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1دورية أكاديمية
المؤلفون: Sydow, Farid FO von, Santiago, Marta A, Neves-Souza, Patricia C, Cerqueira, Denise IS, Gouvea, Adriana S, Lavatori, Maryrose FH, Bertho, Álvaro L, Kubelka, Claire F
المصدر: Memórias do Instituto Oswaldo Cruz. August 2000 95(4)
مصطلحات موضوعية: dengue virus, C6/36, Vero cells, mononuclear leukocytes, flow cytometry
الوصف: Fluorescent activated cell sorter (FACS) analysis is useful for the detection of cellular surface antigens and intracellular proteins. We used this methodology in order to detect and quantify dengue antigens in highly susceptible cells such as clone C6/36 (Aedes albopictus) and Vero cells (green monkey kidney). Additionally, we analyzed the infection in vitro of human peripheral blood mononuclear leukocytes (PBML). FACS analysis turned out to be a reliable technique to quantify virus growth in traditional cell cultures of C6/36 as well as Vero cells. High rates of infection were achieved with a good statistical correlation between the virus amount used in infection and the percentage of dengue antigen containing cells detected in infected cultures. We also showed that human monocytes (CD14+) are preferred target cells for in vitro dengue infection among PBML. Monocytes were much less susceptible to virus infection than cell lines but they displayed dengue antigens detected by FACS five days after infection. In contrast, lymphocytes showed no differences in their profile for dengue specific immunofluorescence. Without an animal model to reproduce dengue disease, alternative assays have been sought to correlate viral virulence with clinical manifestations and disease severity. Study of in vitro interaction of virus and host cells may highlight this relationship.
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2دورية أكاديمية
المؤلفون: Bertho, Álvaro L, Santiago, Marta A, Coutinho, Sérgio G
المصدر: Memórias do Instituto Oswaldo Cruz. June 2000 95(3)
مصطلحات موضوعية: apoptosis, flow cytometry, cell death, necrosis
الوصف: In this report we present a concise review concerning the use of flow cytometric methods to characterize and differentiate between two different mechanisms of cell death, apoptosis and necrosis. The applications of these techniques to clinical and basic research are also considered. The following cell features are useful to characterize the mode of cell death: (1) activation of an endonuclease in apoptotic cells results in extraction of the low molecular weight DNA following cell permeabilization, which, in turn, leads to their decreased stainability with DNA-specific fluorochromes. Measurements of DNA content make it possible to identify apoptotic cells and to recognize the cell cycle phase specificity of apoptotic process; (2) plasma membrane integrity, which is lost in necrotic but not in apoptotic cells; (3) the decrease in forward light scatter, paralleled either by no change or an increase in side scatter, represent early changes during apoptosis. The data presented indicate that flow cytometry can be applied to basic research of the molecular and biochemical mechanisms of apoptosis, as well as in the clinical situations, where the ability to monitor early signs of apoptosis in some systems may be predictive for the outcome of some treatment protocols.
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3دورية أكاديمية
المؤلفون: Santiago, Marta A, Luca, Paula M De, Bertho, Álvaro L, Azeredo-Coutinho, Rilza BG, Coutinho, Sérgio G
المصدر: Memórias do Instituto Oswaldo Cruz. June 2000 95(3)
مصطلحات موضوعية: intracellular cytokines, flow cytometry, leishmaniasis
الوصف: Flow cytometry has been used as a powerful technique for studying cell surface antigen expression as well as intracellular molecules. Its capability of analyzing multiple parameters simultaneously on a single cell has allowed identification and studies of functional cell subsets within heterogeneous populations. In this respect, several techniques have been developed during the past few years to study cytokine-producing cells by flow cytometry in humans and several animal models.
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4دورية أكاديمية
المؤلفون: Gil, José Vicente, Miralles, Alberto, de las Heras, Sandra, Such, Esperanza, Avetisyan, Gayane, Díaz-González, Álvaro, Santiago, Marta, Fuentes, Carolina, Fernández, José María, Lloret, Pilar, Navarro, Irene, Montesinos, Pau, Llop, Marta, Barragán, Eva
المصدر: Frontiers in Molecular Biosciences ; volume 11 ; ISSN 2296-889X
مصطلحات موضوعية: Biochemistry, Genetics and Molecular Biology (miscellaneous), Molecular Biology, Biochemistry
الوصف: Introduction: Acute lymphoblastic leukemia (ALL) is a prevalent childhood cancer with high cure rate, but poses a significant medical challenge in adults and relapsed patients. Philadelphia-like acute lymphoblastic leukemia (Ph-like ALL) is a high-risk subtype, with approximately half of cases characterized by CRLF2 overexpression and frequent concomitant IKZF1 deletions. Methods: To address the need for efficient, rapid, and cost-effective detection of CRLF2 alterations, we developed a novel RT-qPCR technique combining SYBR Green and highresolution melting analysis on a single plate. Results: The method successfully identified CRLF2 expression, P2RY8::CRLF2 fusions, and CRLF2 and JAK2 variants, achieving a 100% sensitivity and specificity. Application of this method across 61 samples revealed that 24.59% exhibited CRLF2 overexpression, predominantly driven by IGH::CRLF2 (73.33%). High Resolution Melting analysis unveiled concurrent CRLF2 or JAK2 variants in 8.19% of samples, as well as a dynamic nature of CRLF2 alterations during disease progression. Discussion: Overall, this approach provides an accurate identification of CRLF2 alterations, enabling improved diagnostic and facilitating therapeutic decision-making.
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المؤلفون: Velasco Santiago, Marta, Zimmermannova, Olga, Hansen, Morten, Fiúza Rosa, Fábio, Pires, Cristiana, Met, Özcan, Pereira, Filipe, Marie Svane, Inge
المصدر: SITC Society for Immunotherapy of Cancer 2022,Boston, United States,-- StemTherapy: National Initiative on Stem Cells for Regenerative Therapy.
مصطلحات موضوعية: Medicin och hälsovetenskap, Klinisk medicin, Cancer och onkologi, Medical and Health Sciences, Clinical Medicine, Cancer and Oncology
الوصف: Background Direct cell reprogramming is characterized by the use of defined factors to rewire the transcriptional and epigenetic network of one cell-type into one of a different lineage. We have recently identified the transcription factors PU.1, IRF8, and BAFT3 (PIB) as sufficient to induce a type 1 conventional dendritic cell (cDC1) fate in both somatic and cancer cells.1,2 cDC1 is a rare dendritic cell subset with unique ability to initiate de novo T cell responses after migrating to the tumor site. Several studies have shown that higher levels of cDC1s within the tumor microenvironment strongly correlate with good prognosis and responsiveness to immunotherapy for patients with melanoma.3 Therefore, we hypothesized that PIB factors could reprogram primary melanoma cells into functional antigen presenting cDC1s capable of presenting tumor antigens and restoring anti-tumor immunity.Methods Primary melanoma cells from eight patients were reprogrammed into cDC1-like cells through transduction with lentivirus constitutively expressing PIB. Reprogrammed cells were profiled at multiple time-points to characterize reprogramming efficiency, phenotype, and functional properties including cytokine secretion and the capacity to prime T cells.Results All eight PIB-transduced melanoma cells progressively acquired a cDC1 surface phenotype characterized by the expression of CD45 and HLA-DR, marking the acquisition of hematopoietic and antigen presentation phenotype. The cell reprogramming process was consistent across all cell lines. Induced cDC1s also expressed CD11c, the cDC1-specific markers CLEC9A and CD141 as well as the costimulatory molecules CD40, CD80 and CD86. Functionally, cDC1-like melanoma cells at day 9 secreted the human cDC1-specific cytokines IL12p70 and IL-29 upon stimulation with Poly(I:C). After pulsing with a 9mer MART-1 peptide restricted to HLA-A2, cDC1-like melanoma cells were able to prime allogeneic HLA-A2 matched naïve CD8+ T cells and resulted in expansion of MART-1-specific T-cells after an eight-day co-culture with IL-2 and IL-7. Moreover, autologous tumor-infiltrating lymphocytes (TILs) were more reactive (higher expression of CD107a, CD137, IFN-gamma, and TNF-alpha) and cytotoxic towards cDC1-like melanoma cells compared to the original tumor cells.Conclusions Here, we demonstrated that melanoma cells from multiple patients can be efficiently reprogrammed into cDC1-like cells and present tumor-associated antigens. These results lay the groundwork for the development of cDC1 reprogramming as an innovative cancer immunotherapy to counteract immune escape and reactivating anti-tumor immunity.
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6دورية أكاديمية
المؤلفون: Mejía, Natalia, Santiago, Marta
المصدر: Journal of Parents and Teachers; No. 394 (2023): Positive Discipline; 46-50 ; Padres y Maestros / Journal of Parents and Teachers; Núm. 394 (2023): Disciplina positiva; 46-50 ; 2255-1042 ; 0210-4679
مصطلحات موضوعية: positive discipline, learning, beliefs, breath, educate for life, social-emotional skills, educational style, disciplina positiva, aprendizaje, creencias, aliento, educar para la vida, habilidades socioemocionales, estilo educativo
الوصف: Positive Discipline is getting to know each other, taking care of oneself to care, establishing links that provide feedback, limits and clear rules with kindness and respect and working with students so that they develop socio-emotional skills. Positive discipline is an educational and social model that starts from an environment based on respect and cooperation, to reach the desired learning. It focuses on the connection between teacher-student and student-student, in such a way that this link allows them to cooperate and develop in society, attending to their own needs and those of others. ; La disciplina positiva es conocerse, cuidarse para cuidar, establecer vínculos que retroalimentan, límites y normas claras con amabilidad y respeto y trabajo con los alumnos para que desarrollen habilidades socioemocionales. La disciplina positiva es un modelo educativo y social que parte de un ambiente basado en el respeto y la cooperación, para llegar al aprendizaje deseado. Se centra en la conexión entre profesoralumno y alumno-alumno, de tal manera que ese vínculo le permita cooperar y desarrollarse en sociedad, atendiendo a sus propias necesidades y las de los demás.
وصف الملف: application/pdf
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7دورية أكاديمية
المؤلفون: Zimmermannova, Olga, Ferreira, Alexandra G., Ascic, Ervin, Velasco Santiago, Marta, Kurochkin, Ilia, Hansen, Morten, Met, Özcan, Caiado, Inês, Shapiro, Ilja E., Michaux, Justine, Humbert, Marion, Soto-Cabrera, Diego, Benonisson, Hreinn, Silvério-Alves, Rita, Gomez-Jimenez, David, Bernardo, Carina, Bauden, Monika, Andersson, Roland, Höglund, Mattias, Miharada, Kenichi, Nakamura, Yukio, Hugues, Stéphanie, Greiff, Lennart, Lindstedt, Malin, Rosa, Fábio F., Pires, Cristiana F., Bassani-Sternberg, Michal, Svane, Inge Marie, Pereira, Carlos-Filipe
المصدر: ISSN: 2470-9468 ; Science immunology, vol. 8, no. 85 (2023) eadd4817.
مصطلحات موضوعية: info:eu-repo/classification/ddc/616.07
الوصف: Decreased antigen presentation contributes to the ability of cancer cells to evade the immune system. We used the minimal gene regulatory network of type 1 conventional dendritic cells (cDC1) to reprogram cancer cells into professional antigen-presenting cells (tumor-APCs). Enforced expression of the transcription factors PU.1, IRF8, and BATF3 (PIB) was sufficient to induce the cDC1 phenotype in 36 cell lines derived from human and mouse hematological and solid tumors. Within 9 days of reprogramming, tumor-APCs acquired transcriptional and epigenetic programs associated with cDC1 cells. Reprogramming restored the expression of antigen presentation complexes and costimulatory molecules on the surfaces of tumor cells, allowing the presentation of endogenous tumor antigens on MHC-I and facilitating targeted killing by CD8 + T cells. Functionally, tumor-APCs engulfed and processed proteins and dead cells, secreted inflammatory cytokines, and cross-presented antigens to naïve CD8 + T cells. Human primary tumor cells could also be reprogrammed to increase their capability to present antigen and to activate patient-specific tumor-infiltrating lymphocytes. In addition to acquiring improved antigen presentation, tumor-APCs had impaired tumorigenicity in vitro and in vivo. Injection of in vitro generated melanoma-derived tumor-APCs into subcutaneous melanoma tumors delayed tumor growth and increased survival in mice. Antitumor immunity elicited by tumor-APCs was synergistic with immune checkpoint inhibitors. Our approach serves as a platform for the development of immunotherapies that endow cancer cells with the capability to process and present endogenous tumor antigens.
العلاقة: info:eu-repo/semantics/altIdentifier/pmid/37418548; https://archive-ouverte.unige.ch/unige:172682Test; unige:172682
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8دورية أكاديمية
المؤلفون: Våbenø, Jon, Oliva-Santiago, Marta, Jørgensen, Astrid S., Karlshøj, Stefanie, Rosenkilde, Mette M.
المصدر: Våbenø , J , Oliva-Santiago , M , Jørgensen , A S , Karlshøj , S & Rosenkilde , M M 2023 , ' Identification of a Salt Bridge That Is Functionally Important for Chemokine Receptor CXCR1 but not CXCR2 ' , ACS Pharmacology and Translational Science , vol. 6 , no. 8 , pp. 1120-1128 . https://doi.org/10.1021/acsptsci.3c00070Test
مصطلحات موضوعية: chemokine receptor, chemokine recognition site, CKR signaling, CXCR1, CXCR2, GPCR, inositol triphosphate, salt bridge
الوصف: CXC chemokine receptors 1 (CXCR1) and 2 (CXCR2) have high sequence similarity and overlapping chemokine ligand profiles. Residue positions 3.32 and 7.39 are critical for signal transduction in the related CXCR4, and in these positions CXCR1 and CXCR2 contain oppositely charged residues (Lys 3.32 and Glu 7.39 ). Experimental and computed receptor structures reveal the possible formation of a salt bridge between transmembrane (TM) helices 3 and 7 via these two residues. To investigate the functional importance of Lys117 3.32 and Glu291 7.39 in CXCR1, along with the flanking Glu118 3.33 , we performed a signaling study on 16 CXCR1 mutants using two different CXCL8 isoforms. While single Ala-mutation (K117 3.32 A, E291 7.39 A) and charge reversal (K117 3.32 E, E291 7.39 K) resulted in nonfunctional receptors, double (K117 3.32 E-E291 7.39 K) and triple (K117 3.32 E-E118 3.33 A-E291 7.39 K) mutants rescued CXCR1 function. In contrast, the corresponding mutations did not affect the CXCR2 function to the same extent. Our findings show that the Lys 3.32 -Glu 7.39 salt bridge between TM3 and −7 is functionally important for CXCR1 but not for CXCR2, meaning that signal transduction for these highly homologous receptors is not conserved.
وصف الملف: application/pdf
الإتاحة: https://doi.org/10.1021/acsptsci.3c00070Test
https://curis.ku.dk/portal/da/publications/identification-of-a-salt-bridge-that-is-functionally-important-for-chemokine-receptor-cxcr1-but-not-cxcr2Test(a540724d-cf91-4969-a4b5-cc200b543396).html
https://curis.ku.dk/ws/files/373669522/v_ben_et_al_2023_identification_of_a_salt_bridge_that_is_functionally_important_for_chemokine_receptor_cxcr1_but_not.pdfTest -
9دورية أكاديمية
المؤلفون: Castilla, Lucia, Aspa, Jose, Nieto, Jorge Martínez, Cabrera, Ángel Zúñiga, Santiago, Marta, Feria, Ana Garcia, Marco Buades, Josefa Esperanza, Marina, María Argüello, Ayala Diaz, Rosa María, Sanchez, Ricardo, Orbe Barreto, Rafael Del, Lobo Olmedo, Ana, Dávila-Valls, Julio, Martín-Pérez, Sonia, Jativa Saez, Cristina, Rodriguez, Inés Hernández, Salido, Eduardo, Recasens, Valle, Fernández, Lucía Guerrero, Moreno Risco, Maria Belén, Fernández, Ataulfo González, Rubio, Montserrat Lopez
المصدر: HemaSphere ; volume 7, issue S3, page e6313050 ; ISSN 2572-9241
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10دورية أكاديمية
المؤلفون: Díaz González, Álvaro, Avetisyan, Gayane, Garcia-Ruiz, Cristian, Vicente Gil, José, Santiago, Marta, José Domínguez-García, Juan, Llop, Marta, Barragan, Eva, Leonor Senent Peris, Maria, DE LA Rubia, Javier, Cervera, José, Such, Esperanza
المصدر: HemaSphere ; volume 7, issue S3, page e8338249 ; ISSN 2572-9241
