Specific and complete human genome amplification with improved yield achieved by phi29 DNA polymerase and a novel primer at elevated temperature

<p>Abstract</p> <p>Backgrounds</p> <p>Whole genome amplification (WGA) is a practical solution to eliminate molecular analysis limitations associated with genomic DNA (gDNA) quantity. Different methods have been developed to amplify the whole genome, including primer extension preamplification (PEP), degenerate oligonucleotide primed PCR (DOP-PCR), and multiple displacement amplification (MDA). Each of these methods has its own merits and limitations.</p> <p>Findings</p> <p>Effects of primer length and composition on amplification quality and quantity were explored in this study at two different temperatures (30°C & 40°C). New primer designs combined with elevated amplification temperature has significantly improved MDA as measured by amplification yield, genome coverage, and allele drop out (ADO) analysis. A remarkable finding was the comprehensive amplification, at 30°C & 40°C, of the human whole genome via the use of GGGCAGGA*N*G hotspot recombination consensus primer. Amplification was characterized by Affymetrix 10K SNP chip analysis. Finally, the use of new primer designs has suppressed the template-independent DNA amplification (TIDA) both at 30°C and 40°C.</p> <p>Conclusion</p> <p>The use of new primers in this study combined with elevated incubation temperatures in MDA has remarkably improved the specificity, amplification yield, and suppressed TIDA.</p

Oxygen Harvesting from Carbon Dioxide: Simultaneous Epoxidation and CO Formation

Due to increasing concentrations in the atmosphere, carbon dioxide has, in recent times, been targeted for utilisation (Carbon Capture Utilisation and Storage, CCUS). In particular, the production of CO from CO2 has been an area of intense interest, particularly since the CO can be utilized in Fischer–Tropsch synthesis. Herein we report that CO2 can also be used as a source of atomic oxygen that is efficiently harvested and used as a waste-free terminal oxidant for the oxidation of alkenes to epoxides. Simultaneously, the process yields CO. Utilization of the atomic oxygen does not only generate a valuable product, but also prevents the recombination of O and CO, thus increasing the yield of CO for possible application in the synthesis of higher-order hydrocarbons

Superior rate capability of high mass loading supercapacitors fabricated with carbon recovered from methane cracking

High mass loading (ca. 30 mg/cm2) electrodes were prepared with carbon recovered from catalytic methane cracking (MC). As-fabricated supercapacitors displayed 74% of capacitance retention from 6 mA/cm2 to 60 mA/cm2 and a Ragone plot’s slope of −7 Wh/kW (compared to 42% and −31 Wh/kW, respectively, for high mass loading devices fabricated with commercial carbon). The high-rate capability of the MC-recovered carbon is attributed to the presence of carbon black and carbon nanotubes produced during the reaction, which likely increased the electronic and ionic conductivity within the electrode. These results suggest that the by-product of this hydrogen generation route might be a suitable active material for supercapacitors.Engineering and Physical Sciences Research Council (EPSRC): EP/R023662/1; EP/S023909/

Genome-wide landscape establishes novel association signals for metabolic traits in the Arab population

While the Arabian population has a high prevalence of metabolic disorders, it has not been included in global studies that identify genetic risk loci for metabolic traits. Determining the transferability of such largely Euro-centric established risk loci is essential to transfer the research tools/resources, and drug targets generated by global studies to a broad range of ethnic populations. Further, consideration of populations such as Arabs, that are characterized by consanguinity and a high level of inbreeding, can lead to identification of novel risk loci. We imputed published GWAS data from two Kuwaiti Arab cohorts (n = 1434 and 1298) to the 1000 Genomes Project haplotypes and performed meta-analysis for associations with 13 metabolic traits. We compared the observed association signals with those established for metabolic traits. Our study highlighted 70 variants from 9 different genes, some of which have established links to metabolic disorders. By relaxing the genome-wide significance threshold, we identified ‘novel’ risk variants from 11 genes for metabolic traits. Many novel risk variant association signals were observed at or borderline to genome-wide significance. Furthermore, 349 previously established variants from 187 genes were validated in our study. Pleiotropic effect of risk variants on multiple metabolic traits were observed. Fine-mapping illuminated rs7838666/CSMD1 rs1864163/CETP and rs112861901/[INTS10,LPL] as candidate causal variants influencing fasting plasma glucose and high-density lipoprotein levels. Computational functional analysis identified a variety of gene regulatory signals around several variants. This study enlarges the population ancestry diversity of available GWAS and elucidates new variants in an ethnic group burdened with metabolic disorders.Peer reviewe

Genome-wide association study identifies novel risk variants from RPS6KA1, CADPS, VARS, and DHX58 for fasting plasma glucose in Arab population

Consanguineous populations of the Arabian Peninsula, which has seen an uncontrolled rise in type 2 diabetes incidence, are underrepresented in global studies on diabetes genetics. We performed a genome-wide association study on the quantitative trait of fasting plasma glucose (FPG) in unrelated Arab individuals from Kuwait (discovery-cohort:n = 1,353; replication-cohort:n = 1,196). Genome-wide genotyping in discovery phase was performed for 632,375 markers from Illumina HumanOmniExpress Beadchip; and top-associating markers were replicated using candidate genotyping. Genetic models based on additive and recessive transmission modes were used in statistical tests for associations in discovery phase, replication phase, and meta-analysis that combines data from both the phases. A genome-wide significant association with high FPG was found at rs1002487 (RPS6KA1) (p-discovery = 1.64E-08, p-replication = 3.71E-04, p-combined = 5.72E-11; beta-discovery = 8.315; beta-replication = 3.442; beta-combined = 6.551). Further, three suggestive associations (p-values <8.2E-06) with high FPG were observed at rs487321 (CADPS), rs707927 (VARS and 2Kb upstream of VWA7), and rs12600570 (DHX58); the first two markers reached genome-wide significance in the combined analysis (p-combined = 1.83E-12 and 3.07E-09, respectively). Significant interactions of diabetes traits (serum triglycerides, FPG, and glycated hemoglobin) with homeostatic model assessment of insulin resistance were identified for genotypes heterozygous or homozygous for the risk allele. Literature reports support the involvement of these gene loci in type 2 diabetes etiology.Peer reviewe

The influence of photoperiod and light intensity on the growth and photosynthesis of Dunaliella salina (chlorophyta) CCAP 19/30

The green microalga Dunaliella salina survives in a wide range of salinities via mechanisms involving glycerol synthesis and degradation and is exploited for large amounts of nutraceutical carotenoids produced under stressed conditions. In this study, D. salina CCAP 19/30 was cultured in varying photoperiods and light intensities to study the relationship of light with different growth measurement parameters, with cellular contents of glycerol, starch and carotenoids, and with photosynthesis and respiration. Results show CCAP 19/30 regulated cell volume when growing under light/dark cycles: cell volume increased in the light and decreased in the dark, and these changes corresponded to changes in cellular glycerol content. The decrease in cell volume in the dark was independent of cell division and biological clock and was regulated by the photoperiod of the light/dark cycle. When the light intensity was increased to above 1000 μmol photons m−2 s−1, cells displayed evidence of photodamage. However, these cells also maintained the maximum level of photosynthesis efficiency and respiration possible, and the growth rate increased as light intensity increased. Significantly, the intracellular glycerol content also increased, >2-fold compared to the content in light intensity of 500 μmol photons m−2 s−1, but there was no commensurate increase in the pool size of carotenoids. These data suggest that in CCAP 19/30 glycerol stabilized the photosynthetic apparatus for maximum performance in high light intensities, a role normally attributed to carotenoids

Effect of Ca2+ Channel Block on Glycerol Metabolism in Dunaliella salina under Hypoosmotic and Hyperosmotic Stresses

The effect of Ca2+ channel blockers on cytosolic Ca2+ levels and the role of Ca2+ in glycerol metabolism of Dunaliella salina under hypoosmotic or hyperosmotic stress were investigated using the confocal laser scanning microscope (CLSM). Results showed that intracellular Ca2+ concentration increased rapidly when extracellular salinity suddenly decreased or increased, but the increase could be inhibited by pretreatment of Ca2+ channel blockers LaCl3, verapamil or ruthenium red. The changes of glycerol content and G3pdh activity in D. salina to respect to hypoosmotic or hyperosmotic stress were also inhibited in different degrees by pretreatment of Ca2+ channel blockers, indicating that the influx of Ca2+ via Ca2+ channels are required for the transduction of osmotic signal to regulate osmotic responses of D. salina to the changes of salinity. Differences of the three blockers in block effect suggested that they may act on different channels or had different action sites, including influx of Ca2+ from the extracellular space via Ca2+ channels localized in the plasma membrane or from intracellular calcium store via the mitochondrial. Other Ca2+-mediated or non-Ca2+-mediated osmotic signal pathway may exist in Dunaliella in response to hypoosmotic and hyperosmotic stresses

Weak or no association of TCF7L2 variants with Type 2 diabetes risk in an Arab population

<p>Abstract</p> <p>Background</p> <p>The rs7903146 and rs12255372 variants of <it>TCF7L2 </it>have been strongly associated with type 2 diabetes (T2D) risk in most populations studied to date. Meta-analysis of 27 different studies has resulted in a global OR of 1.46 [1.42–1.51] (rs7903146 variant). Thus far, despite a high incidence of T2D, the role of this variant in Arabs has not been established.</p> <p>Methods</p> <p>We performed a case-control association study using 522 Saudi T2D patients (WHO criteria), and 346 controls (age > 60; fasting plasma glucose < 7 mmol/L). Genotyping was performed by pyrosequencing. Statistical analyses were performed using SPSS version 13.0 for Windows (SPSS, Chicago, IL, USA).</p> <p>Results</p> <p>For rs7903146, the T allele frequency of the cases (0.415) was not different from that observed in the controls (0.405). The crude odds ratio (OR) was 1.04 with a 95% CI of 0.86–1.27 (P = 0.675). For rs12255372, the T allele frequency of the cases (0.368) was not different from that observed in the controls (0.355). Retrospective power calculations based upon an OR of 1.46 reported in a comprehensive meta-analysis of <it>TCF7L2 </it>risk, indicated this study was sufficiently powered (96.92%; α = 0.05) to detect an effect of similar magnitude to that reported for rs7903146.</p> <p>Conclusion</p> <p>Our study is consistent with weak or no association of T2D in Arabs with the two <it>TCF7L2 </it>variants, however it cannot rule out an effect of other SNPs in this gene. Future studies in this population are required to confirm our findings and may indicate the presence of yet to be defined genetic risk factors for T2D.</p

Highly Efficient and Selective Capture of Pb(II) by New Crosslinked Melamine-Based Polymethyl Methacrylate for Water Treatment

Owing to the recent developments in the polymer’s properties and application, the demands for designing different structure of polymers are greater than ever. Crosslinked polymers (CPs) are a type of porous materials that have a variety of potential applications. Because of simple methods of modification of polymethyl methacrylate (PMMA), the crosslinked PMMA considers the most commonly polymeric adsorbents. A new crosslinked melamine-based polymethyl methacrylate (C-PMMA/Mel) was prepared via a polycondensation reaction between PMMA and melamine used as a crosslinking agent. Different characterization methods were carried out to investigate the molecular structures, thermal stability, and morphology. C-PMMA/Mel was applied for the adsorption behavior toward different metal cations and detected a selective to Pb(II). The evaluation of the new polymers as adsorbent against Pb(II) ion was studied using the contact time, adsorbent dose, initial concentration, and effects of pH. The adsorption efficiency of heavy metals was improved in the presence of melamine in polymeric matrix. The C-PMMA/Mel has high efficacy in the removal of ~94% of Pb at pH 6 for one hour. Noticeably, the adsorption performance of C-PMMA/Mel perfectly suited with Freundlich isotherm and the pseudo-second-order kinetic model. Additionally, the new materials showed no obvious loss in Pb(II) removal after 7 cycles

Contribution à l'intégration monolithique de protections contre les surtensions :<br />application aux convertisseurs de puissance haute tension

A new over-voltage protection circuit has been developed. In this protection circuit, the dissipative part ismonolithically integrated in the same protected power module chip with almost no additional technologicprocess or modifications. This monolithic integration takes advantage of main switch cooling system to extractover heating. At the same time it minimizes interconnections between the protected transistor and its protectionsystem. This gives high performance results in term of over voltage protection time response and thermal powerremoval capabilities. In addition to that, the design of this protection circuit enables to adjust the protectionvoltage threshold. This is useful for series transistor in high voltage applications. The design aims to improvethe performance and to reduce the cost and the volume of the protection circuit.A model of the BJT as a protection transistor is established. This model is distinguished from the existingmodels because of taking into account that the BJT is used in linear mode. A thermal model of the whole of theintegrated transistors evaluates the behavior of these transistors in spite of the difference between the operatingmode. This model gives the better distribution of the protection transistor cells in the chip. Practical resultsstarting from the auto protected MOSFETs components that we manufactured validates the solution suggested.A demonstrator of a buck converter using two auto protected MOSFETs in series shows the effectiveness of oursolution.Un nouveau circuit de protection contre les surtensions a été développé. Dans ce circuit de protection, la partiedissipative est monolithiquement intégrée dans la même puce du transistor à protéger avec aucune modificationtechnologique additionnelle. Cette intégration monolithique tire profit du système de refroidissement dutransistor à protéger pour le refroidissement de la partie intégrée. En même temps, elle réduit au minimum lesproblèmes de connections entre le transistor à protéger et son système de protection. En plus, la conception dece circuit de protection permet d'ajuster le seuil de tension de protection. C'est utile pour la mise en série destransistors pour des applications à haute tension. Un modèle du BJT comme transistor de protection est établi.Ce modèle se distingue des modèles existants car il prend en compte que le BJT fonctionne en mode linéaire.Un modèle thermique de l'ensemble des transistors intégrés évalue le comportement de ces transistors malgré ladifférence entre leur mode de fonctionnement. Ce modèle donne une meilleure distribution des cellules dutransistor de protection dans la puce. Des résultats pratiques à partir des composants MOSFETs autoprotégésque nous avons fabriqués valident la solution proposée. Un démonstrateur de hacheur série utilisant deuxMOSFETs autoprotégés en série montre l'efficacité de notre solution