In Vitro Infectivity Assessment by Drug Susceptibility Comparison of Recombinant Leishmania major Expressing Enhanced Green Fluorescent Protein or EGFP-Luciferase Fused Genes with Wild-Type Parasite

Leishmaniasis is a worldwide uncontrolled parasitic disease due to the lack of effective drug and vaccine. To speed up effective drug development, we need powerful methods to rapidly assess drug effectiveness against the intracellular form of Leishmania in high throughput assays. Reporter gene technology has proven to be an excellent tool for drug screening in vitro. The effects of reporter proteins on parasite infectivity should be identified both in vitro and in vivo. In this research, we initially compared the infectivity rate of recombinant Leishmania major expressing stably enhanced green fluorescent protein (EGFP) alone or EGFP-luciferase (EGFP-LUC) with the wild-type strain. Next, we evaluated the sensitivity of these parasites to amphotericin B (AmB) as a standard drug in 2 parasitic phases, promastigote and amastigote. This comparison was made by MTT and nitric oxide (NO) assay and by quantifying the specific signals derived from reporter genes like EGFP intensity and luciferase activity. To study the amastigote form, both B10R and THP-1 macrophage cell lines were infected in the stationary phase and were exposed to AmB at different time points. Our results clearly revealed that the 3 parasite lines had similar in vitro infectivity rates with comparable parasite-induced levels of NO following interferon-gamma/lipopolysaccharide induction. Based on our results we proposed the more reporter gene, the faster and more sensitive evaluation of the drug efficiency.

In vitro assessment of commercial sunscreens available in Latin America / Valoración in vitro de protectores solares comerciales existentes en Latinoamérica

In Latin America, people have largely abandoned the practice of wearing hats and traditional clothing that provided skin protection. Sunscreen application has therefore become essential to protect against the increased sun exposure. The physician-prescribed medical-grade sunscreens provide sufficient sun protection but the requirement for regular use puts a financial burden on the patient that is often not sustainable. An appropriate sunscreen should provide a high and broad ultraviolet (UV) protection against UVB and UVA. Several over-the-counter (OTC) sunscreens have been developed for sale at affordable prices and are available for purchase in convenient locations, such as local grocery stores. The aim of this study was to assess the in vitro UV protection of 34 popular OTC sunscreens found in the Latin American market. UV absorbance/transmittance was quantified by diffusion transmission spectroscopy using coarse silica plaques. Photostability was tested by irradiating them with simulated solar light and calculating the sun protection factor (SPF), critical length of absorption (C λ ), UVA/UVB ratio, and the spectral uniformity index (SUI). The results indicated that the in vitro SPFs were significantly lower than the value declared on the labels, particularly for those claiming high SPF values; however, the majority of these sunscreens offered high levels of UV protection. Considering the advantages of low cost and ample accessibility, we concluded that this sample of OTC sunscreens can be beneficial to the general public by providing some level of skin protection from solar radiation, and may be promoted to improve compliance with recommended photoprotection behavior.

En Latinoamérica, la población ha abandonado la costumbre de usar sombrero y ropa tradicional para protegerse del sol. En consecuencia, es básico el uso de protectores solares si se realizan actividades bajo sol. Los protectores solares que se usan en la práctica médica son adecuados, pero su uso frecuente condiciona una carga económica que muchos pacientes no pueden solventar debido a sus costos considerables. Un protector apropiado contiene una amplia y elevada protección ultravioleta (UV) A y B. En las tiendas de conveniencia, existen numerosos protectores solares a precios más accesibles. El objetivo del estudio fue determinar la protección UV in vitro de 34 protectores solares con amplia presencia comercial (de venta sin prescripción médica) en el mercado latinoamericano. La absorbancia/transmitancia de la radiación UV se cuantificó mediante espectroscopía de transmisión difusa. Placas de sílice esmerilado fueron recubiertas con el producto y expuestas a radiación solar simulada para conocer su fotoestabilidad. Se calcularon índices como el factor de protección solar (SPF), longitud crítica de absorción (C λ), relación UVA/UVB y el índice de uniformidad espectral (SUI). Se encontró que el SPF in vitro fue inferior al establecido en las etiquetas, especialmente en aquellos con valores altos. No obstante, la mayoría de los protectores incluidos ofrecen niveles de protección UV elevados. Considerando su amplia accesibilidad y menor costo, concluimos que esta muestra comercial de protectores solares podría utilizarse en el entorno clínico para favorecer su apego junto a las otras medidas de fotoprotección sugeridas.