Genotyping, characterization, and imputation of known and novel CYP2A6 structural variants using SNP array data

CYP2A6 metabolically inactivates nicotine. Faster CYP2A6 activity is associated with heavier smoking and higher lung cancer risk. The CYP2A6 gene is polymorphic, including functional structural variants (SV) such as gene deletions (CYP2A6*4), duplications (CYP2A6*1 × 2), and hybrids with the CYP2A7 pseudogene (CYP2A6*12, CYP2A6*34). SVs are challenging to genotype due to their complex genetic architecture. Our aims were to develop a reliable protocol for SV genotyping, functionally phenotype known and novel SVs, and investigate the feasibility of CYP2A6 SV imputation from SNP array data in two ancestry populations. European- (EUR; n = 935) and African- (AFR; n = 964) ancestry individuals from smoking cessation trials were genotyped for SNPs using an Illumina array and for CYP2A6 SVs using Taqman copy number (CN) assays. SV-specific PCR amplification and Sanger sequencing was used to characterize a novel SV. Individuals with SVs were phenotyped using the nicotine metabolite ratio, a biomarker of CYP2A6 activity. SV diplotype and SNP array data were integrated and phased to generate ancestry-specific SV reference panels. Leave-one-out cross-validation was used to investigate the feasibility of CYP2A6 SV imputation. A minimal protocol requiring three Taqman CN assays for CYP2A6 SV genotyping was developed and known SV associations with activity were replicated. The first domain swap CYP2A6-CYP2A7 hybrid SV, CYP2A6*53, was identified, sequenced, and associated with lower CYP2A6 activity. In both EURs and AFRs, most SV alleles were identified using imputation (>70% and >60%, respectively); importantly, false positive rates were <1%. These results confirm that CYP2A6 SV imputation can identify most SV alleles, including a novel SV

Evaluation of sesamum gum as an excipient in matrix tablets

In developing countries modern medicines are often beyond the affordability of the majority of the population. This is due to the reliance on expensive imported raw materials despite the abundance of natural resources which could provide an equivalent or even an improved function. The aim of this study was to investigate the potential of sesamum gum (SG) extracted from the leaves of Sesamum radiatum (readily cultivated in sub-Saharan Africa) as a matrix former. Directly compressed matrix tablets were prepared from the extract and compared with similar matrices of HPMC (K4M) using theophylline as a model water soluble drug. The compaction, swelling, erosion and drug release from the matrices were studied in deionized water, 0.1 N HCl (pH 1.2) and phosphate buffer (pH 6.8) using USP apparatus II. The data from the swelling, erosion and drug release studies were also fitted into the respective mathematical models. Results showed that the matrices underwent a combination of swelling and erosion, with the swelling action being controlled by the rate of hydration in the medium. SG also controlled the release of theophylline similar to the HPMC and therefore may have use as an alternative excipient in regions where Sesamum radiatum can be easily cultivated

Egyptian Satirical Graphics on Social Media after the Arab Spring

This paper investigates Egyptian digital forms of satirical graphics mostly published on Social Media by cartoonists and amateurs after the Arab Spring. These include: webcomics memes, and Graphic Interchange Format (GIF). The paper will focus on two Facebook pages as a case study: Asa7be Sarcasm Society and Islam Gawish's Elwarka/The Paper. With this aim in mind, I will show how these forms of political satire incorporate, cinema, theatre, religion, western elements and pan-Arabism to express a political view in a creative manner to appeal to a more diverse and broad audience. The first part of the paper provides a brief discussion of Arabic/Egyptian satire with specific reference to political cartoons and Egyptian youth activism on social media (specifically Facebook). It will also introduce the concept of satirical graphics and some of its concrete applications. The second part describes the methodology and provides an analysis of the case study

Association of CYP2A6 activity and nicotine reinforcement in daily smokers

Abstrac

Impact of CYP2A6 Activity on Nicotine Reinforcement and Cue Reactivity in Daily Smokers

Introduction: Variation in CYP2A6, the primary enzyme responsible for nicotine metabolism, is associated with nicotine dependence, cigarette consumption, and abstinence outcomes in smokers. The impact of CYP2A6 activity on nicotine reinforcement and tobacco cue-reactivity, mechanisms that may contribute to these previous associations, has not been fully evaluated. Aims and Methods: CYP2A6 activity was indexed using 3 genetic approaches in 104 daily smokers completing forced choice and cue-induced craving tasks assessing nicotine reinforcement and tobacco cue-reactivity, respectively. First, smokers were stratified by the presence or absence of reduced/loss-of-function CYP2A6 gene variants (normal vs. reduced metabolizers). As nicotine metabolite ratio (NMR) is a reliable biomarker of CYP2A6 activity, our second and third approaches used additional genetic variants identified in genome-wide association studies of NMR to create a weighted genetic risk score (wGRS) to stratify smokers (fast vs. slow metabolizers) and calculate a wGRS-derived NMR. Results: Controlling for race and sex, normal metabolizers (vs. reduced) selected a greater proportion of puffs from nicotine-containing cigarettes (vs. denicotinized) on the forced-choice task (p = .031). In confirmatory analyses, wGRS-based stratification (fast vs. slow metabolizers) produced similar findings. Additionally, wGRS-derived NMR, which correlated with actual NMR assessed in a subset of participants (n = 55), was positively associated with the proportion of puffs from nicotine-containing cigarettes controlling for race and sex (p = .015). None of the CYP2A6 indices were associated with tobacco cue-reactivity in minimally deprived smokers. Conclusions: Findings suggest increased nicotine reinforcement is exhibited by smokers with high CYP2A6 activity, which may contribute to heavier smoking and poorer cessation outcomes previously reported in faster metabolizers. Implications: CYP2A6 activity is a key determinant of smoking behavior and outcomes. Therefore, these findings support the targeting of CYP2A6 activity, either therapeutically or as a clinically relevant biomarker in a precision medicine approach, for tobacco use disorder treatment

The synthesis of alkylated or acylated nitroarene cyclopentadienylironcomplexes:

Time-dependent oxidation of η6-alkylaniline-η5-cyclopentadienyliron hexafluorophosphates, 17–32, allows for the preparation of nitrobenzene complexes with alkyl 33–48 or keto 49 substituents. Alkylnitroarene complexes are prepared by the oxidation of their corresponding aniline complexes with H2O2 in CF3CO2H for 20 min. An increase in the reaction time to 24 h gives rise to nitroarene complexes with keto substituents in lower yields. The use of nitroarenes as starting materials in the synthesis of alkanoates is of importance since it allows for the preparation of a large number of this class of compounds with a variety of alkyl substituents. Two different approaches have been utilized to allow for the synthesis of alkanoates. The first approach involves nucleophilic aromatic substitution of alkylnitrobenzene complexes with ethyl alkylacetoacetates followed by demetallation to give the alkanoates. This methodology allows for the preparation of these esters with a variety of alkyl substituents in either the meta or para positions. Another route outlines the reaction of phenylsulfonylacetonitrile with nitroarene complexes to prepare alkanoic acid precursors with alkyl substituents in the ortho, meta and para positions. The preparation of a larger pool of nitroarene complexes clearly demonstrates the advantage of using the cyclopentadienyliron arene complexes in the synthesis of alkanoates or their precursors, arylated phenylsulfonylacetonitriles, over traditional synthetic routes

Anatomical and Physiological Performance of Jojoba Treated with Proline under Salinity Stress Condition

A field trial study was conducted for two consecutive seasons 2020 and 2021 in approximately 8-month-old jojoba plants to evaluate the physiological responses following salt treatment and the role of proline as a foliar application to enhance jojoba tolerance to salinity stress. Jojoba plants were irrigated once a week for four months with diluted seawater in concentrations of 5000, 10,000, and 15,000 ppm and tap water (control). Anti-stress proline was applied four times throughout the experiment, the first at the beginning of the experiment and another three times at 30-day intervals, at concentrations of 0, 300, and 450 ppm. The effect of proline treatments on jojoba plant behavior includes growth vegetative characteristics, namely plant height increase percentage (PHIP), shoot number increase percentage (NSIP), stem diameter increase percentage (SDIP), number of leaves, leaf thickness, leaf area, and fresh and dry weights of leaves, and chemical characteristics, namely chlorophyll a and b, total chlorophyll, carotenoids, leaf mineral contents (N, P, K, Na, and Cl), total phenolic content (TPC), and proline concentration. Moreover, the impacts of proline on hydrogen peroxide (H2O2), superoxide anion (O2&bull;&minus;), malondialdehyde (MDA), and ion leakage (IL) under salinity stress were investigated. Briefly, proline at 450 ppm enhanced all studied growth and physiological characteristics and promoted the antioxidant system of jojoba plants compared with the control and other treatments. The anatomical structure of leaves was also examined, and favorable variations in the anatomical structure were detected in the stressed and proline-treated plants. Exogenous application of proline enhanced most of this anatomical characteristic of jojoba leaf under saline stress. In conclusion, proline as a foliar application at 450 ppm under salinity stress of 10,000 ppm enhances jojoba tolerance to salinity stress by modifying the physicochemical and morphological characteristics of jojoba plants