Succinate dehydrogenase gene as a marker for studying Blastocystis genetic diversity.

Blastocystis has been reported as the most common eukaryotic microorganism residing in the intestines of both humans and animals, with a prevalence of up to 100% in some populations. Since this is a cryptic species, sequence polymorphism are the single strategy to analyses its genetic diversity, being traditionally used the analysis of ssu rRNA gene sequence to determine alleles and subtypes (STs) for this species. This multicopy gene has shown high diversity among different STs, making necessary to explore other genes to assess intraspecific diversity. This study evaluated the use of a novel genetic marker, succinate dehydrogenase (SDHA), for the typing and evaluation of the genetic diversity and genetic population structure of Blastocystis. In total, 375 human fecal samples were collected and subjected to PCR, subtyped using the ssu rRNA marker, and then the SDHA gene was amplified via PCR for 117 samples. We found some incongruences between tree topologies for both molecular markers. However, the clustering by ST previously established for Blastocystis was congruent in the concatenated sequence. SDHA showed lower reticulation (The origination of a lineage through the partial merging of two ancestor lineages) signals and better intra ST clustering ability. Clusters with geographical associations were observed intra ST. The genetic diversity was lower in the marker evaluated compared to that of the ssu rRNA gene (nucleotide diversity = 0.03344 and 0.16986, respectively) and the sequences analyzed showed population expansion with genetic differentiation principally among STs. The ssu rRNA gene was useful to explore interspecific diversity but together with the SDHA gene the resolution power to evaluate intra ST diversity was higher. These results showed the potential of the SDHA marker for studying the intra ST genetic diversity of Blastocystis related with geographical location and the inter ST diversity using the concatenated sequences.

Development of a Multilocus Sequence Typing Scheme for Giardia intestinalis.

Giardia intestinalis is an intestinal protozoan most commonly found in humans. It has been grouped into 8 assemblages (A-H). Markers such as the glutamate dehydrogenase gene, triose phosphate isomerase and beta-giardin (ß-giardin) have been widely used for genotyping. In addition, different genetic targets have been proposed as a valuable alternative to assess diversity and genetics of this microorganism. Thus, our objective was to evaluate new markers for the study of the diversity and intra-taxa genetic structure of G. intestinalis in silico and in DNA obtained from stool samples. We analysed nine constitutive genes in 80 complete genome sequences and in a group of 24 stool samples from Colombia. Allelic diversity was evaluated by locus and for the concatenated sequence of nine loci that could discriminate up to 53 alleles. Phylogenetic reconstructions allowed us to identify AI, AII and B assemblages. We found evidence of intra- and inter-assemblage recombination events. Population structure analysis showed genetic differentiation among the assemblages analysed.

Molecular detection and genotyping of intestinal protozoa from different biogeographical regions of Colombia.

BACKGROUND: Intestinal parasitic protozoa represent a serious problem of public health particularly in developing countries. Protozoa such as Blastocystis, Giardia intestinalis, Entamoeba histolytica and Cryptosporidium spp. are associated with diarrheal symptoms. In Colombia, there is little region-specific data on the frequency and circulating genotypes/species of these microorganisms. Therefore, the main objective of our study was to employ molecular detection and genotyping of G. intestinalis and Blastocystis, Cryptosporidium and Entamoeba spp. in samples from different biogeographical regions of Colombia. METHODS: We collected 649 human fecal samples from five biogeographical regions of Colombia: the Amazon, Andean, Caribbean, Orinoco and Pacific regions. Blastocystis, G. intestinalis, Cryptosporidium spp. and Entamoeba complex were detected by microscopy and conventional PCR. Molecular genotyping was conducted to identify Blastocystis subtypes (STs) (18s), G. intestinalis assemblages (triose phosphate isomerase and glutamate dehydrogenase) and Cryptosporidium species (18s). Genetic diversity indices were determined using dnasp.5. RESULTS: We detected G. intestinalis in 45.4% (n = 280) of samples, Blastocystis in 54.5% (n = 336) of samples, Cryptosporidium spp. in 7.3% (n = 45) of samples, Entamoeba dispar in 1.5% (n = 9) of samples, and Entamoeba moshkovskii in 0.32% (n = 2) of samples. Blastocystis STs 1-4, 8 and 9 and G. intestinalis assemblages AII, BIII, BIV, D and G were identified. The following Cryptosporidium species were identified: C. hominis, C. parvum, C. bovis, C. andersoni, C. muris, C. ubiquitum and C. felis. The Caribbean region had the highest frequency for each of the microorganisms evaluated (91.9% for G. duodenalis, 97.3% for Blastocystis, 10.8% for Cryptosporidium spp., 13.5% for E. dispar and 2.7% for E. moshkovskii). The Orinoco region had a high frequency of Blastocystis (97.2%) and the Andean region had a high frequency of G. intestinalis (69.4%). High and active transmission was apparent in several regions of the country, implying that mechanisms for prevention and control of intestinal parasitosis in different parts of the country must be improved.

Ahorro del sistema de salud por la regulación directa de precios de venta de medicamentos biotecnológicos en Colombia / Savings in the Health System due to the Direct Regulation of the Sale Prices of Biotechnological Medicines in Colombia / Poupança no sistema de saúde pela regulamentação direta de preços de venda de medicamentos biotecnológicos na Colômbia

Resumen Se desarrolló un análisis retrospectivo sobre las ventas de trece biotecnológicos regulados, entre junio del 2010 y junio del 2014, en dos escenarios: valores facturados después de la regulación de precios y valores supuestos, calculados a partir del precio de venta promedio de los seis meses anteriores a la regulación, más el índice de precios al consumidor en salud. La diferencia entre ambos escenarios supone un ahorro al sistema. Los biotecnológicos analizados generaron un ahorro de USD $95 000 000 en dicho periodo; en el primer año el 10.2%, el segundo año el 17.2% y en el tercero el 30.5%. El biotecnológico que más ahorro representó fue el rituximab (40,9%), equivalente a USD $38 510 787. La regulación directa de precios de medicamentos biotecnológicos le ha generado ahorros significativos al sistema de salud, visto desde las ventas registradas en Audifarma. Probablemente, el ahorro económico que ha generado esta política es superior a lo estimado inicialmente por el Gobierno.

Abstract We developed a retrospective analysis on the sales of thirteen biotechnological regulated medicines between June, 2010, and June, 2014, in two scenarios: amounts billed after the price and assumed value regulation, calculated based on the average sale price during the six months prior to the regulation, plus the health consumer price index. The difference between both scenarios entails a saving for the system. The biotechnological medicines analyzed generated savings for US $95 Million in said period; 10.2% during the first year, 17.2% during the second, and 30.5% during the third. The biotechnological medicine that generated the most savings was rituximab (40.9%), equivalent to US $38,510,787. The direct regulation of biotechnological medicines has generated significant savings for the healthcare system, as seen from the sales registered by Audifarma. Probably, the economic savings generated by this policy are above the initial government estimates.

Resumo Análise retrospectiva sobre as vendas de treze biotecnológicos regulados entre junho de 2010 e junho de 2014 desenvolvida em dois cenários: valores faturados após regulamentação de preços e valores supostos, calculados a partir do preço de venda médio dos seis meses prévios à regulamentação, mais o índice de preços ao consumidor em saúde. A diferença entre ambos cenários supõe poupança para o sistema. Os biotecnológicos analisados geraram poupança de USD $95 000 000 nesse período; no primeiro ano o 10.2%, o segundo ano o 17.2% e no terceiro o 30.5%. O biotecnológico que mais poupança gerou foi o rituximab (40,9%), equivalente a USD $38 510 787. A regulamentação direta de preços de medicamentos biotecnológicos gerou poupança significativa ao sistema de saúde, visto desde as vendas registradas em Audifarma. Provavelmente, a poupança econômica gerada por esta política é superior do estimado inicialmente pelo Governo.

Descripción del estado de salud y calidad de vida en pacientes tratados con etanercept dentro de un programa de atención para artritis reumatoide en un centro especializado en Colombi / Description of health status and quality of life in rheumatoid arthritis patients treated with etanercept in a care program in a Colombian specialist center

Resumen Objetivo: Realizar un análisis descriptivo de la actividad de la enfermedad medida por DAS28, de la funcionalidad medida por HAQ (Health Assessment Questionnaire) y escala de depresión, tras un año de seguimiento en un programa de gerenciamiento de la enfermedad «Evaluación día¼, para pacientes colombianos con artritis reumatoide que iniciaron tratamiento con etanercept (Enbrel®). Materiales y métodos: Se diseñó un programa como parte del manejo integral de la enfermedad que comprendía cuatro visitas e incluía paraclínicos, evaluación por reumatólogo, psicólogo, terapeuta físico y ocupacional, y clinimetría. Resultados: Se incluyeron 91 sujetos. Al ingreso del programa, 42 pacientes (80.7%) fueron clasificados con actividad moderada y grave. Al año, la remisión fue alcanzada en 25%. El número de pacientes que presentaron una reducción mayor de 1.2 en el DAS28, respecto al inicio del tratamiento, fue de 19 (37.5%). La medida promedio de HAQ al inicio fue 1,07 y mejoró al disminuir a 0,96 en el último control, aunque en 11 de los individuos (13,5%) no se realizó la medición. Al ingreso del programa, 33.3% de los pacientes, tenía algún grado de depresión mayor de acuerdo con la escala de Zung y al año de la observación sólo la presentaba el 21.9%. Conclusiones: La descripción de estos programas de gerenciamiento es importante para contar con data propia de los pacientes colombianos con artritis reumatoide y brindar el manejo integral que se requiere en las enfermedades crónicas. Los resultados obtenidos parecen corresponder a los descritos en otras observaciones.

Abstract Objective: We conducted a descriptive analysis of rheumatoid arthritis activity according to DAS28, and functionality with HAQ (Health Assessment Questionnaire), and depression scales after one year of follow-up, in a disease management program «Evaluación día¼ for Colombian patients whom started treatment with etanercept. Methods: We designed a program as part of an integral disease management. It involved four visits, including: lab tests, questionnaires, and clinical evaluation by rheumatologist, psychologist, physical and occupational therapist. Results: 91 patients were included. At the beginning of the program, 80.7% of the individuals were classified in moderate and severe disease activity. At the end of follow-up, disease remission by DAS28 was reached in 25% of the patients. The number of patients who experienced DAS28 reduction more than 1.2 points was 19 (37.5%). HAQ average at baseline was 1.07 and improved to 0.96 at the end of the follow-up. At baseline, 33.3% of patients had a mayor depression according to Zung Score and, after one year, this score was present in 21.9% of the patients. Conclusions: The description of this disease management program is important for the understanding of rheumatoid arthritis in Colombia and it may offer an integral treatment, which is necessary in all chronic diseases. The results found in this analysis seem to be similar as described in other observations.