RESUMEN
Low positive predictive values of Neisseria gonorrhoeae nucleic acid amplification tests in low-prevalence populations are a major challenge for accurate diagnostics. It is therefore necessary to verify all positive N. gonorrhoeae results with a different assay and target gene, preferably using the same sample. The BD ProbeTec™ Q(x) Collection Kit for Endocervical or Lesion Specimens, which is recommended for BD Viper™ XTR, is incompatible with other commercial platforms. Therefore, a confirmatory PCR has not been available for samples received on this transport medium. To be able to verify results from these samples with another assay, our objective was to establish a procedure for using the DNA eluates from BD Viper™ XTR for further analysis. DNA eluates from BD Viper™ XTR were collected and analyzed in two in-house confirmatory real-time PCRs targeting the porA pseudogene and the opa multicopy gene. BD Viper™ XTR DNA eluates were analyzed directly and also after purification with the nucleic acid extraction system NucliSENS® easyMag®. Purification of BD Viper™ XTR DNA eluates with the nucleic acid extraction system NucliSENS® easyMag® provided a sensitivity of the in-house PCR comparable to BD Viper™. With the inclusion of two target genes in the confirmatory PCR, specific and reliable verification of results were obtained. This study presents a simple, inexpensive procedure which allows for rapid verification of gonorrhea from samples received on the BD ProbeTec™ Q(x) Collection Kit for Endocervical or Lesion Specimens.
Asunto(s)
ADN Bacteriano/aislamiento & purificación , Gonorrea/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Neisseria gonorrhoeae/aislamiento & purificación , Reacción en Cadena de la Polimerasa/métodos , Manejo de Especímenes/métodos , Proteínas de la Membrana Bacteriana Externa/genética , ADN Bacteriano/genética , Humanos , Neisseria gonorrhoeae/genética , Porinas/genética , Sensibilidad y EspecificidadRESUMEN
The implementation of neural stem cell lines as a source material for brain tissue transplants is currently limited by the ability to induce specific neurochemical phenotypes in these cells. Here, we show that coordinated induction of a ventral mesencephalic dopaminergic phenotype in an immortalized multipotent neural stem cell line can be achieved in vitro. This process requires both the overexpression of the nuclear receptor Nurr1 and factors derived from local type 1 astrocytes. Over 80% of cells obtained by this method demonstrate a phenotype indistinguishable from that of endogenous dopaminergic neurons. Moreover, this procedure yields an unlimited number of cells that can engraft in vivo and that may constitute a useful source material for neuronal replacement in Parkinson's disease.
Asunto(s)
Astrocitos/metabolismo , Proteínas de Unión al ADN , Dopamina/metabolismo , Mesencéfalo/citología , Neuronas/citología , Células Madre/fisiología , Factores de Transcripción/metabolismo , Animales , Astrocitos/citología , Diferenciación Celular , Línea Celular , Cromatografía Líquida de Alta Presión , Técnicas de Cocultivo , Cuerpo Estriado/citología , Mesencéfalo/metabolismo , Ratones , Neuronas/fisiología , Neuronas/trasplante , Miembro 2 del Grupo A de la Subfamilia 4 de Receptores Nucleares , Enfermedad de Parkinson/terapia , Ratas , Factores de Transcripción/genética , Transfección , Transgenes , Tirosina 3-Monooxigenasa/metabolismoAsunto(s)
Lechos , Úlcera por Presión/prevención & control , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana EdadAsunto(s)
Mordeduras y Picaduras , Perros , Traumatismos Faciales/etiología , Adolescente , Adulto , Animales , Niño , Preescolar , Dinamarca , Etnicidad , Femenino , Groenlandia , Humanos , Lactante , Masculino , Infecciones por Pasteurella/prevención & control , Infección de Heridas/prevención & controlRESUMEN
Persephin (PSP) is a neurotrophic factor of the GDNF family that has been found to promote the survival of multiple populations of neurons. In the present study we have examined: (1) the mechanism of action and the function of PSP on nigrostriatal dopamine neurons and (2) the therapeutic potential of PSP, delivered by neural stem cells (NSCs) in a model of Parkinson's disease. Interestingly we found that the prenatal ventral mesencephalon and the newborn striatum express high levels of PSP mRNA. Moreover, midbrain dopamine neurons express its preferred receptor GFRalpha4, allowing a cis type of action of PSP on dopamine neurons. Primary culture studies showed that PSP is as potent and efficacious as GDNF at promoting both survival and neuritogenesis of midbrain dopamine neurons. To study the function and therapeutic potential of PSP in vivo we engineered NSCs to overexpress PSP. PSP-c17.2 cells were found to stably express PSP mRNA and protein for at least 3 months in vivo, to disperse within the striatum, and to give rise to neurons, astrocytes, and a large proportion of oligodendrocytes that integrated within white matter tracts in the striatum. Moreover, PSP-c17.2 cells enhanced dopamine-dependent behavioral parameters in unlesioned mice and prevented the loss of dopamine neurons and the behavioral impairment of mice receiving intrastriatal 6-OHDA injections. Thus, our findings are consistent with a direct action of PSP on developing and adult midbrain dopamine neurons and suggest that the delivery of PSP by NSCs may constitute a very useful strategy in the treatment of Parkinson's disease.