ABSTRACT
The most commonly used Parkinson's disease (PD) treatment is the replacement of dopamine by its levodopa precursor (l-dopa). Monoamine oxidase-B (MAO-B) and catechol-o-methyl transferase (COMT) are enzymes involved in the metabolism and regulation of dopamine availability. In our study we investigated the possible relation among selected single-nucleotide polymorphisms (SNPs) in the MAO-B (rs1799836) and COMT (rs4680) genes and the therapeutic response to levodopa (l-dopa). A total of 162 Brazilian patients from the Pro-Parkinson service of Clinics Hospital of Pernambuco diagnosed with sporadic PD and treated with levodopa were enrolled. PD patients were stratified into 2 groups according to the daily levodopa dose. MAO-B and COMT SNP genotyping was conducted by polymerase chain reaction-restriction fragment length polymorphism. After multivariate analysis, we observed a significant difference between PD groups for the following variables: sex (P = .02), longer duration of disease (P = .02), longer levodopa therapy duration (P = .01), younger onset of PD (P = .01), and use of COMT inhibitor (P = .02). We observed that patients carrying MAO-B (rs1799836) A and AA genotypes and COMT (rs4680) LL genotype suffered more frequently from levodopa-induced-dyskinesia. In addition, we found an increased risk of 2.84-fold for male individuals carrying the MAO-B G allele to be treated with higher doses of levodopa (P = .04). We concluded that before beginning PD pharmacological treatment, it is important to consider the genetic variants of the MAO-B and COMT genes and the sex, reinforcing the evidence that sexual dimorphism in the genes related to dopamine metabolism might affect PD treatment.
Subject(s)
Antiparkinson Agents/administration & dosage , Catechol O-Methyltransferase/genetics , Levodopa/administration & dosage , Monoamine Oxidase/genetics , Parkinson Disease/drug therapy , Parkinson Disease/genetics , Adult , Aged , Antiparkinson Agents/adverse effects , Antiparkinson Agents/pharmacokinetics , Antiparkinson Agents/therapeutic use , Biological Availability , Brazil , Dopamine/metabolism , Dose-Response Relationship, Drug , Dyskinesias , Female , Genotype , Humans , Levodopa/adverse effects , Levodopa/pharmacokinetics , Levodopa/therapeutic use , Male , Middle Aged , Parkinson Disease/enzymology , Pharmacogenomic Variants/drug effects , Polymorphism, Single Nucleotide , Retrospective Studies , Sex Characteristics , Surveys and QuestionnairesABSTRACT
The development of efficient tools for genetic modification of industrial yeast strains is one of the challenges that face the use of recombinant cells in industrial processes. In this study, we examine how the construction of two complementary integrative vectors can fulfill the major requirements of industrial recombinant yeast strains: the use of lactose assimilation genes as a food-grade yeast selection marker, and a system of integration that does not leave hazardous genes in the host genome and involves minimal interference in the yeast physiology. The pFB plasmid set was constructed to co-integrate both LAC4-based and LAC12-based cassettes into the ribosomal DNA (rDNA) locus to allow yeast cells to be selected in lactose medium. This phenotype can also be used to trace the recombinant cells in the environment by simply being plated on X-gal medium. The excisable trait of the LAC12 marker allows the introduction of many different heterologous genes, and makes it possible to introduce a complete heterologous metabolic pathway. The cloned heterologous genes can be highly expressed under the strong and constitutive TPI1 gene promoter, which can be exchanged for easy digestion of enzymes if necessary. This platform was introduced into Saccharomyces cerevisiae JP1 industrial strain where a recombinant with high stability of markers was produced without any change in the yeast physiology. Thus, it proved to be an efficient tool for the genetic modification of industrial strains.
