RESUMEN
Solid-state 19 Fâ NMR is a powerful method to study the interactions of biologically active peptides with membranes. So far, in labelled peptides, the 19 F-reporter group has always been installed on the side chain of an amino acid. Given the fact that monofluoroalkenes are non-hydrolyzable peptide bond mimics, we have synthesized a monofluoroalkene-based dipeptide isostere, Val-Ψ[(Z)-CF=CH]-Gly, and inserted it in the sequence of two well-studied antimicrobial peptides: PGLa and (KIGAKI)3 are representatives of an α-helix and a ß-sheet. The conformations and biological activities of these labeled peptides were studied to assess the suitability of monofluoroalkenes for 19 Fâ NMR structure analysis.
Asunto(s)
Alquenos/química , Péptidos Catiónicos Antimicrobianos/química , Membrana Celular/química , Espectroscopía de Resonancia Magnética , Secuencia de Aminoácidos , Péptidos Catiónicos Antimicrobianos/síntesis química , Flúor/química , Conformación Proteica en Hélice alfa , Coloración y Etiquetado/métodosRESUMEN
An increasing body of evidence suggests that sex steroids play an important role in the development and regulation of vertebrate immune defense. Therefore, compounds with estrogenic activity may influence the immune system via receptor-mediated pathways. The presence of estrogen receptors in immune cells and organs during the early stages of development may indicate that female steroid hormones are involved in the maturation of the fish immune system. This is of particular importance, as some marine fish are probably exposed to sources of exogenous estrogens while they reside in their estuarine nursery grounds. In this study, the influence of 17ß-estradiol (E2) on estrogen receptor and cytokine gene expression was assessed in juvenile sea bass (Dicentrarchus labrax) together with characterization of the head kidney leukocyte populations and corresponding phagocytic activity during organ regionalization from 98 to 239 dph. E2 exposure, beginning at 90 dph resulted in indirect and delayed modifications of interleukin 1ß and estrogen receptor α gene expression, which may affect B-lymphocyte proliferation in the sea bass head kidney. The E2 treatment of 120 dph fish led to an increase in estrogen receptor ß2 and a decrease in transforming growth factor ß1 gene expression, which coincided with decreased phagocytic activity of head kidney lymphocytes and monocytes/macrophages. Additionally, these changes were observed during developmental periods described as critical phases for B-lymphocyte development in mammals. Consequently, exogenous estrogens have the potential to modify the innate immune response in juvenile sea bass and to exert detrimental effects on head kidney development. Copyright © 2015 John Wiley & Sons, Ltd.
Asunto(s)
Lubina , Disruptores Endocrinos/toxicidad , Estradiol/toxicidad , Regulación del Desarrollo de la Expresión Génica/efectos de los fármacos , Sistema Inmunológico/efectos de los fármacos , Riñón/efectos de los fármacos , Contaminantes Químicos del Agua/toxicidad , Adaptación Fisiológica/efectos de los fármacos , Animales , Acuicultura , Lubina/crecimiento & desarrollo , Lubina/inmunología , Lubina/metabolismo , Receptor beta de Estrógeno/agonistas , Receptor beta de Estrógeno/genética , Receptor beta de Estrógeno/metabolismo , Proteínas de Peces/agonistas , Proteínas de Peces/antagonistas & inhibidores , Proteínas de Peces/química , Proteínas de Peces/metabolismo , Explotaciones Pesqueras , Francia , Sistema Inmunológico/crecimiento & desarrollo , Sistema Inmunológico/inmunología , Sistema Inmunológico/metabolismo , Inmunidad Innata/efectos de los fármacos , Riñón/crecimiento & desarrollo , Riñón/inmunología , Riñón/metabolismo , Leucocitos/efectos de los fármacos , Leucocitos/inmunología , Leucocitos/metabolismo , Organogénesis/efectos de los fármacos , Fagocitos/efectos de los fármacos , Fagocitos/inmunología , Fagocitos/metabolismo , Fagocitosis/efectos de los fármacos , Subunidades de Proteína/agonistas , Subunidades de Proteína/genética , Subunidades de Proteína/metabolismo , Factores de Tiempo , Factor de Crecimiento Transformador beta1/antagonistas & inhibidores , Factor de Crecimiento Transformador beta1/genética , Factor de Crecimiento Transformador beta1/metabolismoRESUMEN
Coastal urbanisation exposes surrounding estuarine environments to urban-related contaminants such as polycyclic aromatic hydrocarbons (PAHs), polychlorobiphenyls (PCBs) and pesticide mixtures. Hydrophobic contaminants can adsorb on estuarine sediments. They can subsequently be released on a massive scale in the aquatic environment due to artificial or natural phenomena (e.g. dredging, tides), thereby threatening living organisms. The contamination of sediment is a significant ecological issue in the Seine estuary, France. However, few relevant methods have been developed to assess sediment toxicity and its ecological impacts in a cost-effective way. In this context, we aimed to assess the toxicity of natural sediments from the Seine estuary on the development of the calanoid copepod Eurytemora affinis using a previously developed larval bioassay. This assay involves direct exposure of nauplii to elutriates of sediments for six days. Sediments were collected along the Seine estuary from six polluted sites and one reference site. Pollutants in this estuary included PAHs, PCBs and OCPs (organochlorine pesticides). Nauplius survival was significantly more affected by exposure to all contaminated sediment elutriates, than by exposure to sediment from Yville-sur-Seine (the reference site), whereas nauplius growth was significantly reduced after exposure to contaminated sediment elutriates from four of the six contaminated sites. We identified two distinct site clusters, one including both the sand-rich and the least polluted sediments (Oissel, Quillebeuf-sur-Seine, Caudebec-en-Caux) and the other including both the clay- and silt-rich, and the most polluted sediments (La Bouille, Poses, Pont de Normandie). As expected, survival was significantly more impacted after exposure to elutriates from the second cluster than from the first. This work enables (i) assessment of the toxicity of natural sediments in the Seine estuary and (ii) validation of the larval bioassay previously developed using sorbed sediment with model molecules.
Asunto(s)
Copépodos/efectos de los fármacos , Monitoreo del Ambiente/métodos , Estuarios/estadística & datos numéricos , Sedimentos Geológicos/química , Pruebas de Toxicidad/métodos , Animales , Femenino , Francia , Sustancias Peligrosas/análisis , Sustancias Peligrosas/toxicidad , Hidrocarburos Clorados/análisis , Larva/efectos de los fármacos , Tamaño de la Partícula , Bifenilos Policlorados/análisis , Hidrocarburos Policíclicos Aromáticos/análisis , Análisis de Componente PrincipalRESUMEN
Hydrophobic pollutants, in particular sediment-sorbed organic compounds, are widespread in the aquatic environment and could represent a threat to living organisms. Estuarine species, which live in turbulent ecosystems, are particularly exposed to this mode of contamination. For precise evaluation of the toxicity of hydrophobic contaminants desorbed from particles, a new larval assay using nauplii of the estuarine calanoid copepod Eurytemora affinis was developed. It consists of the direct exposure of copepods during naupliar development to elutriates of an unpolluted sediment spiked with different model contaminants. This bioassay measures the toxicity of the bioavailable fraction of particle-sorbed pollutants on the naupliar stage of copepods. Mortality and growth (non-invasive endpoints) in nauplii were analysed after six days of exposure. This approach was validated using six pollutants with different modes of action: benzo[a]pyrene (BaP), dimethylbenzo[a]anthracene (DMBA), phenanthrene (PHE), polychlorinated biphenyls (PCB 126, PCB 153) and 4-nonylphenol (4-NP). All these compounds induced a dose-dependent increase in toxic effects. Lethal effects only occurred at the highest tested concentrations: 58,541 and 6092 ng g(-1) dry weight sediment (dws), for PHE and DMBA, respectively. Sublethal effects (growth inhibition) were observed at lower concentrations for all tested compounds except PCB 153, from 8, 142, 297, 6092 and 8453 ng g(-1) dws for PCB 126, BaP, PHE, DMBA and 4-NP, respectively.
Asunto(s)
Bioensayo/métodos , Copépodos/fisiología , Monitoreo del Ambiente/métodos , Sedimentos Geológicos/química , Bifenilos Policlorados/toxicidad , Contaminantes Químicos del Agua/toxicidad , Animales , Copépodos/efectos de los fármacos , Larva/efectos de los fármacosRESUMEN
BACKGROUND: Treatment with the atypical antipsychotic risperidone can result in elevated prolactin levels. To date, the relationships between plasma concentrations of prolactin, risperidone and its active 9-hydroxy-metabolite have been little investigated in adolescents with psychosis. METHODS: Prolactin levels were determined at baseline in 16 hospitalized drug-naïve adolescents meeting DSM-IV criteria for schizophreniform disorder. Prolactin, risperidone, 9-hydroxyrisperidone levels were subsequently determined after 3 weeks of oral risperidone treatment. RESULTS: Compared with pretreatment values, prolactin levels at endpoint were significantly increased (p<0.00001) and correlated with risperidone doses (r=0.58, N=16, p<0.02), and plasma levels of risperidone (r=0.60, N=16, p<0.02) and 9-hydroxyrisperidone (r=0.54, N=16, p=0.03). CONCLUSIONS: These data suggest that risperidone's effect on prolactin release is dose-dependent in adolescents and is linked to both plasma risperidone and 9-hydroxyrisperidone concentrations.
Asunto(s)
Antipsicóticos/farmacocinética , Isoxazoles/sangre , Prolactina/sangre , Trastornos Psicóticos/sangre , Pirimidinas/sangre , Risperidona/farmacocinética , Adolescente , Medicina del Adolescente , Factores de Edad , Antipsicóticos/metabolismo , Relación Dosis-Respuesta a Droga , Femenino , Estudios de Seguimiento , Humanos , Masculino , Palmitato de Paliperidona , Prolactina/efectos de los fármacos , Trastornos Psicóticos/tratamiento farmacológico , Risperidona/metabolismoRESUMEN
Evidence suggests that individuals with posttraumatic stress disorder (PTSD) have an enhanced sensitization of the hypothalamic--pituitary--adrenocortical (HPA) axis. However, few studies in adolescents have been performed. Fourteen sexually abused adolescent inpatients with DSM-IV PTSD (12 female, two male; mean +/- SD age, 16.2 +/- 1.9 years) were compared with 14 adolescent hospitalized controls (11 female, three male; mean age, 15.7 +/- 2.0 years). All subjects underwent a standard dexamethasone suppression test (DST, 1 mg given orally at 2300 h) five days after admission. Baseline blood samples were obtained at 0800 h, and the following day, adrenocorticotropin (ACTH) and cortisol levels were measured at 0800, 1600, and 2300 h. Clinical assessment included the Impact of Event Scale, Stanford Acute Stress Reaction Questionnaire, Beck Depression Inventory, and Coping Inventory for Stressful Situations. Post-DST ACTH levels were significantly lower in PTSD than in control adolescents (at 0800 h: P < 0.005; at 1600 h: P < 0.001; at 2300 h: P < 0.05). In patients, post-DST cortisol levels were reduced but not significantly. No correlations were found between ACTH and cortisol levels and time elapsed since trauma. These results demonstrate that sexually abused adolescents with PTSD show ACTH hypersuppression to DST suggesting enhanced glucocorticoid receptor sensitivity in the pituitary.
Asunto(s)
Hormona Adrenocorticotrópica/sangre , Abuso Sexual Infantil/psicología , Hidrocortisona/sangre , Trastornos por Estrés Postraumático/sangre , Adaptación Psicológica/fisiología , Adolescente , Adulto , Factores de Edad , Dexametasona/farmacología , Femenino , Glucocorticoides/farmacología , Humanos , Sistema Hipotálamo-Hipofisario/efectos de los fármacos , Masculino , Sistema Hipófiso-Suprarrenal/efectos de los fármacos , Receptores de Glucocorticoides/efectos de los fármacos , Receptores de Glucocorticoides/metabolismo , Factores Sexuales , Estimulación Química , Trastornos por Estrés Postraumático/psicologíaRESUMEN
Evidence suggests that individuals with posttraumatic stress disorder (PTSD) have enhanced sensitization of the hypothalamic-pituitary-adrenocortical (HPA) axis. Fourteen adolescent inpatients with DSM-IV PTSD were compared with 14 adolescent hospitalized controls without current axis I diagnoses. All patients were drug-naive. The causative trauma had been sexual abuse in all cases. Dexamethasone, 1 mg orally, was given at 11 PM, 5 days after admission. Baseline blood samples were obtained at 8 AM, and on the following day, adrenocorticotropin (ACTH) and cortisol levels were measured at 8 AM, 4 PM, and 11 PM. Clinical assessment included the Impact of Event Scale, Stanford Acute Stress Reaction Questionnaire, Beck Depression Inventory, and Coping Inventory for Stressful Situations. Post-DST ACTH levels were significantly lower in PTSD than in control adolescents (at 8 AM, P <0.005; at 4 PM, P <0.001; and at 11 PM, P <0.05). In patients, post-DST cortisol levels were reduced but not significantly. No correlations were found between ACTH and cortisol levels and time elapsed since trauma. These results demonstrate that sexually abused adolescents with PTSD show ACTH hypersuppression to DST, suggesting enhanced glucocorticoid receptor sensitivity in the pituitary.
Asunto(s)
Hormona Adrenocorticotrópica/antagonistas & inhibidores , Abuso Sexual Infantil/psicología , Dexametasona , Trastornos por Estrés Postraumático/metabolismo , Trastornos por Estrés Postraumático/psicología , Adaptación Psicológica/fisiología , Hormona Adrenocorticotrópica/metabolismo , Adulto , Niño , Relación Dosis-Respuesta a Droga , Femenino , Humanos , Hidrocortisona/sangre , Sistema Hipotálamo-Hipofisario/fisiopatología , Masculino , Escalas de Valoración PsiquiátricaRESUMEN
CONTEXT: High rates of complementary and alternative medicine (CAM) utilization among older adults are of interest because studies suggest relationships among CAM, self-care, and healthy lifestyle. These potential relationships are of particular interest because of the contribution of self-care in maintaining physical, emotional, and spiritual health as key strategies to successful aging. Naturopathic medicine (NM) is a type of primary care CAM used by older adults, yet little is known about older adults' use of NM or its role in healthy aging. OBJECTIVES: We sought to examine perceptions about, and utilization of, NM and the experience of addressing health and wellness as part of the aging process. DESIGN: Phenomenology-based qualitative inquiry. SETTING: A total of 13 community-based clinics affiliated with a naturopathic medical school. PARTICIPANTS: A total of 47 adults aged 58-100 years. METHODS: Qualitative analysis of semi-structured focus groups with quantitative analysis of clinical administrative data. RESULTS: Over 2543 older adults used NM services affiliated with a natural academic clinic in Seattle in 2011 for diagnoses consistent with primary care: fatigue, anxiety, diabetes, diarrhea, and depression, in rank order. Participants reported they sought NM because it offered a different system of care aligned with their values. Themes included (1) feeling heard, (2) a focus on health education including prevention and self-care, and (3) feeling supported. Themes further distilled into optimal descriptive characteristics of patient-centered providers that may best support older adults' healthy aging. A final theme described barriers to further utilization.
Asunto(s)
Envejecimiento , Actitud Frente a la Salud , Estilo de Vida , Naturopatía , Atención Dirigida al Paciente , Relaciones Médico-Paciente , Atención Primaria de Salud/métodos , Anciano , Cultura , Empatía , Femenino , Grupos Focales , Salud , Educación en Salud , Humanos , Masculino , Persona de Mediana Edad , Naturopatía/estadística & datos numéricos , Investigación Cualitativa , Autocuidado , WashingtónRESUMEN
BACKGROUND: P-gp expression has been linked to the efflux of chemotherapeutic drugs in human cancers leading to multidrug resistance. Fluorescence techniques have been widely applied to measure the P-gp activity. In this paper, there is a comparison between the advantages of two fluorescence approaches of commonly available and affordable instruments: the microplate reader (MPR) and the flow cytometer to detect the P-gp efflux activity using calcein-AM. RESULTS: The selectivity, sensibility, and reproducibility of the two methods have been defined. Our results showed that the MPR is more powerful for the detection of small inhibition, whereas the flow cytometry method is more reliable at higher concentrations of the inhibitors. We showed that to determine precisely the inhibition efficacy the flow cytometry is better; hence, to get the correct E max and EC50 values, we cannot only rely on the MPR. CONCLUSION: Both techniques can potentially be used extensively in the pharmaceutical industry for high-throughput drug screening and in biology laboratories for academic research, monitoring the P-gp efflux in specific assays.
Asunto(s)
Subfamilia B de Transportador de Casetes de Unión a ATP/metabolismo , Resistencia a Múltiples Medicamentos/fisiología , Resistencia a Antineoplásicos/fisiología , Fluoresceínas/metabolismo , Línea Celular Tumoral , Fluorescencia , Humanos , Células MCF-7 , Reproducibilidad de los ResultadosRESUMEN
The present study aimed to develop a method of quantification of heat shock protein transcript levels in the estuarine copepod Eurytemora affinis. For that, the full-length cDNA of the 78-kDa glucose-regulated protein (Ea-grp78) and the cytosolic 90-kDa heat shock protein (Ea-hsp90A) from this species have been cloned. These cDNA revealed, respectively, 2,370 and 2,299 bp with 1,971 and 2,124 bp open reading frames encoding 656 and 707 amino acids. Main features, sequence identities and phylogenetic analysis with other species were described. Then, the expression profiles were analysed using reverse transcription/real-time quantitative PCR method from copepods subjected to different thermic and osmotic stresses in laboratory, and from copepods directly sampled into the natural population of the Seine Estuary (France) along a salinity gradient. Thermic shock (7.5°C, 22.5°C and 30°C during 90 min) significantly induced increases of transcript quantities ranged between 1.7- and 19.7-fold the levels observed in control conditions (15°C). Hypo- and hyper-osmotic shocks (salinities of 1 and 30 during 90 min) caused a 2-fold induction of Ea-hsp90A transcript level in comparison to controls (salinity of 15) whereas no significant change was measured for Ea-grp78. On the other hand, similar expression profiles were observed for the two transcripts after 72 h of exposition to salinities of 1 and 25 with a significant 2-fold induction observed for the lower salinity. To finish, strong expression inductions of both Ea-grp78 and Ea-hsp90A genes were observed in field copepods sampled at low salinity during the campaigns of June 2009 and May 2010. These results tend to show that the low salinity and the increase of temperature seem to have a synergic effect on stress condition of copepods.