RESUMEN
BACKGROUND: Epithelial damage, repair and remodelling are critical features of chronic airway diseases including chronic obstructive pulmonary disease (COPD). Interleukin (IL)-33 released from damaged airway epithelia causes inflammation via its receptor, serum stimulation-2 (ST2). Oxidation of IL-33 to a non-ST2-binding form (IL-33ox) is thought to limit its activity. We investigated whether IL-33ox has functional activities that are independent of ST2 in the airway epithelium. METHODS: In vitro epithelial damage assays and three-dimensional, air-liquid interface (ALI) cell culture models of healthy and COPD epithelia were used to elucidate the functional role of IL-33ox. Transcriptomic changes occurring in healthy ALI cultures treated with IL-33ox and COPD ALI cultures treated with an IL-33-neutralising antibody were assessed with bulk and single-cell RNA sequencing analysis. RESULTS: We demonstrate that IL-33ox forms a complex with receptor for advanced glycation end products (RAGE) and epidermal growth factor receptor (EGFR) expressed on airway epithelium. Activation of this alternative, ST2-independent pathway impaired epithelial wound closure and induced airway epithelial remodelling in vitro. IL-33ox increased the proportion of mucus-producing cells and reduced epithelial defence functions, mimicking pathogenic traits of COPD. Neutralisation of the IL-33ox pathway reversed these deleterious traits in COPD epithelia. Gene signatures defining the pathogenic effects of IL-33ox were enriched in airway epithelia from patients with severe COPD. CONCLUSIONS: Our study reveals for the first time that IL-33, RAGE and EGFR act together in an ST2-independent pathway in the airway epithelium and govern abnormal epithelial remodelling and muco-obstructive features in COPD.
Asunto(s)
Interleucina-33 , Enfermedad Pulmonar Obstructiva Crónica , Humanos , Células Epiteliales/metabolismo , Células Epiteliales/patología , Receptores ErbB , Proteína 1 Similar al Receptor de Interleucina-1 , Interleucina-33/genética , Interleucina-33/metabolismo , Oxidación-Reducción , Enfermedad Pulmonar Obstructiva Crónica/genética , Enfermedad Pulmonar Obstructiva Crónica/metabolismo , Enfermedad Pulmonar Obstructiva Crónica/patología , Receptor para Productos Finales de Glicación Avanzada/metabolismoRESUMEN
BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is a devastating interstitial lung disease (ILD) with limited treatment options. Interleukin-33 (IL-33) is proposed to play a role in the development of IPF however the exclusive use of prophylactic dosing regimens means that the therapeutic benefit of targeting this cytokine in IPF is unclear. METHODS: IL-33 expression was assessed in ILD lung sections and human lung fibroblasts (HLFs) by immunohistochemistry and gene/protein expression and responses of HLFs to IL-33 stimulation measured by qPCR. In vivo, the fibrotic potential of IL-33:ST2 signalling was assessed using a murine model of bleomycin (BLM)-induced pulmonary fibrosis and therapeutic dosing with an ST2-Fc fusion protein. Lung and bronchoalveolar lavage fluid were collected for measurement of inflammatory and fibrotic endpoints. Human precision-cut lung slices (PCLS) were stimulated with transforming growth factor-ß (TGFß) or IL-33 and fibrotic readouts assessed. RESULTS: IL-33 was expressed by fibrotic fibroblasts in situ and was increased by TGFß treatment in vitro. IL-33 treatment of HLFs did not induce IL6, CXCL8, ACTA2 and COL1A1 mRNA expression with these cells found to lack the IL-33 receptor ST2. Similarly, IL-33 stimulation had no effect on ACTA2, COL1A1, FN1 and fibronectin expression by PCLS. Despite having effects on inflammation suggestive of target engagement, therapeutic dosing with the ST2-Fc fusion protein failed to reduce BLM-induced fibrosis measured by hydroxyproline content or Ashcroft score. CONCLUSIONS: Together these findings suggest the IL-33:ST2 axis does not play a central fibrogenic role in the lungs with therapeutic blockade of this pathway unlikely to surpass the current standard of care for IPF.
Asunto(s)
Fibrosis Pulmonar Idiopática , Interleucina-33 , Animales , Humanos , Ratones , Bleomicina/toxicidad , Fibroblastos/metabolismo , Fibrosis Pulmonar Idiopática/inducido químicamente , Fibrosis Pulmonar Idiopática/tratamiento farmacológico , Fibrosis Pulmonar Idiopática/metabolismo , Proteína 1 Similar al Receptor de Interleucina-1/genética , Interleucina-33/metabolismo , Pulmón/metabolismo , Ratones Endogámicos C57BL , Factor de Crecimiento Transformador beta/metabolismoRESUMEN
Airway remodeling occurs in chronic asthma leading to increased airway smooth muscle (ASM) mass and extracellular matrix (ECM) deposition. Although extensively studied in murine airways, studies report only selected larger airways at one time-point meaning the spatial distribution and resolution of remodeling are poorly understood. Here we use a new method allowing comprehensive assessment of the spatial and temporal changes in ASM, ECM, and epithelium in large numbers of murine airways after allergen challenge. Using image processing to analyze 20-50 airways per mouse from a whole lung section revealed increases in ASM and ECM after allergen challenge were greater in small and large rather than intermediate airways. ASM predominantly accumulated adjacent to the basement membrane, whereas ECM was distributed across the airway wall. Epithelial hyperplasia was most marked in small and intermediate airways. After challenge, ASM changes resolved over 7 days, whereas ECM and epithelial changes persisted. The new method suggests large and small airways remodel differently, and the long-term consequences of airway inflammation may depend more on ECM and epithelial changes than ASM. The improved quantity and quality of unbiased data provided by the method reveals important spatial differences in remodeling and could set new analysis standards for murine asthma models.
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Asma , Pulmón , Ratones , Animales , Músculo Liso , Matriz Extracelular/fisiología , Remodelación de las Vías Aéreas (Respiratorias)/fisiología , AlérgenosRESUMEN
Autism Spectrum Disorder (ASD) is one of the most common neurodevelopmental disorders with no drugs treating the core symptoms and no validated biomarkers for clinical use. The multi-functional phytochemical sulforaphane affects many of the biochemical abnormalities associated with ASD. We investigated potential molecular markers from three ASD-associated physiological pathways that can be affected by sulforaphane: redox metabolism/oxidative stress; heat shock response; and immune dysregulation/inflammation, in peripheral blood mononuclear cells (PBMCs) from healthy donors and patients with ASD. We first analyzed the mRNA levels of selected molecular markers in response to sulforaphane ex vivo treatment in PBMCs from healthy donors by real-time quantitative PCR. All of the tested markers showed quantifiability, accuracy and reproducibility. We then compared the expression levels of those markers in PBMCs taken from ASD patients in response to orally-delivered sulforaphane. The mRNA levels of cytoprotective enzymes (NQO1, HO-1, AKR1C1), and heat shock proteins (HSP27 and HSP70), increased. Conversely, mRNA levels of pro-inflammatory markers (IL-6, IL-1ß, COX-2 and TNF-α) decreased. Individually none is sufficiently specific or sensitive, but when grouped by function as two panels, these biomarkers show promise for monitoring pharmacodynamic responses to sulforaphane in both healthy and autistic humans, and providing guidance for biomedical interventions.
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Trastorno del Espectro Autista/sangre , Trastorno del Espectro Autista/tratamiento farmacológico , Isotiocianatos/uso terapéutico , Leucocitos Mononucleares/metabolismo , Trastorno del Espectro Autista/metabolismo , Células Cultivadas , Niño , Citocinas/sangre , Citocinas/metabolismo , Humanos , Inflamación/sangre , Inflamación/metabolismo , Leucocitos/metabolismo , Masculino , SulfóxidosRESUMEN
We examined whether gastric acidity would affect the activity of myrosinase, co-delivered with glucoraphanin (GR), to convert GR to sulforaphane (SF). A broccoli seed and sprout extract (BSE) rich in GR and active myrosinase was delivered before and after participants began taking the anti-acid omeprazole, a potent proton pump inhibitor. Gastric acidity appears to attenuate GR bioavailability, as evidenced by more SF and its metabolites being excreted after participants started taking omeprazole. Enteric coating enhanced conversion of GR to SF, perhaps by sparing myrosinase from the acidity of the stomach. There were negligible effects of age, sex, ethnicity, BMI, vegetable consumption, and bowel movement frequency and quality. Greater body mass correlated with reduced conversion efficiency. Changes in the expression of 20 genes in peripheral blood mononuclear cells were evaluated as possible pharmacodynamic indicators. When grouped by their primary functions based on a priori knowledge, expression of genes associated with inflammation decreased non-significantly, and those genes associated with cytoprotection, detoxification and antioxidant functions increased significantly with bioavailability. Using principal components analysis, component loadings of the changes in gene expression confirmed these groupings in a sensitivity analysis.
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Brassica , Suplementos Dietéticos , Glucosinolatos/administración & dosificación , Glicósido Hidrolasas/administración & dosificación , Imidoésteres/administración & dosificación , Isotiocianatos/metabolismo , Omeprazol/administración & dosificación , Extractos Vegetales/administración & dosificación , Inhibidores de la Bomba de Protones/administración & dosificación , Plantones , Semillas , Adulto , Anciano , Disponibilidad Biológica , Brassica/química , Suplementos Dietéticos/efectos adversos , Interacciones Farmacológicas , Femenino , Ácido Gástrico/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Glucosinolatos/efectos adversos , Glucosinolatos/aislamiento & purificación , Glucosinolatos/metabolismo , Glicósido Hidrolasas/efectos adversos , Glicósido Hidrolasas/metabolismo , Humanos , Concentración de Iones de Hidrógeno , Imidoésteres/efectos adversos , Imidoésteres/aislamiento & purificación , Imidoésteres/metabolismo , Leucocitos Mononucleares/efectos de los fármacos , Leucocitos Mononucleares/metabolismo , Masculino , Persona de Mediana Edad , Omeprazol/efectos adversos , Oximas , Proyectos Piloto , Extractos Vegetales/efectos adversos , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/metabolismo , Inhibidores de la Bomba de Protones/efectos adversos , Plantones/química , Semillas/química , Sulfóxidos , Adulto JovenRESUMEN
The tropical tree Moringa oleifera produces high yields of protein-rich leaf biomass, is widely used as a food source, contains an abundance of phytochemicals, and thus has great potential for chronic disease prevention and perhaps, treatment. We have developed and characterized standardized ways of preparing aqueous "teas" from moringa leaves to deliver precisely calibrated levels of phytochemicals for use in clinical trials. These phytochemicals, especially the glucosinolate glucomoringin and the isothiocyanate moringin, produced from it following hydrolysis by the enzyme myrosinase, provide potent anti-inflammatory and cytoprotective indirect antioxidant activity. The taste of both hot and cold teas is palatable without the need for flavor masking. These teas can be easily and reproducibly prepared in underserved tropical regions of the world where moringa is cultivated. Isothiocyanate yield from a cold extraction was rapid and essentially complete after 30 min and its anti-inflammatory potential is comparable to that of equimolar purified moringin. A preparation similar to this may be safe to consume with respect to its bacterial titer even after 48 h without refrigeration. Thus, facile delivery of moringa tea to both adults and children for clinical evaluation of their effects on such conditions as autism, diabetes, and hypertension, is now possible.
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Glucosinolatos/administración & dosificación , Isotiocianatos/administración & dosificación , Moringa oleifera/química , Hojas de la Planta/química , Administración Oral , Animales , Antiinflamatorios/química , Antiinflamatorios/farmacología , Bebidas , Glucosinolatos/química , Isotiocianatos/química , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Ratones , Estructura Molecular , Óxido Nítrico Sintasa de Tipo II/antagonistas & inhibidores , Fitoquímicos/química , Fitoquímicos/farmacología , Extractos Vegetales/química , Extractos Vegetales/farmacología , Células RAW 264.7RESUMEN
Glucosinolates (GS) are metabolized to isothiocyanates that may enhance human healthspan by protecting against a variety of chronic diseases. Moringa oleifera, the drumstick tree, produces unique GS but little is known about GS variation within M. oleifera, and even less in the 12 other Moringa species, some of which are very rare. We assess leaf, seed, stem, and leaf gland exudate GS content of 12 of the 13 known Moringa species. We describe 2 previously unidentified GS as major components of 6 species, reporting on the presence of simple alkyl GS in 4 species, which are dominant in M. longituba. We document potent chemoprotective potential in 11 of 12 species, and measure the cytoprotective activity of 6 purified GS in several cell lines. Some of the unique GS rank with the most powerful known inducers of the phase 2 cytoprotective response. Although extracts of most species induced a robust phase 2 cytoprotective response in cultured cells, one was very low (M. longituba), and by far the highest was M. arborea, a very rare and poorly known species. Our results underscore the importance of Moringa as a chemoprotective resource and the need to survey and conserve its interspecific diversity.
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Quimioprevención/métodos , Enfermedad Crónica/prevención & control , Glucosinolatos , Moringa/química , Moringa/clasificación , Células Cultivadas , Citoprotección/efectos de los fármacos , Glucosinolatos/química , Glucosinolatos/clasificación , Glucosinolatos/aislamiento & purificación , Glucosinolatos/farmacología , Humanos , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Moringa oleifera/química , Moringa oleifera/clasificación , Filogenia , Fitoterapia/métodos , Extractos Vegetales/farmacología , Hojas de la Planta/química , Hojas de la Planta/fisiología , Semillas/químicaRESUMEN
Taste drives consumption of foods. The tropical tree Moringa oleifera is grown worldwide as a protein-rich leafy vegetable and for the medicinal value of its phytochemicals, in particular its glucosinolates, which can lead to a pronounced harsh taste. All studies to date have examined only cultivated, domestic variants, meaning that potentially useful variation in wild type plants has been overlooked. We examine whether domesticated and wild type M. oleifera differ in myrosinase or glucosinolate levels, and whether these different levels impact taste in ways that could affect consumption. We assessed taste and measured levels of protein, glucosinolate, myrosinase content, and direct antioxidant activity of the leaves of 36 M. oleifera accessions grown in a common garden. Taste tests readily highlighted differences between wild type and domesticated M. oleifera. There were differences in direct antioxidant potential, but not in myrosinase activity or protein quantity. However, these two populations were readily separated based solely upon their proportions of the two predominant glucosinolates (glucomoringin and glucosoonjnain). This study demonstrates substantial variation in glucosinolate composition within M. oleifera. The domestication of M. oleifera appears to have involved increases in levels of glucomoringin and substantial reduction of glucosoonjnain, with marked changes in taste.
Asunto(s)
Antioxidantes/análisis , Glucosinolatos/análisis , Glicósido Hidrolasas/análisis , Moringa oleifera , Proteínas de Plantas/análisis , Gusto/fisiología , Antioxidantes/metabolismo , Antioxidantes/farmacología , Desecación , Domesticación , Glucosinolatos/metabolismo , Glicósido Hidrolasas/metabolismo , Humanos , Moringa oleifera/química , Moringa oleifera/clasificación , Moringa oleifera/fisiología , Extractos Vegetales/química , Hojas de la Planta/química , Proteínas de Plantas/metabolismoRESUMEN
BACKGROUND: Increased interferon gamma (IFNγ) release occurs in Chronic Obstructive Pulmonary Disease (COPD) lungs. IFNγ supports optimal viral clearance, but if dysregulated could increase lung tissue destruction. METHODS: The present study investigates which mediators most closely correlate with IFNγ in sputum in stable and exacerbating disease, and seeks to shed light on the spatial requirements for innate production of IFNγ, as reported in mouse lymph nodes, to observe whether such microenvironmental cellular organisation is relevant to IFNγ production in COPD lung. RESULTS: We show tertiary follicle formation in severe disease alters the dominant mechanistic drivers of IFNγ production, because cells producing interleukin-18, a key regulator of IFNγ, are highly associated with such structures. Interleukin-1 family cytokines correlated with IFNγ in COPD sputum. We observed that the primary source of IL-18 in COPD lungs was myeloid cells within lymphoid aggregates and IL-18 was increased in severe disease. IL-18 released from infected epithelium or from activated myeloid cells, was more dominant in driving IFNγ when IL-18-producing and responder cells were in close proximity. CONCLUSIONS: Unlike tight regulation to control infection spread in lymphoid organs, this local interface between IL-18-expressing and responder cell is increasingly supported in lung as disease progresses, increasing its potential to increase tissue damage via IFNγ.
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Interferón gamma/biosíntesis , Interleucina-18/biosíntesis , Pulmón/metabolismo , Linfocitos/metabolismo , Enfermedad Pulmonar Obstructiva Crónica/metabolismo , Índice de Severidad de la Enfermedad , Humanos , Leucocitos Mononucleares/metabolismo , Leucocitos Mononucleares/patología , Estudios Longitudinales , Pulmón/patología , Linfocitos/patología , Estudios Prospectivos , Enfermedad Pulmonar Obstructiva Crónica/patología , Esputo/metabolismoRESUMEN
SCOPE: The isothiocyanate sulforaphane (SF) from broccoli is one of the most potent known inducers of the cytoprotective phase 2 response. Its role in a host of biochemical pathways makes it a major component of plant-based protective strategies for enhancing healthspan. Many nutritional supplements are now marketed that purport to contain SF, which in plants exists as a stable precursor, a thioglucoside hydroxysulfate. However, SF in pure form must be stabilized for use in supplements. METHODS AND RESULTS: We evaluated the stability and bioavailability of two stabilized SF preparations-an α-cyclodextrin inclusion (SF-αCD), and an SF-rich, commercial nutritional supplement. SF-αCD area-under-the-curve peak serum concentrations occurred at 2 h, but six of ten volunteers complained of mild stomach upset. After topical application it was not effective in upregulating cytoprotective enzymes in the skin of SKH1 mice whereas pure SF was effective in doing so. Both of these "stabilized" SF preparations were as potent as pure SF in inducing the cytoprotective response in cultured cells, and they were more stable and as bioavailable. CONCLUSION: Our studies of a stabilized phytochemical component of foods should encourage further examination of similar products for their utility in chronic disease prevention and therapy.
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Anticarcinógenos/farmacología , Brassica/química , Isotiocianatos/farmacología , Tiocianatos/farmacología , Animales , Disponibilidad Biológica , Suplementos Dietéticos , Glucosinolatos/farmacología , Humanos , Imidoésteres/farmacología , Ratones , Oximas , Fitoquímicos/metabolismo , Sulfóxidos , alfa-Ciclodextrinas/metabolismoRESUMEN
OBJECTIVE: To identify biomechanical and clinical parameters that influence knee flexion (KF) angle at initial contact (IC) and during single limb stance phase of gait in children with spastic cerebral palsy (CP) who walk with flexed-knee gait. DESIGN: Retrospective analysis of gait kinematics and clinical data collected from 2010-2013. SETTING: Motion & Gait Analysis Laboratory at Lucile Packard Children's Hospital, Stanford, CA. PARTICIPANTS: Gait analysis data from persons with spastic CP (Gross Motor Function Classification System [GMFCS] I-III) who had no prior surgery were analyzed. Participants exhibiting KF ≥20° at IC were included; the more-involved limb was analyzed. METHODS: Outcome measures were analyzed with respect to clinical findings, including passive range of motion, Selective Motor Control Assessment for the Lower Extremity (SCALE), gait kinematics, and musculoskeletal models of muscle-tendon lengths during gait. MAIN OUTCOME MEASURES: KF at IC (KFIC) and minimum KF during single-limb support (KFSLS) were investigated. RESULTS: Thirty-four participants met the inclusion criteria, and their data were analyzed (20 males and 14 females, mean age 10.1 years, range 5-20 years). Mean KFIC was 34.4 ± 8.4 degrees and correlated with lower SCALE score (ρ = -0.530, P = .004), later peak KF during swing (ρ = 0.614, P < .001), and shorter maximal muscle length of the semimembranosus (ρ = -0.359, P = .037). Mean KFSLS was 18.7 ± 14.9 and correlated to KF contracture (ρ = 0.605, P < .001) and shorter maximal muscle length of the semimembranosus (ρ = -0.572, P < .001) and medial gastrocnemius (ρ = -0.386, P = .024). GMFCS correlated more strongly to KFIC (ρ = 0.502, P = .002) than to KFSLS (ρ = 0.371, P = .031). Linear regression found that both the SCALE score (P = .001) and delayed timing of peak KF during swing (P = .001) independently predicted KFIC. KF contracture (P = .026) and maximal length of the semimembranosus (P = .043) independently predicted KFSLS. CONCLUSION: Correlates of KFIC differed from those for KFSLS and suggest that impaired selective motor control and later timing of swing-phase KF influence knee position at IC, whereas KF contracture and muscle lengths influence minimal KF in single-limb support, findings with important treatment implications.
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Parálisis Cerebral/fisiopatología , Marcha/fisiología , Articulación de la Rodilla/fisiología , Rango del Movimiento Articular/fisiología , Adolescente , Fenómenos Biomecánicos , Parálisis Cerebral/diagnóstico , Niño , Preescolar , Femenino , Humanos , Masculino , Estudios Retrospectivos , Adulto JovenRESUMEN
Glucoraphanin from broccoli and its sprouts and seeds is a water soluble and relatively inert precursor of sulforaphane, the reactive isothiocyanate that potently inhibits neoplastic cellular processes and prevents a number of disease states. Sulforaphane is difficult to deliver in an enriched and stable form for purposes of direct human consumption. We have focused upon evaluating the bioavailability of sulforaphane, either by direct administration of glucoraphanin (a glucosinolate, or ß-thioglucoside-N-hydroxysulfate), or by co-administering glucoraphanin and the enzyme myrosinase to catalyze its conversion to sulforaphane at economic, reproducible and sustainable yields. We show that following administration of glucoraphanin in a commercially prepared dietary supplement to a small number of human volunteers, the volunteers had equivalent output of sulforaphane metabolites in their urine to that which they produced when given an equimolar dose of glucoraphanin in a simple boiled and lyophilized extract of broccoli sprouts. Furthermore, when either broccoli sprouts or seeds are administered directly to subjects without prior extraction and consequent inactivation of endogenous myrosinase, regardless of the delivery matrix or dose, the sulforaphane in those preparations is 3- to 4-fold more bioavailable than sulforaphane from glucoraphanin delivered without active plant myrosinase. These data expand upon earlier reports of inter- and intra-individual variability, when glucoraphanin was delivered in either teas, juices, or gelatin capsules, and they confirm that a variety of delivery matrices may be equally suitable for glucoraphanin supplementation (e.g. fruit juices, water, or various types of capsules and tablets).
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Brassica/química , Glucosinolatos/administración & dosificación , Glicósido Hidrolasas/administración & dosificación , Imidoésteres/administración & dosificación , Isotiocianatos/orina , Adulto , Disponibilidad Biológica , Suplementos Dietéticos , Sistemas de Liberación de Medicamentos , Femenino , Glucosinolatos/farmacocinética , Glicósido Hidrolasas/metabolismo , Humanos , Imidoésteres/farmacocinética , Isotiocianatos/farmacocinética , Masculino , Persona de Mediana Edad , Oximas , Extractos Vegetales/análisis , Extractos Vegetales/farmacocinética , Plantones/metabolismo , Semillas/metabolismo , SulfóxidosRESUMEN
We review herein the basis for using dietary components to treat and/or prevent Helicobacter pylori infection, with emphasis on (a) work reported in the last decade, (b) dietary components for which there is mechanism-based plausibility, and (c) components for which clinical results on H pylori amelioration are available. There is evidence that a diet-based treatment may reduce the levels and/or the virulence of H pylori colonization without completely eradicating the organism in treated individuals. This concept was endorsed a decade ago by the participants in a small international consensus conference held in Honolulu, Hawaii, USA, and interest in such a diet-based approach has increased dramatically since then. This approach is attractive in terms of cost, treatment, tolerability, and cultural acceptability. This review, therefore, highlights specific foods, food components, and food products, grouped as follows: bee products (eg, honey and propolis); probiotics; dairy products; vegetables; fruits; oils; essential oils; and herbs, spices, and other plants. A discussion of the small number of clinical studies that are available is supplemented by supportive in vitro and animal studies. This very large body of in vitro and preclinical evidence must now be followed up with rationally designed, unambiguous human trials.
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Productos Lácteos , Infecciones por Helicobacter/dietoterapia , Helicobacter pylori/crecimiento & desarrollo , Miel , Aceites Volátiles , Plantas Comestibles , Probióticos , Animales , Infecciones por Helicobacter/tratamiento farmacológico , Infecciones por Helicobacter/microbiología , Infecciones por Helicobacter/prevención & control , Humanos , Fitoterapia , EspeciasRESUMEN
OBJECTIVE: To identify clinical and biomechanical parameters that influence swing-phase knee flexion and contribute to stiff-knee gait in individuals with spastic cerebral palsy (CP) and flexed-knee gait. DESIGN: Retrospective analysis of clinical data and gait kinematics collected from 2010 to 2013. SETTING: Motion and gait analysis laboratory at a children's hospital. PARTICIPANTS: Individuals with spastic CP (N=34; 20 boys, 14 girls; mean age ± SD, 10.1±4.1y [range, 5-20y]; Gross Motor Function Classification System I-III) who walked with flexed-knee gait ≥20° at initial contact and had no prior surgery were included; the more-involved limb was analyzed. INTERVENTION: Not applicable. MAIN OUTCOME MEASURES: The magnitude and timing of peak knee flexion (PKF) during swing were analyzed with respect to clinical data, including passive range of motion and Selective Control Assessment of the Lower Extremity, and biomechanical data, including joint kinematics and hamstring, rectus femoris, and gastrocnemius muscle-tendon length during gait. RESULTS: Data from participants demonstrated that achieving a higher magnitude of PKF during swing correlated with a higher maximum knee flexion velocity in swing (ρ=.582, P<0.001) and a longer maximum length of the rectus femoris (ρ=.491, P=.003). In contrast, attaining earlier timing of PKF during swing correlated with a higher knee flexion velocity at toe-off (ρ=-.576, P<.001), a longer maximum length of the gastrocnemius (ρ=-.355, P=.039), and a greater peak knee extension during single-limb support phase (ρ=-.354, P=.040). CONCLUSIONS: Results indicate that the magnitude and timing of PKF during swing were independent, and their biomechanical correlates differed, suggesting important treatment implications for both stiff-knee and flexed-knee gait.
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Parálisis Cerebral/fisiopatología , Parálisis Cerebral/rehabilitación , Trastornos Neurológicos de la Marcha/fisiopatología , Trastornos Neurológicos de la Marcha/rehabilitación , Articulación de la Rodilla/fisiopatología , Adolescente , Fenómenos Biomecánicos , Niño , Preescolar , Femenino , Humanos , Masculino , Músculo Esquelético/fisiopatología , Estudios Retrospectivos , Adulto JovenRESUMEN
Flavonoids are secondary plant products that are well represented in healthy diets because of ingestion of fruit, vegetables, herbs, and teas. Increased consumption is correlated with decreased risks of cardiovascular disease, cancer, and other chronic diseases. Certain flavonoids confer direct antioxidant protection to cells, others induce enzymes that protect cells against oxidative and other insults ("indirect antioxidants"), and others appear to be protective by both mechanisms. Hydroxylated flavones manifest substantial direct antioxidant activity but do not effectively induce cytoprotective enzymes. Methoxylated flavones that potently induce cytoprotective enzymes were evaluated to elucidate the structural prerequisites for effective chemoprotective agents: protecting healthy cells with minimal collateral toxicity. Flavones and flavanones methoxylated at the 5-position of the A-ring were among the most potent inducers of the cytoprotective NAD(P)H:quinone-oxidoreductase 1 (NQO1) in 3 different cell lines. Other flavones were equally potent inducers, but more toxic. Flavanones contain no Michael reaction center, yet some are potent inducers of NQO1, have low cytotoxicity, and inhibit LPS-stimulated iNOS activity, which suggests a redox mechanism of action rather than the Keap1/Nrf2/ARE mechanism by which so many of the classic inducers operate. Evaluation in vivo will reveal whether differential protective advantages support their possible evaluation in a cancer prevention setting.
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Antiinflamatorios/farmacología , Antioxidantes/farmacología , Flavonoides/química , Flavonoides/farmacología , Animales , Benzotiazoles/metabolismo , Células CACO-2 , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Quimioprevención , Humanos , Concentración 50 Inhibidora , Modelos Lineales , Lipopolisacáridos/efectos adversos , Ratones , NAD(P)H Deshidrogenasa (Quinona)/genética , NAD(P)H Deshidrogenasa (Quinona)/metabolismo , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/metabolismo , Oxidación-Reducción , Relación Estructura-Actividad , Ácidos Sulfónicos/metabolismoRESUMEN
Infections by Helicobacter pylori are very common, causing gastroduodenal inflammation including peptic ulcers, and increasing the risk of gastric neoplasia. The isothiocyanate (ITC) sulforaphane [SF; 1-isothiocyanato-4-(methylsulfinyl)butane] derived from edible crucifers such as broccoli is potently bactericidal against Helicobacter, including antibiotic-resistant strains, suggesting a possible dietary therapy. Gastric H. pylori infections express high urease activity which generates ammonia, neutralizes gastric acidity, and promotes inflammation. The finding that SF inhibits (inactivates) urease (jack bean and Helicobacter) raised the issue of whether these properties might be functionally related. The rates of inactivation of urease activity depend on enzyme and SF concentrations and show first order kinetics. Treatment with SF results in time-dependent increases in the ultraviolet absorption of partially purified Helicobacter urease in the 260-320 nm region. This provides direct spectroscopic evidence for the formation of dithiocarbamates between the ITC group of SF and cysteine thiols of urease. The potencies of inactivation of Helicobacter urease by isothiocyanates structurally related to SF were surprisingly variable. Natural isothiocyanates closely related to SF, previously shown to be bactericidal (berteroin, hirsutin, phenethyl isothiocyanate, alyssin, and erucin), did not inactivate urease activity. Furthermore, SF is bactericidal against both urease positive and negative H. pylori strains. In contrast, some isothiocyanates such as benzoyl-ITC, are very potent urease inactivators, but are not bactericidal. The bactericidal effects of SF and other ITC against Helicobacter are therefore not obligatorily linked to urease inactivation, but may reduce the inflammatory component of Helicobacter infections.
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Proteínas Bacterianas/antagonistas & inhibidores , Helicobacter pylori/enzimología , Isotiocianatos/farmacología , Tiocianatos/farmacología , Ureasa/antagonistas & inhibidores , Antibacterianos/química , Antibacterianos/farmacología , Proteínas Bacterianas/metabolismo , Brassica/química , Relación Dosis-Respuesta a Droga , Infecciones por Helicobacter/microbiología , Infecciones por Helicobacter/prevención & control , Helicobacter pylori/efectos de los fármacos , Humanos , Isotiocianatos/química , Cinética , Pruebas de Sensibilidad Microbiana , Estructura Molecular , Espectrofotometría Ultravioleta , Sulfóxidos , Tiocianatos/química , Factores de Tiempo , Ureasa/metabolismoRESUMEN
Sulforaphane [1-isothiocyanato-4-(methylsulfinyl)butane], a naturally occurring isothiocyanate derived from cruciferous vegetables, is a highly potent inducer of phase 2 cytoprotective enzymes and can protect against electrophiles including carcinogens, oxidative stress, and inflammation. The mechanism of action of sulforaphane is believed to involve modifications of critical cysteine residues of Keap1, which lead to stabilization of Nrf2 to activate the antioxidant response element of phase 2 enzymes. However, the dithiocarbamate functional group formed by a reversible reaction between isothiocyanate of sulforaphane and sulfhydryl nucleophiles of Keap1 is kinetically labile, and such modification in intact cells has not yet been demonstrated. Here we designed sulforaphane analogs with replacement of the reactive isothiocyanate by the more gentle electrophilic sulfoxythiocarbamate group that also selectively targets cysteine residues in proteins but forms stable thiocarbamate adducts. Twenty-four sulfoxythiocarbamate analogs were synthesized that retain the structural features important for high potency in sulforaphane analogs: the sulfoxide or keto group and its appropriate distance to electrophilic functional group. Evaluation in various cell lines including hepatoma cells, retinal pigment epithelial cells, and keratinocytes as well as in mouse skin shows that these analogs maintain high potency and efficacy for phase 2 enzyme induction as well as the inhibitory effect on lipopolysaccharide-induced nitric oxide formation like sulforaphane. We further show in living cells that a sulfoxythiocarbamate analog can label Keap1 on several key cysteine residues as well as other cellular proteins offering new insights into the mechanism of chemoprotection.
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Productos Biológicos/química , Productos Biológicos/farmacología , Tiocianatos/química , Tiocianatos/farmacología , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Animales , Línea Celular , Supervivencia Celular/efectos de los fármacos , Proteínas del Citoesqueleto/metabolismo , Electrones , Isotiocianatos , Proteína 1 Asociada A ECH Tipo Kelch , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Ratones , Estructura Molecular , Óxido Nítrico/metabolismo , Piel/efectos de los fármacos , Piel/metabolismo , Relación Estructura-Actividad , Compuestos de Sulfhidrilo/química , SulfóxidosRESUMEN
BACKGROUND: Sarcopenia is a consequence of aging. This atrophic event is responsible for decrease in strength and associated functional deficits seen in the aging adult. PURPOSE: This paper reviews: (1) the mechanisms contributing to sarcopenia, (2) the impact of age-related changes in muscle composition on 3 processes integral to muscle function, (3) the efficacy of pharmaceuticals and over-the-counter nutritional supplements in the management of sarcopenia, (4) experimental use of pharmaceutical regulation of myostatin to increase muscle mass and strength in animal models, and (5) efficacy of resistance training as a means of maintaining or recovering muscle mass and strength. METHODS: PubMed was searched for relevant research articles using the following descriptors: sarcopenia, aging, muscle mass, muscle performance, muscle strength, myostatin, testosterone, growth hormone, dehydroepiandrosterone, hormone replacement, nutrition, resistance training, and endurance training. RESULTS: Sarcopenia is mediated by multiple mechanisms, including alpha-motor neuron death, altered hormone concentrations, increased inflammation, and altered nutritional status. Age-related changes within muscle likely affect processes integral to muscle function. These changes negatively influence muscle performance directly or by contributing to sarcopenia. Pharmaceutical or supplement interventions to treat sarcopenia have not proved encouraging to date, either lacking or providing limited efficacy, along with the potential for negative health consequences. In contrast, resistance training has proven safe and highly effective for increasing muscle mass and strength in aging adults. CONCLUSION: Sarcopenia is a multifactorial consequence of aging that will affect many adults. Resistance training is the most effective and safe intervention to attenuate or recover some of the loss of muscle mass and strength that accompanies aging.
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Envejecimiento/fisiología , Entrenamiento de Fuerza , Sarcopenia/fisiopatología , Sarcopenia/terapia , Anciano , Anciano de 80 o más Años , Anabolizantes/uso terapéutico , Femenino , Humanos , Masculino , Sarcopenia/tratamiento farmacológicoRESUMEN
Participants were recruited from female undergraduate students participating in an ongoing longitudinal study at the time of a campus shooting. Eighty-five percent (N = 691) of the 812 students who were invited to participate in the current study completed questionnaires an average of 27 days following a campus shooting. In a mixed cross-sectional and longitudinal design, the cognitive and the physical concerns dimensions of postshooting anxiety sensitivity accounted for unique variance in posttrauma stress symptom severity (cross-sectional), after controlling for preshooting psychological symptoms (longitudinal). The cognitive concerns dimension showed the strongest relationship. Anxiety sensitivity also appeared to moderate the relationships of hyperarousal symptoms with reexperiencing and numbing symptoms.
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Trastornos de Ansiedad/diagnóstico , Armas de Fuego , Medio Social , Trastornos por Estrés Postraumático/diagnóstico , Estudiantes/psicología , Violencia/psicología , Heridas por Arma de Fuego/psicología , Adolescente , Trastornos de Ansiedad/epidemiología , Trastornos de Ansiedad/psicología , Nivel de Alerta , Estudios Transversales , Femenino , Humanos , Illinois , Estudios Longitudinales , Inventario de Personalidad/estadística & datos numéricos , Psicometría , Psicopatología , Factores de Riesgo , Factores Sexuales , Trastornos por Estrés Postraumático/epidemiología , Trastornos por Estrés Postraumático/psicología , Estudiantes/estadística & datos numéricos , Adulto JovenRESUMEN
Cognitive models of social anxiety suggest that fear of negative evaluation (FNE) is the central cognitive dimension underlying the disorder. The Fear of Positive Evaluation Scale (FPES; Weeks, Heimberg, & Rodebaugh, 2008) was recently developed to assess an additional cognitive dimension purported to underlie social anxiety disorder (SAD), but its psychometric properties have yet to be examined in clinical populations. The present study, with 133 treatment-seeking patients, examined the applicability of the FPES with a clinical sample. Results indicated that the FPES was factorially distinct from a measure assessing FNE, and patients with SAD (n=51) had higher mean scores on the FPES than patients with other anxiety disorders (n=82). The FPES also showed adequate reliability (internal consistency), good convergent and discriminant validity, acceptable criterion-related validity in predicting social interaction anxiety symptoms, and appropriate sensitivity to treatment. The FPES appears to have good psychometric properties and is a promising new assessment tool for better understanding SAD.