Search

Objective: This study aimed to investigate the relationship between follicular fluid Bisphenol A (BPA) concentrations with alterations in the expressions of NOTCH1-3, CASPASE 3/7, HLA-G, and ICAM-1 genes and the number of retrieved mature oocytes (MII oocyte) in the cumulus cells of infertile poor ovarian response stimulates women. Materials and Methods:In this prospective cohort study, 80 infertile unexpected poor ovarian response (POR) subjects were selected on the basis of subgroup 1a of the POSEIDON classification. They were divided into two groups: group 1 consisted of 40 women, each with a higher number of metaphase II (MII) oocytes (G1, 3-4 oocytes retrieved), while group 2 comprised of 40 women, each with a lower number of MII oocytes (G2, ≤2 oocytes retrieved). The expressions of the studied genes were evaluated by quantitative-real time polymerase chain reaction (PCR). The concentration of BPA in follicular fluid was measured with HPLC. Results: The expression levels of NOTCH1-3, HLA-G, and ICAM-1 genes were significantly lower in G2 than G1 (P<0.05). Meanwhile, CASPASE 3/7 expression levels were higher in unexpected POR patients in G2 compared to G1 (P<0.05). There was a significant direct correlation between the levels of NOTCH1-3, HLA-G and ICAM-1 gene expressions and there was also a significant inverse correlation (P<0.05) between the levels of CASPASE 3/7, with the number of MII oocytes and embryo development between the two groups. The concentration of BPA in the follicular fluids of G2 was higher compared to G1 (P<0.05). Conclusion: A higher concentration of BPA was associated with a lower number of mature oocytes and oocyte quality in these patients. Also, alterations of NOTCH1-3, CASPASE 3/7, HLA-G, and ICAM-1 transcript levels in unexpected POR women were associated with BPA concentration. [ABSTRACT FROM AUTHOR]

Background: Two newly identified proteins, EspB and EspC are involved in the pathogenesis of Mycobacterium tuberculosis. The objective of the present study was to evaluate the immunogenicity of recombinant EspC, EspB, and EspC/EspB fusion proteins in mice. Methods: BALB/c mice were immunized subcutaneously with recombinant EspC, EspB, and fusion EspC/EspB proteins, three times with along with Quil-A as an adjuvant. The cellular and humoral immune responses were evaluated by quantifying IFN-γ, IL-4, IgG, IgG1, and IgG2a antibodies against the antigens. Results: The results showed that the mice immunized with recombinant EspC, EspB, and EspC/EspB proteins did not produce IL-4, whereas IFN-γ was secreted in response to all three proteins. EspC/EspB group produced significant amounts of IFN-γ in response to stimulation with all the three recombinant proteins (P<0.001). In mice immunized with EspC, high levels of IFN-γ were detected in response to EspC/EspB, and EspC (P<0.0001); while mice immunized with EspB produced lower levels of IFN-g in response to EspC/EspB, and EspB (P<0.05). Mice immunized with recombinant EspC, EspB, and EspC/EspB proteins exhibited significantly high levels of IgG and IgG2a/IgG1 ratio (P< 0.001). Moreover, high levels of IgG and IgG2a were detected in the sera of mice immunized with EspC/EspB fusion protein. Conclusions: All the three recombinant proteins induced Th1-type immune responses in mice against EspB and EspC; however, EspC/EspB protein is more desirable due to the presence of epitopes from both EspC and EspB proteins and the production of immune responses against both. [ABSTRACT FROM AUTHOR]

β-Thalassemia (β-thal) is an inherited genetic disease that occurs because of the absence or reduction of β-globin chain synthesis. Genetic changes occur in different regions of the β-globin gene, but these mutations are less reported in the 3' untranslated region (3'-UTR). The objective of the present investigation was to evaluate the functional effect of a rare variant in the 3'-UTR of the β-globin gene. A variant at the first nucleotide of the 3'-UTR of the β-globin gene (HBB: c.*1G > A) was identified by DNA sequencing in an individual with low hematological indices and a normal hemoglobin (Hb) electrophoresis pattern. To evaluate the functional effect of this variant, the normal and mutated 3'-UTR of the β-globin gene was synthesized separately and sub cloned in the psiCHEK2 vector. Next, using the calcium phosphate method, the psiCHEK2 vectors containing normal and mutated 3'-UTR were transfected separately into the HEK293T cell line. Finally, the transfected cell line was analyzed by dual luciferase assay. The ratio of Renilla to firefly for the mutant sample was 1.26 ± 0.06, while for normal samples it was 1.12 ± 0.04. The results of the luciferase assay showed that there was no significant difference in the functional effect between the mutant and wild type construct. Therefore, it was concluded that this variant might not reduce the expression of the β-globin gene. Future studies by globin chain synthesis or to evaluate the expression of the gene in erythroid cells, might be necessary to understand the regulatory function of this mutation. [ABSTRACT FROM AUTHOR]

Background: Schizophrenia, a chronic mental disorder, has been recognized as one of the heritable diseases with an increased level of dopamine neurotransmitter. Monoamine oxidase A (MAOA) plays a vital role in the catabolism of dopamine. It is a mitochondrial enzyme which is encoded by the MAOA gene located on the X chromosome. The aim of this study was to detect potential biomarker in order to diagnose schizophrenia. Hence, the association of uVNTR repetitions of the MAOA gene and Schizophrenia was investigated. Method: Blood samples were collected from 102 schizophrenic patients (67 males and 35 females), and 103 volunteers with mental health (65 males and 38 females). Genomic DNA was extracted and the uVNTR polymorphisms were examined using specific primed PCR. Results: The analysis indicated that genotype 3/3repetition (rep) showed the highest frequency in females. While genotype 4 and 3.5 reps revealed the highest frequencies among schizophrenic patients and healthy controls in men, respectively. There were no significant statistical differences in the number of uVNTR repeats of the MAOA gene between control and case individuals neither in women (OR = 0.35, 95% CI = 0.60–1.43 P = 0.845) nor in men (OR = 0.36, 95% CI = 0.80–1.64, P = 0.365). Conclusion: In the current study, the number of uVNTR sequence repetitions located in the promoter of the MAOA gene was not associated with the risk of schizophrenia in Iranian patients. [ABSTRACT FROM AUTHOR]

Bio surfactants are natural surfactants that induce emulsification, displacement, increased solubility, and mobility of hydrophobic organic compounds. In this study, the gene expression of biosurfactant production genes by Pseudomonas aeruginosa in the presence of sodium dodecyl sulfate coated iron nanostructure (Fe/SDS) were evaluated. Emulsification Index and Surface Tension reduction test to check stability and emulsification the rhamnolipid were done. Purification was evaluated using thin layer chromatography (TLC) and expression of rhlA, mvfR, lasR, rhlR genes was determined using q‐PCR technique. Binding of nanoparticles to bio surfactants was confirmed by TEM. The best emulsification index, was by the sample that exposed to 1 mg/L Fe/SDS nanoparticles for 2 days. Rhamnolipid produced in the presence of nanoparticles had an acceptable ability to reduce surface tension. The Rf (retention factor) value obtained was 0.63 by chromatography. q‐PCR results showed that the expression of rhlA, mvfR, lasR, rhlR genes was significantly increased in Fe/SDS treated cells, which indicates the significant positive effect (P < 0.05) of nanoparticles on biosurfactant production of treated cells. While, SDS and Fe alone were not affected significantly (P > 0.05) on the expression of these genes. Our findings indicated the importance of nanoparticles in increasing the expression of genes involved in the bio surfactant production pathway of Pseudomonas aeruginosa. [ABSTRACT FROM AUTHOR]

RNA interference (RNAi) holds immense potential for sequence-specific downregulation of disease-related genes. Small interfering RNA (siRNA) therapy has made remarkable strides, with FDA approval for treating specific human diseases, showcasing its promising future in disease treatment. Designing highly efficient siRNAs is a critical step in this process. Previous studies have introduced various algorithms and parameters for siRNA design and scoring. However, these attempts have often fallen short of meeting all essential criteria or required modifications, resulting in variable and unclear effectiveness of screened siRNAs, particularly against viral mutants with non-conserved short sequences. In this study, we present a fully optimized siRNA screening system considering all necessary parameters. Notably, we highlight the critical role of molecular docking simulations between siRNA and two functional domains of the Argonaute protein (PAZ and PIWI) in identifying the most efficient siRNAs, since the appropriate interaction between the guide siRNA strand and the RISC complex is crucial. Through our stringent method, we designed approximately 50 potential siRNAs targeting the HIV-1 vpr gene. Evaluation through XTT, qRT-PCR, and flow cytometry analysis on RAW 264.7 macrophage stable cells revealed negligible cytotoxicity and exceptional gene-silencing efficiency at both the transcriptional and translational levels for the top-ranked screened siRNAs. Given the growing interest in siRNA-based therapeutics, we anticipate that the insights from this study will contribute to improving treatment strategies against mutant viruses, particularly HIV-1. [Display omitted] • Efficient siRNAs can help control the emergence of drug-resistant viruses. • Conserved regions of HIV-1 vpr gene were identified. • Using a comprehensive strategy, anti -HIV siRNAs were designed and ranked. • Molecular docking simulation appears to be crucial in identifying the most efficient siRNAs. [ABSTRACT FROM AUTHOR]

Background and Aim: Klebsiella pneumoniae is one of the three pathogens that has become a global disease control and treatment problem due to its resistance to common antibiotics. For this reason, it is crucial to study the genes that cause antibiotic resistance in it. Therefore, the aim of this study was to investigate the phenotypic and genotypic frequency of tetracycline resistance in clinical isolates of K. pneumoniae in Tehran, Iran. Materials and Methods: In this study, 100 isolates of K. pneumoniae isolated from clinical samples (urine) during 2018-2019 were studied. In addition to microbial and biochemical phenotypic tests, genotypic tests were conducted to determine the frequency of antibiotic resistance genes tet (A, B, C, 39). Results: Out of 100 isolates of K. pneumoniae, 49 isolates were resistant to tetracyclines. The results of multiplex PCR showed that 31 samples were positive for tetA gene, 8 isolates for tetB gene, 21 samples for tetC and, and 8 isolates for tet39. None of the isolates were positive for all four tetracycline genes. Conclusion: The results of this study showed that the isolates were positive for at least one gene and at most 2 tetracycline resistance genes. The tetA gene showed the highest frequency and the lowest frequency was demonstrated by tetB. The highest binary combination of genes was tetA-tetC, and the lowest was tetA-tet39. [ABSTRACT FROM AUTHOR]

• the role of spatial and climate variables on the genetic structuring of pomegranate plant. • Identify cytogenetic and genetic clines within the pomegranate accessions of Iran. • identify the species distribution modeling and the gene flow corridor. Pomegranate (Punica granatum L.) is an important horticultural plant in Iran and has been a food source since ancient times. Despite several genetic diversity studies conducted on these taxa, there are no reports on the landscape genetics of these important plants in Iran, nor is there any information on the effects of environmental, spatial, and climatic variables on the genetic structure of pomegranate. Therefore, this study used genetics, structural equation modeling (SEM), and species distribution modeling (SDM) approaches to explore the above-mentioned objectives. We used both cytogenetic and molecular analyses (SSR and SCoT markers) of several pomegranate samples along with environmental, spatial, and climatic variables. The redundancy analysis (RDA) analysis showed a significant positive association between terminal chiasmata with the longitudinal distribution of the pomegranate accession, while total chiasma number and interstitial chiasmata have an association with the altitude. The spatial PCA analysis indicated the role played by both global and local spatial variables in the cytogenetic variability like the univalent and terminal chiasmata formation. The Mantel test indicates that the cytogenetic difference in pomegranate plants is not related to their geographical distance, while the Moran I test result shows that geographical regions with similar spatial conditions affect the cytogenetic structuring of pomegranate plants in a similar way. SEM methods showed a strong negative association between bivalent formation and the environmental variables, while they showed both positive and negative associations between some of the environmental variables and different kinds of chiasmata. The sPCA analysis of molecular data showed a significant global, and local, spatial structuring of the molecular data and the genetic cline formation within the pomegranate accessions studied. Similarly, the spatial PCA analysis of climate variables, precipitation, and temperature revealed a significant structuring of genetic diversity and the formation of genetic clines by these factors. RDA analysis identified some of the loci that are significantly associated with temperature and precipitation. Therefore, both geographical/ or spatial variables and climate factors contribute to the genetic structure of the pomegranate accessions studied. SEM models based on genetic data revealed a positive association between environmental and climate variables with genetic diversity parameters and SSR-SCoT molecular loci studied. Climate change is unlikely to hurt pomegranate growing areas due to the high genetic diversity of pomegranate plants and the diverse genetic lineages present in the country. [ABSTRACT FROM AUTHOR]

Objective(s): SLC39A6 (solute carrier family 39) or LIV-1, is a zinc-transporter protein associated with estrogen-positive breast cancer and its metastatic spread. Significantly there is a direct relation between high zinc intake and unregulated cell proliferation and cancers. Blocking SLC39A6 protein may result in reduced metastasis and proliferation in many malignant tumors. This study aimed to develop an anti-SLC39A6 nanobody that is able to detect and block the SLC39A6 protein on the surface of cancerous cells. Materials and Methods: The recombinant SLC39A6 was expressed and used for camel immunization. The VHH library was constructed and screened for SLC39A6-specific nanobody. Then, the strength of nanobody in SLC39A6 detection was evaluated by Western blotting and flow cytometry. Results: We showed the ability of SLC39A6 specific Nanobody (C3) to detect SLC39A6 by Western blotting and flow cytometry. Furthermore, the C3 nanobody potently inhibits cell proliferation in MTT assay. Conclusion: These data show the potential of SLC39A6-specific nanobody for the blockade of zinc transporter protein and provide a basis for the development of novel cancer therapeutics. [ABSTRACT FROM AUTHOR]

Objective(s): Cytotoxic T-lymphocyte-associated protein-4 (CTLA-4) is the most important human immune checkpointthat modulates T cells activity and brings about immune-homeostasis. Accordingly, checkpoint inhibitor cancer therapy has been approved as a growing method to block over-expressed immune checkpoints, such as CTLA-4 receptors. Considering the competitive characteristics of single-domain antibodies with monoclonal antibodies, we tried to develop a camelid Nanobody against human CTLA-4. Materials and Methods: We have constructed the VHH gene library by using immunized-camel peripheral blood mononuclear cells and carrying out the Nested-PCR technique. VHH-library was screened by phage display technique and specific nanobodies against CTLA-4 protein were selected and amplified with bio-panning steps. Stronger binders were screened by Periplasmic Extract-ELISA, followed by estimating the complexity of the library. Specific anti-CTLA-4 Nanobody and 3hCTL55, with longer CDR3 and a higher binding rate, were selected for more assays. Results: Results revealed the existence of two different clones in the library with 108 binders. In comparison with seven different antigens, using the ELISA technique confirmed the specificity of Nanobody 3hCTL55 against human CTLA-4 antigen. We calculated Nanobody 3hCTL55 affinity for human CTLA-4 antigen at 50x10-9 M, approximately. Performing western blot and Flow-cytometry techniques showed that Nanobody 3hCTL5 5 was able to specifically detect and attach both commercial human CTLA-4 protein and human CTLA-4 antigen on the cell surface and in the cell lysate. CONCLUSION: Taken together, this developed camelid-specific anti-CTLA-4 Nanobody 3hCTL5 5, selected from a high-quality immune library by phage display technique, may be effective for further study about cancer diagnosis and cancer-therapy purposes [ABSTRACT FROM AUTHOR]

This study aimed to investigate the relationship between follicular fluid Bisphenol A (BPA) concentrations with alterations of ICAM-1 and HLA-G genes and proteins expression as well as methylation profiles in the cumulus cells of poor ovarian response (POR) women based on their healthy lifestyle habit. Eighty women under the age of 35 were divided into two groups: 1—POR without using plastic containers (n = 40) and 2—POR with using plastic containers (n = 40). The ICAM-1 and HLA-G genes and protein expressions were examined by the quantitative PCR and western blotting technique. The methylation pattern was investigated by the methylation-specific PCR. Total BPA in follicular fluid was measured with high-performance liquid chromatography technique and the detection limit was 1.14 ng/ml. ICAM-1 and HLA-G genes were differentially expressed between the two groups studied. ICAM-1, HLA-G genes, and protein expressions in group 1 were up-regulated compared to the second group (P < 0.05). While DNA methylation status in group 1 were decreased compared to the other group (P < 0.05). The concentration of BPA in the follicular fluid of group 1 was lower compared to the second group (P < 0.05). The oocyte quality and clinical pregnancy ratio showed significantly higher in group 1 than in the other ones (P < 0.05). The alteration of ICAM-1 and HLA-G gene expressions in POR women is probably related to BPA concentration. As a result Lifestyle habits may also affect the methylation pattern and protein levels in the cumulus cells of POR women. Additionally, lifestyle habits may be considered as a marker for ovulation, oocyte maturation, preimplantation, and clinical pregnancy process. [ABSTRACT FROM AUTHOR]

Date palm (Phoenix dactylifera) is an important crop plant both from nutritional and economic points of view. The assessment of genetic diversity and population differentiation of date palms are evaluative for its dynamic conservation and sustainable utilization of its genetic diversity. Estimates of genetic diversity based on molecular markers and fruit characteristics were performed in samples of 23 date palms growing in Ahvaz city (Khuzestan province of Iran). Clustering based on fruit morphological features separated the cultivars in different groups. These cultivars differed significantly in morphological features (P = 0.001). Start codon targeted (SCoT) polymorphism markers revealed a good level of genetic variability (10.17 to 45.76%) in these cultivars. Moreover, STRUCTURE analysis revealed the presence of within-population genetic variability. Analysis of molecular variance revealed a significant genetic difference among date palms, while it showed a higher degree of within-cultivar genetic variability compared with that of among-population diversity. Some degree of common shared alleles occurred between date palm cultivars. Gst versus Nm analysis showed that some of the SCoT markers have a high discrimination power and may have a potential local adaptive value. The Mantel test showed a significant association (r = 0.40, P = 0.001) between morphological and genetic distances. Therefore, both morphological and SCoT molecular data can be used in genetic screening of date palms in the available germplasm. [ABSTRACT FROM AUTHOR]

The salinity caused changes in plant metabolic processes. Cotton is one of the important fiber products which may be affected by salinity stress in Iran. In the present study, the effects of salinity stress and salicylic acid (SA) as improvement of abiotic stress were examined on biochemical traits and retrotransposon based markers profile (IRAP and REMAP) in cotton cultivars. Cotton seeds of two cultivars (GT40 and NNC) were cultivated in pots and different treatments; control plants with normal irrigation, 0.5 and 1mM SA, 100 and 150mM NaCl treatments and, combination of 0.5mM SA and 150mM NaCl were performed. Physiological, morphological, cellular and, genetic diversity parameters were measured after treating and following seven days of irrigation. Activity of antioxidant enzymes; catalase, peroxidase and proline in 150mM NaCl treated plants were more than other treatments. In comparison to two cultivars, GT40 cultivar had a higher activity of antioxidant enzymes. Based on retrotransposon based markers analyses, the highest heterozygosity was shown in 0.5mM SA treatment of GT40 cultivar as well as 0.5 mM SA, 150mM NaCl treatments in NNC cultivar. Totally, GT40 showed the highest genetic variation in retrotransposon profiles. The Salt stress and salicylic acid treatment provided discriminate profiles from retrotransposon movement. [ABSTRACT FROM AUTHOR]

Catharanthus roseus L. (G) Don, commonly known as Madagascar periwinkle, is an important medicinal plant receiving much attention from researchers. This plant is a rich source of terpenoid indole alkaloids (TIAs). The gamma radiation and somaclonal variation may provide suitable methods for genetic changes to increase medicinal components of C. roseus. In the present study, 30 and 60 (Gy) gamma rays were irradiated on seeds. C. roseus seeds were cultured in Murashige and Skoog (MS) medium in three different groups, 0 Gy (control) 30, and 60 Gy gamma irradiations. Morphological characteristics of regenerated plantlets were measured for surface area of leaves, stem length, root length, and fresh weight. The genetic variations were evaluated using nine inter-simple sequence repeats and 25 sequences related amplified polymorphism markers. Morphological results showed a significant difference between studied groups in leaf surface and twisted leaf in 60 Gy irradiated plantlets. The analysis of molecular variance revealed the significant genetic difference among groups. STRUCTURE analysis showed genetic variability both within and among groups. Using the NJ tree analysis, the plants studied were categorized into three distinct groups, which were supported by principal coordinate analysis (PCoA). In conclusion, gamma irradiation produced morphological and genetic changes in C. roseus regenerated in vitro plantlets providing to be a suitable method for further mutation breeding in periwinkle. [ABSTRACT FROM AUTHOR]

Cytotoxic T-lymphocyte-associated protein-4 (CTLA-4) is a critical negative immunomodulatory receptor that is normally expressed in activated T cells and noticeably, in many cancerous cells. Indeed, molecular detection of CTLA-4 protein is crucial in basic research. In this work, the extracellular domain of the human CTLA-4 (hCTLA-4) protein was cloned, expressed, and purified. Subsequently, this protein was used as an antigen for camel (Camelus dromedarius) immunization to obtain polyclonal camelid sera against this protein. Furthermore, we evaluated the benefits of applying camelid hyperimmune sera containing heavy-chain antibodies in different antibody-based techniques. Our results indicated that hCTLA-4 protein was successfully expressed in the prokaryotic system. The polyclonal antibody (pAb) that raised against recombinant hCTLA-4 protein was able to detect the CTLA-4 protein in antibody-based techniques, such as enzyme-linked immunosorbent assay, western blotting, flow cytometry and immunohistochemistry (IHC) staining. This study shows that, due to the advantages such as multi-epitope-binding ability, camelid pAbs are potent to efficiently apply for molecular detection of CTLA-4 receptors in fundamental antibody-based researches such as IHC. [ABSTRACT FROM AUTHOR]

Gene therapy, including small interfering RNA (siRNA) technology, is one of the leading strategies that help to improve the outcomes of the current therapeutic systems against HIV‐1 infection. The successful therapeutic application of siRNAs requires their safe and efficient delivery to specific cells. Here, we introduce a superparamagnetic iron oxide nanoparticle (SPION) for delivering siRNA against HIV‐1 nef (anti‐nef siRNA) into two cell lines, HEK293 and macrophage RAW 264.7. SPIONs were coated with trimethyl chitosan (TMC), and thereafter, different concentrations of SPION–TMC were coated with different ratios of a carboxymethyl dextran (CMD) to modify the physicochemical properties and improve the biological properties of the nanocarriers. The nanoparticles exhibited a spherical shape with an average size of 112 nm. The obtained results showed that the designed delivery route enhanced the uptake of siRNA into both HEK293 and RAW 264.7 cells compared with control groups. Moreover, CMD–TMC–SPIONs containing anti‐nef siRNA significantly reduced the expression of HIV‐1 nef in HEK293 stable cells. The modified siRNA‐loaded SPIONs also displayed no toxicity or apoptosis‐inducing effects on the cells. The CMD–TMC–SPIONs are suggested as potential nanocarriers for siRNA delivery in gene therapy of HIV‐1 infection. [ABSTRACT FROM AUTHOR]

Objective: Schizophrenia is a complicated mental disorder that affects about 1% of the world's population. It is a complex disease and is approximately 80% inherited. One of the candidate genes in schizophrenia is transcription factor 4 (TCF4), which is positioned on chromosome 18 and is a transcription factor that plays a role in the transcription of Neurexin 1(NRXN1) gene, which is one of the candidate genes for developing schizophrenia. This case-control study aimed to investigate the correlation of TCF4 rs13381800 and NRXN1 rs17039988 polymorphisms with the risk of schizophrenia in a sample of Iranian patients with schizophrenia. Method: A total of 200 individuals were included in this study: 100 patients with schizophrenia (65 males and 35 females), with the mean age of 40.80 ± 11.298 years, and 100 as a control group (63 males and 37 females), with the mean age 32.92 ± 7.391 years. Allele specific polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) were done, respectively, for genotyping of rs13381800 (T/C) and rs17039988 (A/C) polymorphisms. Results: The results showed that the frequency of C / C genotype in rs13381800 in patients' group was 9%, while it was 13% in the control group. Also, the frequency of C / C genotype in rs17039988 was 9% in patients and 7% in control groups. Statistical analysis of polymorphisms showed no correlation between patients and controls in rs13381800 (OR = 1.51; CI = 95%; P = 0.366) and rs17039988 (OR = 0.76; CI = 95%; P = 0.602). Conclusion: No significant difference was found between rs13381800 and rs17039988 genotypes between patients and control groups in terms of gender, age and education in the patients group. Our study suggests that there was no correlation between desired polymorphisms with schizophrenia in the studied population. [ABSTRACT FROM AUTHOR]

With the ever-increasing number of GM crops introduced to the market, it is required to respect customers' freedom of choice and their perception toward GMOs. According to obligatory demand by food and drug administration for GM testing on raw and processed materials, efficient and robust analytical approaches are needed for screening and identification of GM crops. This article describes the development and applicability of a PCR-based GMO testing of the most frequent constructs used in transgenic soy, corn and rice varieties which include CaMV35S promoter and the NOS terminator and also plant endogenous sequences Lectin, Zein, SPS as internal controls. A total of 2866 different sample inputs collected during the year 2015 to 2017 including GM and non-GM materials have been tested. Results obtained from qualitative PCR test on rice, maize and soybean-containing food samples showed that in total, 1.59% of maize samples and 7.53% of soya samples were GM positive and no GM positive rice sample was detected. The presented method showed high specificity and sensitivity offering an absolute limit of detection of 0.25% GM construct in the samples and sufficient reproducibility of the method was proved. This method fits the purpose of GMO testing laboratories due to its convenience and robustness. [ABSTRACT FROM AUTHOR]

Flax (Linum usitatissimum subsp. usitatissimum) is an important crop plant cultivated both for its fiber and seed oil content. Uniform cultivation and selection practice may lead to genetic erosion and reduced genetic variability of Linum cultivars, which in turn makes the crop vulnerable to diseases and pathogens. Therefore we performed a genetic diversity analysis of few populations including both Linum usitatissimum subsp. usitatissimum L. and L. Bienne L. We used a combination of single primer amplification reaction markers, including RAPD, ISSR, and RAMPO molecular markers for genetic variability study. In general, all three molecular markers revealed a low degree of genetic variability in the materials studied; however, L. bienne had a higher level of genetic variability. Clustering and network analyses grouped the studied plants inter-mixed and did not separate the studied species or populations. STRUCTURE plot and K-Means clustering also identified these populations as genetically homogenous. These results indicate the need for introducing new linseed cultivars into the country for future hybridization and breeding programmes. [ABSTRACT FROM AUTHOR]

Flax (Linum usitatissimum L.) is one of the most important cultivated oil-producing plants that is highly evaluated for its fatty acid components. Studying and using wild relatives of cultivated crop plants as a spore of gene pool for breeding and hybridization is considered very important now. Therefore, the aim of present investigation was to provide data on saturated and unsaturated fatty acids of three Linum species, Linum usitatissimum, L. austriacum and L. album. We also studied the magnitude of oil composition variability within populations of these species and attempted to find out if oil composition data can be used in the Linum species delimitation. The saturated (C16:0, C18:0 and C20:0) and unsaturated (C18:1, C18:2, C18:3 and C20:1) fatty acids identified by GC represented inter- and intraspecies variations in the linseed and wild linum species. The linolenic (C18:3), linoleic (C18:2) and oleic acid as unsaturated fatty acids had an average of 52.7 %, 12.4 % and 20.6 % respectively in cultivated flax (L. usitatissimum), an average of 51.4 %, 18.8 and 20.3 % in L. austriacum, and 4.3 %, 60.4 %, 22.5 % in L. album respectively. Correlations between saturated and unsaturated fatty acids showed that with increase of linolenic acid in linseed oil, the oleic acid decreased. We found that unsaturated fatty acids (linoleic and linolenic) were higher in wild species as compared to those of the cultivated flaxseed, while precursor saturated fatty acids were lower in the studied wild species. Cluster analysis and a PCoA plot revealed that the Linum species differed in their oil profiles and the populations of each species were placed in a separate group. These data can be used in the Linum species delimitation. [ABSTRACT FROM AUTHOR]

Linum species are of various benefits to mankind, such as fiber and oil source, ornamental as well as medicinal values. Population genetic study by using molecular markers can produce valuable information on genetic structure, genetic variability, genetic fragmentation and gene flow among plant populations. These data can be used in conservation and breeding programs of economically important plants. We performed population genetic study on 6 populations of two Linum species, namely L. usitatissimum L. and wild species L. austriacum L., by using SRAP molecular markers. In general low genetic variability was observed in both studied species, but a little higher genetic diversity occurred in the wild species L. austriacum (6.25-21.88%). AMOVA revealed significant genetic difference among these populations that was supported by Gst and Dest parameters of genetic differentiation. K-Means clustering and STRUCTURE analyses identified three distinct genetic groups within each species. Nm and population assignment test did not show the occurrence of gene flow among the studied populations and suggested genetic fragmentation in both Linum species. Predominant selfing in L. usitatissimum may be the reason for genetic uniformity within each population and genetic differentiation among these populations. L. austriacum shows distyly, which promotes pollination between different floral morphs, and prevents self-pollination. This may be the reason for higher degree of within population genetic variability in this species. [ABSTRACT FROM AUTHOR]

Background: Recurrent spontaneous abortion is one of the diseases that can lead to physical, psychological, and, economical problems for both individuals and society. Recently a few numbers of genetic polymorphisms in kinase insert domain-containing receptor (KDR) gene are examined that can endanger the life of the fetus in pregnant women. Objective: The risk of KDR gene polymorphisms was investigated in Iranian women with idiopathic recurrent spontaneous abortion (RSA). Materials and Methods: A case controlled study was performed. One hundred idiopathic recurrent spontaneous abortion patients with at least two consecutive pregnancy losses before 20 weeks of gestational age with normal karyotypes were included in the study. Also, 100 healthy women with at least one natural pregnancy were studied as control group. Two functional SNPs located in KDR gene; rs1870377 (Q472H), and rs2305948 (V297I) as well as one tag SNP in the intron region (rs6838752) were genotyped by using PCR based restriction fragment length polymorphism (PCR-RFLP) technique. Haplotype frequency was determined for these three SNPs' genotypes. Analysis of genetic STRUCTURE and K means clustering were performed to study genetic variation. Results: Functional SNP (rs1870377) was highly linked to tag SNP (rs6838752) (D' value=0. 214; χ² = 16.44, p<0. 001). K means clustering showed that k = 8 as the best fit for the optimal number of genetic subgroups in our studied materials. This result was in agreement with Neighbor Joining cluster analysis. Conclusion: In our study, the allele and genotype frequencies were not associated with RSA between patient and control individuals. Inconsistent results in different populations with different allele frequencies among RSA patients and controls may be due to ethnic variation and used sample size. [ABSTRACT FROM AUTHOR]

Flax (Linum usitatissimum L.) is one of the most important fiber and oil crop plants cultivated since ancient time. The flax seeds contain high amount of omega- 3-fatty acids and biologically active lignans. In spite of economic importance of cultivated flax, no information is available on its genetic variability and population structure in Iran. Therefore, we used six inter-retrotransposon amplified polymorphism (IRAP) markers and 15 combined IRAP markers to reveal within and among population genetic diversity in this crop plant. We used 30 randomly selected plants in three geographical populations for present investigation. AMOVA test produced significant genetic difference (PhiPT = 0.40, P = 0.010) among the studied populations and also revealed that, 40% of total genetic variability was due to within population diversity while, 60% was due to among population genetic differentiation. Gst (0.78, P = 0.001), Hedrick, standardised fixation index (G'st = 0.83, P = 0.001), revealed that the studied populations are genetically differentiated. STRUCTURE plot based on admixture model revealed that the studied populations differed extensively in their genetic content, but some degree of shared alleles occurred between them. Some adaptive IRAP loci were identified by LFMM analysis. These loci were private alleles restricted to geographical populations. Data obtained may be used in breeding and hybridization program of flax in the country. [ABSTRACT FROM AUTHOR]

A collection of 52 local Iranian olive genotypes (North and south of Iran) and 36 olive cultivars from Mediterranean countries were studied in order to determine the genetic structure and their relationships. Thirteen primer pairs of microsatellite loci were selected that were known to be discriminating from previous study on olive. Polymorphic Information Content values ranged from 0.51 to 0.95. The Gst analysis for SSR loci revealed that some of the loci are more common while, some other loci are present in certain populations and show limited distribution among populations. Analysis of molecular variance (AMOVA) revealed that 11.71% of genetic variation attributed to differences between geographical regions while 88.29% came from variation in within regions ( P < 0.05). Pairwise Fst test revealed significant differences ( P < 0.05) between populations except between West and East Mediterranean populations. Neighbor Joining cluster analysis grouped the olive cultivars in two distinct clusters separating, olive genotypes located in North of Iran from the rest of olive cultivars studied. STRUCTURE and K means analyses showed the presence of distinct groups in olive trees and revealed variability in allelic compositions and their frequency between Iranian and West and East Mediterranean olive cultivars. The results of Bayesian clearly revealed that due to frequent occurrence of gene flow, the West and East Mediterranean olive cultivars had lower Fst values compared to the other population groups. In present study, genetic structure revealed admixture allelic compositions among Mediterranean olive cultivars different from Iran genotypes. [ABSTRACT FROM AUTHOR]

Cultivated cotton species Gossypium hirsutum L. and Gossypium barbadense L. are considered as the highest yield producing cottons, which are cultivated in several regions of Iran. Due to long time selection and cultivation practice, we need to broaden the cotton gene pool by producing new hybrids. Interspecific hybrids ensued after crossing G. hirsutum and G.barbadense. In the present study, cytogenetic charactristics and genome size of parental cotton species (G. hirsutum L. and G. barbadense) and their F1 and F2 hybrid progenies were studied. All cotton genotypes showed 2n = 4x= 52 chromosome number. The genotypes differed significantly in cytogenetic charcaters like number of total intercalary, and terminal chiasmata, mean number of ring chromosomes and quadrivalents (P < 0.05). The genome size of the genotypes varied from 1.345 to 4.37 picograms. This research revealed cytogenetic variability among the cotton offspring and if these are combined with agronomic characteristics, they can be used in cotton breeding. [ABSTRACT FROM AUTHOR]

Cotton (Gossypium hirsutum L.) is an important commercial crop that is cultivated in irrigated lands. Screening and selection of cultivars showing enhanced adaptation to drought is a target of cotton breeding programs. In order to study genetic variability in cotton genotypes under drought stress, eight cotton cultivars were screened using a 17% PEG treatment. Ten pots -each containing 5 seeds from each genotype- were planted using a randomized complete block design. Pots were distributed in two groups: with seedlings facing drought stress, and with well watered seedlings that were taken as controls. Twenty inter simple sequence repeat (ISSR) loci were used to determine genetic diversity. Four of the eight genotypes namely N2G80 and GT40 were rated as showing enhanced adaptation to drought stress while SKT133 and Bakhtegan were drought sensitive. The number of effective alleles -as measured by ISSR- varied from 1.189 (SKT133) to 1.363 (GT40). GT40 and SKT133 genotypes showed the highest and lowest gene diversity and percentage of polymorphism respectively. The highest number of specific alleles occurred in plant samples that were under drought stress. The high number of specific alleles in cotton plants under drought stress supports genetic changes in repeated motif changes within microsatellite regions. This genetic difference may be due to sequence changes in the genome or epigenetics. [ABSTRACT FROM AUTHOR]

Hybridization is one of the main strategies in cotton breeding producing new genotypes and increasing the genetic diversity in the cotton germplasm available. Thirty RAPD and ten ISSR markers were use to assess the genetic diversity in F2 progenies of tetraploid cotton hybrid genotypes (Gossypium hirsutum). Fifteen of 30 RAPD primers produced 126 reproducible bands with 66.44% polymorphism. Among the primers used, OPA09 primer showed highest value of number of effective allele (Ne), Shannon index (I), Nei's genetic diversity (H) (1.814, 0.638 and 0.447 respectively). The value of genetic identity varies from 0.65 between Siokra x No.200 and Tabladila x Siokra to 0.93 between Tabladila x Sindose and Sindose x No.200. Eight of 10 ISSR primers provided 87 reproducible bands with 67.82 % of polymorphism. UBC834 primer with AG repeats as a core sequence showed the highest value of Ne (1.570), I (0.477) and H (0.323) while combined primer UBC807+(GA)9T showed the highest PIC value (0.500). Based on ISSR data obtained, the hybrid cultivars Mehr x Tabladila and Tabladila x Siokra showed the lowest Nei's genetic identity (0.59), while, the hybrid cultivars No.200 x Belilzovar and Tabladila x Sindose showed the highest value. UPGMA Dendrograms obtained in RAPD and ISSR as well as combined RAPD and ISSR markers grouped together the hybrid genotypes which have Mehr cultivar as one of their parents. In conclusion, this study showed that using combined RAPD and ISSR data provides higher resolution of molecular profiles in hybrid genotypes [ABSTRACT FROM AUTHOR]

Sargassum species are one of economically important brown algae in south of Iran. In this study, morphological variation was assessed in three Sargassum species widely distributed species in the southwest of Iran (Oman sea): Sargassum tenerrimum J. Agardh, Sargassum glaucescens J. Agardh and Sargassum ilicifolium C.Agardh. At each site, ten different nonproductive individuals were selected with a minimum distance of 2-3 meters at low tide or snorkeling. We investigated ten measurable and ten categorical morphological parameters from three localities (Chabahar, Quatr and Tang) using ANOVA test as well as factorial analysis. The ANOVA test performed among populations of each species as well as among species showed significant difference (p<0.01) for all quantitative characters used. Different clustering like UPGMA and Neighbor Joining separated populations of each species studied based on measurable characters while both measurable and categorical characters have main rules in differentiation of three species PCA analysis confirmed cluster analysis. In total, both measurable and categorical characters enable to discriminate species while categorical parameters studied did not vary among populations of each species studied. This is the first study on evaluation of inter-population variation of Sargassum species in Iran. [ABSTRACT FROM AUTHOR]

Pomegranate Punica granatum (Lythraceae) is one of the most important horticultural plants of Iran cultivated in different areas with different cultivars having specific fruit characteristic such as size, color, time of ripening, disease resistance, taste, etc. The present study considers morphological and RAPD analysis of eighteen pomegranate cultivars of the country. Agro-morphological characters were studied by using factor analyses. Clustering separated cultivars in distinct groups indicating their morphological differences. Forty RAPD primers used produced 182 bands, out of which 414 bands were polymorphic (about 44%) with average of 4.55 polymorphic bands for each primer. Eight unique bands (cultivar-specific) were observed. Two cultivars of Malas Yazdi and Poost Ghermez Mamooli showed the highest similarity among the cultivars studied [ABSTRACT FROM AUTHOR]

Numerous olive cultivars are cultivated in Iran, mainly in the north. Ninety-two accessions belonging to 10 main olive cultivars were screened by 13 microsatellite markers revealing high genetic variability both within and between cultivars. In total, 72 alleles were detected with a mean number of 5.5 alleles per locus. Twenty-four unique allelic patterns were observed, whereas six genotypes showed 15 unique alleles. Heterozygosity ranged from 0.00 to 0.98, whereas the mean number of discrimination power and polymorphic information content were 0.55 and 0.54, respectively. The combination of 5 simple sequence repeat markers made discrimination of 84% of all accessions included in the study possible. The existence of homonyms, synonyms, or mislabeling as well as intracultivar polymorphism was revealed by allele differences between accessions of the same denomination. The phenogram showed variability among as well as between some cultivars, but most accessions with the same generic names were grouped together. [ABSTRACT FROM AUTHOR]

In spite of the abundance of antifungal therapies, 75% of women in the world suffer from the second most common cause of vaginal infection named vulvovaginal candidiasis. This complication is characterized with overgrowth of Candida albicans. The low efficacy and side effects of current antifungal therapies have convinced the researchers to look for a non-antibiotic based treatment such as cold atmospheric plasmas (CAP). The aim of this research was to evaluate the effects of CAP on C. albicans growth, ergosterol and biofilm formation. In addition, antibiotic resistance, phospholipase and proteinase activity, and structural properties were examined with different exposure duration. Putative critical effect of CAP on the expression of HSP90 as a target of anti-fungal therapy was investigated. ROS production in C. albicans exposed to CAP was assessed. For this purpose, C. albicans subjected to 0, 90, 120, 150, 180 and 210 s of He/O2 (2%), and non-treated cells as control were examined in terms of the mentioned virulence factors. The results showed that CAP had a significant effect on inhibition of C. albicans growth, Inhibition of biofilm formation, ergosterol content, and fluconazole and amphotericin B antibiotic sensitivity were significant in 210 s treatment group. This effect was validated based on changes of the cell architecture and morphology given the microscopy imaging results. The expression of HSP90 in both C. albicans ATCC 10231 and C. albicans PFCC 9362 was inhibited in 210 s of exposition. CAP exposition induced intracellular ROS, which may cause membrane damage and cell death in C. albicans. Taken together, the potential of CAP for therapeutic purposes in C. albicans -induced fungal infections is supported. • Cold atmospheric plasma (CAP) strongly inhibited the growth of C. albicans. • CAP reduced ergosterol content and inhibited biofilm production in C. albicans. • CAP diminished proteinase and phospholipase activity and induced ROS generation. • CAP caused morphological changes in C. albicans by targeting membranous structures. • CAP potently suppressed HSP90 gene expression in C. albicans. [ABSTRACT FROM AUTHOR]

Purpose: Honey is a promising source of bacterial strains producing metabolites with antimicrobial activity. There is a great variety in the antimicrobial activity of honey from different areas of nature. Therefore, the aim of present study was to investigate the antibacterial activity of Iranian honey from different regions and to optimize the culture condition for the highly potent bacterial isolate.Honey samples were collected from ten different regions of Iran and were screened for bacteriocin-producing bacteria. The best bacteriocin-producing strain was characterized and identified by 16S rDNA analysis. One-factor-at-a-time method was used for optimization of culture medium and the yield and time-course of bacteriocin production were compared in both shake flask and bio-reactor.The Bacillus subtilis SB1 that was isolated from Sabalan honey showed potent antibacterial activity with prominent thermal stability. The optimum medium for the bacteiocin production was a yeast extract-based medium. The optimum incubation temperature for bacteriocin production was 34 °C. Bacteriocin production was higher near neutral pH conditions than that produced at acidic or alkaline environment. The results of cell growth and bacteriocin assays revealed that the exponential phase of growth and antibacterial compounds production was started rapidly in bioreactor than flask.Findings of this study supported the folkloric application of honey against some infectious diseases. B.subtilis SB1 that isolated from Sabalan honey was a potential source for bacteriocins-like compounds. Our studies suggested a simple buffered nitrogen-based medium for SB1 growth and bacteriocin activity in both shake flask and bioreactor.Methods: Honey is a promising source of bacterial strains producing metabolites with antimicrobial activity. There is a great variety in the antimicrobial activity of honey from different areas of nature. Therefore, the aim of present study was to investigate the antibacterial activity of Iranian honey from different regions and to optimize the culture condition for the highly potent bacterial isolate.Honey samples were collected from ten different regions of Iran and were screened for bacteriocin-producing bacteria. The best bacteriocin-producing strain was characterized and identified by 16S rDNA analysis. One-factor-at-a-time method was used for optimization of culture medium and the yield and time-course of bacteriocin production were compared in both shake flask and bio-reactor.The Bacillus subtilis SB1 that was isolated from Sabalan honey showed potent antibacterial activity with prominent thermal stability. The optimum medium for the bacteiocin production was a yeast extract-based medium. The optimum incubation temperature for bacteriocin production was 34 °C. Bacteriocin production was higher near neutral pH conditions than that produced at acidic or alkaline environment. The results of cell growth and bacteriocin assays revealed that the exponential phase of growth and antibacterial compounds production was started rapidly in bioreactor than flask.Findings of this study supported the folkloric application of honey against some infectious diseases. B.subtilis SB1 that isolated from Sabalan honey was a potential source for bacteriocins-like compounds. Our studies suggested a simple buffered nitrogen-based medium for SB1 growth and bacteriocin activity in both shake flask and bioreactor.Results: Honey is a promising source of bacterial strains producing metabolites with antimicrobial activity. There is a great variety in the antimicrobial activity of honey from different areas of nature. Therefore, the aim of present study was to investigate the antibacterial activity of Iranian honey from different regions and to optimize the culture condition for the highly potent bacterial isolate.Honey samples were collected from ten different regions of Iran and were screened for bacteriocin-producing bacteria. The best bacteriocin-producing strain was characterized and identified by 16S rDNA analysis. One-factor-at-a-time method was used for optimization of culture medium and the yield and time-course of bacteriocin production were compared in both shake flask and bio-reactor.The Bacillus subtilis SB1 that was isolated from Sabalan honey showed potent antibacterial activity with prominent thermal stability. The optimum medium for the bacteiocin production was a yeast extract-based medium. The optimum incubation temperature for bacteriocin production was 34 °C. Bacteriocin production was higher near neutral pH conditions than that produced at acidic or alkaline environment. The results of cell growth and bacteriocin assays revealed that the exponential phase of growth and antibacterial compounds production was started rapidly in bioreactor than flask.Findings of this study supported the folkloric application of honey against some infectious diseases. B.subtilis SB1 that isolated from Sabalan honey was a potential source for bacteriocins-like compounds. Our studies suggested a simple buffered nitrogen-based medium for SB1 growth and bacteriocin activity in both shake flask and bioreactor.Conclusions: Honey is a promising source of bacterial strains producing metabolites with antimicrobial activity. There is a great variety in the antimicrobial activity of honey from different areas of nature. Therefore, the aim of present study was to investigate the antibacterial activity of Iranian honey from different regions and to optimize the culture condition for the highly potent bacterial isolate.Honey samples were collected from ten different regions of Iran and were screened for bacteriocin-producing bacteria. The best bacteriocin-producing strain was characterized and identified by 16S rDNA analysis. One-factor-at-a-time method was used for optimization of culture medium and the yield and time-course of bacteriocin production were compared in both shake flask and bio-reactor.The Bacillus subtilis SB1 that was isolated from Sabalan honey showed potent antibacterial activity with prominent thermal stability. The optimum medium for the bacteiocin production was a yeast extract-based medium. The optimum incubation temperature for bacteriocin production was 34 °C. Bacteriocin production was higher near neutral pH conditions than that produced at acidic or alkaline environment. The results of cell growth and bacteriocin assays revealed that the exponential phase of growth and antibacterial compounds production was started rapidly in bioreactor than flask.Findings of this study supported the folkloric application of honey against some infectious diseases. B.subtilis SB1 that isolated from Sabalan honey was a potential source for bacteriocins-like compounds. Our studies suggested a simple buffered nitrogen-based medium for SB1 growth and bacteriocin activity in both shake flask and bioreactor. [ABSTRACT FROM AUTHOR]

• Catharanthus roseus seedling growth under Cold Plasma, Mathyl jasmonate, Putrescine and their combinations. • Catalase and peroxidase enzyme activities and alkaloid production. • Genetic variation of treated plants by molecular markers Madagascar Periwinkle (Catharanthus roseus) as a medicinal plant produces more than 100 monoterpenoid indole alkaloids. In present study, six groups including; control, Cold atmospheric plasma, methyl jasmonate hormone, putrecsine, combination of methyl jasmonate and cold plasma, and putrecsine and cold plasma treated plants were studied. Seed germination, morphological traits, catalase and peroxidase enzyme activities and amount of total alkaline were investigated in treated plants. Genetic variation of treated and control plants were evaluated by 21 coding targeted markers; Sequence related amplified polymorphism (SRAP) and Target related amplified polymorphism (TRAP). Cold plasma + putrescine treated plants showed highest amount of stem length as well as leaf length and wide size. Peroxidase and catalase activities were significantly increased in treated plants of putrescine + cold plasma and methyl jasmonate treated plants respectively. Total alkaline was significantly higher in treated plants of putrescine, and putrescine + cold plasma. In both TRAP and SRAP molecular markers, treatments of methyl jasmonate + plasma and putrescine + cold plasma had the highest level of genetic polymorphism and both markers showed moderate discriminating power to differentiate plants treated from each other. The putrescine + cold plasma is one of the best treatments for improved germination rate and seedling growth as well as enzyme activity and amount of alkaloid in C. roseus plants as well as higher genetic variation. [ABSTRACT FROM AUTHOR]

Background: In the present study, we investigate the prevalence of the GJB2 gene mutations, and deletions in the GJB6 gene, namely del (GJB6‐D13S1830) and del (GJB6‐D13S1854), in patients with autosomal recessive non‐syndromic hearing loss (ARNSHL) from the central region of Iran. Methods: One hundred and thirty‐one unrelated ARNSHL cases from the central part of Iran were recruited. Among them, 81% (106 cases) had at least two affected relatives. Coding and noncoding regions of the GJB2 gene were sequenced. Multiplex PCR was used for analysis of del (GJB6‐D13S1830) and del (GJB6‐D13S1854) deletions in GJB6. Results: The GJB2 variants were found in 16.79% (22/131) of the patients. The pathogenic variants were 21/131 (16.03%). The nonpathogenic variants were 1/131 (0. 07%). Allele frequency of the c.35delG as the pathogenic variant was the most common with 59.52% (25/42). The remaining pathogenic variants were c.235delC, p.T8M, p.R32H, p.R143Q, p.R143W, c‐23+1G>A. The only nonpathogenic variant was polymorphism p.V27I. Further segregation analysis showed that variant of p.R143Q might have incomplete penetrance. None of the patients had targeted deletions in the GJB6 gene. Conclusion: In comparison with reports from other areas of Iran, c.35delG demonstrates the highest frequency within the central region (accounting for 57.14% of cases), probably resulting from the founder effect and consanguineous marriage. The pathology of ARNSHL in such patients could be attributed to defects in Connexin 26 encoded by GJB2. [ABSTRACT FROM AUTHOR]

Purpose: Poor response to ovarian stimulation (POR) typically is reflected as decreased follicular response and low estradiol (E2) levels following ovarian stimulation by FSH/HMG. Many genes are involved in oocyte maturation, and demographic features and lifestyle can affect the oocyte maturity and developmental competence. The present study was conducted to investigate the magnitude of gene expression and lifestyle habits in POR women as compared to healthy women, using different statistical and computational methods. Methods: Fifty women in the two groups were studied. The study groups included POR women (n = 25) with 1–9 released oocytes, and the control group (normal women, n = 25) with 9–15 released oocytes. Quantitative PCR was used to estimate the expression of FIGLA, ZAR1, WNT4, LHX8, APC, H1FOO, MOS, and DMC1 genes in granulosa cells. Results: The results showed no significant difference in the magnitude of the studied genes' expression and linear discriminant analysis did not differentiate the studied groups based on all the genes together. Redundancy analysis (RDA) and latent factor mixed model (LFMM) results produce no significant association between the genes' expression magnitude and the geographical variables of the patients' local habitat. Linear discriminant analysis (LDA) of the demographic features differentiated the two groups of women. Conclusion: Our results indicate that demographic features may have an effect on sample gene expression levels. [ABSTRACT FROM AUTHOR]

It is estimated that about 15% of couples concern with the inability to conceive after 1 year of unprotected intercourse. Genetic and environmental factors play important roles in male infertility. MTHFR (Methylene Tetrahydro Folate Reductase) is encoded by a gene which maps to human chromosome 1p36.22. The c.677C>T mutation causes an amino - acid change of Alanine to Valine (Ala222Val) in exon 4 which is related to oligospermia and azoospermia. Our purpose was to investigate the association of rs1801133 in MTHFR gene with azoospermia and also genetic diversity of some Iranian folks. Peripheral blood samples of 100 men suffered from azoospermia and 55 controls collected from Infertility Clinic of Jihad-university of Qom. DNA was extracted by salting-out method and SNP-Genotyping was performed by PCR-RFLP technique and sequencing. Furthermore, statistical and population genetics analysis were done. The rs1801133 showed no significant relation with azoospermia according to statistical analysis. Furthermore, population genetics analysis showed that there is no genetic differentiation between Turk and Fars ethnics and case and control individuals. MTHFR studied SNP has no significant relation with azoospermia infertility, although further studies on more samples are demanded. [ABSTRACT FROM AUTHOR]

CTLA4-Ig (Abatacept) has been produced to suppress immune response by inhibition of T cells functions in autoimmune disease. A new drug, which is called belatacept, has recently been recently developed that is more efficient. The development has been occurred by two substitutions (A29Y, L104E) in the extracellular domain of CTLA4. In the present study, the bioinformatics analysis was used in order to make a new structure that has a better function in comparison with belatacept. Firstly, eight different structures were designed. Thereafter, the secondary and 3D structures, mRNA structure, docking of chimeric proteins with CD80/CD86, antigenicity and affinity of designed chimeric molecules were predicted. Based on the criteria, a new candidate molecule was selected and its gene synthesized. The gene was cloned and expressed in E. coli BL21 (DE3) successfully. The purified rCTLA4-Ig was analyzed by SDS-PAGE, western blotting, and ELISA. Circular dichroism analysis (CD analysis) was used for characterization of the rCTLA4-Ig. Affinity of rCTLA4-Ig was also evaluated by the flow cytometry method. Finally, its biological activity was determined by T cell inhibition test. The results showed rCTLA4-Ig and the belatacept protein have some similarities in structure and function. In addition, rCTLA4- Ig was able to bind CD80/CD86 and inhibit T cell function. Although flow cytomery results showed that the standard protein (CTLA4-Ig), represented better affinity than rCTLA4-Ig, the recombinant protein was able to inhibit T cell proliferation as well as CTLA4-Ig. [ABSTRACT FROM AUTHOR]

BACKGROUND: Prostate cancer (PCa) as the first men's common cancer in the world and the third cancer in Iranian men is a heterogeneous disorder which sometimes several biopsies are needed for its diagnosis. OBJECTIVES: The aim of current study is finding new biomarkers in order to diagnose of PCa at the earliest possible stage. Hence, the relationship between rs1800629 and rs361525 polymorphisms of TNF- α gene with PCa was investigated. MATERIALS AND METHODS: Blood DNA samples were collected from 100 patients with PCa, 110 with BPH, and 110 controls. Collected samples were examined using PCR-RFLP and Tetra-ARMS-PCR techniques to detect the desired polymorphisms. RESULTS: The frequency of rs1800629 genotypes in smokers was significantly different from non-smokers with PCa (p = 0.001). Logistic regression analysis results showed that GA heterozygotes in comparison to GG homozygotes had higher risk of developing Benign Prostatic Hyperplasia (BPH) or prostate cancer. However, no significant correlation was considered between the risk of PCa and the TNF- α gene polymorphisms (rs1800629 and rs361525). CONCLUSIONS: Although, the achieved results of this investigation demonstrated that the two examined genetic variants do not seem to be suitable markers for early diagnosis of prostate cancer in this pilot study; however increased risk for the disease is shown in GA heterozygotes and smokers which is indicative of some epigenetic factors influence on prostate cancer etiology. [ABSTRACT FROM AUTHOR]

The Epidermal Growth Factor Receptor (EGFR) has been of high importance as it is over expressed in a wide diversity of epithelial cancers, promoting cell proliferation and survival pathways. Recombinant immunotoxins (ITs) have emerged as a promising targeted therapy for cancer treatment. In this study, we aimed to investigate the antitumor activity of a novel recombinant immunotoxin designed against EGFR. Using an in silico approach, we confirmed the stability of the RTA-scFv fusion protein. The immunotoxin was successfully cloned and expressed in the pET32a vector, and the purified protein was analyzed by electrophoresis and western blotting. In vitro evaluations were conducted to assess the biological activities of the recombinant proteins (RTA-scFv, RTA, scFv). The novel immunotoxin demonstrated significant anti-proliferative and pro-apoptotic effects against cancer cell lines. The MTT cytotoxicity assay revealed a decrease in cell viability in the treated cancer cell lines. Additionally, Annexin V/Propidium iodide staining followed by flow cytometry analysis showed a significant induction of apoptosis in the cancer cell lines, with half maximal inhibitory concentration (IC50) values of 81.71 nM for MDA-MB-468 and 145.2 nM for HCT116 cells (P < 0.05). Furthermore, the EGFR-specific immunotoxin exhibited non-allergenic properties. The recombinant protein demonstrated high affinity binding to EGFR. Overall, this study presents a promising strategy for the development of recombinant immunotoxins as potential candidates for the treatment of EGFR-expressing cancers. [ABSTRACT FROM AUTHOR]

Aloe belonging to the family Asphodelaceae family, formerly Liliaceae, is a genus of herbaceous and succulents. Shoot tip of 2-3 cm were collected from offshoot of Aloe barbadensis and Aloe littoralis, explants were surface sterilized, after were inoculated with MS medium containing various concentrations of BA (0.5, 1, 2 mg/L), IAA (0.5, 1 mg/L). The cultures were incubated at 25 ± 2°C under a 16 h photoperiod. In A. barbadensis and A. littoralis, the best treatment for highest shoot number and bud proliferation was MS medium containing 2 mg/L BA and 0.5 mg/L IAA. Maximum percentage of proliferated shoot buds (90% and 95%) from a single explant were obtained in MS medium after 4-5 weeks of the second and the first subcultures, respectively. Both of the species were adventitious root induced in MS medium containing 0.5 mg/L BAP and 0.5 mg/L IAA after the second subculture. Genome size of the regenerated plants in A. barbadensis and A. littoralis species were differed in different treatments (from 30.66 pg to 39.62 pg, 2C value). One sample T-test showed significant difference (P=0.001) between genome sizes of two species studied when A. littoralis showed higher C-value in comparison to A. barbadensis. Different genome sizes were also indicated among treatments and subcultures. The mixoploids identified in flow cytometry histograms by the presence of two or three peaks of in A. barbadensis and A. littoralis histograms in apple regenerated plants in different treatments and subcultures in treatment 2 (BA 1 mg/L, IAA 0.5 mg/L). [ABSTRACT FROM AUTHOR]

Several parameters of the in vitro regenerated, grown and adapted to the greenhouse plantlets were studied and compared (P ≤ 0.05). The mean length of stem and leaves of the in vitro regenerated pineapple significantly decreased under greenhouse conditions. The chlorophyll pigments content in the regenerated plantlets was increased and carotenoid, anthocyanins, carbohydrate, total phenols, and bromelain showed a relatively lower amount in the in vitro plantlets. Cluster analysis was performed on the basis of Dice genetic distance matrix for the thirty regenerated plants, as well as the mother plant. On the basis of the Neighbor Joining Cluster the plantlets were grouped into three main clusters. AMOVA test showed that 46 % of the genetic variance is within the groups, while 54 % of the rest belongs in-between the three groups. On the basis of this analysis a significant genetic difference was observed within the regenerated plantlets (P=0.01). [ABSTRACT FROM AUTHOR]

6-Gingerol is the major pungent ingredient of ginger with anti-inflammatory and antioxidant properties. In this study, we evaluate the effects of 6-gingerol on the biochemical parameters and ovarian histological improvements in estradiol valerate (EV) induced PCOS rats. Thirty six female Wistar rats were divided into 4 groups: control, received normal diet, PCOS control, received 4 mg/kg EV injection for 28 days and two experimental groups, received an EV injection for 28 days and followed by 6-gingerol (200 μg/ kg and 400 μg/ kg ) for 14 days. The administration of EV led to increase body and ovarian weights, abnormality in serum sex steroid profile, decrease in antioxidant activity and increase in COX-2 gene expression. 6-gingerol treatments, particularly the 400 μg/ kg dose, markedly attenuated these alterations. 6-gingerol showed beneficial effects in the EV induced PCOS rats via decreased expression of COX-2, restored biochemical parameters to normal and decreased of cysts in the ovaries. [ABSTRACT FROM AUTHOR]

BACKGROUND: Coronary artery disease (CAD) is the one of the most common heart diseases, being the main factor of mortality and morbidity worldwide. CAD has been known as a multifactorial disease and its progression depends on genetic and environmental factors. Numerous studies have shown evidence for association between 12q24.3 locus and CAD. OBJECTIVES: In the present study, the association of two HNF1A polymorphisms, rs2259816 and rs7310409, located on 12q24.3 locus with CAD was investigated in Iranian patients. METHODS: Whole genomic DNA was isolated from peripheral blood obtained from 411 Iranian volunteer. Then, the polymorphisms rs2259816 and rs7310409 located on 12q24.3 locus were genotyped using TaqMan Probe Real Time PCR. Statistical analysis was performed by SPSS software, version 19. RESULTS: rs2259816 (p-value = 0.006) and rs7310409 (p-value = 0.001) showed statistically significant association with CAD risk. Our results indicated that there was a significant correlation between rs2259816 and blood triglyceride. However, no correlation was found between rs2259816, rs7310409 and other CAD risk factors. [ABSTRACT FROM AUTHOR]

There is an increasing interest in the use of nano-fertilizer as an alternative to traditional fertilizers, in order to increase the secondary metabolites in medicinal plants. The present study was conducted to observe the changes in morphological and biochemical properties of three populations of Colchicum speciosum L., from Iran (Filband, Sangdarka, and Kelerd), upon conventional (1.1 and 2.2 mg L-1) and nano-N (1.5 and 3 mg L-1) fertilizers based on completely randomized design (CRD). The anthocyanin and phenolic contents of leaves and corms were determined with a spectrophotometer and corm colchicine was extracted by high-performance liquid chromatography (HPLC). The results represented different responses of morphological and biochemical attributes to populations due to their different origins. Filband with higher altitude showed increased leaf area, plant length, corn weight and diameter when treated with N fertilizers. Increased total phenolic component (TPC) in leaves and corm and also leaf total flavonoids component (TFC) were reported in Filband upon N fertilizers. All fertilizers led to decreased leaf anthocyanin when N fertilizers were applied. A noticeable increase in colchicine was obtained in Kelerd and Sangdarka when plants were treated with 1.5 mg L-1 nano-N fertilizer. Heat map analysis of corm anthocyanin, leaf anthocyanin, leaf TPC, leaf TPC, and colchicine amount showed higher variability under the treatments. According to principal component analysis (PCA), corm TPC, leaf TFC, corm TFC, leaf area, and corm weight were specific traits of Kelerd. In conclusion, different responses of morphological and biochemical traits were reported between populations, and N fertilizers exerted diverse effects on these traits. The population from higher altitudes showed higher phenolic content and lower anthocyanin accumulation under conventional and nano-N fertilizers. [ABSTRACT FROM AUTHOR]

Cirsium aduncum Fisch. & C.A.Mey. Ex DC. and C. haussknechtii Boiss., (Asteraceae) are important medicinal plant species that grow in different geographical regions of Iran. We had no knowledge about population genetic structure, intra-specific and inter-specific gene flow and the presence of hybrid zone for this two species in Iran. Therefore, in order to provide data for conservation of these two medicinally important species, the population genetic analysis and morphometric studies were performed in 18 geographical populations of these species. ANOVA and MDS analyses revealed significant morphological difference among the studied populations in either species, while MDS plot showed morphological overlap in plants of these two species. AMOVA test revealed significant genetic difference among the studied populations. Mantel test showed positive significant correlation between genetic and geographical distances and the occurrence of isolation by distance. Population assignment test and STRUCTURE plot of genetic data revealed inter-specific introgression between these species. [ABSTRACT FROM AUTHOR]

Objective To investigate the association between the polymorphism of miR-146a and The ubiquitin-like with PHD and ring-finger domains 1 ( UHRF1) expression in patients with gastric cancer. Methods MiR-146a rs2910164 was genotyped in 130 patients with gastric cancer and 130 cancer-free individuals using polymerase chain reaction (PCR)-restriction fragment length polymorphism. UHRF1 expression was analyzed in 22 gastric cancer tissues and their adjacent normal tissues using quantitative real-time PCR. Results No significant differences in genotype distributions of miR-146a rs2910164 were found between cases and controls, but we observed that grade II tumors were more frequently detected in patients with CG/CC genotype compared to those with CC genotype. UHRF1 expressions in cancerous tissues were significantly higher than in noncancerous tissues (1.89-fold). Patients with CC genotype showed a significant increase in UHRF1 expression in comparison to the carriers of GG/CG genotype. A higher UHRF1 expression was associated with cancer stage IV and grade III ( P<0.05). Conclusion The overexpression of UHRF1 was correlated with the stage and grade of gastric cancer and is associated with the genotype distribution of rs2910164. [ABSTRACT FROM AUTHOR]

Helichrysum armenium and H. rubicundum are two medicinally important plants of Iran that are distributed in various regions of the country. They are extensively used by locals as medicinal plants and a great negative selection pressure is applied on them. Therefore, due to importance of these plant species, we performed a population genetic study in both species. For this study, we used 66 randomly collected plants from 24 geographical populations of Helichrysum armenium and H. rubicundu. These species had areas of overlap and contact and we found some intermediate plants that were included in our study too. UPGMA and MDS analyses revealed morphological separation of these closely related species and placed intermediate plants in an intermediate position. ISSR analysis revealed inter-population genetic diversity and K-Means clustering and STRUCTURE analyses revealed populations. genetic stratification in both species. Genetic difference of the studied populations was not correlated to geographical distance. Triangle plot of Bayesian analysis and NeighborNet plot showed inter-specific gene flow. The studied populations showed plants with 2n = 2x = 14 and 2n = 4x = 28 chromosomes and differed significantly in their meiotic behavior. Therefore, a combination of genetic stratification, and genetic admixture as well as polyploidy and chromosomes structural changes, have played rule in Helichrysum diversification. [ABSTRACT FROM AUTHOR]

Vitis vinifera L. is one of the economically important plant crops worldwide which is a valuable food source for humans. This precious plant species has several local varieties and accessions which are continuously under selection and cultivation. Due to these human activities, the grape faces genetic homogeneity and erosion. Therefore, it is important to investigate available genetic diversity in grape plants all over the world. We aimed to study the genetic structure and diversity as well as chemical differences of seven grape cultivars in the country. We used SSR, and SRAP molecular markers for genetic diversity analyses, as well as biochemical traits. Both molecular markers showed a medium to moderate genetic variability in the studied grape cultivars (about 20% genetic polymorphism). Similarly, both molecular markers differentiated the studied cultivars into two genetic groups. AMOVA indicated significant genetic difference in these cultivars. ANOVA analysis of flavonols (quercetin, myricetin, kaempferol, and rutin) contents of seeds extract by HPLC indicated the significant difference (P <0.01) among grape cultivars. PCA biplot of cultivars based on chemical features separated these cultivars into two major groups according to their flavone and flavonoid contents. Pairwise Mantel tests performed between molecular and chemical data showed a significant association between SSR and SRAP data, but no significant association was obtained between either SSR or SRAP data with chemical features in grape cultivars studied. A heat-map constructed based on combined molecular and chemical data revealed that some of the studied grape cultivars are distinct in their genetic and chemical features. [ABSTRACT FROM AUTHOR]

Background: Schizophrenia (SCZ) is a severe mental disorder in which people interpret reality abnormally. Different studies indicated a complex polygenic control over SCZ. In the present study, we investigated the potential correlation between ten SNPs among MicroRNA (MIR) and their target genes; rs369770942, rs143525573, rs200982455, rs530404895, rs753764536, rs374732351, rs4680, rs165599, rs340597269, and rs10759, and schizophrenia in the Iranian population. Results: The results revealed that the T allele in rs200982455 increased the risk factor by 3.19 times. We obtained a significant association between rs165599 and schizophrenia in codominant, dominant, and overdominant inheritance models (P = 0.016, P = 0.01, P = 0.004, respectively). Moreover, the risk of schizophrenia increased in the presence of the G allele in rs165599 up to 2.12, 2.35, and 2.28 times, respectively. The A allele in rs10759 increased the risk factor up to 1.05 times. Conclusion: Our finding showed that some of the studied SNPs within the genes and MIRs involved in the dopaminergic pathway may consider as a biomarker in the diagnostic patterns in Schizophrenia. [ABSTRACT FROM AUTHOR]

Human papillomavirus (HPV) L1, L2 and E7 proteins were used as target antigens for development of preventive and therapeutic vaccines. Moreover, linkage of antigens to heat shock proteins (HSPs) could enhance the potency of vaccines. Curcumin and nanocurcumin compounds were suggested as the chemopreventive and chemotherapeutic agents against cancer. In this study, two multiepitope DNA and peptide-based vaccine constructs (L1-L2-E7 and HSP70-L1-L2-E7) were used along with curcumin and nanocurcumin to evaluate immune responses, and protective/therapeutic effects in tumor mouse model. At first, the multiepitope L1-L2-E7 and HSP70-L1-L2-E7 fusion genes were subcloned in eukaryotic and prokaryotic expression vectors. The recombinant multiepitope peptides were generated in E. coli strain. Then, the cytotoxic effects of curcumin and nanocurcumin were evaluated on HEK-293 T non-cancerous and C3 cancerous cells. Finally, mice vaccination was performed using different regimens. Curcumin and nanocurcumin compounds were administered alone or along with different vaccine constructs. Our data indicated that the use of nanocurcumin along with the multiepitope HSP70-L1-L2-E7 vaccine construct could completely protect mice against HPV-related C3 tumor cells, and eradicate tumors in a therapeutic test. Furthermore, nanocurcumin showed higher protection than curcumin alone. Generally, curcumin and nanocurcumin compounds could reduce tumor growth synergistically with the multiepitope vaccine constructs, but they did not influence the immune responses in different regimens. These data demonstrated that the designed multiepitope vaccine constructs along with curcumin and nanocurcumin can be used as a promising method for HPV vaccine development. [ABSTRACT FROM AUTHOR]