RESUMO
BACKGROUND: Family-centered care is a valued approach to improving child and family outcomes in early intervention (EI), yet there is need to implement interventions that support information exchange for shared decision-making when planning and monitoring EI care. This study aims at estimating the feasibility, acceptability, and value of implementing the Young Children's Participation and Environment Measure (YC-PEM), a valid electronic patient-reported outcome (e-PRO) that is designed to support family engagement when planning care and monitoring outcomes of care. METHODS: Data were gathered from caregivers (N = 139) that were enrolled in a Phase 1 trial of the YC-PEM e-PRO as implemented within 1 month of their child's next EI evaluation of progress. YC-PEM e-PRO feasibility was estimated according to enrollment and completion rates, and mean completion time. Chi-square tests were used to examine parent perceptions of YC-PEM e-PRO acceptability by caregiver education and family income. Caregiver feedback via open-ended responses were content coded to inform intervention and protocol optimizations. YC-PEM e-PRO value was estimated via composite and item-level scores to capture the extent of participation difficulty in home and community activities, and common areas of need regarding caregivers desired change in their child's participation. RESULTS: Feasibility of implementing the YC-PEM e-PRO in routine EI care was mixed, as evidenced by low enrollment rates (21.0-29.2%), a high completion rate (85.3%), and limited missing data (80.6% of completed cases contained no missing data). More than half of the participants reported that the completion of the YC-PEM e-PRO was at least somewhat helpful, regardless of family income or caregiver education, providing support for its acceptability. As for its value, the YC-PEM e-PRO results were viewed by 64% of caregivers, whose desire for change most often pertained to the child's participation in non-discretionary activities at home and structured activities in the community. CONCLUSIONS: Results may support the implementation of YC-PEM e-PRO as a feasible, acceptable, and valued option for engaging families in planning the child's EI care. Results also inform select intervention and protocol optimizations prior to undertaking a multi-site pragmatic trial of its effectiveness on family engagement and shared decision-making within an EI clinical workflow. TRIAL REGISTRATION: Trial number: NCT03904797 . Trial registered at Clinicaltrials.gov . Registered 22 March 2019. Retrospectively registered.
Assuntos
Cuidadores , Família , Medidas de Resultados Relatados pelo Paciente , Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Pais , Inquéritos e Questionários , Adulto JovemRESUMO
BACKGROUND: Young children with developmental disabilities and delays spend significant amounts of time at home, show decreased participation in home-based activities, and receive home-based early intervention services to improve participation in activities. Yet, knowledge about the relationship between EI service use and children's home participation in activities remains poorly understood but needed for program improvement. The purpose of this study was to understand the relationships between EI service use and children's home participation. METHODS: In a cross-sectional design, data were gathered from caregivers (N = 139) who enrolled in a pilot trial of the Young Children's Participation in Environment Measure (YC-PEM) electronic patient-reported outcome (e-PRO), as implemented within 1 month of their child's next EI progress evaluation. A series of path analytic models were used to estimate EI service intensity as a predictor of parent-reported young children's home participation 1) frequency, 2) level of involvement, and 3) desired change, adjusting for family and child social and functional characteristics. Models included caregiver perceptions of home environmental support to test its indirect (i.e., mediation) effects on the relationship between EI service intensity and each of the three home participation dimensions. RESULTS: All three models fit the data well (comparative fit index = 1.00). EI service intensity was not a significant predictor of participation frequency. However, EI service intensity had a significant direct effect on a child's participation according to level of involvement and desired change, explaining between 13.3-33.5% of the variance in home participation. Caregiver perceptions of environmental support had a small yet significant indirect effect on the relationship between EI service intensity and level of involvement and desired change; these models explained between 18.5-38.1% of the variance in home participation. CONCLUSIONS: EI service intensity has important links with involvement in and desired change for home-based activities. Caregiver perceptions of environmental support appears to be a factor in the relationship between EI service intensity and home participation. Results warrant longitudinal replication with a control group, which would be possible with the implementation of the YC-PEM e-PRO in a routine EI clinical workflow. TRIAL RETROSPECTIVELY REGISTERED: NCT03904797 .
Assuntos
Cuidadores , Intervenção Educacional Precoce , Criança , Pré-Escolar , Estudos Transversais , Família , Instalações de Saúde , Humanos , Participação SocialRESUMO
BACKGROUND: Infants and toddlers with developmental difficulties represent a heterogeneous group who often receives early intervention (EI). Notable population heterogeneity exists and complicates unmet need and effectiveness research. However, a mix of relatively homogeneous clinically policy relevant 'subgroups' may create the apparent heterogeneity. To date, methodological challenges have impeded identifying these potential groups and their policy-relevance. METHODS: From the 2005-2006 National Survey of Children with Special Health Care Needs, we derived a sample (n = 965) of infants and toddlers with parent-reported developmental difficulties. We used latent class analysis (LCA) to identify subgroups of developmental vulnerability based upon functional, social and biological characteristics that would make children eligible for EI. Mixture modelling estimated the likelihood of each subgroup receiving parent-reported EI, controlling for race/ethnicity, child's age, and state of residence. RESULTS: LCA identified four distinct subgroups of developmental vulnerability: developmental disability (Group 1), mild developmental delay (Group 2), socially at risk with behaviour problems (Group 3), and socially at risk with functional vision difficulties (Group 4). Black, non-Hispanic children are significantly more likely than their white counterparts to be in Group 3 (ß = 1.52, P = 0.001) or group 4 (ß = 1.83, P < 0.001). Compared with children with a mild developmental delay (Group 2), children in group 1 (ß = -0.61, P < 0.001), group 3 (ß = -0.47, P = 0.001) and group 4 (ß = -0.38, P = 0.009) are significantly less likely to receive EI. CONCLUSIONS: Racial and ethnic differences exist with regard to membership in developmental vulnerability subgroups. Observed inconsistencies in access to EI suggest the need for improved surveillance, referral and outreach.
Assuntos
Negro ou Afro-Americano , Serviços de Saúde da Criança , Transtornos Cognitivos/diagnóstico , Deficiências do Desenvolvimento/diagnóstico , Intervenção Educacional Precoce , Disparidades em Assistência à Saúde , População Branca , Serviços de Saúde da Criança/estatística & dados numéricos , Serviços de Saúde da Criança/provisão & distribuição , Pré-Escolar , Transtornos Cognitivos/epidemiologia , Transtornos Cognitivos/terapia , Deficiências do Desenvolvimento/epidemiologia , Deficiências do Desenvolvimento/terapia , Intervenção Educacional Precoce/estatística & dados numéricos , Intervenção Educacional Precoce/provisão & distribuição , Etnicidade , Feminino , Política de Saúde , Acessibilidade aos Serviços de Saúde , Necessidades e Demandas de Serviços de Saúde , Humanos , Lactente , Masculino , Fatores Socioeconômicos , Estados UnidosRESUMO
BACKGROUND: Among families of infants born preterm, the association between post-natal depression and children's cognitive function is not well understood, but thought to be compromised. The purpose of this study is to investigate maternal depressive symptoms and perceived social support as predictors of children's cognitive function trajectories. METHODS: This is a longitudinal study of a sample of infants born preterm (less than 37 weeks) in Wisconsin. This study includes 130 infants who were hospitalized in one of three Wisconsin neonatal intensive care units in 2002-2005 and followed until 36 months of age. Maternal depressive symptoms were measured using the Center for Epidemiologic Studies Depression Scale. Social support was measured using the Maternal Support Scale. Children's cognitive function was measured using the Bayley Scales of Infant Development, 2nd Edition, and the Stanford-Binet Intelligence Scale, 5th Edition. RESULTS: Children's cognitive function trajectories declined initially and then increased. Being female (coefficient = 5.14, SE = 1.89) and non-poor (coefficient = 11.26, SE = 5.78), and having a mother who has a graduate degree (coefficient = 7.67, SE = 3.37) was associated with higher levels of cognition initially. Being white was associated with a more optimal cognitive trajectory. Although depression did not predict children's cognitive trajectories, the presence of clinically elevated depressive symptoms at 9 months post term was associated with lower cognitive functioning at 16 months when mothers reported low social support. CONCLUSION: Post-natal depressive symptoms appear to have a meaningful, dynamic influence on the cognitive outcomes of children born preterm, above and beyond family socio-demographic risk when the presence and timing of perceived social support are considered. Interventions to ameliorate developmental risk associated with preterm birth should include repeated assessments of maternal social support and post-natal depression and be targeted towards socially disadvantaged families.
Assuntos
Desenvolvimento Infantil , Depressão Pós-Parto/psicologia , Recém-Nascido Prematuro , Bem-Estar Materno/psicologia , Apoio Social , Adulto , Pré-Escolar , Cognição , Escolaridade , Feminino , Humanos , Lactente , Recém-Nascido , Estudos Longitudinais , Masculino , Programas de Rastreamento , Grupos Minoritários , Percepção , Fatores Socioeconômicos , Wisconsin/epidemiologia , Adulto JovemRESUMO
Granzyme B (GZMB) is a proapoptotic serine protease that is released by cytotoxic lymphocytes. However, GZMB can also be produced by other cell types and is capable of cleaving extracellular matrix (ECM) proteins. GZMB contributes to abdominal aortic aneurysm (AAA) through an extracellular, perforin-independent mechanism involving ECM cleavage. The murine serine protease inhibitor, Serpina3n (SA3N), is an extracellular inhibitor of GZMB. In the present study, administration of SA3N was assessed using a mouse Angiotensin II-induced AAA model. Mice were injected with SA3N (0-120 µg/kg) before pump implantation. A significant dose-dependent reduction in the frequency of aortic rupture and death was observed in mice that received SA3N treatment compared with controls. Reduced degradation of the proteoglycan decorin was observed while collagen density was increased in the aortas of mice receiving SA3N treatment compared with controls. In vitro studies confirmed that decorin, which regulates collagen spacing and fibrillogenesis, is cleaved by GZMB and that its cleavage can be prevented by SA3N. In conclusion, SA3N inhibits GZMB-mediated decorin degradation leading to enhanced collagen remodelling and reinforcement of the adventitia, thereby reducing the overall rate of rupture and death in a mouse model of AAA.
Assuntos
Proteínas de Fase Aguda/metabolismo , Aneurisma da Aorta Abdominal/patologia , Ruptura Aórtica/patologia , Decorina/metabolismo , Granzimas/metabolismo , Serpinas/metabolismo , Proteínas de Fase Aguda/genética , Proteínas de Fase Aguda/farmacologia , Angiotensina II/toxicidade , Animais , Aneurisma da Aorta Abdominal/metabolismo , Ruptura Aórtica/metabolismo , Apolipoproteínas E/genética , Apolipoproteínas E/metabolismo , Colágeno/metabolismo , Modelos Animais de Doenças , Granzimas/antagonistas & inibidores , Granzimas/genética , Camundongos , Camundongos Knockout , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacologia , Serpinas/genética , Serpinas/farmacologiaRESUMO
Peritransplant ischemia and reperfusion (I/R) injury contributes to posttransplant vascular dysfunction and cardiac allograft vasculopathy (CAV). We have previously shown that cytochrome p450 (CYP) 2C inhibition significantly reduces I/R-induced myocardial infarction and postischemic vascular dysfunction. In the latter study, pretreatment with sulfaphenazole (SP), a specific inhibitor of CYP 2C, restored postischemic NO-mediated, endothelium-dependent vasodilation and reduced vascular superoxide production. Given the association between I/R injury, early vascular dysfunction and CAV, we hypothesized that CYP 2C may also contribute to the onset of CAV. Lewis-to-Fisher rat heterotopic heart transplants were performed. Donors and recipients were treated with 5 mg/kg SP or vehicle control 1 h prior to surgery. SP did not affect posttransplant morbidity, mortality or weight gain. Coronary blood vessels from rats treated with SP exhibited significantly reduced luminal narrowing and demonstrated a corresponding decrease in smooth muscle cell (SMC) proliferation compared to controls. SP did not reduce diffuse, focal, epicardial, endocardial or perivascular immune infiltration nor did it significantly alter TUNEL positivity in myocardial, endothelial or SMC populations. In conclusion, CYP 2C contributes to SMC proliferation CAV without affecting general immune infiltration.
Assuntos
Proliferação de Células/efeitos dos fármacos , Vasos Coronários , Sistema Enzimático do Citocromo P-450/metabolismo , Músculo Liso Vascular/efeitos dos fármacos , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Animais , Anti-Infecciosos/administração & dosagem , Inibidores Enzimáticos/administração & dosagem , Transplante de Coração , Masculino , Músculo Liso Vascular/enzimologia , Traumatismo por Reperfusão Miocárdica/enzimologia , Ratos , Ratos Endogâmicos Lew , Sulfafenazol/administração & dosagem , Transplante HomólogoRESUMO
The purpose of this study was to characterize tissue esterase activity and blood fenitrothion concentrations in the rat dam and foetus following in-utero exposure to the organophosphate insecticide fenitrothion. Time-mated, 8-week-old rats were gavaged on gestation day 19 with 0, 5, or 25 mg fenitrothion kg-1. Fenitrothion was absorbed rapidly from the gastrointestinal tract, with peak maternal and foetal blood levels observed 0.5-1.0 h after dosing. Fenitrothion concentrations in maternal and foetal blood were virtually identical and demonstrated a non-linear dose-response relationship. Acetylcholinesterase and carboxylesterase activities in maternal liver and blood and in foetal liver and brain decreased within 30-60 min of fenitrothion exposure. Esterase inhibition occurred at a fenitrothion dose (5 mg kg-1) that has not been previously associated with reproductive toxicity, suggesting that esterase inhibition should be considered as the critical effect in risk assessments for this pesticide.
Assuntos
Inibidores da Colinesterase/farmacologia , Colinesterases/metabolismo , Fenitrotion/farmacologia , Feto/efeitos dos fármacos , Feto/enzimologia , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/enzimologia , Inibidores da Colinesterase/administração & dosagem , Inibidores da Colinesterase/sangue , Inibidores da Colinesterase/farmacocinética , Feminino , Fenitrotion/administração & dosagem , Fenitrotion/sangue , Fenitrotion/farmacocinética , Fígado/efeitos dos fármacos , Fígado/enzimologia , Gravidez , RatosRESUMO
BNip (formerly known as Nip) proteins, including homologues isolated from human, mouse and Caenorhabditis. elegans, are a relatively new subgroup of the Bcl-2 family. These proteins are classified into this family based on limited sequence homology with the Bcl-2 homology domain 3 and carboxyl terminal transmembrane domain. BNip proteins were first discovered based on their interaction with the adenovirus E1B 19 kDa/Bcl-2 family protein and since then, their roles in cell death pathways have been actively studied. However, the precise mechanisms by which the BNip proteins induce apoptosis and/or necrosis remain to be determined. To advance our knowledge, we have provided a summary and review of current literature regarding BNip proteins including comparative sequence analysis, mutational mapping of the functional domains, and cell death mechanisms involving disruption of mitochondrial homeostasis. Since BNip proteins are expressed at high levels in the heart as compared to other organs, their roles in cardiomyocyte injury during hypoxia or viral infection is a focus of this review. Finally, we discuss potential directions for further study on this increasingly important group of pro-apoptotic proteins.
Assuntos
Apoptose/fisiologia , Proteínas de Membrana/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Animais , Cardiomiopatias/genética , Cardiomiopatias/metabolismo , Cardiomiopatias/fisiopatologia , Humanos , Proteínas de Membrana/genética , Mitocôndrias/metabolismo , Miócitos Cardíacos/citologia , Miócitos Cardíacos/metabolismo , Estrutura Terciária de Proteína/fisiologia , Proteínas Proto-Oncogênicas c-bcl-2/genética , Homologia de Sequência de AminoácidosRESUMO
Apolipoprotein E (apo E) deficiency (or its abnormalities in humans) is associated with a series of pathological conditions including dyslipidemia, atherosclerosis, Alzheimer's disease, and shorter life span. The purpose of this study was to characterize these conditions in apo E-deficient C57BL/6J mice and relate them to human disorders. Deletion of apo E gene in mice is associated with changes in lipoprotein metabolism [plasma total cholesterol (TC) (>+400%), HDL cholesterol (-80%), HDL/TC, and HDL/LDL ratios (-93% and -96%, respectively), esterification rate in apo B-depleted plasma (+100%), plasma triglyceride (+200%), hepatic HMG-CoA reductase activity (-50%), hepatic cholesterol content (+30%)], decreased plasma homocyst(e)ine and glucose levels, and severe atherosclerosis and cutaneous xanthomatosis. Hepatic and lipoprotein lipase activities, hepatic LDL receptor function, and organ antioxidant capacity remain unchanged. Several histological/immunohistological stainings failed to detect potential markers for neurodegenerative disease in the brain of 37-wk-old male apo E-KO mice. Apo E-KO mice may have normal growth and development, but advanced atherosclerosis and xanthomatosis may indirectly reduce their life span. Apo E plays a crucial role in regulation of lipid metabolism and atherogenesis without affecting lipase activities, endogenous antioxidant capacity, or appearance of neurodegenerative markers in 37-wk-old male mice.
Assuntos
Apolipoproteínas E/deficiência , Animais , Antioxidantes/metabolismo , Apolipoproteínas E/genética , Glicemia/metabolismo , Peso Corporal/fisiologia , Encéfalo/metabolismo , Colesterol/metabolismo , Esterificação , Genótipo , Proteína Glial Fibrilar Ácida/análise , Homocisteína/sangue , Humanos , Hidroximetilglutaril-CoA Redutases/metabolismo , Hiperlipidemias/sangue , Hiperlipidemias/fisiopatologia , Imuno-Histoquímica , Rim/metabolismo , Lipase/sangue , Lipídeos/sangue , Lipoproteínas HDL/metabolismo , Fígado/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Proteínas de Neurofilamentos/análise , Receptores de LDL/fisiologia , Análise de Sobrevida , Fatores de Tempo , Distribuição TecidualRESUMO
Photodynamic therapy (PDT) is clinically approved for the treatment of several types of cancer as well as age-related macular degeneration, the leading cause of blindness in the elderly. PDT using the photosensitizer verteporfin has been previously shown to induce rapid apoptosis via a mitochondrial-caspase activation pathway. The impact of PDT on other cellular organelles such as the endoplasmic reticulum (ER) is undefined. The effect of PDT on intracellular Ca2+ ([Ca2+]i) in control and Bcl-2-overexpressing HeLa cells was assessed. A greater [Ca2+]i transient was observed for Bcl-2 overexpressing cells in response to PDT. The PDT-induced Ca2+ release was due to the emptying of Ca2+ from ER and possibly mitochondrial stores and was not due to an influx of Ca2+ from the medium. For Bcl-2-transfected cells, the release of Ca2+ was incomplete as determined by a further [Ca2+]i transient produced by the addition of the Ca2+ ionophore ionomycin after PDT. Furthermore, extrusion of Ca2+ was not hindered while ER-mediated sequestration of Ca2+ was impaired after PDT. Impairment of ER-mediated sequestration of Ca2+ may be due to the immediate caspase-independent depletion of sarco/endoplasmic reticulum Ca2+ ATPase-2 (SERCA2) that occurred in response to PDT in birth HeLa/Neo and Bcl-2 overexpressed HeLa cells. In summary, PDT induced the rapid degradation of SERCA2 and release of ER and mitochondrial Ca2+ stores. Although overexpression of Bcl-2 did not protect against SERCA2 degradation, it may influence the release of Ca2+ from ER and mitochondrial stores in PDT-treated cells.
Assuntos
Apoptose/efeitos dos fármacos , Cálcio/metabolismo , Retículo Endoplasmático/metabolismo , Células HeLa/metabolismo , Fotoquimioterapia , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Transporte Biológico/efeitos dos fármacos , Transporte Biológico/efeitos da radiação , Sinalização do Cálcio/fisiologia , ATPases Transportadoras de Cálcio/metabolismo , Grupo dos Citocromos c/metabolismo , Células HeLa/efeitos dos fármacos , Humanos , Mitocôndrias/metabolismo , Fármacos Fotossensibilizantes/farmacologia , Porfirinas/farmacologia , ATPases Transportadoras de Cálcio do Retículo Sarcoplasmático , VerteporfinaRESUMO
Studies of the molecular basis of insulin resistance have focused on the peroxisome proliferator activated receptor gamma (PPARgamma, gamma1 and gamma2). The aim of this study was to determine whether the insulin resistance in liver of diabetic animals is associated with abnormal expression of these receptors. PPARgamma mRNA and protein expression levels were quantified in liver of 9-week-old male ob/ob mice as a model of diabetes and compared to age- and gender-matched wild type control animals of the same genetic background. Semi-quantitative reverse transcription-polymerase chain reaction, using 18S rRNA as an internal standard, indicated that PPARgamma2 mRNA was significantly upregulated in ob/ob liver vs. that in wild type mice. Western blotting revealed greater immunoreactivity of PPARgamma2 in liver from ob/ob mice relative to that in wild type mice. An index of insulin resistance (product of serum glucose and insulin concentration) was correlated with liver PPARgamma2 mRNA expression (r = 0.776; p < 0.001). The findings that liver PPARgamma2 expression is (1) significantly elevated in the ob/ob model of diabetes and (2) positively associated with an index of insulin resistance, suggests a possible compensatory response through which type II diabetic and obese organisms strive to maintain insulin sensitivity of the liver.
Assuntos
Diabetes Mellitus Tipo 2/metabolismo , Fígado/metabolismo , Obesidade/metabolismo , Receptores Citoplasmáticos e Nucleares/metabolismo , Fatores de Transcrição/metabolismo , Tecido Adiposo/metabolismo , Tecido Adiposo/patologia , Envelhecimento , Animais , Western Blotting , Diabetes Mellitus Tipo 2/genética , Diabetes Mellitus Tipo 2/patologia , Modelos Animais de Doenças , Expressão Gênica , Resistência à Insulina/genética , Fígado/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Obesos , Músculo Esquelético/metabolismo , Músculo Esquelético/patologia , Obesidade/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores Citoplasmáticos e Nucleares/genética , Fatores de Transcrição/genéticaRESUMO
Among the models of dyslipidemia and atherosclerosis, a number of wild-type, naturally defective, and genetically modified animals (rabbits, mice, pigeons, dogs, pigs, and monkeys) have been characterized. In particular, their similarities to and differences from humans in respect to relevant biochemical, physiologic, and pathologic conditions have been evaluated. Features of atherosclerotic lesions and their specific relationship to plasma lipoprotein particles have been critically reviewed and summarized. All animal models studied have limitations: the most significant advantages and disadvantages of using a specific animal species are outlined here. New insights in lipid metabolism and genetic background with regard to variations in pathogenesis of dyslipidemia-associated atherogenesis have also been reviewed. Evidence suggests that among wild-type species, strains of White Carneau pigeons and Watanabe Heritable Hyperlipidemic and St. Thomas's Hospital rabbits are preferable to the cholesterol-fed wild-type animal species in dyslipidemia and atherosclerosis research. Evidence for the usefulness of both wild-type and transgenic animals in studying the involvement of inflammatory pathways and Chlamydia pneumoniae infection in pathogenesis of atherosclerosis has also been summarized. Transgenic mice and rabbits are excellent tools for studying specific gene-related disorders. However, despite these significant achievements in animal experimentation, there are no suitable animal models for several rare types of fatal dyslipidemia-associated disorders such as phytosterolemia and cerebrotendinous xanthomatosis. An excellent model of diabetic atherosclerosis is unavailable. The question of reversibility of atherosclerosis still remains unanswered. Further work is needed to overcome these deficiencies.
Assuntos
Arteriosclerose , Hiperlipidemias , Animais , Modelos Animais de DoençasRESUMO
Tumor necrosis factor-related apoptosis inducing ligand (TRAIL) and Fas ligand (FasL) trigger apoptosis by stimulating the formation of a death inducing signaling complex at the cytoplasmic terminus of their respective receptors. Photodynamic therapy (PDT) is an approved treatment for several types of cancer as well as for age-related macular degeneration and is under investigation for different cancer, ocular, autoimmune and cardiovascular indications. The effect of low dose PDT in combination with TRAIL and FasL on Jurkat lymphoma cell apoptosis was examined. Individually, TRAIL, FasL, and PDT could induce apoptosis in these cells. However, at suboptimal levels of PDT, the number of cells undergoing apoptosis was increased when recombinant FasL and/or TRAIL were added. Additive effects of these treatments were evident for different apoptosis parameters including DNA fragmentation, caspase processing and activity and caspase substrate degradation. Overall, these results provide evidence that PDT-treated cells may be more likely to undergo apoptosis when also exposed to receptor-mediated signals delivered by factors such as TRAIL or FasL. For PDT, immune cell-mediated death receptor ligation may represent a way whereby tumor cells that have withstood the direct effects of photosensitization may be eliminated.
Assuntos
Apoptose/efeitos dos fármacos , Glicoproteínas de Membrana/farmacologia , Fotoquimioterapia , Fármacos Fotossensibilizantes/farmacologia , Fator de Necrose Tumoral alfa/farmacologia , Proteínas Reguladoras de Apoptose , Western Blotting , Caspases/metabolismo , Fragmentação do DNA/efeitos dos fármacos , Endopeptidases/química , Proteína Ligante Fas , Citometria de Fluxo , Humanos , Células Jurkat , Células Matadoras Naturais/efeitos dos fármacos , Porfirinas/farmacologia , Linfócitos T/efeitos dos fármacos , Ligante Indutor de Apoptose Relacionado a TNF , VerteporfinaRESUMO
The process of inflammation and immune response is regulated by proinflammatory cytokines. Interleukin-6 (IL-6), one of the proinflammatory cytokines, plays a potentially critical role in viral-induced myocarditis. Our previous work demonstrates that exogenous IL-6 administration, given at the time of encephalomyocarditis virus (EMCV) inoculation in C3H/HeJ mice, has a protective effect on myocardium and improves survival rates. In the present study, we examined whether overexpression of IL-6 modified viral myocarditis. On day 3 and 10 after inoculation with EMCV, the ratio of heart weight to body weight and myocardial injury were significantly increased in IL-6 transgenic mice (IL-6TG). On day 3, a reduction of viral clearance was shown by the presence of elevated viral titers and viral replication in the heart of IL-6TG. The concentrations of serum tumor necrosis factor- alpha (TNF alpha) were dramatically increased in wild-type mice on day 1, in contrast, this change was not observed in IL-6TG. Treatment with recombinant human TNF (2 microg) significantly improved viral clearance in the IL-6TG hearts. Thus, overexpression of IL-6 promotes myocardial injury by interrupting both the cytokine network and viral clearance. These experiments suggest the possibility that IL-6 is one of the factors that accelerates tissue damage, including myocardial injury, in the viral myocarditis.
Assuntos
Infecções por Cardiovirus/imunologia , Vírus da Encefalomiocardite/fisiologia , Interleucina-6/metabolismo , Miocardite/imunologia , Fator de Necrose Tumoral alfa/metabolismo , Animais , Peso Corporal , Infecções por Cardiovirus/patologia , Infecções por Cardiovirus/virologia , Vírus da Encefalomiocardite/genética , Coração/virologia , Humanos , Hibridização In Situ , Interleucina-6/genética , Interleucina-6/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Miocardite/patologia , Miocardite/virologia , Miocárdio/patologia , Tamanho do Órgão , Replicação ViralRESUMO
The high turnover of endothelial cells (EC) in atherosclerosis suggests that an increase in the frequency of both cell proliferation and cell death is important in the pathogenesis of this common disorder. Further, increased apoptosis of EC, smooth muscle cells (SMC) and immune cells has been observed in atheromatous plaques. Many pro-atherogenic factors, including oxidized low-density lipoproteins, angiotensin II and oxidative stress, can induce EC apoptosis. Such damage to the endothelium may be an initiating event in atherogenesis since EC apoptosis may compromise vasoregulation, increase SMC proliferation, SMC migration and blood coagulation. In addition, EC overlying vascular lesions have been shown to increase their expression of pro-apoptotic proteins, such as Fas and Bax, while decreasing levels of anti-apoptotic factors. Therefore, understanding EC apoptotic pathways that are altered in atherosclerosis may enable a greater understanding of disease pathogenesis and foster the development of new therapies. The present discussion outlines the biochemical characteristics of EC apoptosis and the role that altered regulation of apoptosis plays in vasculopathy.
Assuntos
Apoptose , Arteriosclerose/fisiopatologia , Endotélio Vascular/citologia , Proteínas Serina-Treonina Quinases , Angiotensina II/metabolismo , Cálcio/metabolismo , Fatores de Crescimento Endotelial/metabolismo , Proteína Ligante Fas , Humanos , Lipoproteínas LDL/metabolismo , Linfocinas/metabolismo , Glicoproteínas de Membrana/metabolismo , Mitocôndrias/metabolismo , Modelos Biológicos , Óxido Nítrico/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Proto-Oncogênicas c-akt , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Estresse Mecânico , Fator de Necrose Tumoral alfa/metabolismo , Fator A de Crescimento do Endotélio Vascular , Fatores de Crescimento do Endotélio VascularRESUMO
Photodynamic therapy (PDT) is under investigation for the treatment of intimal hyperplastia in conditions such as atherosclerosis and restenosis. Although smooth muscle cells (SMCs) may be a key target for treatment, the effects of PDT on these cells are poorly characterized. In the present study, apoptosis was induced in primary human aortic SMCs by the combination of the photosensitizer verteporfin and visible light. After PDT, an increase in mitochondrial cytochrome c (cyt c) and apoptosis-inducing factor (AIF) levels were detected in the cytosol immediately and their levels increased steadily up to 2 hours. Cytosolic levels of the pro-apoptotic Bcl-2 family member Bax decreased reciprocally throughout this period, but this change did not occur before cyt c release. Confocal microscopy revealed a diffuse staining pattern of cyt c within apoptotic cells as compared to a distinct mitochondrial staining in normal cells. AIF translocated from mitochondria to the nucleus during the progression of apoptosis. After cyt c release, caspase-9 and caspase-3 processing was visible by 1 hour and caspase-6, -7, and -8 processing was apparent by 2 hours after PDT. In summary, these results demonstrate for the first time the cellular redistribution of mitochondrial AIF during SMC apoptosis, as well as the early release of cyt c and the subsequent activation of multiple caspases during PDT-induced SMC apoptosis.
Assuntos
Apoptose/fisiologia , Grupo dos Citocromos c/metabolismo , Flavoproteínas/metabolismo , Proteínas de Membrana/metabolismo , Mitocôndrias Musculares/metabolismo , Músculo Liso Vascular/fisiologia , Proteínas Proto-Oncogênicas c-bcl-2 , Aorta/citologia , Aorta/fisiologia , Fator de Indução de Apoptose , Caspases/metabolismo , Células Cultivadas , Fragmentação do DNA , Ativação Enzimática , Humanos , Luz , Músculo Liso Vascular/citologia , Fotoquimioterapia , Fármacos Fotossensibilizantes/uso terapêutico , Porfirinas/farmacologia , Proteínas Proto-Oncogênicas/metabolismo , Distribuição Tecidual , Verteporfina , Proteína X Associada a bcl-2RESUMO
The 5' and 3' untranslated regions (UTRs) of coxsackievirus B3 (CVB3) RNA form highly ordered secondary structures that have been confirmed to play important regulatory roles in viral cap-independent internal translation initiation and RNA replication. We previously demonstrated that deletions in different regions of the 5' UTR significantly reduced viral RNA translation and infectivity. Such observations suggested strongly that viral RNA translation and replication could be blocked if highly specific antisense oligodeoxynucleotides (AS-ODNs) were applied to target crucial sites within the 5' and 3' UTRs. In this study, seven phosphorothioate AS-ODNs were synthesized, and the antiviral activity was evaluated by Lipofectin transfection of HeLa cells with AS-ODNs followed by infection of CVB3. Analysis by Western blotting, reverse transcription-PCR, and viral plaque assay demonstrated that viral protein synthesis, genome replication, and infectivity of CVB3 were strongly inhibited by the AS-ODNs complementary to different regions of the 5' and 3' UTRs. The most effective sites are located at the proximate terminus of the 5' UTR (AS-1), the proximate terminus of the 3' UTR (AS-7), the core sequence of the internal ribosome entry site (AS-2), and the translation initiation codon region (AS-4). These AS-ODNs showed highly sequence-specific and dose-dependent inhibitory effects on both viral protein synthesis and RNA replication. It is noteworthy that the highest inhibitory activities were obtained with AS-1 and AS-7 targeting the termini of the 5' and 3' UTRs. The percent inhibition values of AS-1 and AS-7 for CVB3 protein VP1 synthesis and RNA replication were 70.6 and 79.6 for AS-1 and 73.7 and 79.7 for AS-7, respectively. These data suggest that CVB3 infectivity can be inhibited effectively by AS-ODNs.
Assuntos
Enterovirus Humano B/efeitos dos fármacos , Oligodesoxirribonucleotídeos Antissenso/farmacologia , Biossíntese de Proteínas/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos , Enterovirus Humano B/genética , Células HeLa , Humanos , RNA Viral/efeitos dos fármacos , Tionucleotídeos/farmacologia , Transfecção , Regiões não Traduzidas , Ensaio de Placa Viral , Proteínas Estruturais Virais/biossínteseRESUMO
PURPOSE: To determine whether perivascular delivery of paclitaxel prevents luminal narrowing after balloon injury by inhibiting intimal hyperplasia. MATERIALS AND METHODS: Immediately after balloon injury of the entire left common carotid artery, three slow-release formulations of paclitaxel or control formulations without drug were applied around a distal segment of the artery. The noninjured right carotid arteries were evaluated as a control. The animals were maintained for 14 and 28 days (n = 5 in each group at each time interval). Histology, immunohistochemistry, and morphometric analysis were performed. RESULTS: Injured nontreated arteries exhibited a pronounced intimal hyperplasia (0.185 +/- 0.01 mm2 at 14 days and 0.189 +/- 0.01 mm2 at 28 days) and a marked reduction in luminal area (44% at 14 days and 43% at 28 days). Medial area and the number of medial cells increased by 44% and 45%, respectively, at 14 days, and by 22% and 37%, respectively, at 28 days. Injured arteries treated with perivascular paclitaxel did not show any intimal hyperplasia, and luminal area was increased in five of six groups and was unchanged in one group. These arteries had an increased medial area but they had fewer medial cells than noninjured arteries. Injured arteries treated with control implants without paclitaxel exhibited intimal hyperplasia and luminal narrowing. CONCLUSION: Perivascular slow release of paclitaxel totally inhibits intimal hyperplasia and prevents luminal narrowing after balloon injury. Because of its efficacy, perivascular paclitaxel represents a possible approach for prevention of restenosis in humans.