Long interspersed nuclear elements (LINE-1 or L1) comprise 17% of the human genome, although only 80–100 L1s are considered retrotransposition-competent (RC-L1). Despite their small number, RC-L1s are still potential hazards to genome integrity through insertional mutagenesis, unequal recombination and chromosome rearrangements. In this study, we provide several lines of evidence that the LINE-1 retrotransposon is susceptible to RNA interference (RNAi). First, double-stranded RNA (dsRNA) generated in vitro from an L1 template is converted into functional short interfering RNA (siRNA) by DICER, the RNase III enzyme that initiates RNAi in human cells. Second, pooled siRNA from in vitro cleavage of L1 dsRNA, as well as synthetic L1 siRNA, targeting the 5′-UTR leads to sequence-specific mRNA degradation of an L1 fusion transcript. Finally, both synthetic and pooled siRNA suppressed retrotransposition from a highly active RC-L1 clone in cell culture assay. Our report is the first to demonstrate that a human transposable element is subjected to RNAi.
There are significant statistical differences between the measurements of free testosterone by direct and mathematical methods. Although certain correlation is observed, this is very low. In conclusion, free testosterone measurement by enzymoimmunoassay is not reliable.
Starting from 50 years of age, a significant age-related decrease in total testosterone, free testosterone, dehydroepiandrosterone sulfate and bioavailable estradiol is observed. The frequency of biochemical hypogonadism is higher when free testosterone levels are used for diagnosis. Total testosterone levels were related to age, diabetes mellitus and obesity. Free testosterone was related to age. diabetes mellitus, obesity and hyperlipemia. The probability of suffering low free testosterone levels increases with age, diabetes mellitus and/or obesity.
RESUMENEVALUACIÓN DE UNA TÉCNICA DE INMUNOANÁLISIS PARA LA DETERMINACIÓN DE TESTOSTERONA LIBRE. Introducción y objetivos: Los mejores indicadores bioquímicos para el diagnóstico del hipogonadismo son la testosterona libre y la biodisponible. La determinación de ambas fracciones de testosterona es compleja, aunque se han desarrollado técnicas de inmunoanálisis que permiten la determinación de la testosterona libre de manera directa y sencilla. Valoramos la utilidad de una técnica no evaluada de enzimoinmunoanálisis para la determinación de testosterona libre.Material y métodos: En el estudio se incluyeron 133 varones sanos en los que se determinó la testosterona total, la SHBG, la albúmina y la testosterona libre. La testosterona libre fue determinada mediante 2 métodos diferentes: determinación directa mediante enzimoinmunoanálisis (DSL 10-49100 ACTIVE ® Free Testosterone EIA, Diagnostics System Laboratories) y mediante cálculo matemático utilizando la fórmula desarrollada por Vermeulen, que utiliza la concentración de albúmina, testosterona total y SHBG para calcular la testosterona libre (método recomendado por la "International Society for the Study of the Aging Male"). Comparamos los valores obtenidos mediante un test de comparación de medias y realizamos un análisis de regresión para observar el grado de correlación existente entre ambos tipos de determinaciones.Resultados: La edad media fue de 37 ± 11 años. Los niveles medios de testosterona total fue de 21,43 ± 6,8 nmol/L y los de testosterona libre de 0,0508 ± 0,0118 nmol/L para el método directo y de 0,474 ± 0,123 nmol/L para el méto-do matemático (p<0,05). En el estudio de regresión lineal se observa que aunque existe una correlación positiva entre ambos métodos (p<0,05) el coeficiente de correlación es bajo (r = 0,25).Conclusiones: Existen diferencias significativas entre los valores de testosterona libre determinados por el método directo de enzimoinmunoanálisis y la testosterona libre calculada. Aunque se observa una cierta correlación, esta es tan baja que hace poco fiable la utilización del enzimoinmunoanálisis para la determinación de la testosterona libre.Palabras clave: Andropausia. Andrógenos. Testosterona. Inmunoanálisis.
ABSTRACT EVALUATION OF AN IMMUNOASSAY KIT TO MEASURE FREE TESTOSTERONE Introduction and objetives:The best indicators to the diagnosis of hypogonadism are free and bioavailable testosterone circulating levels. Free and bioavailable testosterone measurements are complex. However, simple kits for direct measurement of free testosterone by analog immunoassay are available. We examined the utility of an enzymoimmunoassay kit for free testosterone measurement.Material and method: One hundred thirty-three healthy males were included. Total testosterone, SHBG, albumin and free testosterone was measured. We used two different methods to free testosterone estimation: direct measurement by enzymoimmunoassay and mathematical calculation with Vermeulen's formula, which uses albumin concentration, total testosterone and SHBG to calculate...
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