Genetic polymorphisms associated to folate transport as predictors of increased risk for acute lymphoblastic leukemia in Mexican children

Acute lymphoblastic leukemia (ALL) is a frequent neoplasia occurring in children. The most commonly used drug for the treatment of ALL is methotrexate (MTX), an anti-folate agent. Previous studies suggest that folate transporters play a role in ALL prognosis and that genetic polymorphism of genes encoding folate transporters may increase the risk of ALL. Therefore, the main goal of this study was to determine the associations among six genetic polymorphisms in four genes related with the folate transporter pathway to determine a relationship with the occurrence of ALL in Mexican children. A case-control study was performed in 73 ALL children and 133 healthy children from Northern and Northwestern Mexico. COL18A1 (rs2274808), SLC19A1 (rs2838956), ABCB1 (rs1045642 and rs1128503), and ABCC5 (rs9838667 and rs3792585). Polymorphisms were assayed through qPCR. Our results showed an increased ALL risk in children carrying CT genotype (OR = 2.55, CI 95% 1.11-5.83, p = 0.0001) and TT genotype (OR = 21.05, CI 95% 5.62-78.87, p < 0.0001) of COL18A1 rs2274808; in SLC19A1 rs2838956 AG carriers (OR = 44.69, CI 95% 10.42-191.63, p = 0.0001); in ABCB1 rs1045642 TT carriers (OR = 13.76, CI 95% 5.94-31.88, p = 0.0001); in ABCC5 rs9838667 AC carriers (OR = 2.61, CI 95% 1.05-6.48, p < 0.05); and in ABCC5 rs3792585 CC carriers (OR = 9.99, CI 95% 3.19-31.28, p = 0.004). Moreover, several combinations of genetic polymorphisms were found to be significantly associated with a risk for ALL. Finally, two combinations of ABCC5 polymorphisms resulted in protection from this neoplasia. In conclusion, certain genetic polymorphisms related to the folate transport pathway, particularly COL18A1 rs2274808, SLC19A1 rs2838956, ABCB1 rs1045642, and ABCC5 rs3792585, were associated with an increased risk for ALL in Mexican children

Impact of genetic polymorphism of metabolic pathway of methotrexate and survival rate of Mexican children will all [Impacto de polimorfismos genéticos de la vía metabólica del metotrexato sobre la sobrevida de niños Mexicanos con leucemia linfoblástica aguda (LLA)]

Background: Acute lymphoblastic leukemia (ALL) is a major cancer disease in Mexican pediatric population, were the genotype could affect the effectiveness of chemotherapy in which the methotrexate (MTX) is involved and consequently the time of disease free survival and overall survival. Objective: Determine the association of 10 genetic polymorphisms of the folate pathway: in cellular carriers (COL18A1, SLC19A1, ABCB1 and ABCC5) and in enzymes such as folylpolyglutamate synthetase (FPGS) and xanthine oxidase (XO), with survival of children with acute lymphoblastic leukemia. Methods: Thirtynine children with acute lymphoblastic leukemia from the State Cancer Center in Durango (Mexico) treated with MTX and 102 healthy controls, were qPCR analyzed for 10 polymorphisms in the folate pathway. During 5 years of follow up, the disease-free survival and overall survival rates were investigated in relation with their genotypes. Results: Four polymorphisms were not found in Hardy-Weinberg Equilibrium COL18A1 (rs2274808), ABCC5 (rs9838667 and rs3792585) and XO (rs170113685). Only XO (rs170113685) was associated with risk of being present in patients with ALL whose odds ratio was 9.771 (95% 4.974-19.196, p=0.001). The polymorphism rs1544105 for FPGS affected disease free survival and overall survival (Log Rank test p<0.05). Conclusion: Polymorphism (rs17011368) of XO showed risk association for acute lymphoblastic leukemia; likewise, an important association was found between carriers of the FPGS (rs1544105) and increased survival times of patients treated with methotrexate

Asociación de los Polimorfismos Genéticos de Folilpoliglutamil sintetasa (FPGS), RFC1, ABCB1, ABCC5 y Xantina Oxidasa (XO) con la Farmacocinética de Metotrexate (MTX) y la Respuesta al Tratamiento en Pacientes Pediátricos con LLA

Antecedentes: La leucemia linfoblástica aguda (LLA) es uno de los padecimientos oncológicos más frecuentes en pediatría. Para su tratamiento farmacológico se emplean varias fármacos como el metotrexate (MTX), cuya farmacocinética y farmacodinamia (efectividad y toxicidad) está asociado con el genotipo de enzimas y transportadores de la vía del folato. El objetivo de esta investigación fue determinar la asociación de los polimorfismos genéticos (SNPs) de folilpoliglutamil sintetasa (FPGS), COL18A1, RFC1 (SLC19A1), ABCB1, ABCC5 y xantina oxidasa (XO) con la farmacocinética de MTX, la susceptibilidad a la enfermedad y la respuesta adversa al MTX en niños con LLA. Métodos: se evaluaron 73 niños con LLA, en quienes se identificó el genotipo de 10 SNPs por qPCR. En un grupo de pacientes en fase de mantenimiento se determinó el MTX plasmático por HPLC, luego de una dosis i.m. de 40mg/m2. La farmacocinética poblacional (PopPk) se realizó por modelado de efectos mixtos. Finalmente, se evaluó la respuesta al tratamiento con el estudio de farmacovigilancia y del análisis de supervivencia. Resultados: se encontró una asociación entre COL18A1-rs2274808(OR=2.55, IC95%, 1.11-5.83, p=0.0001); SLC19A1-rs2838956(OR=44.69, IC95%, 10.42-191.63, p=0.0001); ABCB1-rs1045642(OR=13.76, IC95%,5.94-31.88, p=0.0001); ABCC5- rs9838667 (OR=2.61, IC95% 1.05-6.48, p=0.005) y ABCC5-rs3792585 (OR=9.99, IC 95%, 3.19-31.28 p=0.001) con la ocurrencia de LLA. En cuanto a la PopPk, ésta respondió a un modelo bicompartimental (CL/F=6.09L/h y Vc/F=6.78L), con una variabilidad interindividual de 40.7% y 31.5% respectivamente y residual de 31.2%. La covariable FPGS-rs4451422 explicó el modelo final, incrementando el CL/F. Los SNPs ABCB1-rs1128503 (OR 0.19, IC95% 0.03-0.9, p<0.05) y ABCC5-rs3792585(OR 0.12, IC95%, 0.027-0.58) se manifestaron protectores para mielosupresión. Finalmente los portadores del FPGS-rs1544105 aumentaron el tiempo de sobrevida libre de enfermedad a 16.67 meses vs a los no portadores.Conclusión. Los portadores de los SNPs (COL18A1-rs2274808, SLC19A1-rs2838956, ABCB1-rs1045642 y ABCC5-rs3792585) mostraron una susceptibilidad mayor para la ocurrencia de LLA. El modelo farmacocinético poblacional obtenido de MTX a dosis bajas para los pacientes pediátricos con LLA fue bicompartimental y el efecto de los FPGS- rs1544105 y rs4451422 no influyó en el modelo final. Sin embargo, el peso fue la única covariable que estadísticamente si entró en el modelo final. El ABCB1- rs1128503 y ABCC5-rs3792585 se asociaron como factores de protección frente a mielosupresión. Finalmente el FPGS-rs1544105 se asoció con un incremento de la supervivencia

Association of ABCB1, ABCC5 and xanthine oxidase genetic polymorphisms with methotrexate adverse reactions in Mexican pediatric patients with ALL

Background: Acute lymphoblastic leukemia (ALL) is one of the most frequent oncological disorders in pediatric populations. To date, the drug of choice for the treatment of ALL is methotrexate, a drug associated with a high risk of adverse reactions (ADRs). The xanthine oxidase (XO) polymorphisms, 1936A>G and 2107A>G, as well as the polymorphic variants derived from ATP-binding cassette transporter gene subfamilies, ABCB1 and ABCC5, of drug resistant codifying genes, are implicated as precursors of drug-related neurologic, hepatic, and renal toxicities. Our aim was to determine whether the mentioned polymorphisms are risk or protective factors for the development of adverse reactions by methotrexate in our pediatric population with ALL. Methods: A total of 35 Mexican children from Centro Estatal de Cancerología-Durango, Mexico, with ALL and the previously noted polymorphisms as determined qPCR were studied. At the same time, a 12-month drug monitoring program was conducted in accordance with WHO-PAHO guidelines for pharmacovigilance. Results: The ABCB11936A>G and 2107A>G and ABCC5 3414+434A>C polymorphisms were not associated with methotrexate ADRs. Single nucleotide polymorphisms (SNPs) of ABCB1 1236C>T (OR 0.19, 95% CI: 0.03-0.9, pC (OR 0.12, 95% CI: 0.027-0.58, pT of ABCB1 and ABCC5 3933+313T>C are not associated with the development of typical ADRs by methotrexate, rather, they showed a protective factor for myelosuppression in the studied sick population

Viperidae snakebites in Ecuador: a review of epidemiological and ecological aspects

Snakebite envenoming is a neglected disease of public health concern. Most snakebite accidents occur in developing countries. In Ecuador, 17 viper species are responsible for 99% of official accidents, and ten species are in critical conservation states. This report analyzes the snakebite incident cases and mortality rates in Ecuador between 2014 and 2019. The data obtained from the national surveillance system suggests that the incidence and mortality rates remained constant. The geographic region with the highest incidence rates is the Amazonian region. National policies are urgently needed to prevent snakebite accidents and to protect snakes in danger of extinction

Venomics of the poorly studied hognosed pitvipers Porthidium arcosae and Porthidium volcanicum

We report the first proteomics analyses of the venoms of two poorly studied snakes, the Manabi hognosed pitviper Porthidium arcosae endemic to the western coastal province of Manabí (Ecuador), and the Costa Rican hognosed pitviper P. volcanicum with distribution restricted to South Pacific Costa Rica and western Panamá. These venom proteomes share a conserved compositional pattern reported in four other congeneric species within the clade of South American Porthidium species, P. nasutum, P. lansbergii, P. ophryomegas, and P. porrasi. The paraspecific immunorecognition profile of antivenoms produced in Costa Rica (ICP polyvalent), Perú (Instituto Nacional de Salud) and Brazil (soro antibotrópico pentavalente, SAB, from Instituto Butantan) against the venom of P. arcosae was investigated through a third-generation antivenomics approach. The maximal venom-binding capacities of the investigated antivenoms were 97.1 mg, 21.8 mg, and 25.7 mg of P. arcosae venom proteins per gram of SAB, ICP, and INS-PERU antibody molecules, respectively, which translate into 28.4 mg, 13.1 mg, and 15.2 mg of total venom proteins bound per vial of SAB, ICP, and INS-PERU AV. The antivenomics results suggest that 21.8%, 7.8% and 6.1% of the SAB, ICP, and INS-PERU antibody molecules recognized P. arcosae venom toxins. The SAB antivenom neutralized P. arcosae venom's lethality in mice with an ED50 of 31.3 mgV/g SAB AV. This preclinical neutralization paraspecificity points to Brazilian SAB as a promising candidate for the treatment of envenomings by Ecuadorian P. arcosae.Fundação Rondônia de Amparo ao Desenvolvimento das Ações Científicas e Tecnológicas e à Pesquisa do Estado de Rondônia/[EFP-00021744]/FAPERO/BrasilMinisterio de Ciencia e Innovación/[BFU2017-89103-P]//EspañaUniversidad de Costa Rica/[741-C0-071]/UCR/Costa RicaUniversidad de Costa Rica/[ED-3248]/UCR/Costa RicaUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Instituto Clodomiro Picado (ICP)UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Básicas::Centro de Investigación en Biodiversidad y Ecología Tropical (CIBET)UCR::Vicerrectoría de Docencia::Salud::Facultad de Microbiologí

Corrigendum to: “venomics of the poorly studied hognosed pitvipers porthidium arcosae and porthidium volcanicum”

We report the first proteomics analyses of the venoms of two poorly studied snakes, the Manabi hognosed pitviper Porthidium arcosae endemic to the western coastal province of Manabí (Ecuador), and the Costa Rican hognosed pitviper P. volcanicum with distribution restricted to South Pacific Costa Rica and western Panamá. These venom proteomes share a conserved compositional pattern reported in four other congeneric species within the clade of South American Porthidium species, P. nasutum, P. lansbergii, P. ophryomegas, and P. porrasi. The paraspecific immunorecognition profile of antivenoms produced in Costa Rica (ICP polyvalent), Perú (Instituto Nacional de Salud) and Brazil (soro antibotrópico pentavalente, SAB, from Instituto Butantan) against the venom of P. arcosae was investigated through a third-generation antivenomics approach. The maximal venom-binding capacities of the investigated antivenoms were 97.1 mg, 21.8 mg, and 25.7 mg of P. arcosae venom proteins per gram of SAB, ICP, and INS-PERU antibody molecules, respectively, which translate into 28.4 mg, 13.1 mg, and 15.2 mg of total venom proteins bound per vial of SAB, ICP, and INS-PERU AV. The antivenomics results suggest that 21.8%, 7.8% and 6.1% of the SAB, ICP, and INS-PERU antibody molecules recognized P. arcosae venom toxins. The SAB antivenom neutralized P. arcosae venom's lethality in mice with an ED50 of 31.3 mgV/g SAB AV. This preclinical neutralization paraspecificity points to Brazilian SAB as a promising candidate for the treatment of envenomings by Ecuadorian P. arcosae. BIOLOGICAL SIGNIFICANCE: Assessing the preclinical efficacy profile of antivenoms against homologous and heterologous medically relevant snake venoms represents an important goal towards defining the biogeographic range of their clinical utility. This is particularly relevant in regions, such as Mesoamerica, where a small number of pharmaceutical companies produce antivenoms against the venoms of a small number of species of maximum medical relevance among the local rich herpetofauna, leaving a wide range of snakes of secondary medical relevance, but also causing life-threatening human envenomings without nominal clinical coverage. This work is part of a larger project aiming at mapping the immunological characteristics of antivenoms generated in Latin American countries towards venoms of such poorly studied snakes of the local and neighboring countries' herpetofauna. Here we report the proteomics characterization of the Manabi hognosed pitviper Porthidium arcosae endemic to the western coastal province of Manabí (Ecuador), and the Costa Rican hognosed pitviper P. volcanicum with distribution restricted to southwestern Costa Rica, the antivenomics assessment of three bothropoid commercial antivenoms produced in Costa Rica, Perú, and Brazil against the venom components of P. arcosae, and the in vivo capacity of the Brazilian soro antibotrópico pentavalente (SAB) from Instituto Butantan to neutralize the murine lethality of P. arcosae venom. The preclinical paraspecific ED50 of 31.3 mg of P. arcosae venom per gram of antivenom points to Brazilian SAB as a promising candidate for the treatment of envenomings by the Manabi hognosed pitviper P. arcosae

Assessing the stability of historical and desiccated snake venoms from a medically important ecuadorian collection

Bothrops asper y Bothrops atrox son serpientes venenosas importantes de Ecuador responsables de la mayoría de los accidentes de serpientes, que en el pasado fueron tratados con un antiveneno nacional multipropósito. Durante años, se recolectaron piscinas de veneno y se almacenaron a temperatura ambiente en un laboratorio. Considerando la controvertida capacidad de las muestras secas para retener sus efectos biológicos y actividades enzimáticas, investigamos las propiedades bioquímicas y toxicológicas de los venenos después de años de almacenamiento. Los perfiles proteómicos de los venenos históricos analizados por cromatografía líquida de alto rendimiento y electroforesis son muy similares. Los nuevos lotes de veneno fueron más letales que los almacenados durante años, al igual que los valores iniciales y actuales de LD50 para estas muestras. Se mostraron diferencias significativas en la actividad miotóxica y hemorrágica de algunos grupos de venenos, mientras que no se encontraron diferencias estadísticamente significativas para la actividad del edema. Los ensayos enzimáticos revelaron una variación en la actividad proteolítica sobre la actividad de azocaseína y fosfolipasa A2, y se informaron diferencias bajas para la actividad de serina proteasa similar a la trombina. Mantener el perfil proteómico y ciertas actividades toxicológicas hacen de esta biblioteca de venenos una fuente valiosa para fines de investigación. Sin embargo, una reducción significativa en las actividades toxicológicas, como la actividad de sangrado, no es factible utilizando estas muestras para la producción de antiveneno.Bothrops asper and Bothrops atrox are important venomous snakes from Ecuador responsible for the most of ophidic accidents, which in the past were treated with a national polyvant antivenom. For years, the venom pools were collected and stored at room temperature in a laboratory. Taking into account the controversial ability of desiccated samples to retain their biological effects and enzymatic activities, we investigated the biochemical and toxicological properties of venoms after years of storage. The proteomic profiles of historical venoms analyzed by high-performance liquid chromatography and electrophoresis are very similar. The fresh batches of venom were more lethal than those stored for years, just as the initial and current LD50 values of these samples changed. Significant differences were showed in the myotoxic and hemorrhagic activity of some venom pools, while no significant statistical differences were found for the edema activity. The enzymatic assays revealed a variation in proteolytic activity on azocasein and phospholipase A2 activity, and low differences were reported for thrombin-like serine protease activity. The maintenance of the proteomic profile and certain toxicological activities convert this venom library in a valuable source for research purposes. Nonetheless, the significative reduction of toxicological activities, such as hemorrhagic activity not feasible using these samples for the antivenom production

Acute oral toxicity of a novel functional drink based on Ilex guayusa, Vernonanthura patens, and cocoa husk

Se formuló una nueva bebida funcional con propiedades nutracéuticas a partir de extractos acuosos de Ilex guayusa, hojas de Vernonanthura patens y cáscaras de cacao. Este jugo contiene diversos compuestos bioactivos, como compuestos fenólicos y metilxantinas, con propiedades antioxidantes y estimulantes de interés farmacológico. Sin embargo, se sabe si la interacción de los extractos de hierbas puede tener efectos tóxicos adversos sobre la salud humana. Para evaluar la inocuidad de esta bebida funcional, estimamos la toxicidad oral aguda (AOT) en ratones experimentales. En este trabajo se presenta la evaluación AOT de dos formulaciones de una bebida funcional (preformulación y microencapsulación) a una dosis única de 2000 mg / kg de peso corporal (p.v.). No se observaron signos de toxicidad adversa y mortalidad después de una dosis oral única de 2000 mg / kg de peso corporal. Asimismo, no se observaron cambios significativos de peso corporal y de órganos, comportamiento de consumo de alimentos y agua, ni cambios histopatológicos en los principales órganos evaluados. En conclusión, esta bebida funcional se puede catalogar como de baja toxicidad "según el Sistema de Clasificación Globalmente Armonizado (GHS), lo que la convierte en una bebida potencial con alto valor nutricional y farmacológico.A novel functional drink with nutraceutical properties was formulated from the aqueous extracts of Ilex guayusa, and Vernonanthura patens leaves, and cocoa husks. This juice contains various bioactive compounds, such as phenolic compounds and methylxanthines, with antioxidant and stimulant properties of pharmacological interest. However, it is known whether herbal extracts' interaction may have adverse toxic effects on human health. To evaluate this functional drink's innocuity, we estimated the acute oral toxicity (AOT) in experimental mice. This paper presents the AOT evaluation of two formulations of a functional drink (pre-formulation and microencapsulation) at a single dose of 2000 mg/kg of body weight (b.w.). No signs of adverse toxicity and mortality were observed after a single oral dose of 2000 mg/kg b.w. Likewise, no significant body and organ weight changes, food and water consumption behavior, and no histopathological changes were observed in the main organs evaluated. In conclusion, this functional drink can be categorized as low toxicity " according to the Globally Harmonized Classification System (GHS), making it a potential beverage with high nutritional and pharmacological value