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Understanding of cellular responses to ionizing radiation (IR) is essential for the development of predictive markers useful for assessing human exposure. Biological markers of exposure to IR in human populations are of great interest for assessing normal tissue injury in radiation oncology and for biodosimetry in nuclear incidents and accidental radiation exposures. Traditional radiation exposure biomarkers based on cytogenetic assays (biodosimetry), are time-consuming and do not provide results fast enough and requires highly trained personnel for scoring. Hence, the development of rapid biodosimetry methods is one of the highest priorities. Exposure of cells to IR activates multiple signal transduction pathways, which result in complex alterations in gene-expression. Real-time quantitative reverse transcription-polymerase chain reaction (RT-qPCR) has become the benchmark for the detection and quantification of RNA targets and is being utilized increasingly in monitoring the specific genes with more accurately and sensitively. This review evaluates the RT-qPCR as a biodosimetry method and we investigated the papers from 2000 up to now, which identified the genes-expression related the DNA repair, cell cycle checkpoint, and apoptosis induced by ionization radiation in peripheral blood and determined as biodosimeters. In conclusion, it could be say that RT-qPCR technique for determining the specific genes as biodosimeters could be a fully quantitative reliable and sensitive method. Furthermore, the results of the current review will help the researchers to recognize the most expressed genes induced by ionization radiation.

In this article, various methods including soxhlet, Bligh & Dyer (B&D), and ultrasonic-assisted B&D were investigated for the extraction of lipid from algal species Chlorella vulgaris. Relative polarity/water content and impolar per polar ratios of solvents were considered to optimize the relative proportions of each triplicate agent by applying the response surface method (RSM). It was found that for soxhlet, hexane-methanol (54-46%, respectively) with total lipid extraction of 14.65% and chloroform-methanol (54-46%, respectively) with the extraction of 19.87% lipid were the best set of triplicate where further addition of acetone to the first group and ethanol to the second group did not contributed to further extraction. In B&D, however, chloroform-methanol-water (50%-35%-15%, respectively) reached the all-time maximum of 24%. Osmotic shock as well as ultrasonication contributed to 3.52% of further extraction, which is considered to promote the total yield up to almost 15%. From the growth data and fatty acid analysis, the applied method was assessed to be appropriate for biodiesel production with regard to selectivity and extraction yield., (© 2014 International Union of Biochemistry and Molecular Biology, Inc.)

Background: Gestational diabetes mellitus (GDM) affects nearly 5% of pregnancies. Significant proportion of the women with previous GDM develops type 2 diabetes mellitus (T2DM) in the next years, which indicates a higher risk in them than in the general population., Objectives: We conducted this study to determine the risk factors and incidence of abnormal glucose level and metabolic syndrome (MetS) in women with a history of GDM in a long period after delivery in our region., Patients and Methods: We extracted the demographic characteristics of 110 women with GDM who had delivered during 2004 - 2010 in three main hospitals of Zanjan City, Iran. The patients were recalled to perform oral glucose tolerance test (OGTT) and other necessary tests for MetS diagnosis. Anthropometric measurements were recorded of all the participants., Results: In this study, 110 women with a history of GDM were studied at one to six years since delivery. Among these women, 36 (32.7%) developed T2DM and 11 (10%) had impaired fasting glucose (IFG) or impaired glucose tolerance (IGT). Moreover, 22 women (20%) had developed MetS. among those with abnormal results in glycemic test, 93.6% had fasting blood sugar (FBS) ≥ 95 mg/dL (≥ 5.27 mmol/L)at the time of GDM diagnosis in the index pregnancy that was significantly higher than the normal glycemic test (NGT) group with 42.9% being affected (OR, 19.55; P < 0.0001). There was a significant difference between those with abnormal results and NGT group in interval between delivery and performing laboratory tests (27 ± 18.8 and 18.5 ± 17.7 months, respectively; OR, 1.02; P = 0.02). No insulin use during pregnancy was discovered as a protective factor in women with a history of GDM (OR, 0.35; P = 0.01). Those with abnormal results were significantly different from NGT group in the number of parities (2.61 ± 1.4 vs. 2.05 ± 1.1, respectively; OR, 1.4; P = 0.03). The most common component of MetS among women with a history of GDM was FBS > 100 mg/dL (> 5.55 mmol/L)., Conclusions: Regarding the high incidence of the T2DM and MetS among women with a history of GDM, they should be screened at a regular interval for diabetes and other cardiovascular risk factors.

Sulfur mustard (SM) affects divergent cellular pathways including cell cycle, apoptosis, necrosis, and inflammatory responses. SM-induced lesions in skin include late-onset hyper-pigmentation, xerosis, and atrophy. It seems that TGF-b signaling pathway is a major player for SM pathogenesis. Here, we have employed a real-time polymerase chain reaction (PCR) approach to evaluate the expression alterations of all TGF-b variants and their receptors in skin biopsies obtained from 10 Iran-Iraq war veterans. Using specific LNA primers, the expression alteration of a TGF-bR2 regulator, miR-20a, and TGF-b downstream target, miR-21, was also assessed in the same samples Our real-time PCR data revealed a significant down-regulation of TGF-b1 and TGF-bR2, the major mediators of TGF-b signaling pathway, in skin biopsies of SM-exposed patients (p = 0.0015 and p = 0.0115, respectively). Down-regulation of TGF-b signaling pathway seems to contribute in severe inflammation observed in SM-exposed patients' tissues. MiR-20a and miR-21, as two important TGF-b associated microRNAs (miRNAs), were also down-regulated in SM-exposed skin lesions, compared to those of control group (p = 0.0003). Based on our findings, these miRNAs could be directly or indirectly involve in the pathogenesis of SM. Altogether, our data suggest the suitability of TGF-b1, TGF-bR2, as well as miR-20a and miR-21 as potential biomarkers for diagnosis and treatment of SM-exposed patients.

BACKGROUND Furazolidone has been used as an alternative for clarithromycin or metronidazole in Helicobacterpylori (H.pylori) eradication regimens. In Iran, 14-day Furazolidone-containing quadruple regimens have shown promising eradication rates, but short-course, low dose therapies are always attractive. Therefore, we designed a study to compare the efficacy of two 10-day triple regimens containing moderate and high dose furazolidone for H.pylori eradication. METHODS Two hundred and ten patients with peptic ulcer disease who were naïve to H.pylori treatment were included. They were randomized into 2 groups: 105 patients received omeprazole 20mg, amoxicillin 1000mg, and furazolidone 200mg(OAF-400), all twice a day for ten days.And the remaining 105 patients received omeprazole 20mg twice a day, amoxicillin 1000mg twice a day and furazolidone 200mg three times a day for ten days(OAF-600). Urease breath test was performed 8 weeks after the treatment to confirm H. pylori eradication. RESULTS The intention-to-treat eradication rate was 76.19% in group OAF-400 and 80.95% in group OAF-600 (pp=0.38). Per protocol eradication rates were 81.63% and 89.47%, respectively (p= 0.11).Severe adverse effects were reported by 8.6% of the patients in group OAF-400 and 5.7% of the patient in group OAF-600 (p=0.1). However, the total side effects (including mild, moderate, and severe ones) were significantly more prevalent in the OAF-600 group (p=0.001). CONCLUSION None of our triple furazolidone-based regimens (moderate- and high-dose) could achieve the standard eradication rate, and therefore, cannot be considered as a suitable option for first-line treatment.

Objective: To assess the effect of ginger consumption on glycemic status, lipid profile and some inflammatory markers in patients with type 2 diabetes mellitus., Methods: In a double-blinded, placebo-controlled clinical trial, 70 type 2 diabetic patients were enrolled. They allocated randomly into ginger group and control group. They consumed 1600 mg ginger versus 1600 mg wheat flour placebo daily for 12 weeks. Serum sugar, lipids, CRP, PGE2 and TNFα were measured before and after intervention., Results: Ginger reduced fasting plasma glucose, HbA1C, insulin, HOMA, triglyceride, total cholesterol, CRP and PGE₂ significantly compared with placebo group (p < 0.05). There were no significant differences in HDL, LDL and TNFα between two groups (p > 0.05)., Conclusion: Ginger improved insulin sensitivity and some fractions of lipid profile, and reduced CRP and PGE₂ in type 2 diabetic patients. Therefore ginger can be considered as an effective treatment for prevention of diabetes complications.

Productivity of sunflower (Helianthus annuus L.), the fourth most important oilseed crop, is strongly dependent on water availability. To search for genetic variation in the ability of roots to grow into drying soil, 16 sunflower lines were screened in 2 years field experiments by imposing drought stress at flowering stage. The results differentiated RGK 21 and BGK 329 as the most sensitive and tolerant lines respectively. The time course physiological assay of these lines at seedling stage revealed roots as the most affected organ 6 days after imposing drought stress. A proteomics approach was adapted for investigating of differential changes in roots proteome under contrasting moisture regimes. Protein spots with significant changes in protein abundance were identified by nano LC-MS/MS. The results indicated that under drought stress relative abundance of metabolism related proteins were decreased in both sensitive and tolerant lines. Abundance of energy and disease/defence related proteins were decreased in the sensitive but increased in the tolerant line. The results indicate that changes in energy usage, water transport and ROS scavenging are important mechanisms for maintaining root growth as the soil dries.

Background: Various risk of mortality due to hip fracture has been reported by different studies. There is scarce controlled study on hip fracture mortality from developing countries and no data from Middle East region. The objective of this study is to determine mortality and its risk factors one year after low trauma hip fracture., Methods: One hundred and two patients after hip fracture not caused by high impact injuries or local bone diseases followed up prospectively for one year. Control group consisted of sex and age matched patients admitted to ophthalmology ward for eye surgery. Data about comorbidity obtained from both groups at baseline. Functional state and health-related quality of life for the participants were measured using RDRS-2 and SF-36 questionnaires, respectively., Results: The overall survival was 83% in cases and 92% in controls (log rank test 3.62, df = 1, P = 0.057). Early mortality within the first 6 months of observation was significantly higher in patients than controls (13 in patients vs. 2 in controls) (log rank test 8.84, df = 1, P = 0.003). The risk of mortality in the first year after fracture was significantly and independently associated with age and baseline RDRS score. By the end of follow-up, in the patient group, 55.4% of survivors were able to walk without any assistance and 10.8% were not able to walk., Conclusions: The risk of mortality within the first 6 months of observation was significantly and independently associated with low trauma hip fracture. However, age and baseline RDRS score were independent predictors of mortality in the first year following hip fracture.

The development of meiotic division and associated genetic recombination paved the way for evolutionary changes. However, the secondary and tertiary structure and functional domains of many of the proteins involved in genetic recombination have not been studied in detail. We used the human Dmc1 gene product along with secondary and tertiary domain structures of Escherichia coli RecA protein to help determine the molecular structure and function of maize Dmc1, which is required for synaptonemal complex formation and cell cycle progression. The maize recombinase Dmc1 gene was cloned and characterized, using rice Dmc1 cDNA as an orthologue. The deduced amino acid sequence was used for elaborating its 3-D structure, and functional analysis was made with the CDD software, showing significant identity of the Dmc1 gene product in Zea mays with that of Homo sapiens. Based on these results, the domains and motives of WalkerA and WalkerB as ATP binding sites, a multimer site (BRC) interface, the putative ssDNA binding L1 and L2 loops, the putative dsDNA binding helix-hairpin-helix, a polymerization motif, the subunit rotation motif, and a small N-terminal domain were proposed for maize recombinase Dmc1.

Leaf and internode explants of Withania coagulans Dunal were used to evaluate the effect of growth regulators on the in vitro callus induction and plant regeneration. Two independent experiments were performed to establish callus and plant regeneration. In the first experiment, leaf segments were cultured on MS basal medium fortified with 2, 4-dichlorophenoxyacetic acid (2, 4-D, 2-4 mg L(-1)) with combination of 6-benzyladenine (BA, 0.5-1 mg L(-1)) or kinetin (Kin, 0.5-1 mg L(-1)). Callus initiation was observed best in all media after 14-16 days (100%). Highest callus growth in terms of dry weight (76 +/- 5.34 mg) was observed in MS medium fortified with 2 mg L(-1) 2, 4-D and 0.5 mg L(-1) Kin. In the second experiment, callusing also obtained from internode segments cultured on MS medium with the presence of 2, 4-D (2-4 mg L(-1)) in association with BA (0.25-0.5 mg L(-1)). Best callusing rates (42%) and dry weight (86 +/- 3.68 mg) occurred in the presence of 4 mg L(-1) 2, 4-D with combination of 0.25 mg L(-1) BA. Shoot induction was obtained just from callus induced from internode explants on MS medium containing BA (2 mg L(-1)) with IBA (0.5 mg L(-1)), but the yield of shoot regeneration was unsatisfactory (18-33%). Regenerated shoots were rooted best (100%) on half strength MS medium containing IBA (2 mg L(-1)) and produced 23 roots with an average root length of 6.5 +/- 0.41 cm. Rooted plantlets transferred to soil and sand mixture (2:1) showed 75% survival when transferred to outdoor. The procedure reported here in vitro culture of W. coagulans for first time.

Objectives: To examine the efficacy of combination therapy with levothyroxine and liothyronine in improvement of general health, psychological problems, and metabolic status in primary hypothyroidism., Methods: Seventy-one patients diagnosed with primary hypothyroidism were randomly allocated into two study groups: the first group received usual dose of levothyroxine and the second group received combination of levothyroxine and liothyronine for at least 4 months. The main outcomes were psychosocial problems (Goldberg's General Health Questionnaire, GHQ-28), bodyweight, heart rate, blood pressure, and serum lipid levels., Results: In both groups serum thyroid-stimulating hormone levels remained unchanged compared with baseline. Psychosocial scores, body weight, heart rate, blood pressure, and lipid profile in the two groups remained constant. The only exception was a small but significant reduction in anxiety/insomnia in combined treatment group as compared with monotherapy., Conclusions: The data do not support the hypothesis that combined therapy improves the well-being and general health of patients.

The cultivated potato Solanum tuberosum L. is an autotetraploid with Endosperm Balance Number (EBN) of 4. There are some allotetraploid 2EBN wild species with desirable traits for potato improvement. However, it is very difficult to cross between these two groups due to differences in EBN. Eleven genotypes of autotetraploid cultivated potato including the six cultivars of subsp. Tuberosum (tbr) and five clones of subsp. Andigena (adg) were crossed with two wild allotetraploid species: S. acaule (acl) and S. stoloniferum (sto). For asses the pollen-pistil incompatibility, some ofpollinated flowers were fixed in Carnoy's solution and stained with aniline blue. Evaluation with fluorescent microscope revealed that pre-zygotic pollen-pistil incompatibility in these crosses is frequent. The pistils of both subsp. of cultivated potato were incompatible with pollens of acl but when sto used as staminate parent it was compatible with five cultivars of subsp. Tuberosum, but incompatible with most ofsubsp. Andigena (4 clones). From 739, 4x, 4EBN x 4x, 2EBN crosses we couldn't get any viable seed. In reciprocal crosses, allotetraploid species were pollinated with a mix of pollens from Tuberosum or Andigena clones. The crosses of sto x adg, acl x adg and acl x tbr were compatible but in the case of sto x tbr the situation was not clear. From these crosses 157 fruits were obtained but most of them contained only shrink seed and we find just one hybrid from the sto x tbr combination. The hybrid was tetraploid (2n = 4x = 48) and vigorous with profuse flowering and good fertility. When this hybrid was used as the pistillate parent it was compatible with both parental species.

To date enormous attempts have been devoted to improve Taxol production exploiting various methodologies from bioprocess engineering to biotechnological and synthetic approaches. We have developed a 2-stage suspension cell culture of Taxus baccata L. using modified B5 medium in order to improve cell growth as well as productivity. After callus induction and cell line selection, B5 medium was supplemented with vanadyl sulfate (0.1 mg/l), silver nitrate (0.3 mg/l) and cobalt chloride (0.25 mg/l) at the first day of stage I culture to maximize cell growth. This medium was further supplemented with sucrose (1%) and ammonium citrate (50 mg/l) on day 10 and sucrose (1%) and phenylalanine (0.1 mM) on day 20 (i.e., biomass growth medium). At stage II (day 25), two different concentrations of several elicitors such as methyl jasmonate (10 or 20 mg/l), salicylic acid (50 or 100 mg/l) and fungal elicitor (25 or 50 mg/l) were added to the biomass growth medium with the aim of improving cellular productivity. For morphological analysis, microscopic inspection was carried out during cultivation. Cell-associated and extracellular amount of Taxol were detected and measured using HPLC methodology. At stage I, overall Taxol amount of biomass growth medium was 13.75 mg/l (i.e., 5.6-fold higher than that of untreated B5 control). At stage II, treated cells with methyl jasmonate (10 mg/l), salicylic acid (100 mg/l) and fungal elicitor (25 mg/l) produced the highest amount of Taxol (39.5 mg/l), which is 16-fold higher than that of untreated B5 control (2.45 mg/l). Microscopic analyses of Taxus cells in suspension cultures showed various positional auto-fluorescence showing direct correlation with Taxol production. Our studies revealed that intervallic supplementation of B5 medium with combination of biomass growth factors at stage I and mixture of elicitors at stage II could significantly increase Taxol production. Thus, we suggest that the exploitation of this methodology may improve the production of Taxol since demands for Taxol pharmaceuticals are increasingly growing and resource paucities have limited its direct harvesting from Taxus trees.

Proteolytic cleavage in an exposed loop of human tartrate-resistant acid phosphatase (TRAcP) with trypsin leads to a significant increase in activity. At each pH value between 3.25 and 8.0 the cleaved enzyme is more active. Substrate specificity is also influenced by proteolysis. Only the cleaved form is able to hydrolyze unactivated substrates efficiently, and at pH >6 cleaved TRAcP acquires a marked preference for ATP. The cleaved enzyme also has altered sensitivity to inhibitors. Interestingly, the magnitude and mode of inhibition by fluoride depends not only on the proteolytic state but also pH. The combined kinetic data imply a role of the loop residue D158 in catalysis in the cleaved enzyme. Notably, at low pH this residue may act as a proton donor for the leaving group. In this respect the mechanism of cleaved TRAcP resembles that of sweet potato purple acid phosphatase.

Purple acid phosphatases (PAPs) are a family of binuclear metalloenzymes that catalyze the hydrolysis of phosphoric acid esters and anhydrides. A PAP in sweet potato has a unique, strongly antiferromagnetically coupled Fe(III)-Mn(II) center and is distinguished from other PAPs by its increased catalytic efficiency for a range of activated and unactivated phosphate esters, its strict requirement for Mn(II), and the presence of a mu-oxo bridge at pH 4.90. This enzyme displays maximum catalytic efficiency (k(cat)/K(m)) at pH 4.5, whereas its catalytic rate constant (k(cat)) is maximal at near-neutral pH, and, in contrast to other PAPs, its catalytic parameters are not dependent on the pK(a) of the leaving group. The crystal structure of the phosphate-bound Fe(III)-Mn(II) PAP has been determined to 2.5-A resolution (final R(free) value of 0.256). Structural comparisons of the active site of sweet potato, red kidney bean, and mammalian PAPs show several amino acid substitutions in the sweet potato enzyme that can account for its increased catalytic efficiency. The phosphate molecule binds in an unusual tripodal mode to the two metal ions, with two of the phosphate oxygen atoms binding to Fe(III) and Mn(II), a third oxygen atom bridging the two metal ions, and the fourth oxygen pointing toward the substrate binding pocket. This binding mode is unique among the known structures in this family but is reminiscent of phosphate binding to urease and of sulfate binding to lambda protein phosphatase. The structure and kinetics support the hypothesis that the bridging oxygen atom initiates hydrolysis.

Purple acid phosphatases are metal-containing hydrolases. While their precise biological role(s) is unknown, the mammalian enzyme has been linked in a variety of biological circumstances (e.g., osteoporosis) with increased bone resorption. Inhibition of the human enzyme is a possible strategy for the treatment of bone-resorptive diseases such as osteoporosis. Previously, we determined the crystal structure of pig purple acid phosphatase to 1.55A and we showed that it is a good model for the human enzyme. Here, a study of the pH dependence of its kinetic parameters showed that the pig enzyme is most efficient at pH values similar to those encountered in the osteoclast resorptive space. Based on the observation that phosphotyrosine-containing peptides are good substrates for pig purple acid phosphatase, peptides containing a range of phosphotyrosine mimetics were synthesized. Kinetic analysis showed that they act as potent inhibitors of mammalian and plant purple acid phosphatases, with the best inhibitors exhibiting low micromolar inhibition constants at pH 3-5. These compounds are thus the most potent organic inhibitors yet reported for the purple acid phosphatases.

The genetics of two enzymatic loci, esterase (Est-D) and acid phosphatase (AcP-A), were studied by means of polyacrylamide gel electrophoresis in the fig tree (Ficus carica L.). Two codominant alleles are described at the Est-D locus and four codominant alleles at the AcP-A locus. Heterozygotes at the AcP-A locus have a hybrid band, thus showing that the AcP-A allozymes, are at least dimer molecules. Both loci are independent of the male sterility factor in F. carica. The polymorphism in four natural populations was investigated for both loci. A significant deficiency of heterozygotes was observed.