CYP3A4 and CYP3A5 genotyping by Pyrosequencing

BACKGROUND: Human cytochrome P450 3A enzymes, particularly CYP3A4 and CYP3A5, play an important role in drug metabolism. CYP3A expression exhibits substantial interindividual variation, much of which may result from genetic variation. This study describes Pyrosequencing assays for key SNPs in CYP3A4 (CYP3A4*1B, CYP3A4*2, and CYP3A4*3) and CYP3A5 (CYP3A5*3C and CYP3A5*6). METHODS: Genotyping of 95 healthy European and 95 healthy African volunteers was performed using Pyrosequencing. Linkage disequilibrium, haplotype inference, Hardy-Weinberg equilibrium, and tag SNPs were also determined for these samples. RESULTS: CYP3A4*1B allele frequencies were 4% in Europeans and 82% in Africans. The CYP3A4*2 allele was found in neither population sample. CYP3A4*3 had an allele frequency of 2% in Europeans and 0% in Africans. The frequency of CYP3A5*3C was 94% in Europeans and 12% in Africans. No CYP3A5*6 variants were found in the European samples, but this allele had a frequency of 16% in the African samples. Allele frequencies and haplotypes show interethnic variation, highlighting the need to analyze clinically relevant SNPs and haplotypes in a variety of ethnic groups. CONCLUSION: Pyrosequencing is a versatile technique that could improve the efficiency of SNP analysis for pharmacogenomic research with the ultimate goal of pre-screening patients for individual therapy selection

Genetic polymorphisms in MDR1, CYP3A4 and CYP3A5 genes in a Ghanaian population: a plausible explanation for altered metabolism of ivermectin in humans?

<p>Abstract</p> <p>Background</p> <p>Ivermectin, a substrate of multidrug resistance (MDR1) gene and cytochrome P450 (CYP) 3A4, has been used successfully in the treatment of onchocerciasis in Ghana. However, there have been reports of suboptimal response in some patients after repeated treatment. Polymorphisms in host MDR1 and CYP3A genes may explain the observed suboptimal response to ivermectin. We genotyped relevant functional polymorphisms of MDR1 and CYP3A in a random sample of healthy Ghanaians and compared the data with that of ivermectin-treated patients with a view to exploring the relationship between suboptimal response to ivermectin and MDR1 and CYP3A allelic frequencies.</p> <p>Methods</p> <p>Using PCR-RFLP, relevant polymorphic alleles of MDR1 and CYP3A4 genes were analysed in 204 randomly selected individuals and in 42 ivermectin treated patients.</p> <p>Results</p> <p>We recorded significantly higher MDR1 (3435T) variant allele frequency in suboptimal responders (21%) than in patients who responded to treatment (12%) or the random population sample (11%). <it>CYP3A4*1B</it>, <it>CYP3A5*3 </it>and <it>CYP3A5*6 </it>alleles were detected at varied frequencies for the sampled Ghanaian population, responders and suboptimal responders to ivermectin. <it>CYP3A5*1/CYP3A5*1 </it>and <it>CYP3A5*1/CYP3A5*3 </it>genotypes were also found to be significantly different for responders and suboptimal responders. Haplotype (*1/*1/*3/*1) was determined to be significantly different between responders and suboptimal responders indicating a possible role of these haplotypes in treatment response with ivermectin.</p> <p>Conclusion</p> <p>A profile of pharmacogenetically relevant variants for MDR1, CYP3A4 and CYP3A5 genes has been generated for a random population of 204 Ghanaians to address the scarcity of data within indigenous African populations. In 42 patients treated with ivermectin, difference in MDR1 variant allele frequency was observed between suboptimal responders and responders.</p

Quality Evaluation of Lemon Cordial Stored at Different Times with Microwave Heating (Pasteurization)

Consumer interest in food quality and safety has shifted over time, as consumers increasingly prefer minimally processed items. As a result, numerous non-thermal approaches have been implemented due to their potential to preserve the nutritional profile of products along with lengthening their storability. Microwaving, a green processing technique, volumetrically heats the product because of the interactions developed between charged ions, polar water molecules of foodstuff and the incoming electromagnetic waves. The study was mapped out to investigate the effect of microwave exposure time (60, 90 and 120 s) at fixed power (1000 W) and frequency (2450 MHz) on physicochemical properties, phytochemical constituents, antioxidant potential and microbial counts of lemon cordial stored at refrigerated temperature (4 ± 2 °C). The mentioned parameters were analyzed after an interval of 30–90 days. Statistical findings illustrated a highly significant (p ≤ 0.01) impact of microwave treatment and storage on titratable acidity, pH, total soluble solids, total phenolic contents, total flavonoids contents, antioxidant potential and total plate count. Sample microwaved for 120 s showed the highest pH values (2.45 ± 0.050), total soluble solids (56.68 ± 2.612 °B) and antioxidant activity (1212.03 ± 716.5 µg—equivalent of ascorbic acid per 100 mL of cordial); meanwhile, it exhibited the lowest total plate counts (1.75 ± 0.144 Log 10 CFU/mL). Therefore, microwaving can be suggested as a suitable alternate to traditional pasteurization techniques as well as to chemical preservatives

Utilization of Active Edible Films (Chitosan, Chitosan Nanoparticle, and CaCl2) for Enhancing the Quality Properties and the Shelf Life of Date Palm Fruits (Barhi Cultivar) during Cold Storage

Interest in edible coatings applications has progressively developed towards improving the quality and shelf life of climacteric fruits. This study aimed to investigate the influence of pre-harvest treatments (chitosan, chitosan nanoparticle, and CaCl2) on the physicochemical and quality attributes of Barhi date palm fruits during storage periods. Different pre-harvest treatments (control, chitosan 1, 2, and 3 g/L, CaCl2 1, 2, and 3 g/L, nano-chitosan 1, 2, and 3 cm3 /L) were applied. The results showed that all treatments were effective for enhancing the fruit quality, with increasing total soluble solids and total sugars, decreasing weight loss, discarded total acidity, and total soluble tannins compared to the control treatment. Additionally, the results revealed that the highest percentage of TSS was obtained in control fruits (35.78%). Meanwhile, the lowest mean values were obtained from chitosan nanoparticle at 3 cm3/L (33.91%). Treatments with chitosan nanoparticle at 3 cm3/L and CaCl2 at 3 g/L gave the statistically highest values of total tannins (0.225 and 0.220, respectively). The optimal treatment involved spraying the fruit with 3 cm3/L of nano-chitosan or 3 g/L of CaCl2 to increase the fruit quality and the shelf life of Barhi dates. The results indicated that weight loss was negatively linked with the moisture content and firmness, while decay had a strong positive relationship with the Rutab index and a negative correlation with the moisture content. Furthermore, the Rutab index was negatively associated with the total tannins and total chlorophyll