A comprehensive review on fructosyl peptide oxidase as an important enzyme for present hemoglobin A1c assays.

Glycated proteins are generated by binding of glucose to the proteins in blood stream through a nonenzymatic reaction. Hemoglobin A1c (HbA1c) is a glycated protein with glucose at the N-terminal of ß-chain. HbA1c is extensively used as an indicator for assessing the blood glucose concentration in diabetes patients. There are different conventional clinical methods for the detection of HbA1c. However, enzymatic detection method has newly obtained great attention for its high precision and cost-effectiveness. Today, fructosyl peptide oxidase (FPOX) plays a key role in the enzymatic measurement of HbA1c, and different companies have marketed HbA1c assay systems based on FPOX. Recent investigations show that FPOX could be used in assaying HbA1 without requiring HbA1c primary digestion. It could also be applied as a biosensor for HbA1c detection. In this review, we have discussed the recent improvements of FPOX properties, different methods of FPOX purification, solubility, and immobilization, and also the use of FPOX in HbA1c biosensors.

Engineering erucic acid biosynthesis in camelina (Camelina sativa) via FAE1 gene cloning and antisense technology.

Oil seeds now make up the world's second-largest food source after cereals. In recent years, the medicinal- oil plant Camelina sativa has attracted much attention for its high levels of unsaturated fatty acids and low levels of saturated fatty acids as well as its resistance to abiotic stresses. Improvement of oil quality is considered an important trait in this plant. Erucic acid is one of the fatty acids affecting the quality of camelina oil. Altering the fatty acid composition in camelina oil through genetic manipulation requires the identification, isolation, and cloning of genes involved in fatty acid biosynthesis. The Fatty Acid Elongase 1 (FAE1) gene encodes the enzyme ß-ketoacyl CoA synthase (KCS), a crucial enzyme in the biosynthesis of erucic acid. In this study, the isolation and cloning of the FAE1 gene from Camelina sativa were conducted to construct an antisense structure. The molecular homology modeling of DFAE1 proteins using the SWISS-MODEL server on ExPASy led to the generation of the 3D structures of FAE1 and DFAE1 proteins. The GMQE values of 0.44 for FAE1 and 0.08 for DFAE1 suggest high accuracy in the structural estimation of these genes. The fragments were isolated from the DNA source of the genomic Soheil cultivar with an erucic acid content of about 3% (in matured seeds) using PCR. After cloning the FAE1 gene into the Bluescript II SK+ vector and sequencing, the resulting fragments were utilized to construct the antisense structure in the pBI121 plant expression vector. The approved antisense structure was introduced into the Camelina plant using the Agrobacterium-mediated method, with optimization of tissue culture and gene transfer conditions. This approach holds potential to advance our knowledge of fat biosynthesis, leading to potential improvements in oil quality in Camelina sativa.

Control of erucic acid biosynthesis in Camelina (Camelina sativa) by antisense technology.

Oil seeds now make up the world's second-largest food source after cereals. In recent years, the medicinal- oil plant Camelina sativa has attracted much attention for its high levels of unsaturated fatty acids and low levels of saturated fatty acids as well as its resistance to abiotic stresses. Improvement of oil quality is considered an important trait in this plant. Erucic acid is one of the fatty acids affecting the quality of camelina oil. Altering the fatty acid composition in camelina oil through genetic manipulation requires the identification, isolation, and cloning of genes involved in fatty acid biosynthesis. The Fatty Acid Elangase 1 (FAE1) gene encoded the enzyme ß-ketoacyl COA synthase (KCS), which is a key factor in the biosynthesis of erucic acid. In this study, isolation and cloning of the FAE1 from the Camelina sativa were performed to prepare an antisense structure.  The fragments were isolated from the DNA source of the genomic Soheil cultivar with an erucic acid content of about 3% (in matured seeds) using PCR. After cloning FAE1 into the Bluescriprt II SK+ vector and sequencing, these fragments were used for the preparation of antisense structure in the pBI121 plant expression vector. The approved structure was transferred to the camelina plant via the Agrobacterium-mediated method. Also, the conditions of tissue culture and gene transfer were optimized. Moreover, the erucic acid content of the immature seeds of T0 transgenic plants was analyzed with gas chromatography (GC). Results showed significant changes in erucic acid levels of two control plants (0.88%), while two lines of the RFAE1 transgenic plants showed a decrease of approximately 0% in erucic acid level. It can be concluded that the antisense structure can be effective in reducing erucic acid.

Identification of Thiobacillus bacteria in agricultural soil in Iran using the 16S rRNA gene.

Thiobacillus, as useful soil bacteria, plays an important role in sulfur cycling. The purpose of this study was to identify the species Thiobacillus thioparus, Thiobacillus novellas and Thiobacillus denitrificans in rainfed and irrigated lands soil in Ajabshir, Ilam, Qorveh, Rojintaak, Sonqor, Kermanshah and Research Farm of Razi University in Iran. Sampling was performed as randomized completely with three replications at depth of 0-30 cm. The Thiobacillus species were determined via 16S rRNA characteristics. The results of agarose gel electrophoresis indicated that T. thioparus was the highest amount in the irrigated land in Research Farm and its lowest amount was in the Rojintaak rainfed land. These species not found in four locations and conditions including the Ajabshir irrigated, Qorveh rainfed, Research Farm rainfed and Rojintaak irrigated lands. The results of the T. novellas indicated that this was found in Ilam irrigated, Qorveh rainfed, Research Farm irrigated, Rojintaak irrigated and Rojintaak rainfed lands. The highest and lowest amount of T. novellas was indicated in the Rojintaak and Ilam irrigated lands respectively. The T. denitrificans gene showed that this bacterium was observed only in both samples of Ajabshir. Our study showed that Thiobacillus was not detected in all of the soils. If sulfur fertilizer is given to the soil without this bacterium, it is necessary to use sulfur fertilizer with Thiobacillus bacteria inoculation for better sulfur oxidation.

Study of association between interleukin-8 - 845 T/C and + 781 C/T polymorphisms with periodontitis disease among population from Western Iran.

Periodontitis is a chronic inflammatory disease that influences the protective tissues of teeth. IL-8, a member of the chemokine super-family, plays vital roles in the pathogenesis of periodontitis with activation and migration of neutrophils in inflammatory regions. The purpose of present study was to evaluate the association of interleukin-8 - 845 T/C and + 781 C/T polymorphisms with periodontitis in an Iranian population. A total of 65 patients with periodontitis including 18 patients with chronic periodontitis and 47 patients with aggressive periodontitis and 55 controls were enrolled into our study. Interleukin-8 - 845 T/C and + 781 C/T polymorphisms were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. For + 781C/T locus, in the dominant genetic model there was a significant difference between TT vs. CC + CT genotypes that significantly had a protective role against periodontitis disease with a value of 0.38 (95% CI 0.16-0.90, p = 0.02). Also, the analysis of results showed a significant positive association between the distribution of IL-8 - 845 T/C alleles and the risk of periodontitis disease (χ2 = 6.2, p = 0.01) that presence of C allele of IL-8 - 845 increased the risk of periodontitis disease by 9.08-fold [OR 9.08 (95% CI 1.14-72.12, p = 0.03)]. In conclusion, our results demonstrate a positive association between distribution of IL-8 - 845 T/C alleles and risk of periodontitis disease.

Development of PCR primer systems for amplification of 16S-rDNA to detect of Thiobacillus spp.

Thiobacillus is a genus of Gram-negative, rod-shaped and autotrophic Betaproteobacteria. They catalyze the dissimilatory oxidation of elemental sulfur and reduced inorganic sulfur compounds. Whereas more than 30 species have been known in this genus, most were never reliably or effectively published. The rest were either reclassified into Thiomonas, Paracoccus, Starkeya, Sulfuriferula, Halothiobacillus, Thermithiobacillus or Acidithiobacillus, were lost from culture. Most of Thiobacillus species are obligate autotrophs via elementary sulfur, thiosulfate or polythionates as energy sources. Based on 16S ribosomal RNA sequence analysis, many members of Thiobacillus have been reclassified. A system was developed for the detection of Thiobacillus bacteria by the amplification of specific 16S ribosomal RNA sequence gene (16S rDNA) fragments with PCR. Primer sequences were designed for the amplification of fragments of 16S rDNA.