are presented including a few case studies that show how quick historical analysis of environmental and climate parameters can be performed and research questions *** examples demonstrate that Earth Map is a comprehensive and user-friendly tool for land monitoring and climate assessment and that it has the potential to be used to assess land use,land use change,climate change impacts,and natural disasters.

关键词： Earth Map thereby

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Therapies targeting cancer stem cells: Current trends and future challenges

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World Journal of Stem Cells 2015年第9期7卷 1185-1201页

作者： Denisa L Dragu laura G Necula Coralia Bleotu carmen C Diaconu Mihaela Chivu-Economescu Cellular and Molecular Department Stefan S. Nicolau Institute of Virology Nicolae Cajal Institute Titu Maiorescu University Biochemistry-Proteomics Department Victor Babes National Institute of Pathology

Traditional therapies against cancer, chemo- and radiotherapy, have multiple limitations that lead to treatment failure and cancer recurrence. These limitations are related to systemic and local toxicity, while treatment failure and cancer relapse are due to drug resistance and self-renewal, properties of a small population of tumor cells called cancer stem cells(CSCs). These cells are involved in cancer initiation, maintenance, metastasis and recurrence. Therefore, in order to develop efficient treatments that can induce a longlasting clinical response preventing tumor relapse it is important to develop drugs that can specifically target and eliminate CSCs. Recent identification of surface markers and understanding of molecular feature associated with CSC phenotype helped with the design of effective treatments. In this review we discuss targeting surface biomarkers, signaling pathways that regulate CSCs self-renewal and differentiation, drug-efflux pumps involved in apoptosis resistance, microenvironmental signals that sustain CSCs growth, manipulation of mi RNA expression, and induction of CSCs apoptosis and differentiation, with specific aim to hamper CSCs regeneration and cancer relapse. Some of these agents are under evaluation in preclinical and clinical studies, most of them for using in combination with traditional therapies. The combined therapy using conventional anticancer drugs with CSCs-targeting agents, may offer a promising strategy for management and eradication of different types of cancers.

关键词： Cancer stem cells Targeted therapy Anticancer drug

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Diversity of Saxicolous Lichens along an Aridity Gradient in Central México

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Phyton-International Journal of Experimental Botany 2022年第4期91卷 827-840页

作者： Josécarmen Soto-Correa Abraham Saldaña-Vega Víctor Hugo Cambrón-Sandoval laura Concostrina-zubiri Mariela Gómez-Romero Facultad de Ciencias Naturales Universidad Autónoma de QuerétaroQuerétaro73230México Departamento de Biología y Geología Física y Química Inorgánica Tulipán s/nUniversidad Rey Juan CarlosMóstolesMadrid28933España Cátedra CONACYT–Facultad de Biología Universidad Michoacana de San Nicolás de HidalgoMoreliaMichoacán58030México

Lichens are symbiotic organisms that comprise a fungus and a photosynthetic partner wich are recognized as a good indicator of climate ***,our understanding of how aridity affects the diversity of saxicolous lichens in drylands is still *** evaluate the relationship between saxicolous lichen diversity and aridity in a central México dryland,a geographical transect was established of 100 km to build an aridity gradient in the semiarid zone of the State of Querétaro,Mexico,comprising ten sampling sites with a 10 km *** richness,abundance and diversity of soil lichen species were recorded using two sampling methods:the quadrat-intercept and the line-intercept method,to compare their performance in assessing soil lichen diversity in *** number of species and Shannon diversity of saxicolous lichens were higher at intermediate values of the aridity index(AI=0.10-0.34).Quadrat intercept and point intercept methods gave quite similar results,which means that the selected method does not influence the results in a significant *** study confirms the role of saxicolous lichens as climate change indicators and reveals the importance of the sampling method selection in the evaluation of different parameters of soil lichen diversity in drylands.

关键词： Climate change drylands ecological indicators environmental stress precipitation temperature

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Residual activity of cetrimide and chlorhexidine on Enterococcus faecalis-infected root canals

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International Journal of Oral Science 2014年第1期6卷 46-49页

作者： carmen María Ferrer-Luque María Teresa Arias-Moliz Matilde Ruíz-Linares María Elena Martínez García Pilar Baca Department of Stomatology School of Dentistry University of Granada Department of Microbiology School of Dentistry University of Granada

Effective final irrigation regimen is an important step in order to achieve better disinfection and ensure residual antimicrobial effects after root canal preparation. The aim of this study was to compare the residual antimicrobial activity of 0.2% cetrimide, and 0.2% and 2% chlorhexidine in root canals infected with Enterococcus faecalis. Biofilms of E. faecalis were grown on uniradicular roots for 4 weeks. After root canal preparation, root canals were irrigated with 17% ethylenediaminetetraacetic acid （EDTA） to remove the smear layer. The roots were randomly divided into three experimental groups （n=26） according to the final irrigating solution： Group I, 5 mL 0.2% cetrimide; Group II, 5 mL 0.2% chlorhexidine; and Group III, 5 mL 2% chlorhexidine. Samples were collected for 50 days to denote the presence of bacterial growth. The proportion of ungrown specimens over 50 days was evaluated using the nonparametric Kaplan-Meier survival analysis. Differences among groups were tested using the log-rank test and the level of statistical significance was set at P〈0.05. The highest survival value was found with 2% chlorhexidine, showing statistically significant differences from the other two groups. At 50 days, E. faecalisgrowth was detected in 69.23% specimens in Groups I and II, and in 34.61% specimens of Group III. There were no significant differences between 0.2% cetrimide and 0.2% chlorhexidine. Final irrigation with 2% chlorhexidine showed greater residual activity than 0.2% chlorhexidine and 0.2% cetrimide in root canals infected with E. faecalis.

关键词： antimicrobial effects biofilms cetrimide chlorhexidine Enterococcus faecalis final irrigation residual activity

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