Production of human recombinant phenylalanine hydroxylase in Lactobacillus plantarum for gastrointestinal delivery.

Phenylketonuria (PKU) is an autosomal recessive disorder caused by a defective phenylalanine hydroxylase (PAH), which catalyzes the hydroxylation of l-phenylalanine (l-Phe) to l-tyrosine (l-Tyr) in presence of the cofactor tetrahydrobiopterin (BH4). Defective PAH causes accumulation of phenylalanine, which has neurotoxic effects and leads to dermatological, behavioral, and neurocognitive problems. Treatments for this disease consist in life-long diets that are hard for patients to keep, or supplementation with BH4. In this study, we propose a system where a probiotic lactic acid bacteria (LAB) can be used as vehicle to express in situ an engineered human PAH. Engineered PAHs contain a secretion peptide, a gastrointestinal signal (GI), the human PAH, and a flexible glycine linker followed by the fluorescence protein mEGFP. Engineered constructs were successfully transformed, expressed, and secreted in Lactobacillus plantarum CM_PUJ411. PAH construct containing either the signal peptide GI1 or GI2 were transported through a Caco-2 cell monolayer. Nevertheless, the one containing GI1 allowed the highest transport through the cell monolayer. Co-culture of L. plantarum and Caco-2 cells showed that engineered PAH is produced in-situ and transported through the cell monolayer. Finally, the activity test showed that the engineered PAH secreted by L. plantarum CM_PUJ411 is active, leading to a reduction in l-Phe and an increase in l-Tyr levels, respectively. These results show the potential of this system as a new therapeutic alternative for the treatment of PKU patients.

Diseño de dispositivo para detección de alteraciones del sistema vestibular / Design device for tamper detection of the vestibular system / Design de dispositivo para detecção de distúrbios do sistema vestibular

Los desórdenes vestibulares como el vértigo con mayor prevalencia en la población afectada pueden diagnosticarse con diferentes herramientas incluido el test de agudeza visual dinámica. Test que a nivel clínico únicamente incluye la monitorización de velocidad de movimiento de rotación de la cabeza sin mayores herramientas que permitan el diagnóstico puntual del canal semicircular horizontal acotado en la dinámica completa del movimiento. En el presente artículo se presenta una herramienta tecnológica completa para el desarrollo del test que permite el sensado de los movimientos de la cabeza en los ejes x, y, z y la interacción con una interfaz gráfica para la evaluación de la agudeza visual con errores encontrados al final de la implementación inferiores al 2% en ángulos de inclinación y flexión y del 4% al 15% para el ángulo máximo de rotación.

Vestibular disorders as the vertigo, with higher prevalence in the affected population, can be diagnosed using different tools including the dynamic vestibular acuity test. Test including at clinical level only the speed motorization of head without more tools to allow the specific diagnose of the horizontal semicircular canal bounded in the complete dynamic of movement. This article present a complete technologic tool to the development of the test, allowing the sensing of head movements in the x, y, z axis and the interaction with a graphical interface to evaluate the visual acuity, with found differences at the end of the implementation lower of 2% in flexion and inclination angles and of 4% to 15% to the maximum rotation angle.

Distúrbios vestibulares como a vertigem, com maior prevalência na população afetada, pode ser diagnosticada através de diferentes ferramentas, incluindo o teste de acuidade vestibular dinâmico. Incluindo o teste a nível clínico somente a motorização velocidade da cabeça sem mais ferramentas para permitir o diagnóstico específico do canal semicircular horizontal delimitada a dinâmica de movimentos. Este artigo apresenta uma ferramenta tecnológica completa para o desenvolvimento do teste, Permitindo que o sensor de movimentos da cabeça nos x, y, eixo z ea interação com uma interface gráfica para avaliar a acuidade visual, com diferenças encontradas no final da Implementação inferior de 2% em ângulos de flexão e de inclinação e de 4% a 15% para o ângulo de rotação máxima.