Three-Dimensional paladar shutter fisurated in pediatric patient / Obturador tridimensional de paladar fisurado en paciente pediátrico

ABSTRACT: Palatal fissure is one of the congenital craniofacial malformations with the highest incidence worldwide, which compromises both the aesthetic and functional part, thus altering the vital functions: chewing, phonation, hearing and swallowing. The literature indicates that this anomaly occurs in 25% individually and 50% when associated with the cleft lip. One of the characteristics of these patients is the presence of notable malnutrition, caused by difficulty feeding, because they cannot exert enough pressure to suck breast or bottle milk; In addition, oral-nasal communication will lead to food entering the nostrils, causing regurgitation until possible bronchial aspiration. Given this problem, it is necessary to use an acrylic device capable of momentarily sealing the palatal fissure, in order to prevent the diversion of food and restore the sucking reflex in the infant. The objective of this study is to determine the effectiveness of the Three-dimensional Cleft Palate Shutter in a pediatric patient who attended the National Foundation of Maxillofacial Rehabilitation (FUNARMAF) in June 2019. The methodological design corresponds to an analytical - synthetic study since it was based on the analysis of a single case of a 5-month-old female patient, with a Unilateral Palatal Fissure (right), giving her the treatment. Results show that a closure of the oral-nasal communication was obtained, re-establishing the suction reflex. In all, it was observed that the use of this device is effective in patients with lip-palatal fissure since it acts as an adjuvant in breastfeeding and feeding the baby.

RESUMEN: La Fisura Palatina es una de las malformaciones congénitas craneofacial con mayor incidencia a nivel mundial, que compromete tanto la parte estética como funcional, alterando así las funciones vitales: masticación, fonación, audición y deglución. La literatura nos indica que esta anomalía se presenta en un 25 % de forma individual y en un 50 % cuando se asocia al labio leporino. Una de las características de estos pacientes es la presencia de una notable desnutrición, ocasionado por la dificultad para alimentarse, debido a que no pueden ejercer la suficiente presión para succionar la leche materna o del biberón; además la comunicación buco-nasal dará lugar a que los alimentos ingresen a las fosas nasales, ocasionando regurgitaciones hasta provocar una posible bronco-aspiración. Ante esta problemática es necesario el uso de un dispositivo de acrílico capaz de sellar momentáneamente la fisura palatina, con el fin de impedir el desvió de los alimentos y reestablecer el reflejo de succión en el lactante. Objetivo: Determinar la eficacia del Obturador Tridimensional de Paladar Fisurado en un paciente pediátrico que asistió a la Fundación Nacional de Rehabilitación Maxilo-Facial (FUNARMAF) en junio del 2019. Metodología: Analítico - Sintético, ya que se basó en el análisis de un solo caso de una paciente de sexo femenino de 5 meses, con un Fisura Labio Palatina Unilateral (derecha), brindándole el tratamiento. Resultados: Se obtuvo un cierre de la comunicación buco-nasal, reestableciendo el reflejo de succión. Conclusión: se observó que es eficaz el uso de este dispositivo en pacientes con Fisura Labio Palatina ya que actúa como coadyuvante en la lactancia materna y alimentación del bebé.

Recombinant human N-acetylgalactosamine-6-sulfate sulfatase (GALNS) produced in the methylotrophic yeast Pichia pastoris.

Mucopolysaccharidosis IV A (MPS IV A, Morquio A disease) is a lysosomal storage disease (LSD) produced by mutations on N-acetylgalactosamine-6-sulfate sulfatase (GALNS). Recently an enzyme replacement therapy (ERT) for this disease was approved using a recombinant enzyme produced in CHO cells. Previously, we reported the production of an active GALNS enzyme in Escherichia coli that showed similar stability properties to that of a recombinant mammalian enzyme though it was not taken-up by culture cells. In this study, we showed the production of the human recombinant GALNS in the methylotrophic yeast Pichia pastoris GS115 (prGALNS). We observed that removal of native signal peptide and co-expression with human formylglycine-generating enzyme (SUMF1) allowed an improvement of 4.5-fold in the specific GALNS activity. prGALNS enzyme showed a high stability at 4 °C, while the activity was markedly reduced at 37 and 45 °C. It was noteworthy that prGALNS was taken-up by HEK293 cells and human skin fibroblasts in a dose-dependent manner through a process potentially mediated by an endocytic pathway, without any additional protein or host modification. The results show the potential of P. pastoris in the production of a human recombinant GALNS for the development of an ERT for Morquio A.

Human recombinant lysosomal enzymes produced in microorganisms.

Lysosomal storage diseases (LSDs) are caused by accumulation of partially degraded substrates within the lysosome, as a result of a function loss of a lysosomal protein. Recombinant lysosomal proteins are usually produced in mammalian cells, based on their capacity to carry out post-translational modifications similar to those observed in human native proteins. However, during the last years, a growing number of studies have shown the possibility to produce active forms of lysosomal proteins in other expression systems, such as plants and microorganisms. In this paper, we review the production and characterization of human lysosomal proteins, deficient in several LSDs, which have been produced in microorganisms. For this purpose, Escherichia coli, Saccharomyces cerevisiae, Pichia pastoris, Yarrowia lipolytica, and Ogataea minuta have been used as expression systems. The recombinant lysosomal proteins expressed in these hosts have shown similar substrate specificities, and temperature and pH stability profiles to those produced in mammalian cells. In addition, pre-clinical results have shown that recombinant lysosomal enzymes produced in microorganisms can be taken-up by cells and reduce the substrate accumulated within the lysosome. Recently, metabolic engineering in yeasts has allowed the production of lysosomal enzymes with tailored N-glycosylations, while progresses in E. coli N-glycosylations offer a potential platform to improve the production of these recombinant lysosomal enzymes. In summary, microorganisms represent convenient platform for the production of recombinant lysosomal proteins for biochemical and physicochemical characterization, as well as for the development of ERT for LSD.