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The Qui-Bot H2O project involves developing four educational sustainable robots and their associated software. Robots are equipped with HRI features such as voice recognition and color sensing, and they possess a humanoid appearance. The project highlights the social and ethical aspects of robotics applied to chemistry and industry 4.0 at an early age. Here, we report the results of an interactive study that involved 212 students aged within the range of 3-18. Our educational robots were used to measure the backgrounds, impact, and interest of students, as well as their satisfaction after interacting with them. Additionally, we provide an ethical study of the use of these robots in the classroom and a comparison of the interactions of humanoid versus non-humanoid educational robots observed in early childhood learning. Our findings demonstrate that these robots are useful in teaching technical and scientific concepts in a playful and intuitive manner, as well as in increasing the number of girls who are interested in science and engineering careers. In addition, major impact measures generated by the project within a year of its implementation were analyzed. Several public administrations in the area of gender equality endorsed and participated in the Qui-Bot H2O project in addition to educational and business entities.
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Robótica , Preescolar , Femenino , Humanos , Robótica/educación , Niño , AdolescenteRESUMEN
We report a new learning approach in science and technology through the Qui-Bot H2O project: a multidisciplinary and interdisciplinary project developed with the main objective of inclusively increasing interest in computer science engineering among children and young people, breaking stereotypes and invisible social and gender barriers. The project highlights the social aspect of robotics applied to chemistry, at early ages. We successfully tested the project activities on girls between 3 to 13 years old. After taking part in the project, the users rated their interest in science and technology to be higher than before. Data collected during experiences included background information on students, measurements of the project's impact and students' interest in it, and an evaluation of student satisfaction of this STEM activity. The Qui-Bot H2O project is supported by the actions of territorial public administrations towards gender equality and the contributions of humanistic and technological universities and entities which specialize in education and business.
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Ingeniería , Robótica , Adolescente , Niño , Preescolar , Computadores , Investigación Empírica , Ingeniería/educación , Femenino , Humanos , TecnologíaRESUMEN
Poor access and quality of intrapartum and postpartum health care contribute to high global maternal and neonatal mortality rates and intracountry inequity. We examined barriers to careseeking and health care utilization for obstetric and immediate neonatal care in Chiapas, a state with one of the largest indigenous populations and poorest health indicators in Mexico. We conducted 74 in-depth interviews with recently delivered women, their male partners, and traditional birth attendants, and 27 interviews with health facility and hospital staff in rural Chiapas. Interviews were conducted and recorded in Tzeltal and Ch'ol; data were transcribed, coded and analyzed in Spanish using thematic analysis techniques. Barriers to utilization of facility delivery that were reported in order of frequency were: (1) economic and geographic barriers; (2) traditions incompatible with facility policies; (3) fear or previous experience of mistreatment or abuse; (4) perceived poor quality care at facilities; (5) language and political barriers. Commonly reported barriers included distance, cost, lack of vehicles, and poor perceived quality of care, as well as linguistic barriers, lack of space, and fears of surgery or mistreatment. Some women reported obstetric violence and rights violations, including two cases of possible forced sterilizations, an unauthorized transfer of a newborn to another facility without consent or accompaniment of a guardian, and one failure to discharge a newborn because the family could not pay. Political conflict in the region contributed to additional barriers such as reduced trust in government facilities, and physical roadblocks during political activities. Improving geographic and economic access to obstetric and neonatal care can contribute to improved service utilization, but uptake of services can only be improved if quality of care, including communication and consent, are addressed. Historical and current relationships between various stakeholder and political groups should be considered when planning programs, which should be created as collaboratively as possible.
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Servicios de Salud Materna , Parto Obstétrico/métodos , Femenino , Accesibilidad a los Servicios de Salud , Humanos , Recién Nacido , Masculino , México , Embarazo , Investigación Cualitativa , ConfianzaRESUMEN
Despite global efforts, postnatal care utilization remains low. There is almost no research on factors influencing postnatal care utilization in Latin America. Chiapas has one of the highest rates of maternal and neonatal mortality in Mexico. In 2014, we conducted 101 interviews with recently delivered women, male partners, traditional birth attendants (TBAs), and health staff, to assess factors influencing postnatal care utilization. Data underwent content analysis and thematic coding. Travel costs, postnatal seclusion practices, and a low perceived need for postnatal care were found to be disincentives to seek care. Providers ascribed low care adherence to cultural beliefs and low health literacy, while families indicated that their decision to seek facility care was mediated by the TBA's recognition of danger signs and perceived ability to manage complications. TBAs should be leveraged as gatekeepers, health literacy programs should emphasize the importance of primary care, and structural barriers to care should be reduced.
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Servicios de Salud Materna , Partería , Femenino , Humanos , Lactante , Recién Nacido , Masculino , México , Atención Posnatal , EmbarazoRESUMEN
OBJECTIVES: Apolipoprotein A-1 (ApoA-1) is a protein fraction of the high-density lipoproteins with anti-inflammatory and antioxidant properties that play a major role in reverse cholesterol transport. The presence of anti-ApoA-1 IgG has been reported in SLE to be variably associated with disease activity or cardiovascular events (CVEs). We assessed the clinical performance of anti-ApoA-1 IgG and of antibodies directed against its immunodominant F3L1 peptide (F3L1 IgG) in a well-characterized Swiss SLE cohort study. METHODS: A total of 354 biological samples and interviews from 176 individuals were studied. SLEDAI, clinical characteristics, anamnestic CVEs and therapy details were recorded. Sera were tested for the presence of anti-ApoA-1 IgG, anti-F3L1 IgG, anti-dsDNA IgG and aPL. RESULTS: Anti-ApoA-1 and anti-F3L1 IgG positivity was associated with higher SLEDAI, mostly due to concomitant positivity of dsDNA IgG and low complement. Variations in time of anti-ApoA-1 IgG correlated positively with variations of anti-dsDNA IgG and inversely to variations of C3 levels. No cross-reactivity was found between anti-ApoA-1 and anti-dsDNA IgG. Positivity for anti-Apo-A1 IgG was more frequent in individuals receiving 10 mg/day or more of prednisone. We did not find any significant association between anti-ApoA-1 IgG positivity and CVEs. CONCLUSION: Anti-ApoA-1 and anti-F3L1 IgG in SLE correlate strongly with laboratory markers of activity, particularly with the presence and titre of dsDNA IgG. These results confirm and extend previous findings and support the use of anti-ApoA1 IgG in the clinical setting. Their role in CVEs deserves further investigation.
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Apolipoproteínas A/inmunología , Autoanticuerpos/sangre , Lupus Eritematoso Sistémico/diagnóstico , Adolescente , Adulto , Anciano , Niño , Femenino , Humanos , Lupus Eritematoso Sistémico/sangre , Lupus Eritematoso Sistémico/inmunología , Masculino , Persona de Mediana Edad , Índice de Severidad de la Enfermedad , Adulto JovenRESUMEN
BACKGROUND: The biology of some hematological diseases varies among different populations. No previous studies have evaluated the clinical behavior of mantle cell lymphoma (MCL) in México. OBJECTIVE AND METHODS: This is a retrospective review of MCL cases seen in Mexico from January 2003 to June 2020. A total of 12 cases were identified. RESULTS: There were nine males and three females; median age was 56 years. Eight patients had a high MCL international prognostic index score, one was intermediate, and three were low. Five patients had circulating malignant monoclonal cells. Initial treatment included rituximab, cyclophosphamide, daunorubicin, vincristine, and prednisone (R-CHOP) and CHOP. Subsequent treatment included hematopoietic stem cell transplantation in five patients; two were given maintenance therapy. Splenectomy was done in four patients. Median overall survival (OS) for all the patients has not been reached and exceeds 162 mos: OS at 162 mos was 56%. Achieving a complete remission (CR) after the first treatment was a significant prognostic factor, with a median OS exceeding 141 mos in patients achieving CR, and 16 mos among those not achieving CR (p = 0.0006). CONCLUSION: Some of MCL patients in Mexico have an indolent clinical course, particularly patients who achieve a CR to initial treatment and who undergo splenectomy.
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OBJECTIVES: Interleukin (IL) 22 mRNA in systemic sclerosis (SSc) skin and Th22 cells in SSc peripheral blood are increased, but the role of IL-22 in fibrosis development remains poorly understood. METHODS: Biopsies were obtained from the involved skin of 15 SSc, 4 morphea and 8 healthy donors (HD). The presence of IL-22+ cells in the skin was determined by immunostaining. The in vitro response of HD and SSc fibroblasts to IL-22, IL-22 in conjunction with tumour necrosis factor (TNF) or keratinocyte conditioned medium was assessed by ELISA, radioimmunoassay (RIA), real-time PCR and western blot. The in vivo response in mice was assessed by histomorphometry. RESULTS: IL-22+ cells were over-represented in the dermis and epidermis of morphea and in the epidermis of SSc compared with HD. The majority of dermal IL-22+ cells were T cells. Dermal fibroblasts expressed both IL-22 receptor subunits IL-10RB and IL-22RA, expression of which was enhanced by TNF and reduced by transforming growth factor (TGF)-ß. IL-22 induced rapid phosphorylation of p38 and ERK1/2 in fibroblasts, but failed to induce the synthesis of chemokines and extracellular matrix components. However, IL-22 enhanced the production of monocyte chemotactic protein 1, IL-8 and matrix metalloproteinase 1 induced by TNF. Fibroblast responses were maximal in the presence of conditioned medium from keratinocytes activated by IL-22 in conjunction with TNF. Dermal thickness was maximal in mice injected simultaneously with IL-22 and TNF. CONCLUSIONS: IL-22 capacitates fibroblast responses to TNF and promotes a proinflammatory fibroblast phenotype by favouring TNF-induced keratinocyte activation. These results define a novel role for keratinocyte-fibroblast interactions in the context of skin fibrosis.
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Fibroblastos/metabolismo , Interleucinas/metabolismo , Esclerodermia Sistémica/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Adulto , Anciano , Animales , Estudios de Casos y Controles , Epidermis/metabolismo , Femenino , Fibrosis , Humanos , Queratinocitos/metabolismo , Masculino , Ratones , Persona de Mediana Edad , Esclerodermia Localizada/metabolismo , Esclerodermia Localizada/patología , Esclerodermia Sistémica/patología , Piel/patología , Adulto Joven , Interleucina-22RESUMEN
Although immune-based therapies have revolutionized the management of cancer, novel approaches are urgently needed to improve their outcome. We investigated the role of endogenous steroids in the resistance to cancer immunotherapy, as these have strong immunomodulatory functions. Using a publicly available database, we found that the intratumoral expression of 11 beta-hydroxysteroid dehydrogenase type 1 (HSD11B1), which regenerates inactive glucocorticoids into active glucocorticoids, was associated with poor clinical outcome and correlated with immunosuppressive gene signatures in patients with renal cell carcinoma (RCC). HSD11B1 was mainly expressed in tumor-infiltrating immune myeloid cells as seen by immunohistochemistry in RCC patient samples. Using peripheral blood mononuclear cells from healthy donors or immune cells isolated from the tumor of RCC patients, we showed that the pharmacological inhibition of HSD11B1 improved the response to the immune checkpoint inhibitor anti-PD-1. In a subcutaneous mouse model of renal cancer, the combination of an HSD11B1 inhibitor with anti-PD-1 treatment increased the proportion of tumor-infiltrating dendritic cells. In an intrarenal mouse tumor model, HSD11B1 inhibition increased the survival of mice treated with anti-PD-1. In addition, inhibition of HSD11B1 sensitized renal tumors in mice to immunotherapy with resiquimod, a Toll-like receptor 7 agonist. Mechanistically, we demonstrated that HSD11B1 inhibition combined with resiquimod increased T cell-mediated cytotoxicity to tumor cells by stimulating the antigen-presenting capacity of dendritic cells. In conclusion, these results support the use of HSD11B1 inhibitors to improve the outcome of immunotherapy in renal cancer and highlight the role of the endogenous glucocorticoid metabolism in the efficacy of immunotherapy.
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Carcinoma de Células Renales , Neoplasias Renales , Humanos , Animales , Ratones , Glucocorticoides/farmacología , Glucocorticoides/uso terapéutico , Glucocorticoides/metabolismo , Carcinoma de Células Renales/tratamiento farmacológico , Leucocitos Mononucleares/metabolismo , Neoplasias Renales/tratamiento farmacológico , Inmunidad , 11-beta-Hidroxiesteroide Deshidrogenasa de Tipo 1/genética , 11-beta-Hidroxiesteroide Deshidrogenasa de Tipo 1/metabolismoRESUMEN
Murine immune effector cells express three different stimulatory FcγRs (FcγRI, FcγRIII and FcγRIV) and one inhibitory receptor, FcγRIIB. Competitive engagement of stimulatory and inhibitory FcγRs has been shown to be critical for the development of immune complex-mediated inflammatory disorders. Because of the previous demonstration that FcγRIIB was unable to inhibit FcγRIII-mediated autoimmune hemolytic anemia induced by 105-2H IgG1 anti-RBC mAb, we reevaluated the regulatory role of FcγRIIB on the development of anemia using two additional IgG1 anti-RBC mAbs (34-3C and 3H5G1) and different 34-3C IgG subclass-switch variants. We were able to induce a more severe anemia in FcγRIIB-deficient mice than in FcγRIIB-sufficient mice after injection of 34-3C and 3H5G1 IgG1, but not 105-2H IgG1. Structural analysis of N-linked oligosaccharides attached to the CH2 domain revealed that 105-2H was poorly galactosylated as compared with the other mAbs, while the extent of sialylation was comparable between all mAbs. In addition, we observed that a more galactosylated 105-2H variant provoked more severe anemia in FcγRIIB-deficient mice than FcγRIIB-sufficient mice. In contrast, the development of anemia induced by three non-IgG1 subclass variants of the 34-3C mAb was not down-regulated by FcγRIIB, although they were more galactosylated than its IgG1 variant. These data indicate that FcγRIIB-mediated inhibition of autoimmune hemolytic anemia is restricted to the IgG1 subclass and that galactosylation, but not sialylation, of IgG1 (but not other IgG subclasses) is critical for the interaction with FcγR, thereby determining the pathogenic potential of IgG1 autoantibodies.
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Anemia Hemolítica Autoinmune/inmunología , Inmunoglobulina G/inmunología , Receptores de IgG/inmunología , Animales , Anticuerpos Monoclonales/inmunología , Autoanticuerpos/inmunología , Eritrocitos/inmunología , Galactosa/química , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Receptores de IgG/antagonistas & inhibidores , Receptores de IgG/deficienciaRESUMEN
The delivery of drugs is a great challenge, since most of active pharmaceutical ingredients developed today are hydrophobic and poorly water soluble. From this perspective, drug encapsulation on biodegradable and biocompatible polymers can surpass this problem. Poly(γ-glutamic acid) (PGGA), a bioedible and biocompatible polymer has been chosen for this purpose. Carboxylic side groups of PGGA have been partially esterified with 4-phenyl-butyl bromide, producing a series of aliphatic-aromatic ester derivatives with different hydrophilic-lipophilic balances. Using nanoprecipitation or emulsion/evaporation methods, these copolymers were self-assembled in a water solution, forming nanoparticles with average diameters between 89 and 374 nm and zeta potential values between -13.1 and -49.5 mV. The hydrophobic core containing 4-phenyl-butyl side groups was used for the encapsulation of an anticancer drug, such as Doxorubicin (DOX). The highest encapsulation efficiency was reached for a copolymer derived from PGGA, with a 46 mol% degree of esterification. Drug release studies carried out for 5 days at different pHs (4.2 and 7.4) indicated that DOX was released faster at pH 4.2, revealing the potential of these nanoparticles as chemotherapy agents.
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Engineered gold nanoparticles (GNPs) have become a useful tool in various therapeutic and diagnostic applications. Uncertainty remains regarding the possible impact of GNPs on the immune system. In this regard, we investigated the interactions of polymer-coated GNPs with B cells and their functions in mice. Surprisingly, we observed that polymer-coated GNPs mainly interact with the recently identified subpopulation of B lymphocytes named age-associated B cells (ABCs). Importantly, we also showed that GNPs did not affect cell viability or the percentages of other B cell populations in different organs. Furthermore, GNPs did not activate B cell innate-like immune responses in any of the tested conditions, nor did they impair adaptive B cell responses in immunized mice. Together, these data provide an important contribution to the otherwise limited knowledge about GNP interference with B cell immune function, and demonstrate that GNPs represent a safe tool to target ABCs in vivo for potential clinical applications.
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Oro , Nanopartículas del Metal , Ratones , Animales , Supervivencia Celular , Polietilenglicoles , PolímerosRESUMEN
INTRODUCTION: We have previously shown that some patients present thrombocytopenia (less than 100 × 109/L platelets) in non-alcoholic fatty liver disease (NAFLD). To further explore the nature of this association, we have now analyzed the association of thrombocytopenia with neutropenia (less than 0.5 × 109/L granulocytes) in NAFLD. MATERIAL AND METHODS: Persons with NAFLD were prospectively accrued in the study after February 2018. The presence of NAFLD was defined by both serologic determinations (Fibromax ®) and liver transient elastography (TE/Fibroscan ®). RESULTS: In 123 consecutive patients with NAFLD without cirrhosis, thrombocytopenia was identified in 20 (16%), whereas neutropenia was identified in 9 (7%). In the subset of 20 patients with NAFLD and thrombocytopenia, granulocytopenia was identified in 5 (25%), whereas in the subset of 9 patients with granulocytopenia, thrombocytopenia was identified in 5 (55%). We found a significant association between thrombocytopenia and both leukopenia and granulocytopenia (OR 8.25, 95% CI 1.9-34.2, p = 0.004). CONCLUSIONS: Both thrombocytopenia and neutropenia were identified in persons with NAFLD and, as there is a significant relationship between these two variables, we speculate that this finding may support the possibility of hypersplenism being involved in the cytopenias found in NAFLD without cirrhosis.
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Circulating monocytes are divided into two major, phenotypically and functionally distinct subsets: Gr-1(hi) "inflammatory" and Gr-1(lo) "resting" monocytes. One of the unique cellular abnormalities in lupus-prone mice is monocytosis, which is characterized by a selective expansion of Gr-1(lo) monocytes and dependent on the expression of stimulatory IgG Fc receptors (FcγR). We speculated that IgG immune complexes containing nuclear antigens could stimulate Gr-1(hi) monocytes through interaction with FcγRs and then TLR7 and TLR9, thereby promoting the maturation towards Gr-1(lo) monocytes. In the present study, we assessed this hypothesis by analyzing effects of TLR9 or TLR7 agonist on monocytes in vivo. The analysis of various surface markers differentially expressed on both subsets of monocytes in combination with selective depletion of either subset revealed that within 48 h after injection of the TLR9 agonist CpG, approximately one third of Gr-1(hi) monocytes became phenotypically identical to Gr-1(lo) monocytes. In addition, we observed approximately two-fold increases in the total monocyte population 8-24 h after injection of CpG. Moreover, the activation of TLR9 resulted in an increased expression of stimulatory FcγRIV relative to inhibitory FcγRIIB on monocytes, thereby enhancing their responsiveness to IgG immune complexes. Essentially identical results were obtained after stimulation of TLR7 with a synthetic agonist (1V136). Our results indicate that the activation of TLR7 and TLR9 not only induced the maturation of a fraction of Gr-1(hi) monocytes towards Gr-1(lo) monocytes but also promoted the overall generation of monocytes, thereby supporting the critical role of TLR7 and TLR9 for the development of monocytosis in lupus-prone mice.
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Inflamación/patología , Lupus Eritematoso Sistémico/patología , Monocitos/inmunología , Oligodesoxirribonucleótidos/administración & dosificación , Receptores de IgG/inmunología , Receptor Toll-Like 7/inmunología , Receptor Toll-Like 9/inmunología , Animales , Recuento de Células Sanguíneas , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Modelos Animales de Enfermedad , Femenino , Citometría de Flujo , Eliminación de Gen , Inflamación/inmunología , Inflamación/metabolismo , Procedimientos de Reducción del Leucocitos , Lupus Eritematoso Sistémico/inmunología , Lupus Eritematoso Sistémico/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Monocitos/citología , Monocitos/efectos de los fármacos , Monocitos/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Receptores de IgG/deficiencia , Receptores de IgG/genética , Receptor Toll-Like 7/agonistas , Receptor Toll-Like 7/deficiencia , Receptor Toll-Like 7/genética , Receptor Toll-Like 9/agonistas , Receptor Toll-Like 9/deficiencia , Receptor Toll-Like 9/genéticaRESUMEN
BACKGROUND: The outcomes of Hodgkin´s lymphoma (HL) in México have not been widely reported. Simplified and affordable treatments have been adopted in middle-income countries. AIM: The aim was to evaluate long-used therapies for HL in México in a long-term basis. METHODS: In a 34-year time period, 88 patients with HL were treated at a single institution in México. Patients were treated with adriamycin bleomycin vinblastine and dacarbazine (ABVD) or mechlorethamine, vincristine, procarbazine, and prednisone (MOPP). Relapsed or refractory patients were given ifosfamide, carboplatin, and etoposide (ICE) followed by autologous or allogeneic stem cell transplants. RESULTS: Thirty-seven women and 51 men were included; the median age was 29 years. Patients were followed for a mean of 128 mo. The 310-mo overall survival (OS) was 83% for patients treated with MOPP and 88% for those treated with ABVD. The OS of patients who received autologous stem cell transplantation was 76% (330 mo) vs 93% (402 mo) in those who did not. CONCLUSION: HL may be less aggressive in Mexican population than in Caucasians. Combined chemotherapy renders acceptable results, regardless of clinical stage.
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Microenvironment molecular cues direct T helper (Th) cell differentiation; however, Th17 fate determination is still imprecisely understood in humans. To assess the role of prostaglandin E(2) (PGE(2)) in Th expansion, we activated peripheral blood mononuclear cells by CD3 cross-linking. In the presence of exogenous PGE(2), peripheral blood mononuclear cells produced higher interleukin-17 (IL-17), C-C chemokine ligand 20 (CCL20)/macrophage inflammatory protein 3alpha (MIP-3alpha), CXC chemokine ligand 8 (CXCL8)/IL-8, and lower interferon-gamma and IL-22 levels than in control cultures. Exogenous PGE(2) and IL-23 synergized in inducing IL-17, whereas indomethacin and IL-23 blockade drastically reduced IL-17 but not interferon-gamma production. Furthermore, IL-1 but not tumor necrosis factor was absolutely required for IL-17 production. PGE(2) doubled the frequency of CD4+ T cells producing IL-17 and within the CD4+ subset enhanced C-C chemokine receptor 6 (CCR6) and CCR4 while decreasing CXC chemokine receptor 3 (CXCR3) expression. Furthermore, in CD4+ T-cell lines, the production of IL-17 segregated with the CCR6+ subset. In the presence of CCR6+ compared with CXCR3+ Th cells, monocytes/macrophages produced much higher levels of matrix metalloproteinase-1, -3, and -9 but similar levels of CXCL10 and IL-1beta. These results identify PGE(2) and IL-23 as participating in the expansion of CD4+ T cells endowed with high IL-17 production capacity, which in turn favors monocyte production of mediators important for host defense and tissue destruction.
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Dinoprostona/administración & dosificación , Interleucina-17/biosíntesis , Interleucina-23/administración & dosificación , Subgrupos de Linfocitos T/efectos de los fármacos , Subgrupos de Linfocitos T/inmunología , Linfocitos T Colaboradores-Inductores/efectos de los fármacos , Linfocitos T Colaboradores-Inductores/inmunología , Línea Celular , Técnicas de Cocultivo , Dinoprostona/metabolismo , Sinergismo Farmacológico , Humanos , Interferón gamma/metabolismo , Interleucina-1/antagonistas & inhibidores , Interleucina-1/metabolismo , Interleucina-23/metabolismo , Macrófagos/inmunología , Macrófagos/metabolismo , Metaloproteinasas de la Matriz/biosíntesis , Monocitos/inmunología , Monocitos/metabolismo , Receptores CCR4/metabolismo , Receptores CCR6/metabolismo , Proteínas Recombinantes/administración & dosificación , Subgrupos de Linfocitos T/citología , Subgrupos de Linfocitos T/metabolismo , Linfocitos T Colaboradores-Inductores/citología , Linfocitos T Colaboradores-Inductores/metabolismoRESUMEN
IL-17A is abundant in scleroderma but its role in fibrogenesis is controversial. We interrogated the role of IL-17A in extracellular matrix deposition and inflammation by investigating its effects on keratinocytes and fibroblasts cross-talk and in organotypic skin cultures. Keratinocyte-conditioned media of resting, IL-17A-, and/or transforming growth factor-ß-primed primary keratinocytes were used to stimulate healthy donors and scleroderma fibroblasts. Alternatively, organotypic cultures of full human skin were challenged with these cytokines. Keratinocyte-conditioned media tilted the balance of col-I to matrix metalloproteinase-1 production by fibroblasts in favor of matrix metalloproteinase-1, significantly more so in healthy donors than in scleroderma, resulting in enhanced extracellular matrix turnover, further increased by IL-17A. In organotypic skin, transforming growth factor-ß induced an extensive pro-fibrotic gene signature, including the enhanced expression of several collagen genes associated with Wnt signaling. IL-17A strongly promoted the expression of pro-inflammatory genes, with no direct effects on collagen genes, and attenuated Wnt signaling induced by transforming growth factor-ß. In this model, at the protein level, IL-17A significantly decreased col-I production. Our data strongly support a pro-inflammatory and antifibrogenic activity of IL-17A in the context of keratinocyte-fibroblast interaction and in full skin. These data help in directing and interpreting targeted therapeutic approaches in scleroderma.
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Fibroblastos/fisiología , Inflamación/inmunología , Interleucina-17/metabolismo , Queratinocitos/fisiología , Esclerodermia Localizada/inmunología , Esclerodermia Sistémica/inmunología , Piel/patología , Células Cultivadas , Colágeno Tipo I/metabolismo , Matriz Extracelular/metabolismo , Fibrosis , Regulación de la Expresión Génica , Redes Reguladoras de Genes , Humanos , Metaloproteinasa 1 de la Matriz/metabolismo , Técnicas de Cultivo de Órganos , Piel/metabolismo , Factor de Crecimiento Transformador beta/genética , Factor de Crecimiento Transformador beta/metabolismo , Vía de Señalización WntRESUMEN
Systemic sclerosis (SSc) is a chronic autoimmune disease characterized by fibrosis and vasculopathy. CXCL4 represents an early serum biomarker of severe SSc and likely contributes to inflammation via chemokine signaling pathways, but the exact role of CXCL4 in SSc pathogenesis is unclear. Here, we elucidate an unanticipated mechanism for CXCL4-mediated immune amplification in SSc, in which CXCL4 organizes "self" and microbial DNA into liquid crystalline immune complexes that amplify TLR9-mediated plasmacytoid dendritic cell (pDC)-hyperactivation and interferon-α production. Surprisingly, this activity does not require CXCR3, the CXCL4 receptor. Importantly, we find that CXCL4-DNA complexes are present in vivo and correlate with type I interferon (IFN-I) in SSc blood, and that CXCL4-positive skin pDCs coexpress IFN-I-related genes. Thus, we establish a direct link between CXCL4 overexpression and the IFN-I-gene signature in SSc and outline a paradigm in which chemokines can drastically modulate innate immune receptors without being direct agonists.
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ADN Bacteriano/metabolismo , Interferón-alfa/metabolismo , Factor Plaquetario 4/metabolismo , Esclerodermia Sistémica/inmunología , Receptor Toll-Like 9/metabolismo , Adulto , Anciano , Biopsia , Estudios de Casos y Controles , ADN Bacteriano/inmunología , Células Dendríticas/inmunología , Células Dendríticas/metabolismo , Femenino , Voluntarios Sanos , Humanos , Interferón-alfa/inmunología , Cristales Líquidos , Masculino , Persona de Mediana Edad , Factor Plaquetario 4/inmunología , Receptores CXCR3/inmunología , Receptores CXCR3/metabolismo , Esclerodermia Sistémica/microbiología , Esclerodermia Sistémica/patología , Piel/citología , Piel/inmunología , Piel/microbiología , Piel/patología , Receptor Toll-Like 9/inmunologíaRESUMEN
ApoA-1 (apolipoprotein A-1) is the main component of HDL (high-density lipoprotein) and stabilizes PON-1 (paraoxonase-1), which prevents lipid peroxidation and oxLDL (oxidized low-density lipoprotein) formation. Autoantibodies against apoA-1 [anti-(apoA-1) IgG] have been found in antiphospholipid syndrome and systemic lupus erythematosous, two diseases with an increased risk of thrombotic events, as well as in ACS (acute coronary syndrome). OxLDL levels are also elevated in these diseases. Whether anti-(apoA-1) IgGs exist in other prothrombotic conditions, such as APE (acute pulmonary embolism) and stroke, has not been studied and their potential association with oxLDL and PON-1 activity is not known. In the present study, we determined prospectively the prevalence of anti-(apoA-1) IgG in patients with ACS (n=127), APE (n=58) and stroke (n=34), and, when present, we tested their association with oxLDL levels. The prevalance of anti-(apoA-1) IgG was 11% in the ACS group, 2% in the control group and 0% in the APE and stroke groups. The ACS group had significantly higher median anti-(apoA-1) IgG titres than the other groups of patients. Patients with ACS positive for anti-(apoA-1) IgG had significantly higher median oxLDL values than those who tested negative (226.5 compared with 47.7 units/l; P<0.00001) and controls. The Spearman ranked test revealed a significant correlation between anti-(apoA-1) IgG titres and serum oxLDL levels (r=0.28, P<0.05). No association was found between PON-1 activity and oxLDL or anti-(apoA-1) IgG levels. In conclusion, anti-(apoA-1) IgG levels are positive in ACS, but not in stroke or APE. In ACS, their presence is associated with higher levels of oxLDL and is directly proportional to the serum concentration of oxLDL. These results emphasize the role of humoral autoimmunity as a mediator of inflammation and coronary atherogenesis.
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Síndrome Coronario Agudo/sangre , Apolipoproteína A-I/inmunología , Autoanticuerpos/sangre , Inmunoglobulina G/sangre , Lipoproteínas LDL/sangre , Síndrome Coronario Agudo/inmunología , Enfermedad Aguda , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores/sangre , Ensayo de Inmunoadsorción Enzimática/métodos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Embolia Pulmonar/inmunología , Accidente Cerebrovascular/inmunología , Adulto JovenRESUMEN
Functional cytokine networks have been poorly characterized in systemic sclerosis (SSc). While interleukin-17A (IL-17A) is increased in SSc skin and other organs, its role is still debated, particularly considering fibrogenesis. We uncover here a dual function of IL-17A in the presence of transforming growth factor-ß 1 (TGF-ß), the master pro-fibrotic cytokine. In the one hand, we report an unexpected synergic activity resulting in enhanced production of IL-6 by dermal fibroblasts; in the other hand, a substantial inhibition of type I collagen (col-I) production. IL-17A or TGF-ß enhanced the production of IL-6 by 8- to 16-folds when compared to control in healthy donors (HD) and SSc cultures. However, the joint presence of IL-17A and TGF-ß resulted in robustly exuberant responses with levels of IL-6 up to 100-folds higher than those observed in untreated cells. Inhibition of NFκB signaling pathway preferentially inhibited the production of IL-6 driven by IL-17A in HD fibroblasts, while inhibition of PI3K preferentially inhibited the production of IL-6 driven by TGF-ß. Interestingly, when p38 MAPK was inhibited, substantial reduction of IL-6 production was observed for both IL-17A and TGF-ß. Consistently with the inhibition experiments, the combined stimulation of fibroblasts by IL-17A and TGF-ß resulted in 1.8-fold increase in p38 MAPK phosphorylation (P = 0.025), when compared to levels of phosphorylated p38 MAPK induced by IL-17A alone. Furthermore, the enhanced phosphorylation of p38 MAPK in the joint presence of IL-17A and TGF-ß was unique among the signaling molecules we examined. As expected, TGF-ß induced SMAD2 phosphorylation and col-I production. However, in fibroblasts cultured in the joint presence of TGF-ß and IL-17A, SMAD2 phosphorylation was decreased by 0.6-folds (P = 0.022) when compared to that induced by TGF-ß alone. Remarkably, in this condition, the production of col-I and fibronectin was significantly decreased in both HD and SSc. Thus, IL-17A and TGF-ß reciprocally influence each other effector functions in fibroblasts. Intracellular molecular switches may favor synergic or antagonistic activities, which are revealed by specific readouts. The implications of these data in the context of SSc are far reaching, particularly in terms of therapeutic approaches since IL-6, IL-17A, and TGF-ß are all putative targets of treatment.
Asunto(s)
Colágeno Tipo I/metabolismo , Fibroblastos/fisiología , Interleucina-17/metabolismo , Interleucina-6/metabolismo , Esclerodermia Sistémica/inmunología , Piel/patología , Factor de Crecimiento Transformador beta1/metabolismo , Adulto , Anciano , Células Cultivadas , Femenino , Fibrosis , Regulación de la Expresión Génica , Humanos , Sistema de Señalización de MAP Quinasas , Masculino , Persona de Mediana Edad , FN-kappa B/metabolismoRESUMEN
The hydrolytic degradability and erythromycin release from stoichiometric ionic complexes of biotechnological poly(beta,L-malic acid)s and poly(gamma,D-glutamic acid)s with alkyltrimethylammonium surfactants were investigated. The influence of pH, temperature and antibiotic load on hydrolysis rate was examined. It was found that poly(malic acid) complexes degraded by a surface erosion mechanism at a higher rate than poly(glutamic acid) complexes, which eroded in bulk. Erythromycin was lodged in the paraffinic subphase of the complexes and upon aging it was delivered according to a sigmoidal profile that appeared to be independent on the antibiotic load.