Haptoglobin transport into human ovarian follicles and its binding to apolipoprotein A-1.

Controlled ovarian stimulation was induced in 19 women for IVF-ET. After ovum pick up, Haptoglobin titers were determined by ELISA in sera and homologous follicular fluids. The Haptoglobin phenotype of each subject was assessed and the penetration of the protein forms through the blood-follicle barrier was predicted on the basis of their molecular weight. The penetration threshold compatible with the barrier integrity was calculated 92, 73 and 57% of the blood level for phenotypes Hpt 1-1, Hpt 1-2 and Hpt 2-2 respectively. Penetration values comparable-lower or higher than threshold were found associated with 46/49 or 3/49 fertilized oocytes respectively. Complexes of Haptoglobin with Apoliprotein A-1 were isolated from follicular fluids by affinity chromatography with Hemoglobin. The Haptoglobin beta chain, after Western blotting and incubation with Apolipoprotein A-1, was found involved in the protein-protein interaction as detected by anti-Apoliprotein A-1 antibodies. Complexes from separate fluids were analysed by electrophoresis and densitometry : the beta chain/Apoliprotein A-1 stoichiometric ratio was found 0.75 and 1.40 in fluids associated with fertilized and unfertilised oocytes respectively. The results suggest that the Haptoglobin transport in the follicle depends on the integrity of the blood-follicle and might be associated with the oocyte quality, possibly by interfering with the role of Apoliprotein A-1 in cholesterol or vitamin E exchange between HDLs and granulosa cells

Potentiation of cisplatin-induced antiproliferative and apoptotic activities by the antiarrhythmic drug procainamide hydrochloride

Background We describe the potentiation of antIProliferative and apoptotic activities triggered by cis-diamminedichloroplatinum(II) (DDP), and obtained in vitro by the co-administration of procainamide hydrochloride (PdHCl) in murine P388, and human A2780 and A549 cells. Methods We determined the antIProliferative and apoptotic activities of DDP and PdHCl combinations by different techniques. Moreover, cell cycle analysis, restriction enzyme inhibition followed by agarose gel electrophoresis, and TUNEL analysis of tumour cells in vivo were also used to strengthen our hypothesis. Results Our results show that PdHCl may significantly increase the inhibition of cell proliferation and apoptosis. Experiments in vivo showed that the co-administration of DDP and PdHCl increased the percentage of apoptotic cells compared to DDP alone treatment, both in subcutaneous (sc) and intraperitoneal (IP) P388 tumours. We finally demonstrated that the co-administration of PdHCl prevents DNA digestion accounting for a restriction enzyme inhibition that in some cases was greater than that obtained by DDP alone. Moreover, when PdHCl was mixed with the reaction products (RP) of DDP (RP-PdHCl) we obtained a restriction enzyme inhibition greater for some enzymes (Bsp1407I, Hin1II, and Psp1406I) than that obtained by the DDP-PdHCl solution. Conclusions On the whole our data demonstrate that the class I antiarrhythmic drug PdHCl may increase the antIProliferative activity of DDP by improving its triggering of apoptosis, and that this phenomenon may be likely linked to the formation of a new Pt compound

Association between SCN1A gene polymorphisms and drug resistant epilepsy in pediatric patients

Purpose: “Single Nucleotide Polymorphisms (SNPs)” could be an important explanation of drug resistance in epilepsy. The aim of this study was to investigate if genetic polymorphisms (SNPs) of the SCN1A gene could influence the response to anti – epileptic drugs (AED) and if they could predispose to a drug resistant epilepsy in pediatric patients. Methods: We investigated SNPs in exon and intronic regions of the SCN1A gene in a sample of 120 pediatric patients, in both drug-resistant and drug-responsive patients. Association between polymorphisms and refractory epilepsy were investigated by comparing SNPs in exon and intronic regions between the two groups. The genotypes of each intronic polymorphism in the drug-resistant group was analyzed. Odds ratios and confidence intervals were calculated. Results: None of the SNPs identified in exons of the SCN1A gene were associated with drug-resistance. In the intronic regions, a statistically significant difference was found in the prevalence of three polymorphisms was found between the two patient groups (rs6730344A/C, rs6732655A/T, rs10167228A/T). The analysis of the genotypes of each intronic polymorphism in the drug-resistant group revealed that the AA and AT genotypes for the rs1962842 polymorphism are associated with an increased risk of developing drug resistance compared to TT genotype. Conclusion: The intronic rs6730344, rs6732655 and rs10167228 polymorphisms of the SCN1A gene are a potential risk factors for drug resistance. AA e AT genotype of the rs1962842 intronic polymorphism also emerged as a risk factor in the drug resistant group. Therefore, polymorphisms of the SCN1A gene could play a role in the response to AED in patients with drug-resistant epilepsy, with important implications for clinical practice

A Combination of Aqueous Extraction and Ultrafiltration for the Purification of Phycocyanin from Arthrospira maxima

The purification of phycocyanin (PC) from Spirulina generally involves a combination of different techniques. Here, we report the results on PC yields from a combined aqueous extraction-ultrafiltration (UF) process of a strain of Arthrospira maxima cultivated in a farm devoted to producing PC with food-grade purity. Samples optimized from different biomass/solvent ratios were purified by using a polyethersulphone (PES) membrane with a molecular weight cut-off (MWCO) of 20 kDa. The UF system was operated at 2.0 ± 0.1 bar and at 24 ± 2 °C up to a volume concentration factor (VCF) of 5. A diafiltration (DF) process was conducted after UF in order to increase the PC recovery in the retentate. Samples were collected during both UF and DF processes in order to evaluate membrane productivity and PC purity. The average permeate fluxes of about 14.4 L/m2h were measured in the selected operating conditions and more than 96% of PC was rejected by the UF membrane independently ofthe extraction yields and times. The concentration of PC in the final retentate was 1.17 mg/mL; this confirmed the observed rejection and the final VCF of the process (about 5-fold when compared to the concentration of PC in the crude extract). In addition, the combination of UF and diafiltration allowed the removal of about 91.7% of the DNA from the crude extract, thereby improving the purity of the phycocyanin in the retentate fraction