RESUMEN
Catecholamines and their metabolites act as neurotransmitters in the brain and are important for nervous system function. In the current work, a highly selective and sensitive UPLC-MS/MS assay was developed for quantitation of six catecholamines and their metabolites, including dopamine, norepinephrine, serotonin, 3,4-dihydroxyphenylacetic acid, homovanillic acid and 5-hydroxyindolacetic acid from rat and mouse striatum as pharmacodynamic biomarkers to support neuroscience and pharmaceutical research. A fit-for-purpose strategy for method development, assay qualification and study support were adopted for this assay. A surrogate matrix (brain homogenizing solution absent of targeted analytes) was used for preparation of calibration samples and certain levels of quality control samples to avoid interference from endogenous baselines. Homogenized rodent striatum was derivatized by dansyl chloride to enhance the sensitivity, followed by liquid-liquid extraction with ethyl acetate in 96-well plate format. The lower limit of quantitation (LLOQ) was 0.2 ng/mL in tissue homogenate, equivalent to 3.2 pg/mg in brain tissue, which could be further reduced to ten times lower by changing the re-dissolving and injecting volume in the last sample purification step. Acceptable accuracy, precision, linearity, specificity, recovery, and matrix effect was obtained. Bench-top stability (2 h), freeze-thaw stability (3 cycles at -20 °C), and - 80 °C storage stability (up to 51 days) in both tissue homogenate and surrogate matrix along with autosampler stability (60 h at 4°C) all met acceptance criteria. This assay was successfully applied to measure the six analytes in striatum from mice treated with the neurotoxin 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), an animal model of Parkinsonism for which dosing protocols can vary widely, and further confirmed the metabolic pathway of neurotoxicity by the quantification of catecholamine metabolites. Our study is the first detailed the step-by-step recovery and pointed out the key factors for the assay to simultaneously quantify these six neurotransmitters in rodent striatum with superior sensitivity.
Asunto(s)
Catecolaminas , Espectrometría de Masas en Tándem , Animales , Cromatografía Líquida de Alta Presión/métodos , Cromatografía Liquida/métodos , Ratones , Neurotransmisores , Ratas , Reproducibilidad de los Resultados , Espectrometría de Masas en Tándem/métodosRESUMEN
While GM1 may interact with α-synuclein in vitro to inhibit aggregation, the ability of GM1 to protect against α-synuclein toxicity in vivo has not been investigated. We used targeted adeno-associated viral vector (AAV) overexpression of human mutant α-synuclein (A53T) in the rat substantia nigra (SN) to produce degeneration of SN dopamine neurons, loss of striatal dopamine levels, and behavioral impairment. Some animals received daily GM1 ganglioside administration for 6 weeks, beginning 24 hours after AAV-A53T administration or delayed start GM1 administration for 5 weeks beginning 3 weeks after AAV-A53T administration. Both types of GM1 administration protected against loss of SN dopamine neurons and striatal dopamine levels, reduced α-synuclein aggregation, and delayed start administration of GM1 reversed early appearing behavioral deficits. These results extend prior positive results in MPTP models, are consistent with the results of a small clinical study of GM1 in PD patients that showed slowing of symptom progression with chronic use, and argue for the continued refinement and development of GM1 as a potential disease modifying therapy for PD.
Asunto(s)
Gangliósido G(M1)/farmacología , Fármacos Neuroprotectores/farmacología , Enfermedad de Parkinson/tratamiento farmacológico , alfa-Sinucleína/genética , Animales , Conducta Animal/efectos de los fármacos , Cuerpo Estriado/efectos de los fármacos , Cuerpo Estriado/metabolismo , Dependovirus/efectos de los fármacos , Modelos Animales de Enfermedad , Dopamina/metabolismo , Neuronas Dopaminérgicas/efectos de los fármacos , Neuronas Dopaminérgicas/patología , Regulación de la Expresión Génica/efectos de los fármacos , Vectores Genéticos/genética , Humanos , Enfermedad de Parkinson/genética , Enfermedad de Parkinson/patología , Ratas , Sustancia Negra/efectos de los fármacos , Sustancia Negra/metabolismoRESUMEN
Fetal Alcohol Spectrum Disorders (FASD) collectively describes the constellation of effects resulting from human alcohol consumption during pregnancy. Even with public awareness, the incidence of FASD is estimated to be upwards of 5% in the general population and is becoming a global health problem. The physical, cognitive, and behavioral impairments of FASD are recapitulated in animal models. Recently rodent models utilizing voluntary drinking paradigms have been developed that accurately reflect moderate consumption, which makes up the majority of FASD cases. The range in severity of FASD characteristics reflects the frequency, dose, developmental timing, and individual susceptibility to alcohol exposure. As most rodent models of FASD use C57BL/6 mice, there is a need to expand the stocks of mice studied in order to more fully understand the complex neurobiology of this disorder. To that end, we allowed pregnant Swiss Webster mice to voluntarily drink ethanol via the drinking in the dark (DID) paradigm throughout their gestation period. Ethanol exposure did not alter gestational outcomes as determined by no significant differences in maternal weight gain, maternal liquid consumption, litter size, or pup weight at birth or weaning. Despite seemingly normal gestation, ethanol-exposed offspring exhibit significantly altered timing to achieve developmental milestones (surface righting, cliff aversion, and open field traversal), as analyzed through mixed-effects Cox proportional hazards models. These results confirm Swiss Webster mice as a viable option to study the incidence and causes of ethanol-induced neurobehavioral alterations during development. Future studies in our laboratory will investigate the brain regions and molecules responsible for these behavioral changes.
Asunto(s)
Modelos Animales de Enfermedad , Etanol/efectos adversos , Trastornos del Espectro Alcohólico Fetal/fisiopatología , Efectos Tardíos de la Exposición Prenatal/fisiopatología , Modelos de Riesgos Proporcionales , Factores de Edad , Consumo de Bebidas Alcohólicas , Animales , Animales Recién Nacidos , Peso Corporal , Etanol/sangre , Conducta Exploratoria/fisiología , Femenino , Trastornos del Espectro Alcohólico Fetal/sangre , Estimación de Kaplan-Meier , Tamaño de la Camada/fisiología , Masculino , Ratones , Embarazo , Reflejo/fisiologíaRESUMEN
Projection of cancer incidence is essential for planning cancer control actions, health care and allocation of resources. Here we project the cancer burden at the National and State level to understand the magnitude of cancer problem for the various calendar years from 2011 to 2026 at 5-yearly intervals. The age, sex and site-wise cancer incidence data along with populations covered by the registries were obtained from the report of National Cancer Registry Programme published by Indian Council of Medical Research for the period 2001-2004. Pooled age sex specific cancer incidence rates were obtained by taking weighted averages of these seventeen registries with respective registry populations as weights. The pooled incidence rates were assumed to represent the country's incidence rates. Populations of the country according to age and sex exposed to the risk of development of cancer in different calendar years were obtained from the report of Registrar General of India providing population projections for the country for the years from 2001 to 2026. Population forecasts were combined with the pooled incidence rates to estimate the projected number of cancer cases by age, sex and site of cancer at various 5-yearly periods Viz. 2011, 2016, 2021 and 2026. The projections were carried out for the various leading sites as well as for 'all sites' of cancer. In India, in 2011, nearly 1,193,000 new cancer cases were estimated; a higher load among females (603,500) than males (589,800) was noted. It is estimated that the total number of new cases in males will increased from 0.589 million in 2011 to 0.934 million by the year 2026. In females the new cases of cancer increased from 0.603 to 0.935 million. Three top most occurring cancers namely those of tobacco related cancers in both sexes, breast and cervical cancers in women account for over 50 to 60 percent of all cancers. When adjustments for increasing tobacco habits and increasing trends in many cancers are made, the estimates may further increase. The leading sites of cancers in males are lung, oesophagus, larynx, mouth, tongue and in females breast and cervix uteri. The main factors contributing to high burden of cancer over the years are increase in the population size as well as increase in proportion of elderly population, urbanization, and globalization. The cancer incidence results show an urgent need for strengthening and augmenting the existing diagnostic/treatment facilities, which are inadequate even to tackle the present load.
Asunto(s)
Neoplasias/epidemiología , Sistema de Registros , Adolescente , Adulto , Anciano , Niño , Preescolar , Femenino , Estudios de Seguimiento , Humanos , Incidencia , India/epidemiología , Lactante , Recién Nacido , Masculino , Persona de Mediana Edad , Pronóstico , Factores de Tiempo , Adulto JovenRESUMEN
Projection of load of cancer mortality helps in quantifying the burden of cancer and is essential for planning cancer control activities. As per our knowledge, there have not been many attempts to project the cancer mortality burden at the country level in India mainly due to lack of data on cancer mortality at the national and state level. This is an attempt to understand the magnitude of cancer mortality problem for the various calendar years from 2011 to 2026 at 5-yearly intervals. Age, sex and site-wise specific cancer mortality data along with populations covered by the registries were obtained from the report of National Cancer Registry Programme published by Indian Council of Medical Research for the period 2001-2004. Pooled age sex specific cancer mortality rates were obtained by taking weighted average of these six registries with respective registry populations as weights. The pooled mortality rates were assumed to represent the country's mortality rates. Populations of the country according to age and sex exposed to the risk of cancer mortality in different calendar years were obtained from the report of Registrar General of India providing population projections for the country for the years from 2011 to 2026. Population forecasts were combined with the pooled mortality rates to estimate the projected number of cancer mortality cases by age, sex and site of cancer at various 5-yearly periods Viz. 2011, 2016, 2021 and 2026. The projections were carried out for the various cancer-leading sites as well as for 'all sites' of cancer. The results revealed that an estimated 0.44 million died due to cancer during the year 2011, while 0.51 million and 0.60 million persons are likely to die from cancer in 2016 and 2021. In the year 2011 male mortality was estimated to be 0.23 million and female mortality to be 0.20 million. The estimated cancer mortality would increase to 0.70 million by the year 2026 as a result of change in size and composition of population. In males increase will be to 0.38 millions and in females to 0.32 millions. Among women, cancer of the breast, cervical and ovary account for 34 percent of all cancer deaths. The leading sites of cancer mortality in males are lung, oesophagus, prostrate and stomach. The above results show a need for commitment for tackling cancer by reducing risk factors and strengthening the existing screening and treatment facilities.
Asunto(s)
Mortalidad/tendencias , Neoplasias/epidemiología , Neoplasias/mortalidad , Sistema de Registros , Adolescente , Adulto , Factores de Edad , Anciano , Niño , Preescolar , Femenino , Estudios de Seguimiento , Humanos , Incidencia , India/epidemiología , Lactante , Recién Nacido , Masculino , Persona de Mediana Edad , Pronóstico , Factores de Riesgo , Factores Sexuales , Factores de Tiempo , Adulto JovenRESUMEN
A community-based, longitudinal interventional study was conducted in a slum in north-east (NE) Mumbai, using a pretested, semi-structured proforma and a pre- and follow-up interview schedule to assess the male health clinic (MHC) strategy as an approach for the control of sexually transmitted infections (STIs) in males. The focus groups that emerged for behavior change communication (BCC) activities were clients in second and third decades of life, unmarried and married but not staying with wife, clients with no permanent place of occupation, clients with habit of alcohol, and illiterate clients. Postintervention, there was an overall increase in STI knowledge score, alcohol habit score, and high-risk sexual activity score. Less than 50% of the clients received advice on substance abuse and information on consequences of STI/human immunodeficiency virus (HIV). The study concluded that MHC strategy is an effective approach for control of STIs in males and recommended strengthening of the weaker components of the strategy with focus on the important client groups.
Asunto(s)
Instituciones de Atención Ambulatoria/organización & administración , Infecciones por VIH/prevención & control , Enfermedades de Transmisión Sexual/prevención & control , Adolescente , Adulto , Distribución de Chi-Cuadrado , Niño , Demografía , Infecciones por VIH/epidemiología , Humanos , India/epidemiología , Estudios Longitudinales , Masculino , Persona de Mediana Edad , Evaluación de Programas y Proyectos de Salud , Enfermedades de Transmisión Sexual/epidemiología , Estadísticas no Paramétricas , Población UrbanaRESUMEN
Caspases, a family of cysteine proteases, are widely activated in neurons and glia in the injured brain, a response thought to induce apoptosis. However, caspase activation in astrocytes following injury is not strongly associated with apoptosis. The present study investigates the potential role of caspase activation in astrocytes with another characteristic response to neural injury, astrogliosis. Caspase activity and morphological and biochemical indices of astrogliosis and apoptosis were assessed in (i) cultured neonatal rat astrocytes treated with astrogliosis-inducing stimuli (dibutryl cAMP, ß-amyloid peptide), and (ii) cultures of adult rat hippocampal astrocytes generated from control and kainate-lesioned rats. The effects of broad spectrum and specific pharmacological caspase inhibitors were assessed on indicators of astrogliosis, including stellate morphology and expression of glutamine synthetase and fibroblast growth factor-2. Reactive neonatal and adult astrocytes demonstrated an increase in total caspase activity with a corresponding increase in the expression of active caspase-3 in the absence of cell death. Broad spectrum caspase inhibition with zVAD significantly attenuated increases in glutamine synthetase and fibroblast growth factor-2 in the reactive astrocytes. In the reactive neonatal astrocyte cultures, specific inhibition of caspases-3 and -11 also attenuated glutamine synthetase and fibroblast growth factor-2 expression, but did not reverse the morphological reactive phenotype. Astrogliosis is observed in all forms of brain injury and despite extensive study, its molecular triggers remain largely unknown. While previous studies have demonstrated active caspases in astrocytes following acute brain injury, here we present evidence functionally implicating the caspases in astrogliosis.
Asunto(s)
Apoptosis/fisiología , Astrocitos/metabolismo , Caspasas/metabolismo , Gliosis/metabolismo , Hipocampo/metabolismo , Animales , Apoptosis/efectos de los fármacos , Astrocitos/efectos de los fármacos , Astrocitos/patología , Células Cultivadas , Factor 2 de Crecimiento de Fibroblastos/metabolismo , Gliosis/patología , Glutamato-Amoníaco Ligasa/metabolismo , Hipocampo/efectos de los fármacos , Hipocampo/patología , Ácido Kaínico/farmacología , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Neuronas/patología , Ratas , Ratas Sprague-DawleyRESUMEN
BACKGROUND: Social marketing is an important tool in the delivery of healthcare services. For any healthcare programme or project to be successful, community/consumer participation is required. The four principles of social marketing can guide policymakers and healthcare providers to successfully plan and implement health programmes. AIM: To review the existing literature in order to project the benefits of social marketing in healthcare. METHOD: A search of periodical literature by the author involving social marketing and marketing concepts in health was carried out. Items were identified initially through health-oriented indexing services such as Medline, Health STAR and Cinahl, using the identifiers "social marketing" and "marketing in health". An extensive search was also carried out on educational database ERIC. RESULTS: A literature review of various studies on social marketing indicated that the selection of the right product (according to the community need) at the right place, with the right strategy for promotion and at the right price yields good results. However, along with technical sustainability (product, price, promotion and place), financial sustainability, institutional sustainability and market sustainability are conducive factors for the success of social marketing. CONCLUSION: The purpose of this literature review was to ascertain the likely effectiveness of social marketing principles and approaches and behaviour change communication towards health promotion. It is important for all healthcare workers to understand and respond to the public's desires and needs and routinely use consumer research to determine how best to help the public to solve problems and realise aspirations. Social marketing can optimise public health by facilitating relationship-building with consumers and making their lives healthier.
RESUMEN
Toxic organic cations can damage nigrostriatal dopaminergic pathways as seen in most parkinsonian syndromes and in some cases of illicit drug exposure. Here, we show that the organic cation transporter 3 (Oct3) is expressed in nondopaminergic cells adjacent to both the soma and terminals of midbrain dopaminergic neurons. We hypothesized that Oct3 contributes to the dopaminergic damage by bidirectionally regulating the local bioavailability of toxic species. Consistent with this view, Oct3 deletion and pharmacological inhibition hampers the release of the toxic organic cation 1-methyl-4-phenylpyridinium from astrocytes and protects against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced dopaminergic neurodegeneration in mice. Furthermore, Oct3 deletion impairs the removal of the excess extracellular dopamine induced by methamphetamine and enhances striatal dopaminergic terminal damage caused by this psychostimulant. These results may have far-reaching implications for our understanding of the mechanism of cell death in a wide range of neurodegenerative diseases and may open new avenues for neuroprotective intervention.