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Fluoropyrimidines, the major chemotherapeutic agents in various malignancies treatment, are metabolized by dihydropyrimidine dehydrogenase (DPD). DPD deficiency can lead to severe and sometimes fatal toxicity. In the present study, we developed a simple protocol to detect the DPYD*2A variant. Common side effects in patients treated with these drugs were also evaluated in a Kurdish population.We established a reverse-transcriptase polymerase chain reaction (RT-PCR) technique for detection of DPYD*2A. Sanger sequencing was used to confirm the results. 121 Kurdish patients receiving fluoropyrimidine derivatives were enrolled, and clinical information regarding the dosage and toxicity was analyzed.Our RT-PCR method was able to detect one patient with heterozygous state for DPYD*2A (0.8%). The most observed adverse drug reactions were tingling, nausea, and hair loss. The frequency of patients with the toxicity of grade 3 or worse was 6.6%.This was the first study that detect DPYD*2A polymorphism in the Kurdish population. Our method was successfully able to detect the DPYD*2A variant and, due to its simplicity and cost-effectiveness, it may be considered as an alternative to the current methods, especially in developing countries. Our detected polymorphism rate at 0.8% is comparable with other studies. Despite the low rate of DPYD*2A polymorphism, pharmacogenetics assessment before beginning the treatment process is highly recommended due to its association with a high risk of severe toxicity.

Aims: The aim of the present study was to assess and compare the effect of different methods of cooking on B-group vitamins (B1, B2, and B6), As, Cd, Ni, Pb, and some essential elements (Cu, Co, and Mn) in the main local varieties of Iranian rice. Background: Different cooking procedures have been shown to have contradictory benefits in terms of nutritional value by affecting the bioavailability of vitamins and microminerals. Objective: The aim of the present study was to assess and compare the effect of different methods of cooking on B-group vitamins (B1, B2 and B6), As, Cd, Ni, Pb, and some essential elements (Cu, Co, and Mn) in the main local varieties of Iranian rice. Method: The effectiveness of different cooking procedures, including direct cooking (Dampokht) and cooking by washing the rice with water and draining off the water (Abkesh), on B1, B2, and B6 vitamins contents and Cd, As, Pb, Ni, Cu, Co, Mn elements in the different local varieties of Iranian rice was evaluated. For the Dampokht method, the non-rinsed cooked samples were directly used for evaluation, but for the Abkesh method, the cooked rice samples were first rinsed and then evaluated. Results: Generally, our results showed decreased levels of vitamins and trace elements in cooking with the Abkesh method in comparison to that of Dampokht. Interestingly, the amount of B1 vitamin increased after the Dampokht method. Conclusion: The results indicate that the rice cooking procedure is very important in the maintenance of B vitamins and trace elements in the foodstuff, and the Abkesh method leads to the loss of a great deal of water-soluble vitamins and essential elements. However, it is good for decreasing poisonous heavy metals and should be recommended for populations that consume rice as the main staple food and who live in the areas with a high level of toxic heavy metals in drinking water.

Triple-negative breast cancer (TNBC) is the most aggressive type of breast cancer (BC) that hardly responds to common treatment. Recent studies show that circ-ELP3 (Elongator Acetyltransferase Complex Subunit 3 or hsa-circ-0001785) is involved in the pathogenesis of several malignancies. The present study aimed to evaluate the possible role of this circRNA in the progression of TNBC cells and the possible relation between the circular and linear forms of the ELP3. We evaluated the circ-ELP3 and its host gene expression level in clinical samples and breast cancer cell lines. Using an expression vector, hsa-circ-0001785 was upregulated to investigate its role on cancer cell progression. After a transient transfection, we evaluated possible alterations in the cell cycle progression, cell viability, and cell proliferation. Quantitative real-time polymerase chain reaction analyses verified that circ-ELP3 and its host gene were significantly upregulated in TNBC tissues and breast cancer cells. Overexpression of circ-ELP3 markedly increases the cell viability and proliferation and also the formation of colonies in transfected cells compared to the controls. Briefly, our results showed that Circ-ELP3 and its host gene were significantly upregulated in TNBC. Circ-ELP3 is involved in TNBC progression and may exert its effects by indirectly regulating of ELP3 expression.

SummaryMouse embryonic fibroblast (MEF) cells are commonly used as feeder cells to maintain the pluripotent state of stem cells. MEFs produce growth factors and provide adhesion molecules and extracellular matrix (ECM) compounds for cellular binding. In the present study, we compared the expression levels of Fgf2, Bmp4, ActivinA, Lif and Tgfb1 genes at the mRNA level and the level of Fgf2 protein secretion and Lif cytokine secretion at passages one, three and five of MEFs isolated from 13.5-day-old and 15.5-day-old embryos of NMRI and C57BL/6 mice using real-time PCR and enzyme-linked immunosorbent assay. We observed differences in the expression levels of the studied genes and secretion of the two growth factors in the three passages of MEFs isolated from 13.5-day-old and 15.5-day-old embryos, respectively. These differences were also observed between the NMRI and C57BL/6 strains. The results of this study suggested that researchers should use mice embryos that have different genetic backgrounds and ages, in addition to different MEF passages, when producing MEFs based on the application and type of their study.

Rotifers are widely used in hatcheries to feed small-sized aquatic larvae although one of their disadvantages is the lack of zinc and selenium 5- and 30-fold lower than in copepods, respectively. To improve the rotifers quality, different concentrations of zinc and selenium (2, 4, 5 and 10 mg L−1 of each mineral) were added to the medium of the microalgae Isochrysis aff. galbana and Nannochloropsis oculata for 4 days, and then the microalgae were harvested and concentrated to feed the rotifers. N. oculata accumulated a greater amount of Zn and Se into cells than I. galbana. The cell size of algae given 0, 2, and 4 mg L−1 of minerals did not change in both microalgae, but enrichment of the microalgae with the 5 and 10 mg L−1 decreased the sizes and paled the colour of cells and increased cell division. The 2 mg L−1 was the best group for rotifers in terms of growth (population density, number of eggs, egg ratio, Specific growth rate, the maximum number and doubling time), and contained the second-highest level of Zn (69.26 ± 0.60) and Se (103.5 ± 5.0) content within a safe limit. Thus, rotifers enriched with Se and Zn can be used as a mineral delivery method to cover the nutritional requirements of marine larvae. info:eu-repo/semantics/acceptedVersion

Rotifers are widely used as a live prey for finfish larvae, although their content in minerals such as selenium and zinc is lower than in fish. In the present study, mixed microalgae (i.e., Isochrysis aff. galbana and Nannochloropsis oculata) were enriched with different amounts of Se and Zn either alone (45, 90, and 135 mg L−1 of Zn, 60, 120, and 180 mg L−1 of Se) or mixed (22.5, 45, and 90 mg Zn L−1 and 30, 60, and 120 mg Se L−1). The content of minerals in the microalgae was analysed after 1 and 3 h enrichment and their effect on the growth and mineral composition of the rotifer. Based on the results, 1 h enrichment time and 90 and 120 mg L−1 of Zn and Se, respectively, resulted in the highest amount of Se and Zn in enriched microalgae. The maximum Zn content was detected in the rotifers fed 90 mg L−1 Zn and 90 mg L−1 Zn +120 mg L−1 of Se, but the maximum Se was observed in rotifers fed with 120 and 180 mg L−1 Se. Thus, Se- and Zn-enriched rotifers can be used as a mineral delivery method to cover marine fish larvae nutritional requirements. info:eu-repo/semantics/acceptedVersion

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Background Oxidative stress has been identified as a predisposing factor for dental caries. Saliva, as a rich source of antioxidants, plays an essential role in the protection against dental caries. Salivary enzymatic antioxidants include superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx). Objectives The aim of this study was to evaluate the correlation of salivary enzymatic antioxidant activity with different levels of dental caries in children. Material and methods In this cross-sectional study, 90 healthy children aged 7-12 years (36 girls, 54 boys) were investigated. Demographic information was gathered and dental examinations were provided for all participants. Then, unstimulated whole saliva samples were collected in the morning. The salivary SOD, CAT and GPx activity was measured spectrophotometrically. For statistical analysis, Spearman's correlation test, the Mann-Whitney U test and the Kruskal-Wallis test were used with the SPSS for Windows software, v. 16. Results Our results showed no significant correlation between SOD, CAT and GPx and the decayed, missing, filled teeth index for permanent/primary dentition (DMFT/dmft). The CAT activity was elevated in proportion to the number of decayed teeth. The SOD activity showed a positive correlation with the frequency of tooth brushing. The activity of SOD, CAT and GPx was higher in boys than in girls. An inverse relationship between enzymatic antioxidant activity and age was also observed. Conclusions Although enzymatic antioxidants had no positive correlation with DMFT/dmft, they were positively correlated with the number of decayed teeth and the improvement of oral hygiene.

Retinal degeneration is the major and principal cause behind many incurable blindness diseases. Several studies indicated the neuroprotective effect of Curcuma longa in eye pathologies, specifically retinopathy. However, the molecular mechanism behind its effect has not been completely elucidated. Using an

Nonalcoholic fatty liver disease (NAFLD) is a common chronic liver disease that is rapidly becoming a public health problem. An imbalance in lipid distribution to the hepatocytes and metabolism causes hepatocyte steatosis. Vaspin is a newly discovered adipokine that has been linked to a variety of metabolic disorders. The effects of vaspin on steatosis and fibrosis pathogenesis and related mechanisms are unclear. Thus, this study investigated the molecular mechanism of vaspin on hepatocyte steatosis and fibrosis. HepG2 cells were treated with 1.2 mM free fatty acid and the intracellular lipid values were measured by flow cytometry and Nile red assay. RT-qPCR was used to assess the effect of vaspin and blocking of the GRP78 receptor on the expression of lipogenesis, oxidation, uptake, and secretion of fatty acid (FA), as well as AMPK activity. In co-cultured HepG2 and LX-2 cell lines, the expression of main proteins of hepatocyte fibrosis was analyzed using Western blot analysis. In the HepG2 cell line, we discovered that vaspin increased oxidation, FA secretion and gene expression, and AMPK activity and decreased lipogenesis and FA uptake and gene expression. Western blot analysis in co-cultured HepG2 and LX-2 cell lines showed that α-SMA and TGF-β1 protein expression decreased. The data demonstrated that vaspin acts as a novel regulator of hepatocyte steatosis through the GRP78 receptor, effectively reducing hepatocyte fibrosis through AMPK activation and decreasing NF-κB gene expression.

Introduction Aerobic training and green tea extract can be used to reduce the risk of prostate cancer. The goal of this study was to evaluate the effects of 8-wk aerobic exercise training and administration of green tea extract on the level of nuclear factor kappa B (NF-kB), cyclooxygenase-2 (COX-2) and p53 tumor suppressor protein (p53) in prostate of rats which were stimulated by N-methyl-N-nitrosourea to induce the prostate cancer. Methods Sixty adult male Wistar rats were assigned into six groups including healthy control, cancer control (CCt), cancer training (CTr: 45 min·d at low to moderate intensity, five times per week, 8 wk), cancer extract (CEx: 1.34 mL of green tea extract, three times per week, 8 wk), cancer training+ cancer extract (CTr + CEx) and sham groups. Rats were sacrificed 48 h after the last intervention session, and the prostate tissue was isolated to measure the levels of NF-kB, COX-2, and p53. Results The NF-kB level in CCt group was increased significantly compared to the healthy control (P = 0.02). In the CTr group, NF-kB level was decreased significantly compared to the CCt and CEx groups (P = 0.001 and 0.05, respectively). In addition, the levels of P53 protein were reduced in CTr, CEx, and CTr + CEx groups compared to CCt group (P = 0.001, 0.02 and 0.004, respectively). No significant changes were found in the level of COX-2 between groups. Conclusions These results suggest that a long-term exercise training combined with the intake of green tea extract may reduce levels of NF-kB and p53 in rats with prostate cancer. Given the importance of recognizing complementary therapies in this regard, future studies are warranted.

The aim of the present study is to assess the effect of L-carnitine and Coenzyme Q10 (CoQ10) on human sperm motility, DNA fragmentation, chromatin structure, and reactive oxygen species (ROS) during, before and after freezing in oligospermia men.Semen was collected from 30 oligospermic men, who referred to infertility clinic of Beasat Hospital in Sanandaj, Iran. The samples of each individual were divided into 8 equal parts: 1. control group before freezing; 2. incubated with L-carnitine; 3. incubated with coenzyme Q10; 4. incubated with the combination of L-carnitine + CoQ10; 5. control freezing group; 6. the experimental freezing group with L-carnitine; 7. the experimental freezing group with coenzyme Q10 and 8. the experimental freezing with the combination of L-c + CoQ10. Sperm motility was assessed by WET MOUNT method. DNA fragmentation was evaluated by SCD (Sperm Chromatin Desperation), ROS, was evaluated by quantitative fluorescence reaction, and chromatin deficiency was determined by chromatin staining (CMA3).Antioxidant treatments, significantly reduced the number of ROS + in the pre and post freezing groups. Significant improvement was seen in the sperm motility of class B in the pre freezing groups with L-carnitine. Antioxidants also reduced the percentage of DNA fragmentation and protamine deficiency in pre-and post-freezing.Addition of Coq10 and L-carnitine to human sperm medium significantly reduced the number of ROS. This reduction in ROS reduced sperm damage during cryopreservation.

Trimethylamine N-oxide (TMAO) is emerging as a new generation of metabolites related to the activation of inflammatory reactions in the macrophages during atherosclerosis. Stress-activation of cell surface toll-like receptors (TLRs) as well as nicotinamide adenine dinucleotide phosphate (NADPH) oxidases (NOX) is also assumed to be involved in TMAO-induced inflammatory reaction in the macrophages. To elucidate the possible contribution of TLRs and NOX to the mentioned signaling pathway, we aimed to simultaneously evaluate the expression level of TLR2, TLR6, and NOX2 in TMAO-treated macrophages.2.5 × 106 cells of U937-derived macrophages were treated in triplicates with different concentrations (37.5, 75, 150, and 300 μM) of TMAO for 24 hours. The cells were also treated with tunicamycin (TUN), as a positive control of stress. Normal control group (CTR) cells received no treatment. The viability of treated cells was checked by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, a tetrazole (MTT) assay. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was also used to evaluate the relative expression (fold change) of TLR2, TLR6, and NOX2 at messenger ribonucleic acid (mRNA) levels. One-way analysis of variance (ANOVA) with post-hoc Dunnett's test was performed to compare every mean with that of the control.No cell death occurred because of treatments. Dose of 300 μM of TMAO significantly increased the relative expression of both TLR2 and NOX2 compared to the CTR cells (P0.001 for both). The elevation of TLR6 was not statistically significant in all groups of TMAO-treated cells (P0.050).Our results provide documentation supporting contribution of TLR2 and NOX2 to previously described inflammatory reactions induced by TMAO in macrophages. In addition, they may clarify the proatherogenic role of TMAO in foam cell formation as well as abnormal activation of macrophages during atherosclerosis.

Background: There are promising data that exercise training and dietary polyphenols intake may alter the tumor microenvironment to facilitate conventional treatment. Objectives: The purpose of this study was to investigate the effect of eight weeks of aerobic training (AT) and green tea extract (GTE) consumption on matrix metalloproteinases-2 and -9 (MMP-2/-9) and vascular endothelial growth factor (VEGF) in normal and prostate cancer tissue of rats. Methods: Ninety Wistar rats were assigned to two healthy and cancer groups, then the healthy group was divided into four subgroups, including healthy control (Ctr), healthy AT (treadmill exercise for 5 days/week), healthy GTE (three times of week for eight weeks by gavage) and healthy AT + GTE, and after tumor induction, the cancer group was divided into five subgroups, including cancer control (CaCtr), cancer AT (CaAT), cancer GTE (CaGTE), cancer AT + GTE and sham groups. Results: The results showed that there was no significant difference between healthy and CaCtr groups in the MMP-2 levels (P = 0.07), but MMP-2 levels in the CaAT group were lower than the healthy AT (P = 0.01). No significant changes were found in the levels of MMP-9 and VEGF between the healthy and cancer groups (P = 0.23 and P = 0.08, respectively). Conclusions: The present study demonstrated that low to moderate-intensity aerobic training and treatment with a low dose of green tea extract did not change angiogenesis and metastasis markers. These results pave the way for further researches on modulation approaches for tumor metastasis and vascularization in responses to exercise training and antioxidant supplementation.

Background: Previous studies showed the role of vitamin D (Vit D) on the progression of chronic urticaria. To the best of our knowledge, there are no other results regarding the contribution of single nucleotide polymorphisms (SNPs) in the vitamin D receptor (VDR) and vitamin D binding protein (VDBP) genes in chronic urticaria (CU). Aim: In the present study, we investigated the Vit pathway and the association between VDR and VDBP gene polymorphisms and CU risk in Iranian population. Methods: All participating individuals in the present study were evaluated for serum Vit D and VDBP concentration VDR rs1544410 and rs2228570 and VDBP rs7041using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. The associations of studied analytes and three SNPs with clinical and laboratory outcomes were investigated in CU patients. Results: Patients with CU showed lower Vit D compared to controls (19.2661.26 vs. 31.7267.14 ng/ml, P-value=0.006). There was a significant correlation between Vit D levels and urticaria activity score. Serum VDBP was significantly higher in CU patients than controls (1317.36183.71 vs. 395.77612.96 I¼g/ml, P-value

Family with sequence similarity 83 member H (FAM83H) protein-coding geneplay an essential role in the structural organization, calcification of developing enamel, and keratin cytoskeleton disassembly by recruiting Casein kinase 1 alpha (CSNK1A1) to keratin filaments. In this study, we have applied CRISPR Cas9 nickase (D10A) to knockout (KO) the Fam83h gene in NMRI outbred mice. We generated homozygous Fam83h KO mice (Fam83h Ko/Ko) through a premature termination codon, which was validated by Sanger sequencing in F0 generation. Next, we also bred the FAM83H KO for two generations. Reverse-transcription polymerase chain reaction and Western blot analysis approved the Fam83h KO mice. The Fam83h KO mice had evidence of normal morphology at the cervical loops, secretory and maturation stages, and mandibular molars. In comparison with the normal wild-type mice (Fam83h W/W), the F2 homozygous KO (Fam83h Ko/Ko) had sparse, scruffy coats with small body size and decreased general activity. Also, they had the natural reproductive ability and natural lifespan. In addition, delay in opening the eyes and dry eyes among infant mice were seen. The F1 heterozygous mice looked comparable to the normal wild-type mice (Fam83h W/W), which showed autosomal recessive inheritance of these phenotypes. The KO of FAM83H had controversial effects on the development of teeth and the formation of enamel. The phenotype defect in dental development and the enamel formation were seen in three mice among four generations. It can be concluded that null FAM83H in outbred mice not only showed the reported phenotypes in null inbred mouse but also showed normal lifespan and reproductive ability; dental deficiency in three homozygous mice; and the symptoms that were similar to the symptoms of dry eye syndrome and curly coat dog syndrome in all four evaluated KO generations.

Background: Mesenchymal stem cells (MSCs) can be used to treat premature ovarian failure (POF). Different methods have already been applied to detect MSCs in tissues. This study aimed to investigate the quantitative distribution of CM-DiI-labeled human umbilical cord vein MSCs (hUCV-MSCs) in different regions of the ovarian tissue of the cyclophosphamide (CTX)-induced POF in mice. Methods: Adult female C57BL/6 mice (n = 40) were divided into four groups: (1) Mice receiving PBS as control (Ctrl) group; (2) mice receiving hUCV-MSCs intravenously as Ctrl + hUCV-MSCs group; (3) mice receiving CTX intraperitoneally (i.p.) as CTX group; (4) mice receiving CM-DiI-labeled hUCV-MSCs after CTX injection as CTX + hUCV-MSCs group. Histological changes and CM-DiI-labeled hUCV-MSCs distribution were analyzed in the ovarian tissues. Quantitative real-time PCR was performed to detect human mitochondrial cytochrome b (MTCYB) gene in the ovarian tissues of the mice. Results: The mean number of the fluorescent hUCV-MSCs was 20 ± 2.5 (57.1%) in the medulla, 11.3 ± 2.8 (32.2%) in the cortex, and 5.5 ± 1 (15%) in the germinal epithelium of the ovarian tissue (p < 0.05). Moreover, MTCYB gene was detected in the mice ovaries of the CTX + hUCV-MSCs group, but not in other groups. Conclusion: Our findings suggest that the distribution of the transplanted hUCV-MSCs in different regions of the ovarian tissue is not equal, and it is greater in the medulla than the cortex and germinal epithelium. This is the first report of quantitative distribution of MSCs in different regions of ovarian tissue in the POF model.

To investigate the effect of lecithin on FMO3 and FMO5 gene expression, L-α- lecithin or ethanol 30% (as control) was administrated to female N-mary mice as intraperitoneal injection and animals were sacrificed 48h after treatment. The relative expression of mRNA and protein were assessed by real-time PCR and western blotting analysis. In this study, FMO5 expression was also evaluated in HepG2 cell line. FMO3 mRNA expression in mice liver was increased to 1.8-fold in animal received lecithin compared to control group, While FMO5 expression did not showed any changes as in vivo and in vitro experiments. The our results suggest to induce of FMO3 expression by lecithin.

Background and Aim: Nonalcoholic fatty liver disease (NAFLD) and gallstone disease (GD) are both highly prevalent in the general population and have many risk factors in common. The aim of this study was to evaluate the prevalence of GD in the patients with NAFLD. Materials and Methods: In this case-control study, our case group included 145 patients with NAFLD and control group consisted of 215 age-, sex-, and BMI-matched healthy subjects. NAFLD and gallstone disease were diagnosed by sonography. The prevalence and risk factors for gallstone disease were evaluated in the groups and compared between the 2 groups. Results: The frequency rates of gallstones in the case and control groups were 10/135 (6.9 %) and 5/210 (2.3 %) respectively (p=0.04). The frequency rates of gallstones in the female participants in the case and control groups were 6.8% and 3.4%, respectively, and in the men were 6.9% in the case and 1% in the control groups. We found no significant relationship between sex and frequency of gallstones in both groups (P> 0.05). In the control group, the prevalence rate of GD in the subjects over 50 years of age were significantly higher than that in the subjects under 50 years of age (P = 0.04). However, there was no significant difference between ag e and GD in the case group (P = 0.51). Also, in both groups, the prevalence of GD was significantly higher in obese subjects (BMI> 30) (P> 0.05). There was a significant difference between the grade of fatty liver and the prevalence of GD (P = 0.01). Comparison of the risk factors associated with the GD, showed no significant difference between the two groups. Conclusion: The prevalence of gallstones was more in NAFLD than in normal population and was associated with grade of fatty liver disease.