Research, diagnosis and education in inborn errors of metabolism in Colombia: 20 years' experience from a reference center.

The use of specialized centers has been the main alternative for an appropriate diagnosis, management and follow up of patients affected by inborn errors of metabolism (IEM). These centers facilitate the training of different professionals, as well as the research at basic, translational and clinical levels. Nevertheless, few reports have described the experience of these centers and their local and/or global impact in the study of IEM. In this paper, we describe the experience of a Colombian reference center for the research, diagnosis, training and education on IEM. During the last 20 years, important advances have been achieved in the clinical knowledge of these disorders, as well as in the local availability of several diagnosis tests. Organic acidurias have been the most frequently detected diseases, followed by aminoacidopathies and peroxisomal disorders. Research efforts have been focused in the production of recombinant proteins in microorganisms towards the development of new enzyme replacement therapies, the design of gene therapy vectors and the use of bioinformatics tools for the understanding of IEM. In addition, this center has participated in the education and training of a large number professionals at different levels, which has contributed to increase the knowledge and divulgation of these disorders along the country. Noteworthy, in close collaboration with patient advocacy groups, we have participated in the discussion and construction of initiatives for the inclusion of diagnosis tests and treatments in the health system.

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.

Systems biology study of mucopolysaccharidosis using a human metabolic reconstruction network.

Mucopolysaccharidosis (MPS) is a group of lysosomal storage diseases (LSD), characterized by the deficiency of a lysosomal enzyme responsible for the degradation of glycosaminoglycans (GAG). This deficiency leads to the lysosomal accumulation of partially degraded GAG. Nevertheless, deficiency of a single lysosomal enzyme has been associated with impairment in other cell mechanism, such as apoptosis and redox balance. Although GAG analysis represents the main biomarker for MPS diagnosis, it has several limitations that can lead to a misdiagnosis, whereby the identification of new biomarkers represents an important issue for MPS. In this study, we used a system biology approach, through the use of a genome-scale human metabolic reconstruction to understand the effect of metabolism alterations in cell homeostasis and to identify potential new biomarkers in MPS. In-silico MPS models were generated by silencing of MPS-related enzymes, and were analyzed through a flux balance and variability analysis. We found that MPS models used approximately 2286 reactions to satisfy the objective function. Impaired reactions were mainly involved in cellular respiration, mitochondrial process, amino acid and lipid metabolism, and ion exchange. Metabolic changes were similar for MPS I and II, and MPS III A to C; while the remaining MPS showed unique metabolic profiles. Eight and thirteen potential high-confidence biomarkers were identified for MPS IVB and VII, respectively, which were associated with the secondary pathologic process of LSD. In vivo evaluation of predicted intermediate confidence biomarkers (ß-hexosaminidase and ß-glucoronidase) for MPS IVA and VI correlated with the in-silico prediction. These results show the potential of a computational human metabolic reconstruction to understand the molecular mechanisms this group of diseases, which can be used to identify new biomarkers for MPS.

Factores de riesgo y protectores del bullying escolar en estudiantes con bajo rendimiento de cinco instituciones educativas de santa marta, colombia / Protection and risk factors related to bullying in students with low performance, in five schools from santa marta, colombia

El presente artículo de investigación tiene como objetivo caracterizar factores del bullying escolar en estudiantes de 10° con bajo rendimiento, de cinco colegios del Núcleo Educativo N° 5 del Distrito de Santa Marta. El tipo de investigación es descriptiva, con diseño transversal y se trabajó con una población de 582 estudiantes y una muestra de 186, a quienes se les aplicó la Escala MESSY (The Matson Evaluation of Social Skills in Youngsters, de Matson, Rotatori y Helsel, 1983), previa validación en población escolar para este estudio. Los datos fueron procesados mediante el SPSS 17.0v. Se halló una concentración de altos porcentajes en el rango medio de los seis factores que mide la Escala: habilidades sociales adecuadas (59 %), asertividad inadecuada (77 %), impulsividad (75 %), sobreconfianza (49 %), celos/soledad (81 %), y Varios (83 %). Se concluyó que no existe bullying en esta población, pero se hallaron dos factores de riesgo: impulsividad (19 %) y soledad (16 %); y, un factor protector (habilidades sociales adecuadas, 31 %). Se recomienda aplicar programas de prevención centrados en estos tres últimos factores.

To characterize factors related to bullying in tenth grade students with low performance, taken as reference 5 from schools district No. 5 in Santa Marta. A cross-sectional study is conducted; 582 students and 186 students sampled, to whom a MESSY (Matson Evaluation of Social Skills in Youngsters de Matson Rotatori y Helsel 1983). Scale was applied, previous validity in school population for this research. Data was processed using SPSS 17.0v a highest percentages of 6 factors measured by Adequate Social Skills Scale in average level (59 %), inadequate assertiveness (77 %), impulsivity (75 %), overconfidence (49 %), jealousy/loneliness and others (83 %): As a conclusion, there is no bullying in this population, but two risk factors were found: impulsivity (19 %) and loneliness (16 %); and protective factor (adequate social skills 31 %). It is suggested to apply prevention programs focused on the last three factors.