Abstract
Cigarette smoke extract (CSE) decreases the expression of the CFTR. Furthermore, CSE has profound effects on epithelial cells but different results are reported with cell lines and primary cells. This study aimed to compare the effects of CSE on spontaneous and induced IL-8 release from primary nasal epithelial cells isolated from CF patients possessing two different classes of CFTR mutation (F508del homozygous; R1 17H heterozygote) and healthy volunteers. Nasal epithelial cells were obtained via nasal brushings of the inferior turbinate of CF patients; F508del homozygote (n=5), R1 17H heterozygote (n=7) and healthy volunteers (n=6). The cells were expanded in culture and used at passage 3. Monolayer cultures were pre-exposed for 4h in the presence or absence of 5% CSE. The cells were then stimulated with LPS from P. aeruginosa (LPS-PA 50 or 100 ?g/ml); Cytomix 1 (TNF-? 10 ng/ml, IL-1? 5 ng/ml, LPS-PA 5 ?g/ml) or Cytomix 2 (TNF-? 10 ng/ml, IL-1? 10 ng/ml, IFN-? 1000 U/ml) for 24 h. Culture supernatants were harvested and IL-8 release measured using ELISA (R & D systems). LPS (100 ?g/ml) induced IL-8 release in control cells was increased by CSE (1188?292 vs. 1774?501.6, p< 0.05). In cells from F508del homozygous patients CSE increased cytomix 2 and LPS 50 ?g/ml induced IL-8 secretion. In the R1 17H heterozygotes only the spontaneous IL-8 secretion was significantly increased by CSE exposure. Comparing the responses from the various genetic backgrounds of the subjects; the only significant differences between the groups were with cytomix 2 where the releases followed R1 17H< F508del< control (8157?1746, vs 16031?3729 vs 21296?2576). R1 17H cells were also significantly less responsive to cytomix 2 compared to F508del cells in the presence of CSE.In contrast to our data using cell lines, where CSE consistently decreased IL-8 secretion from the CF cell line (CFBE41o-), preincubation of primary cells with CSE only caused sporadic increases in IL-8 release. Cytomix 2 induced significantly different levels of IL-8 secretion depending on the genetic background of the cells. As yet, we have no explanation for these data but further studies will investigate the action of IFN-? alone on primary cells. Williams MTS et al. Ped Pulmon 2008;287:246

Abstract
Parkinsons disease (PD) is the second most common neurodegenerative disorder next to Alzheimers disease. It accounts for about 0.2 to 1% of the age of 65 ? 69 and raises to 1 ? 2% of the age of 80 and above. It is mainly characterised by the involvement of the four classical symptoms - tremor, bradykinesia, postural instability and the rigidity. It may be due to the cause of the environmental toxins and also due to the genetic factors. There is high relationship for the PD with the environmental toxins. The enzyme CYP2D6 is one which acts on the chemical 1-methyl-4-phenyll, 2,3,6- tetrahydropyridine (MPTP), 1,2,3,4-tetrahydroisoquinoline (TIQ) and many other neurotoxins that cause the PD. An exciting possibility is that chemical transformation to produce PD toxin in the course of the detoxification mediated by CYP2D (cytochrome P450). The CYP2D comprises many isoenzymes which are involved in the metabolism of exogenous and endogenous toxins. The CYP2D subfamily comprise of genes which are highly specific for the debrisoquine- 4 hydroxylase activity. In this, the CYP2D6 is widely studied polymorphism. In the present study, we analyzed this CYP2D6*4 polymorphism in both the PD cases and controls using polymerase chain reaction (PCR) and Restriction Fragment Length polymorphism (RFLP). Bearers of the CYP2D6 gene variants may be classified as the extensive metabolizers (EM) and poor metabolizers (PM). PM allele carriers have lower gene activity and they are more prone to the environmental toxins. In this study we determined the CYP2D6*4 allele in both PD and control to evaluate the relation of this allele with the PD.

Abstract
Cystic fibrosis transmembrane conductance regulator (CFTR) potentiators are compounds with a double effect. At lower concentration they increase the activity of CFTR, whereas at higher concentrations they inhibit it. The binding site for potentiators is hypothesized to be located in the interface of the dimer formed by the two nucleotide binding domains (NBDs), interacting mostly with NBD1 and to a lesser extent with NBD2. This binding site seems to involve at least two protonable residues, Cys491 and His1348. The binding of potentiators to the NBDs probably relies on electrostatic interaction, as electrostatic interactions between oppositely charged amino acids might be involved in the formation and stabilization of the dimer. Here we have analysed the electrostatic contribution to the binding of genistein by modifying intracellular pH. We have analysed the effect of pH on both, the activating and the inhibitory dissociation constants. The study was done on polarized epithelia expressing high levels of wild type CFTR after establishing that modifications in the pH of the basolateral solution, between pH 6 and 8, change the intracellular pH to the same extent. We found that at pH 6 it was necessary a lower concentration of CPTcAMP to activate CFTR. In addition, we found that the maximal CFTR current was significantly lower at this pH value. Regarding genistein binding, we found that at alkaline pH the apparent dissociation constant for the activating site was shifted to higher concentrations. In contrast, we found that the inhibitory apparent dissociation constant was affected by alkaline and acidic pH values to the same extent. In fact, at pH 6 and at pH 8 the affinity of genistein for the inhibitory site was similarly increased. This study suggests that cysteine, possibly Cys491, is involved in the activating binding site. In contrast, an histidine (e.g. His1348) does not seem to be part of it. Besides, our data indicate that the inhibitory site, whose location is still unknown, may contain both, a cysteine and a histidine, the only two amino acids with a pKa in the range of the pH analysed. This work was supported by the Italian Cystic Fibrosis Foundation grant FFC#2/2008 with the contribution of Mille bambini a Via Margutta onlus, Blunotte, and Lega Italiana FC - Associazione Toscana.

Abstract
The advent of 2nd generation, low-cost, DNA sequencing technologies is leading to dramatic increases in the rate of genome data generation. The utilization of these new technologies to sequence ever increasing numbers of human genomes for personalized diagnostics and medicines will generate even more data. Current database and software technologies are becoming progressively ineffective for managing, processing and analyzing these new levels of information. Synamatix addresses this need by leveraging SynaBASE?, which is an innovative database solution based upon indexing patterns in data. Rapid cross referencing of clinical patient sequence data back to the human genome is a critical step in SNP discovery and identifying potential disease states. Performance improvements of between 100 and 10,000 fold for comparative and differential genomics and for mapping-based genome assembly from 2nd generation sequencers will be reviewed.

Abstract
Oman is a country with a population comprising of a wide range of ethnic groups, high rates of consanguinity and increased incidences of inter-cousin marriages. There is an increased prevalence of hemoglobinopathies which is of growing importance as knowledge of a population structure can be a unique aid in planning genetic services. The aim of this study was to establish neonatal cord blood screening in the Sultanate of Oman, in an effort to determine the prevalence of hemoglobinopathies by a costeffective method. High performance liquid chromatography [HPLC] is a powerful tool to screen newborns for hemoglobinopathies. Neonatal screening includes cord blood samples collection, screening, and follow up of all newborns with abnormal results. A total of 7837 consecutive cord blood samples were screened for presence of possible hemoglobinopathies by HPLC using Biorad Variant ?? program between April 2005 and March 2007. Complete blood counts [CBC] were also obtained on Cell Dyn 4000 automated blood cell counter. All samples were then processed to isolate and store mononuclear leukocytes for subsequent molecular diagnostics. The findings indicated a 47.07% incidence of ??thalassemia, based on low mean cell volume [MCV] and mean cell hemoglobin [MCH] on the CBC and significant amounts of Hb Barts on HPLC. The overall incidence of other hemoglobinopathies was 9.87%, with 5.47% incidence of sickle hemoglobin. On HPLC, D-window, E-window and C-window were present in 0.93%, 0.77% and 0.06% of the samples respectively. Since HPLC cannot diagnose beta thalassemia major at birth, in samples with HbA below 10%, the beta globin gene was directly sequenced including the promoter, all exons and introns in these samples. Amongst 206 (2.62%) samples sequenced, beta thalassemia trait was confirmed in 201 cases and 5 cases were found to be homozygous for beta thalassemia major. Additionally, direct sequencing of all abnormal samples with HbS [n=429], HbD [n=73], HbE [n=42], and HbC [n=5] was also performed on ABI Prism 3100 genetic analyzer to assign the correct genotype status to these subjects and use the same to validate the HPLC results. The significantly high incidence of hemoglobinopathies in newborns in the Sultanate of Oman emphasizes the value of neonatal cord blood screening to be implemented as the first step in the national strategy towards total management of hemoglobinopathies including early diagnosis, comprehensive clinical care and counseling of the affected families. The results of this large study wound indicate that using HPLC [<2 USD/sample] is a cost effective method. Moreover, prescreening of both parents and selecting only samples of neonatal cord blood from newborns with either parent having an underlying genetic trait for hemoglobinopathy would result in the a huge cost saving to the tune of over 90% as compared to universal neonatal cord blood screening and can be recommended as a highly cost-effective method targeted to screen only the abnormal samples.