Epistasis between IL1A, IL1B, TNF, HTR2A, 5-HTTLPR and TPH2 Variations Does Not Impact Alcohol Dependence Disorder Features

We assessed a set of biological (HDL, LDL, SGOT, SGPT, GGT, HTc, Hb and T levels) and psychometric variables (investigated through HAM-D, HAM-A, GAS, Liebowitz Social Anxiety Scale, Mark & Mathews Scale, Leyton scale, and Pilowski scale) in a sample of 64 alcohol dependent patients, at baseline and after a detoxification treatment. Moreover, we recruited 47 non-consanguineous relatives who did not suffer alcohol related disorders and underwent the same tests. In both groups we genotyped 11 genetic variations (rs1800587; rs3087258; rs1799724; 5-HTTLPR; rs1386493; rs1386494; rs1487275; rs1843809; rs4570625; rs2129575; rs6313) located in genes whose impact on alcohol related behaviors and disorders has been hypothesized (IL1A, IL1B, TNF, 5-HTTLPR, TPH2 and HTR2A). We analyzed the epistasis of these genetic variations upon the biological and psychological dimensions in the cases and their relatives. Further on, we analyzed the effects of the combined genetic variations on the short – term detoxification treatment efficacy. Finally, being the only not yet investigated variation within this sample, we analyzed the impact of the rs6313 alone on baseline assessment and treatment efficacy. We detected the following results: the couple rs6313 + rs2129575 affected the Leyton -Trait at admission (p = 0.01) (obsessive-compulsive trait), whilst rs1800587 + 5-HTTLPR impacted the Pilowski test at admission (p = 0.01) (hypochondriac symptoms). These results did not survive Bonferroni correction (p ≤ 0.004). This lack of association may depend on the incomplete gene coverage or on the small sample size which limited the power of the study. On the other hand, it may reflect a substantial absence of relevance of the genotype variants toward the alcohol related investigated dimensions. Nonetheless, the marginal significance we detected could witness an informative correlation worth investigating in larger samples

Localisation of sex steroid receptors and effect of Tamoxifen upon glioma cell lines in vitro

Gliomas are the most common form of primary tumour of the central nervous system with a lethality rate approaching 80% in the first year of diagnosis. Their highly invasive nature renders local therapies such as surgery and radiation ineffective, whereas novel approaches such as immunotherapy are being actively studied as possible adjuncts in the treatment of patients with malignant gliomas. A number of factors have been proposed to play a role in glioma immune escape. These include their poor immunogenicity since they do not express specific glioma antigens; their location with in the CNS, which is considered to be an immune-privileged organ and some indications that glioma cells are capable of releasing various immunosuppressive cytokines, which inhibit the cytotoxic function of T cells. Glioma brain tumours differ from most other cancers by their diffusive invasion of the surrounding normal tissue and frequent recurrence following all forms of therapy. It is this invasive nature that ultimately contributes to their poor 6-12 month prognosis. Because of their profound devastating effect much research is currently being carried out to investigate gliomas and it seems that the chemosensitivity of gliomas, i.e. the analysis of glioma response to a specific drug, is a focal point of developing treatment, and more recently an idea of prevention. Tamoxifen is a member of the selective estrogen receptor modulator (SERM) family, is widely used in the treatment of estrogen receptor (ER)-expressing breast cancer. It has been shown that a high dose of Tamoxifen has cytotoxic activity against gliomas, but whether this effect is drug specific or represents a general property of SERMs is unknown. Tamoxifen is considered to exert its anitproliferative effect in ER-positive breast cancer neoplasms through blocking the estrogen receptor, thus, inhibiting gene transcription and cellular proliferation. In vitro studies have shown that Tamoxifen can inhibit the growth of a brain tumour cell lines, but whether the mechanism involves oestrogen blockade remains a matter of some debate. As well as its effect in blocking estrogen receptors, Tamoxifen is also known to exert antiproliferative effects via non-receptor mediated mechanisms including the blockage of Protein Kinase C, inhibition of the Mitogen-7 Activated Protein Kinase (MAPK) family among a vast array of cellular signalling pathways. This study investigated the expression of sex steroid receptors in glioma cell lines using molecular biology technique (RT-PCR), and immunological tecimique (Dot blot, and Western blot). The cells were then separated in different groups of treatment (Tamoxifen, or estradiol and a combination of the two drugs) to obtain any differences in their morphological characteristics, the COX-2 immunoreactivity levels, and the levels of saturated and unsaturated fatty acids. The results showed a molecular expression of sex steroid receptors on the glioma cells, i finding that was later confirmed employing immunological techniques. The cells later showed significant differences on the treatment groups with the most changes obtaining on the projection (length and number) of the cells. The levels of COX-2 were higher on the treatment group (Tamoxifen) indicating more chances of tumour development in later stages. Furthermore, an increase of saturated fats in comparison with the unsaturated was shown in all the groups of treatment (Tamoxifen, estradiol, and Tamoxifen plus estradiol) but with no significant differences with the control group. In conclusion, the results of this study showed an expression of Estrogen and Progesterone receptors on the glioma cell lines, as well as morphological differences with significant difference on the projection length and number of projections between the different treatments of drugs (Tamoxifen and estradiol). The tamoxifen treated cell lines showed an increased COX-2 immunoreactivity, and high levels of saturated fatty acids in comparison with the very low levels of the unsaturated fatty acids