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Background: Hypertension is a common disease globally that accounts for the highest number of lost healthy life years and strongly associated with sequelae such as stroke and myocardial infarction. Early detection of individuals with high blood pressure can be ensured by screening also those who consider themselves "healthy". Screening has both positive and negative effects where a diagnosis of hypertension can lead to worry about the future. These effects need to be elucidated in order to balance between benefit and harm before screening is introduced. The aim of the study was to describe individuals' experiences of being screened for hypertension in dental health care. Methods: Data from individual semi-structured interviews, with twenty participants screened for cardiovascular risk factors in connection with dental examination and aged 55-80 years, were analysed by means of qualitative content analysis. Results: The results describe individuals' experiences of blood pressure screening in dental health care by means of the following theme: "No big deal" based on two categories: "Convenient way of measuring blood pressure" and "Increased awareness of health". Conclusion: The overall message from the interviews was that having one's blood pressure measured when visiting the dentist was convenient, easy and "No big deal". Blood pressure screening did not create any major concerns and contributed to an increased awareness of health.
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OBJECTIVE: To study white-coat hypertension (WCHT, blood pressure ≥140/90 mmHg in a clinic and normal blood pressure <135/85 mmHg at home), with blood pressure screening of a healthy population during their dental healthcare visit and the associated risk factors. DESIGN: A multicentre observational study. SETTING: A healthy general population at four dental clinics in a region in southern Sweden. SUBJECTS: 2025 individuals aged 40-75 years were screened for high blood pressure at their annual regular check-up dental visit. MAIN OUTCOME MEASURES: Frequencies of normal and elevated blood pressure (BP) in dental clinics, with home BP as a reference. According to BP results, the population was divided into three groups: normotension (NT), WCHT and suspected hypertension (HT). Background and life style factors were measured: sex, age, family history of hypertension, body mass index (BMI kg/m2), education level, tobacco use, and physical activity level. RESULTS: The overall prevalence of WCHT in the study was 17.7%, and the prevalence was 57.2% among those with clinically high blood pressure. Compared with NT, WCHT was associated with male sex (OR 1.56, CI 1.18-2.06), older age group (OR 2.33, CI 1.66-3.26), family history of hypertension (OR 1.61, CI 1.24-2.10), high BMI kg/m2 (OR 2.36, CI 1.80-3.10), daily snuff use (OR 1.74, CI 1.19-2.53). In comparison with WCHT, HT was associated with male sex (OR 2.16, CI 1.44-3.25), older age group (OR 2.85, CI 1.75-4.65), daily smoking (OR 2.10, CI 1.14-3.85), less daily snuff use (OR 0.59, CI 0.34-0.99). CONCLUSIONS: The prevalence of WCHT in a healthy population was 17.7%. Regarding cardiovascular risk factors, WCHT seems to be in the middle of NT and HT. Individuals with WCHT can be identified and given lifestyle advice in connection with a dental check-up, but follow-up and assessment of their cardiovascular risk should take place in primary care.Key pointsScreening in dental practice can detect white-coat hypertension (WCHT) (17.7%) and suspected hypertension (HT) (12.4%).Individuals with WCHT have more cardiovascular risk factors than normotensive individuals.Individuals with WCHT could be given lifestyle advice in dental clinics according to current guidelines.
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Hipertensión , Hipertensión de la Bata Blanca , Adulto , Anciano , Presión Sanguínea , Atención a la Salud , Femenino , Humanos , Hipertensión/diagnóstico , Hipertensión/epidemiología , Masculino , Persona de Mediana Edad , Factores de Riesgo , Hipertensión de la Bata Blanca/diagnóstico , Hipertensión de la Bata Blanca/epidemiologíaRESUMEN
BACKGROUND: Hypertension is one of the largest contributors to the disease burden and a major economic challenge for health-care systems. Early detection of persons with high blood pressure can be achieved through screening and has the potential to reduce morbidity and mortality. We evaluate the cost-effectiveness of an opportunistic hypertension screening programme in a dental-care facility for individuals aged 40-75 in comparison to care as usual (the no-screening baseline scenario). METHODS: A cost-effectiveness analysis (CEA) was carried out from the payer and societal perspectives, and the short-term (from screening until diagnosis has been established) cost per identified case of hypertension and long-term (20 years) cost per quality-adjusted life year (QALY) were reported. Data on the short-term cost were based on a real-world screening programme in which 2025 healthy individuals were screened for hypertension. Data on the long-term cost were based on the short-term outcomes combined with modelling in a Markov cohort model. Deterministic and probabilistic sensitivity analyses were carried out to assess uncertainty. RESULTS: The short-term analysis showed an additional cost of 4,800 SEK (470) per identified case of hypertension from the payer perspective and from the societal perspective 12,800 SEK (1,240). The long-term analysis showed a payer cost per QALY of 2.2 million SEK (210,000) and from the societal perspective 2.8 million SEK per QALY (270,000). CONCLUSION: The long-term model results showed that the screening model is unlikely to be cost-effective in a country with a well-developed health-care system and a relatively low prevalence of hypertension.
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Monitoreo Ambulatorio de la Presión Arterial/economía , Análisis Costo-Beneficio , Servicios de Salud Dental/estadística & datos numéricos , Tamizaje Masivo/economía , Monitoreo Ambulatorio de la Presión Arterial/métodos , Servicios de Salud Dental/economía , Femenino , Humanos , Masculino , Tamizaje Masivo/métodos , Persona de Mediana Edad , Modelos EstadísticosRESUMEN
New robust and reproducible differentiation approaches are needed to generate induced pluripotent stem cell (iPSC)-derived cardiomyocytes of specific subtypes in predictable quantities for tissue-specific disease modeling, tissue engineering, and eventual clinical translation. Here, we assessed whether powdered decellularized extracellular matrix (dECM) particles contained chamber-specific cues that could direct the cardiac differentiation of human iPSCs toward an atrial phenotype. Human hearts were dissected and the left ventricle (LV) and left atria (LA) were isolated, minced, and decellularized using an adapted submersion decellularization technique to generate chamber-specific powdered dECM. Comparative proteomic analyses showed chamber-specific dECM segregation, with atrial- and ventricle-specific proteins uniquely present in powdered dECM-hA and dECM-hV, respectively. Cell populations differentiated in the presence of dECM-hA showed upregulated atrial molecular markers and a two-fold increase in the number of atrial-like cells as compared with cells differentiated with dECM-hV or no dECM (control). Finally, electrophysiological data showed an increase in action potentials characteristic of atrial-like cells in the dECM-hA group. These findings support the hypothesis that dECM powder derived from human atria retained endogenous cues to drive cardiac differentiation toward an atrial fate.
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Células Madre Pluripotentes Inducidas , Diferenciación Celular , Señales (Psicología) , Matriz Extracelular , Humanos , Miocitos Cardíacos , Proteómica , Ingeniería de TejidosRESUMEN
Coastal global oceans are expected to undergo drastic changes driven by climate change and increasing anthropogenic pressures in coming decades. Predicting specific future conditions and assessing the best management strategies to maintain ecosystem integrity and sustainable resource use are difficult, because of multiple interacting pressures, uncertain projections, and a lack of test cases for management. We argue that the Baltic Sea can serve as a time machine to study consequences and mitigation of future coastal perturbations, due to its unique combination of an early history of multistressor disturbance and ecosystem deterioration and early implementation of cross-border environmental management to address these problems. The Baltic Sea also stands out in providing a strong scientific foundation and accessibility to long-term data series that provide a unique opportunity to assess the efficacy of management actions to address the breakdown of ecosystem functions. Trend reversals such as the return of top predators, recovering fish stocks, and reduced input of nutrient and harmful substances could be achieved only by implementing an international, cooperative governance structure transcending its complex multistate policy setting, with integrated management of watershed and sea. The Baltic Sea also demonstrates how rapidly progressing global pressures, particularly warming of Baltic waters and the surrounding catchment area, can offset the efficacy of current management approaches. This situation calls for management that is (i) conservative to provide a buffer against regionally unmanageable global perturbations, (ii) adaptive to react to new management challenges, and, ultimately, (iii) multisectorial and integrative to address conflicts associated with economic trade-offs.
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Ecosistema , Océanos y Mares , Países Bálticos , Cambio Climático , Economía , Geografía , Biología Marina , Modelos TeóricosRESUMEN
AIM: The purpose of this study was to evaluate a two-step screening method for hypertension in dentistry regarding the number needed to screen (NNS) and positive predictive value (PPV) and to risk-classify those with newly diagnosed hypertension. METHODS: In connection with their regular dental care check-up, 2025 subjects aged 40-75 years were screened for high blood pressure. Via a health questionnaire, data were collected concerning risk factors. Blood pressure was screened comprehensively in two steps, which included screening in a dental clinic and home measurements for one week. Recently discovered hypertensive participants were assessed for 10-year risk of cardiovascular mortality according to the guidelines of the European Society of Hypertension (ESH) and the European Society of Cardiology (ESC). RESULTS: A total of 170 new hypertensive participants were found (NNS = 12; 95% confidence interval (CI): 11-13). The method yielded a PPV of 0.73 (95% CI: 0.68-0.78) and eliminated 84.8% of the false-positive participants. The results also showed that based on ESH/ESC risk estimation, 76.5% of those newly diagnosed hypertensive participants had a moderate or high risk of cardiovascular mortality within 10 years. CONCLUSIONS: The study shows that a two-step method for blood pressure screening in a dental setting including home measurement resulted in a high PPV and eliminated most of those with a false high blood pressure reading. The findings also show that two-step screening for hypertension is feasible in a larger population with more screening providers involved.
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Presión Sanguínea , Atención Odontológica , Hipertensión/diagnóstico , Tamizaje Masivo/métodos , Adulto , Anciano , Femenino , Humanos , Hipertensión/epidemiología , Masculino , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Factores de Riesgo , Suecia/epidemiologíaRESUMEN
Reducing anthropogenic nutrient inputs is a major policy goal for restoring good environmental status of coastal marine ecosystems. However, it is unclear to what extent reducing nutrients would also lower fish production and fisheries yields. Empirical examples of changes in nutrient loads and concurrent fish production can provide useful insights to this question. In this paper, we investigate to what extent a multi-fold increase in nutrient loads from the 1950s to 1980s enhanced forage fish production in the Baltic Sea. We use monitoring data on fish stock dynamics covering the period of the nutrient increase, combined with nutrient concentrations from a 3-dimensional coupled physical-biogeochemical ocean model. The results suggest that nutrient enrichment enhanced the biomass level of forage fish by up to 50 % in some years and areas due to increased body weight of fish. However, the trends in fish biomasses were generally decoupled from changes in nutrient concentrations.
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Monitoreo del Ambiente/métodos , Eutrofización , Peces/crecimiento & desarrollo , Agua de Mar/química , Contaminación del Agua/análisis , Animales , Países Bálticos , Nitrógeno/análisis , Océanos y Mares , Fósforo/análisis , Estaciones del AñoRESUMEN
Acute lung injury (ALI) and its most severe manifestation, acute respiratory distress syndrome (ARDS), is a clinical syndrome defined by acute hypoxemic respiratory failure and bilateral pulmonary infiltrates consistent with edema. In-hospital mortality is 38.5% for AL, and 41.1% for ARDS. Activation of alveolar macrophages in the donor lung causes the release of pro-inflammatory chemokines and cytokines, such as TNF-α. To determine the relevance of TNF-α in disrupting bronchial endothelial cell function, we stimulated human THP-1 macrophages with lipopolysaccharide (LPS) and used the resulting cytokine-supplemented media to disrupt normal endothelial cell functions. Endothelial tube formation was disrupted in the presence of LPS-activated THP- 1 conditioned media, with reversal of the effect occurring in the presence of 0.1µg/ml Enbrel, indicating that TNF-α was the major serum component inhibiting endothelial tube formation. To facilitate lung conditioning, we tested liposomal and porous silicon (pSi) delivery systems for their ability to selectively silence TNFR1 using siRNA technology. Of the three types of liposomes tested, only cationic liposomes had substantial endothelial uptake, with human cells taking up 10-fold more liposomes than their pig counterparts; however, non-specific cellular activation prohibited their use as immunosuppressive agents. On the other hand, pSi microparticles enabled the accumulation of large amounts of siRNA in endothelial cells compared to standard transfection with Lipofectamine(®) LTX, in the absence of non-specific activation of endothelia. Silencing of TNFR1 decreased TNF-α mediated inhibition of endothelial tube formation, as well as TNF-α-induced upregulation of ICAM-1, VCAM, and E-selection in human lung microvascular endothelial cells.
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Lesión Pulmonar Aguda/fisiopatología , ARN Interferente Pequeño/administración & dosificación , Receptores Tipo I de Factores de Necrosis Tumoral/genética , Síndrome de Dificultad Respiratoria/fisiopatología , Animales , Cationes/metabolismo , Citocinas/metabolismo , Selectina E/genética , Células Endoteliales/metabolismo , Silenciador del Gen , Humanos , Molécula 1 de Adhesión Intercelular/genética , Lipopolisacáridos/farmacología , Liposomas , Macrófagos/metabolismo , Microvasos/citología , Microvasos/metabolismo , Especificidad de la Especie , Porcinos , Factor de Necrosis Tumoral alfa/metabolismo , Regulación hacia Arriba/genética , Molécula 1 de Adhesión Celular Vascular/genéticaRESUMEN
Modulation of the cytokine milieu is one approach for vaccine development. However, therapy with pro-inflammatory cytokines, such as IL-12, is limited in practice due to adverse systemic effects. Spatially-restricted gene expression circumvents this problem by enabling localized amplification. Intracellular co-delivery of gold nanorods (AuNR) and a heat shock protein 70 (HSP70) promoter-driven expression vector enables gene expression in response to near infrared (NIR) light. AuNRs absorb the light, convert it into heat and thereby stimulate photothermal expression of the cytokine. As proof-of-concept, human HeLa and murine B16 cancer cells were transfected with a HSP70-Enhanced Green Fluorescent Protein (EGFP) plasmid and polyethylenimine (PEI)-conjugated AuNRs. Exposure to either 42 °C heat-shock or NIR light induced significant expression of the reporter gene. In vivo NIR driven expression of the reporter gene was confirmed at 6 and 24 h in mice bearing B16 melanoma tumors using in vivo imaging and flow-cytometric analysis. Overall, we demonstrate a novel opportunity for site-directed, heat-inducible expression of a gene based upon the NIR-absorbing properties of AuNRs and a HSP70 promoter-driven expression vector.
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We present a multi-model ensemble study for the Baltic Sea, and investigate the combined impact of changing climate, external nutrient supply, and fisheries on the marine ecosystem. The applied regional climate system model contains state-of-the-art component models for the atmosphere, sea ice, ocean, land surface, terrestrial and marine biogeochemistry, and marine food-web. Time-dependent scenario simulations for the period 1960-2100 are performed and uncertainties of future projections are estimated. In addition, reconstructions since 1850 are carried out to evaluate the models sensitivity to external stressors on long time scales. Information from scenario simulations are used to support decision-makers and stakeholders and to raise awareness of climate change, environmental problems, and possible abatement strategies among the general public using geovisualization. It is concluded that the study results are relevant for the Baltic Sea Action Plan of the Helsinki Commission.
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Cambio Climático , Ecosistema , Países Bálticos , Océanos y MaresRESUMEN
Multi-model ensemble simulations using three coupled physical-biogeochemical models were performed to calculate the combined impact of projected future climate change and plausible nutrient load changes on biogeochemical cycles in the Baltic Sea. Climate projections for 1961-2099 were combined with four nutrient load scenarios ranging from a pessimistic business-as-usual to a more optimistic case following the Helsinki Commission's (HELCOM) Baltic Sea Action Plan (BSAP). The model results suggest that in a future climate, water quality, characterized by ecological quality indicators like winter nutrient, summer bottom oxygen, and annual mean phytoplankton concentrations as well as annual mean Secchi depth (water transparency), will be deteriorated compared to present conditions. In case of nutrient load reductions required by the BSAP, water quality is only slightly improved. Based on the analysis of biogeochemical fluxes, we find that in warmer and more anoxic waters, internal feedbacks could be reinforced. Increased phosphorus fluxes out of the sediments, reduced denitrification efficiency and increased nitrogen fixation may partly counteract nutrient load abatement strategies.
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Cambio Climático , Ecología , Modelos Teóricos , Países Bálticos , Geología , Océanos y Mares , Fitoplancton/crecimiento & desarrollo , Fitoplancton/aislamiento & purificaciónRESUMEN
Porous silicon microparticles presenting pathogen-associated molecular patterns mimic pathogens, enhancing internalization of the microparticles and activation of antigen presenting dendritic cells. We demonstrate abundant uptake of microparticles bound by the TLR-4 ligands LPS and MPL by murine bone marrow-derived dendritic cells (BMDC). Labeled microparticles induce concentration-dependent production of IL-1ß, with inhibition by the caspase inhibitor Z-VAD-FMK supporting activation of the NLRP3-dependent inflammasome. Inoculation of BALB/c mice with ligand-bound microparticles induces a significant increase in circulating levels of IL-1ß, TNF-α, and IL-6. Stimulation of BMDC with ligand-bound microparticles increases surface expression of costimulatory and MHC molecules, and enhances migration of BMDC to the draining lymph node. LPS-microparticles stimulate in vivo C57BL/6 BMDC and OT-1 transgenic T cell interactions in the presence of OVA SIINFEKL peptide in lymph nodes, with intact nodes imaged using two-photon microscopy. Formation of in vivo and in vitro immunological synapses between BMDC, loaded with OVA peptide and LPS-microparticles, and OT-1 T cells are presented, as well as elevated intracellular interferon gamma levels in CD8(+) T cells stimulated by BMDC carrying peptide-loaded microparticles. In short, ligand-bound microparticles enhance (1) phagocytosis of microparticles; (2) BMDC inflammasome activation and upregulation of costimulatory and MHC molecules; (3) cellular migration of BMDC to lymphatic tissue; and (4) cellular interactions leading to T cell activation in the presence of antigen.
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Movimiento Celular/inmunología , Células Dendríticas/inmunología , Inflamasomas/inmunología , Ganglios Linfáticos/inmunología , Activación de Linfocitos/inmunología , Animales , Células de la Médula Ósea/inmunología , Linfocitos T CD8-positivos/inmunología , Células Cultivadas , Interferón gamma/inmunología , Interleucina-1beta/inmunología , Interleucina-6/inmunología , Ligandos , Lipopolisacáridos/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Fagocitosis/inmunología , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
The selective estrogen receptor modulator tamoxifen is used for the prevention and treatment of breast cancer. The adverse effects of tamoxifen include vaginal endometrial bleeding, endometrial hyperplasia, and cancer, conditions associated with angiogenesis. The aim of this study was to examine the effects of tamoxifen on cell migration and angiogenesis-related gene expression in human endometrial endothelial cells (HEECs). The regulatory effects of tamoxifen on endometrial stromal cells and HEECs were also examined. HEECs and stromal cells were isolated and grown in monocultures or co-cultures, and incubated with 0.1 to 100 µmol/L tamoxifen for 48 hours. Quantitative PCR demonstrated that tamoxifen decreased the mRNA expression of vascular endothelial growth factor-A (VEGF-A) and increased the mRNA expression of VEGF receptor-1 and placental growth factor (PLGF) in HEECs. Tamoxifen's effects on VEGF-A were inhibited when HEECs were co-cultured with stromal cells. In addition, tamoxifen reduced VEGF-induced HEEC migration. The tamoxifen-metabolizing enzymes CYP1A1 and CYP1B1 were detected by immunohistochemistry in and around endometrial blood vessels and by quantitative PCR in HEECs. Our data suggest that tamoxifen changes the regulation of angiogenesis in the endometrium, likely by reducing angiogenic activity. The results also indicate that endometrial stromal cells regulate some of tamoxifen's effects in HEECs, and the presence of tamoxifen-metabolizing enzymes suggests tamoxifen bioactivation in the endometrial vasculature in vivo. These findings may help to elucidate the mechanism of the bleeding disturbances associated with tamoxifen treatment.
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Antineoplásicos Hormonales/farmacología , Endometrio/irrigación sanguínea , Neovascularización Fisiológica/efectos de los fármacos , Tamoxifeno/farmacología , Antineoplásicos Hormonales/administración & dosificación , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Técnicas de Cocultivo , Sistema Enzimático del Citocromo P-450/biosíntesis , Sistema Enzimático del Citocromo P-450/genética , Relación Dosis-Respuesta a Droga , Endometrio/citología , Endometrio/efectos de los fármacos , Células Endoteliales/efectos de los fármacos , Células Endoteliales/metabolismo , Estradiol/farmacología , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Neovascularización Fisiológica/genética , ARN Mensajero/genética , Receptores de Endotelina/biosíntesis , Receptores de Endotelina/genética , Células del Estroma/metabolismo , Células del Estroma/fisiología , Tamoxifeno/administración & dosificación , Factor A de Crecimiento Endotelial Vascular/farmacologíaRESUMEN
Bisphenol A (BPA) is widely used in the manufacturing of consumer products such as plastic food containers and food cans. Experimental studies suggest a relationship between exposure to BPA and changes in metabolic processes and reproductive organs. Also, epidemiological studies report an association between elevated exposure to BPA and cardiovascular disease and diabetes. Although alterations in the vascular endothelium are implicated in pathological conditions associated with BPA, little is known about the effects of BPA in the human endothelium. This study aimed to investigate the effects of 0.1 nM-1 µM of BPA on selected biomarkers of endothelial dysfunction, inflammation, and angiogenesis in human umbilical vein endothelial cells (HUVEC). The mRNA expression of biomarkers was assayed using qRT-PCR, and the production of nitric oxide and reactive oxygen species was measured using the H(2)DCFDA and the DAF-FM assays. The effect of BPA on phosphorylated eNOS was examined using Western blot and immunofluorescence, and the endothelial tube formation assay was used to investigate in vitro angiogenesis. BPA (≤1 µM) increased the mRNA expression of the proangiogenic genes VEGFR-2, VEGF-A, eNOS, and Cx43 and increased the production of nitric oxide in HUVEC. Furthermore, BPA increased the expression of phosphorylated eNOS and endothelial tube formation in HUVEC. These studies demonstrate that environmentally relevant levels of BPA have direct proangiogenic effects on human primary endothelial cells in vitro suggesting that the human endothelium may be an important target for BPA.
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Células Endoteliales/efectos de los fármacos , Contaminantes Ambientales/toxicidad , Neovascularización Fisiológica/efectos de los fármacos , Fenoles/toxicidad , Compuestos de Bencidrilo , Células Cultivadas , Conexina 43/genética , Células Endoteliales/metabolismo , Humanos , Óxido Nítrico/biosíntesis , Óxido Nítrico Sintasa de Tipo III/genética , ARN Mensajero/análisis , Factor A de Crecimiento Endotelial Vascular/genética , Receptor 2 de Factores de Crecimiento Endotelial Vascular/genéticaRESUMEN
The serine proteases, neutrophil elastase (HNE) and proteinase 3 (PR3), are aberrantly expressed in human myeloid leukemias. T-cell responses to these proteins have been correlated with remission in patients with chronic myeloid leukemia (CML). Human PR3/HNE-specific CD8(+) T cells predominantly recognize a nonameric HLA-A2-restricted T-cell epitope called PR1 which is conserved in both Ags. However, CML patients have CD8(+) T cells in peripheral blood recognizing an additional HLA-A2 epitope termed PR2. To assess immunologic properties of these Ags, novel recombinant vaccinia viruses (rVV) expressing PR3 and HNE were evaluated in HLA-A2 transgenic (Tg) mice (HHDII). Immunization of HHDII mice with rVV-PR3 elicited a robust PR3-specific CD8(+) T-cell response dominated by recognition of PR2, with minimal recognition of the PR1 epitope. This result was unexpected, because the PR2 peptide has been reported to bind poorly to HLA. To account for these findings, we proposed that HHDII mice negatively selected PR1-specific T cells because of the presence of this epitope within murine PR3 and HNE, leading to immunodominance of PR2-specific responses. PR2-specific splenocytes are cytotoxic to targets expressing naturally processed PR3, though PR1-specific splenocytes are not. We conclude that PR2 represents a functional T-cell epitope recognized in mice and human leukemia patients. These studies are registered at www.clinicaltrials.gov as NCT00716911.
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Epítopos de Linfocito T/inmunología , Antígeno HLA-A2/inmunología , Leucemia Mielógena Crónica BCR-ABL Positiva/inmunología , Secuencia de Aminoácidos , Animales , Presentación de Antígeno , Linfocitos T CD8-positivos/inmunología , Gránulos Citoplasmáticos/enzimología , Gránulos Citoplasmáticos/inmunología , Epítopos de Linfocito T/genética , Antígeno HLA-A2/genética , Antígeno HLA-A2/metabolismo , Trasplante de Células Madre Hematopoyéticas , Humanos , Epítopos Inmunodominantes/genética , Epítopos Inmunodominantes/inmunología , Leucemia Mielógena Crónica BCR-ABL Positiva/terapia , Elastasa de Leucocito/genética , Elastasa de Leucocito/inmunología , Ratones , Ratones Transgénicos , Datos de Secuencia Molecular , Mieloblastina/genética , Mieloblastina/inmunología , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Homología de Secuencia de Aminoácido , Linfocitos T Citotóxicos/inmunología , Virus Vaccinia/genética , Virus Vaccinia/inmunologíaRESUMEN
Epidemiological and experimental studies suggest an association between elevated serum levels of co-planar PCBs and hypertension, and one study indicate that this effect is dependent on the level of oestrogen. This study investigated the effects of 3,3',4,4',5-pentachlorobiphenyl (PCB126) and 17ß-oestradiol (E2) on vasoactive factors in human umbilical vein endothelial cells (HUVEC). The results reveal that PCB126 stimulated the vasoconstriction factors COX-2 and PGF(2α) in HUVEC. An up-regulation of COX-2 expression was demonstrated using qRT-PCR, western blot and immunofluorescence and increased production of PGF(2α) was demonstrated using LC/MS² and enzyme immunoassay. Also, PCB126 slightly increased ROS production and decreased NO production in HUVEC. The addition of E2 enhanced PCB126-induced transcription of CYP1A1, CYP1B1 and COX-2 in HUVEC whereas an increased transcription of eNOS only occurred following combined treatment with E2 and PCB126. Immunofluorescence demonstrated that HUVEC expressed AHR and ERß but lacked ERα and the involvement of AHR and ERß on the effects of PCB126 was examined by the addition of AHR and ER antagonists. The binding of PCB126 to AHR was critical for the effects of PCB126 whereas the role of ERß was equivocal. In conclusion, these studies suggest that PCB126 induced changes in human endothelial cells that are characteristic for endothelial dysfunction in human hypertension and that PCB126-induced transcription of genes important for vascular function in human endothelial cells can be elevated by increased oestrogen levels. These findings may help understanding the mechanism for the association between PCB126 exposure and hypertension reported in human subjects and experimental animals.
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Células Endoteliales/efectos de los fármacos , Estradiol/farmacología , Antagonistas de Estrógenos/toxicidad , Bifenilos Policlorados/toxicidad , Células Cultivadas , Ciclooxigenasa 2/efectos de los fármacos , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/metabolismo , Dinoprost/metabolismo , Células Endoteliales/metabolismo , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/metabolismo , Técnica del Anticuerpo Fluorescente , Humanos , Óxido Nítrico/biosíntesis , Unión Proteica , Especies Reactivas de Oxígeno/metabolismo , Receptores de Hidrocarburo de Aril/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transcripción Genética/efectos de los fármacos , Venas Umbilicales/citología , Regulación hacia Arriba/efectos de los fármacosRESUMEN
Survivin is a universal tumor antigen that is being currently targeted in vaccine approaches against cancer. Our study here examined the immunogenicity of a novel variant of an HLA-A0201-binding decamer peptide from region 95 to 104 of Survivin (ELMLGEFLKL) with a TâM modification at position 3 in the peptide. We found that this new modified 10-mer peptide had enhanced HLA-A0201 binding and induced a stronger T-cell response over its wild type counterpart peptide (ELTLGEFLKL) in select HLA-A0201(+) normal donors. In addition, when compared to the previously characterized altered 96-104 peptide (LMLGEFLKL) from the same region of Survivin currently used in vaccine trials, we found that both peptides had similar immunogenicity, but donor T cells preferentially reacted strongly to either one or the other, but not strongly to both. These results suggest that these two closely related Survivin peptides yield distinct T-cell responses and that most individuals dominantly respond to one or the other altered peptide. We also found a novel association between positive reactivity to the new altered decamer Survivin peptide in some individuals and their expression of the HLA-C0701 allele along with HLA-A0201. Thus, vaccinating with both the 10-mer and 9-mer peptides would be required to immunize a maximum number of individuals in the HLA-A0201(+) population and could lead to more consistent T-cell responses against this region of Survivin.
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Antígenos de Neoplasias/inmunología , Epítopos de Linfocito T/inmunología , Antígenos HLA-A/inmunología , Proteínas Inhibidoras de la Apoptosis/inmunología , Linfocitos T/inmunología , Línea Celular Tumoral , Reacciones Cruzadas , Antígeno HLA-A2 , Humanos , Modelos Moleculares , Péptidos/inmunología , Unión Proteica , SurvivinRESUMEN
The beneficial effects of tamoxifen in the prevention and treatment of breast cancer are compromised by an increased risk of endometrial polyps, hyperplasia, and cancer. Tamoxifen is metabolized to an array of metabolites with estrogenic effects but also to reactive intermediates that may form protein and DNA adducts. The aim of this study was to investigate cellular [(3)H]tamoxifen adduct formation by light microscopic autoradiography and cell stress by immunohistochemical analysis of glucose-regulating protein 78 (GRP78), nuclear factor kappaB (NF-kappaB), and caspase 3 in human endometrial explants after short-term incubation with tamoxifen. The cellular expression of tamoxifen-metabolizing enzymes in human endometrial biopsy samples was also determined by immunohistochemistry. The results showed selective [(3)H]tamoxifen adduct formation in glandular and surface epithelia after incubation with a nontoxic concentration of [(3)H]tamoxifen (6 nM). There was also a selective expression of the endoplasmic reticulum stress chaperone GRP78 and activated caspase 3 at these sites after incubation with cytotoxic concentrations of tamoxifen (10-100 microM). The cell stress was preferentially observed in samples from women in the proliferative menstrual phase. No treatment-related expression of NF-kappaB was observed. Constitutive expression of the tamoxifen-metabolizing enzymes CYP1B1, CYP2A6, CYP2B6, CYP2C8/9/19, CYP2D6, and SULT2A1 in glandular and surface epithelia was shown, but there was a large interindividual variation. The colocalization of [(3)H]tamoxifen adducts, expression of GRP78, caspase 3, and tamoxifen-metabolizing enzymes in human glandular and surface epithelia suggest a local bioactivation of tamoxifen at these sites and that epithelial cells are early target sites for tamoxifen-induced cell stress.
Asunto(s)
Endometrio/efectos de los fármacos , Endometrio/metabolismo , Estrés Fisiológico/efectos de los fármacos , Tamoxifeno/farmacología , Adulto , Hidrocarburo de Aril Hidroxilasas/metabolismo , Biotransformación , Caspasa 3/metabolismo , Citocromo P-450 CYP1B1 , Citocromo P-450 CYP2A6 , Citocromo P-450 CYP2B6 , Citocromo P-450 CYP2D6/metabolismo , Sistema Enzimático del Citocromo P-450/metabolismo , Endometrio/patología , Chaperón BiP del Retículo Endoplásmico , Epitelio/efectos de los fármacos , Epitelio/metabolismo , Femenino , Fase Folicular/metabolismo , Expresión Génica/efectos de los fármacos , Proteínas de Choque Térmico/metabolismo , Humanos , Fase Luteínica/metabolismo , FN-kappa B/metabolismo , Oxidorreductasas N-Desmetilantes/metabolismo , Células del Estroma/efectos de los fármacos , Células del Estroma/metabolismo , Sulfotransferasas/metabolismo , Tamoxifeno/farmacocinéticaRESUMEN
Type I IFNs, including IFN-alpha, enhance Ag presentation and promote the expansion, survival, and effector function of CD8(+) CTL during viral infection. Because these are ideal characteristics for a vaccine adjuvant, we examined the efficacy and mechanism of exogenous IFN-alpha as an adjuvant for antimelanoma peptide vaccination. We studied the expansion of pmel-1 transgenic CD8(+) T cells specific for the gp100 melanocyte differentiation Ag after vaccination of mice with gp100(25-33) peptide in IFA. IFN-alpha synergized with peptide vaccination in a dose-dependent manner by boosting relative and absolute numbers of gp100-specific T cells that suppressed B16 melanoma growth. IFN-alpha dramatically increased the accumulation of gp100-specific, IFN-gamma-secreting, CD8(+) T cells in the tumor through reduced apoptosis and enhanced proliferation of Ag-specific CD8(+) T cells. IFN-alpha treatment also greatly increased the long-term maintenance of pmel-1 CD8(+) T cells with an effector memory phenotype, a process that required expression of IFN-alpha receptor on the T cells and IL-15 in the host. These results demonstrate the efficacy of IFN-alpha as an adjuvant for peptide vaccination, give insight into its mechanism of action, and provide a rationale for clinical trials in which vaccination is combined with standard-of-care IFN-alpha therapy for melanoma.