Multivariate Analysis of Dopaminergic Gene Variants as Risk Factors of Heroin Dependence

BACKGROUND: Heroin dependence is a debilitating psychiatric disorder with complex inheritance. Since the dopaminergic system has a key role in rewarding mechanism of the brain, which is directly or indirectly targeted by most drugs of abuse, we focus on the effects and interactions among dopaminergic gene variants. OBJECTIVE: To study the potential association between allelic variants of dopamine D2 receptor (DRD2), ANKK1 (ankyrin repeat and kinase domain containing 1), dopamine D4 receptor (DRD4), catechol-O-methyl transferase (COMT) and dopamine transporter (SLC6A3) genes and heroin dependence in Hungarian patients. METHODS: 303 heroin dependent subjects and 555 healthy controls were genotyped for 7 single nucleotide polymorphisms (SNPs) rs4680 of the COMT gene; rs1079597 and rs1800498 of the DRD2 gene; rs1800497 of the ANKK1 gene; rs1800955, rs936462 and rs747302 of the DRD4 gene. Four variable number of tandem repeats (VNTRs) were also genotyped: 120 bp duplication and 48 bp VNTR in exon 3 of DRD4 and 40 bp VNTR and intron 8 VNTR of SLC6A3. We also perform a multivariate analysis of associations using Bayesian networks in Bayesian multilevel analysis (BN-BMLA). FINDINGS AND CONCLUSIONS: In single marker analysis the TaqIA (rs1800497) and TaqIB (rs1079597) variants were associated with heroin dependence. Moreover, -521 C/T SNP (rs1800955) of the DRD4 gene showed nominal association with a possible protective effect of the C allele. After applying the Bonferroni correction TaqIB was still significant suggesting that the minor (A) allele of the TaqIB SNP is a risk component in the genetic background of heroin dependence. The findings of the additional multiple marker analysis are consistent with the results of the single marker analysis, but this method was able to reveal an indirect effect of a promoter polymorphism (rs936462) of the DRD4 gene and this effect is mediated through the -521 C/T (rs1800955) polymorphism in the promoter

Analysis and quantification of GPCR allosteric receptor\u2013receptor interactions using radioligand binding assays: the a2ar-d2r heteroreceptor complex example

There is a large body of biochemical and biophysical experimental evidences which establishes the existence of G protein-coupled receptors (GPCRs) as homo- and heteroreceptor complexes. The results indicate that there are allosteric interactions across the receptor\u2013receptor interface of homo- and heteroreceptor complexes that modulate the binding properties of their receptor protomer components in terms of affinity and density, and thereby change their pharmacology. In the adenosine A2A-dopamine D2 heteroreceptor complexes (A2AR-D2R), the activation of the A2AR protomer by its standard receptor agonist CGS21680 causes a conformational change in the A2AR-D2R heteroreceptor complex. The allosteric wave passes over the receptor interface, invades the orthostatic dopamine binding site of the dopamine D2R protomer, and reduces the affinity of the high but not the low affinity D2R agonist binding site. In view of the complex nature of allosteric mechanisms, the detection, analysis, and quantification of the effects of this phenomenon rely on the use of competition radioligand binding assays to ensure proper demonstration of the high and low affinity D2R agonist binding sites. Outlined in this chapter is simple but useful experimental approaches for measuring the allosteric receptor\u2013receptor interactions at GPCR heteroreceptor complexes. The readers will also find tips and discussion on the pitfalls of these assay and instructions for data analysis

Effect of maintenance immunosuppressive drugs on virus pathobiology: evidence and potential mechanisms

Recent evidence suggesting a potential anti-CMV effect of mTORis is of great interest to the transplant community. However, the concept of an immunosuppressant with antiviral properties is not new, with many accounts of the antiviral properties of several agents over the years. Despite these reports, to date, there has been little effort to collate the evidence into a fuller picture. This manuscript was developed to gather the evidence of antiviral activity of the agents that comprise a typical immunosuppressive regimen against viruses that commonly reactivate following transplant (HHV1 and 2, VZV, EBV, CMV and HHV6, 7, and 8, HCV, HBV, BKV, HIV, HPV, and parvovirus). Appropriate immunosuppressive regimens posttransplant that avoid acute rejection while reducing risk of viral reactivation are also reviewed. The existing literature was disparate in nature, although indicating a possible stimulatory effect of tacrolimus on BKV, potentiation of viral reactivation by steroids, and a potential advantage of mammalian target of rapamycin (mTOR) inhibition in several viral infections, including BKV, HPV, and several herpesviruses