Purification and characterization of genetic variants of 6-phosphogluconate dehydrogenase

This paper describes the physicochemical characteristics in partially purified enzyme on subjects with the Pd A, Pd AB, and Pd B variants of 6-phosphogluconic dehydrogenase (6PGD). For these studies, whole blood was purified about 225-fold using ion exchange chromatography on DEAE cellulose column and fractionation with ammonium sulfate. 6PGD emerges as a single peak between 0.01 m and 0.1 m phosphate buffer on the column and is precipitated in the 55–80% fraction of ammonium sulfate. This purified enzyme can be stored frozen for several months without appreciable loss of activity and contains no detectable activity of glucose 6-phosphate dehydrogenase and glutathione reductase. The three variants of partially purified 6PGD varied from each other in two respects. The transitional temperature is 47.8 C for Pd A, 45.4 C for Pd AB, and 41.1 C for Pd B. The K _m for 6PGA is 30 m for Pd A, 21 m for Pd AB, and 15 mfor Pd B. These observations add further strength to the concept that the polymorphism in 6PGD represents alterations in either the configuration or structure of the protein molecule itself.Supported by grants from the Chicago Community Trust and the U.S. Public Health Service (Tl-AM-5186).     

Association of the <Emphasis Type="Italic">rs1870634</Emphasis> Variant in Long Intergenic Non-protein Coding RNA 841 with Coronary Artery Disease: A GWAS-Replication Study in an Iranian Population

Recent genome-wide association studies (GWAS) identified a list of single-nucleotide polymorphisms (SNPs) associated with coronary artery disease (CAD). Replication of GWAS findings in different population corroborated the observed association in the parent GWAS. In this study, we aimed to replicate the association of rs1870634, a GWAS identified SNP, to CAD in an Iranian population. The study population consisted of 267 subjects undergoing coronary angiography coronary angiography including 155 CAD patients and 112 non-CAD age- and gender-matched controls. The genotype determination of rs1870634 SNP performed using high-resolution melting analysis (HRM) technique. Our results revealed that the GG genotype frequency was significantly higher in CAD patients compared with controls (P?=?0.03). The results of binary logistic regression suggested that this genotype was significantly associated with CAD risk adjustment for age, BMI, sex, TC, and LDL-C lipid levels (OR of 2.78, 95% CI (1.10–7.01), P?=?0.03). Moreover, our results showed that the GG+TG genotypes were 2.52 times more likely to develop CAD (95% CI 1.05–6.03) than TT genotype carriers after adjusting for age, sex, and lipid profiles (P?=?0.037). These data showed that the GG genotype could be associated with increased risk of CAD in a sample of Iranian population.     

Deferoxamine Preconditioning of Neural-Like Cells Derived from Human Wharton’s Jelly Mesenchymal Stem Cells as a Strategy to Promote Their Tolerance and Therapeutic Potential: An In Vitro Study

Transplantation of neural-like cells is considered as a promising therapeutic strategy developed for neurodegenerative disease in particular for ischemic stroke. Since cell survival is a major concern following cell implantation, a number of studies have underlined the protective effects of preconditioning with hypoxia or hypoxia mimetic pharmacological agents such as deferoxamine (DFO), induced by activation of hypoxia inducible factor-1 (HIF-1) and its target genes. The present study has investigated the effects of DFO preconditioning on some factors involved in cell survival, angiogenesis, and neurogenesis of neural-like cells derived from human Wharton’s jelly mesenchymal stem cells (HWJ-MSCs) in presence of hydrogen peroxide (H₂O₂). HWJ-MSCs were differentiated toward neural-like cells for 14 days and neural cell markers were identified using immunocytochemistry. HWJ-MSC-derived neural-like cells were then treated with 100 µM DFO, as a known hypoxia mimetic agent for 48 h. mRNA and protein expression of HIF-1 target genes including brain-derived neurotrophic factors (BDNF) and vascular endothelial growth factor (VEGF) significantly increased using RT-PCR and Western blotting which were reversed by HIF-1α inhibitor, while, gene expression of Akt-1, Bcl-2, and Bax did not change significantly but pAkt-1 was up-regulated as compared to poor DFO group. However, addition of H₂O₂ to DFO-treated cells resulted in higher resistance to H₂O₂-induced cell death. Western blotting analysis also showed significant up-regulation of HIF-1α, BDNF, VEGF, and pAkt-1, and decrease of Bax/Bcl-2 ratio as compared to poor DFO. These results may suggest that DFO preconditioning of HWJ-MSC-derived neural-like cells improves their tolerance and therapeutic potential and might be considered as a valuable strategy to improve cell therapy.     

Detection of adulteration in Iranian saffron samples by <Superscript>1</Superscript>H NMR spectroscopy and multivariate data analysis techniques

Introduction

The high market value of saffron (Crocus sativus L.) has made it an attractive candidate for adulteration. Safflower (Carthamus tinctorius L.) and tartrazine are among the most common herbal and synthetic foreign materials that may be added to pure saffron for the purpose of adulteration. In spite of encouraging advances achieved in the identification of adulteration in saffron samples, the lack of a simple method with sufficient power for discrimination of pure high grade saffron from meticulously adulterated saffron samples persuaded us to perform this study.

Objectives

In this work, we show that ¹H NMR spectroscopy together with chemometric multivariate data analysis methods can be used for the detection of adulteration in saffron.

Methods

Authentic Iranian saffron samples (n?=?20) and adulterated samples that were prepared by adding either different quantities of natural plant materials such as safflower, or synthetic dyes such as tartrazine or naphthol yellow to pure saffron (n?=?22) composed the training set. This training set was used to build multivariate Principal Component Analysis (PCA) and Partial Least Squares Discriminant Analysis (PLS-DA) models. The predictive power of the PLS-DA model was validated by testing the model against an external dataset (n?=?13).

Results

PCA and PLS-DA models could both discriminate between the authentic and adulterated samples, and the external validation showed 100% sensitivity and specificity for predicting the authenticity of suspicious samples. Peaks specific to authentic and adulterated samples were also characterized. Proximity of samples with unknown adulteration status to the samples adulterated with known compounds in the PCA provided insight regarding the identity of the adulterant in the suspicious samples. Furthermore, the authentic samples could be distinguished based on their cultivation site.

Conclusion

The present study demonstrates that the application of ¹H NMR spectroscopy coupled with multivariate data analysis is a suitable approach for detection of adulteration in saffron specimens. Outstanding sensitivity and specificity of the PLS-DA model in discriminating the authentic from adulterated samples in external validation confirmed the high predictive power of the model. The advantage of the present method is its power for detecting a wide spectrum of adulterants, ranging from synthetic dyes to herbal materials, in a single assay.

     

Variation of DAT1 VNTR alleles and genotypes among old ethnic groups in Mesopotamia to the Oxus region

Variation of a VNTR in the DAT1 gene in seven ethnic groups of the Middle East was used to infer the history and affinities of these groups. The populations consisted of Assyrian, Jewish, Zoroastrian, Armenian, Turkmen, and Arab peoples of Iran, Iraq, and Kuwait. Three hundred forty subjects from these seven ethnic groups were screened for DAT1. DAT1 VNTR genotyping showed 3, 6, 7, 8, 9, 10, 11, and 12 alleles in the samples. Analysis of these data revealed differentiation and relationship among the populations. In this region, which covers an area of 2-2.5 million km2, the influence of geography and especially of linguistic characteristics has had potentially major effects on differentiation. Religion also has played a major role in imposing restrictions on some ethnic groups, who as a consequence have maintained their community. Overall, these ethnic groups showed greater heterogeneity compared to other populations.