فهرست مطالب kheirollah yari

Infertility is one of the most common health problems in the world, affecting about 15% of couples. Abnormal DNA methylation in sperm can be affective in male infertility. 5 and 10-methylene tetrahydrofolate reductase (MTHFR) is a vital enzyme for regulating DNA methylation patterns in sperm and the folate cycle, playing a crucial role in spermatogenesis processes. A Change in the expression level of this gene can lead to male infertility. This study aimed to investigate the level of methylation of the MTHFR gene in different methylated regions (DMRs) and the correlation between sperm DNA methylation patterns with semen quality parameters in fertile and infertile individuals. Semen samples were collected from 30 infertile men and 15 fertile men. After DNA treatment with sodium bisulfite, the methylation status of the MTHFR gene in DMRs was evaluated by qMSP method. There is a significant correlation between DMR methylation of the MTHFR gene in the infertile group compared to the fertile group (p = 0.036) and an increase was observed in infertile men. Additionally, a significant decrease in sperm parameters (sperm concentration, count, and morphology) was observed in the men of the infertile group. Also, there was a negative correlation between the methylation of DMRs of the MTHFR gene and semen parameters in patients. Hypermethylation of the MTHFR gene in DMRs in infertile individuals is associated with a decrease in sperm parameters.

Chimeric antigen receptor (CAR) T cell therapy has recently emerged as a promising approach for the treatment of different types of cancer. Improving CAR T cell manufacturing in terms of costs and product quality is an important concern for expanding the accessibility of this therapy. One proposed strategy for improving T cell expansion is to use genetically engineered artificial antigen presenting cells (aAPC) expressing a membrane-bound anti-CD3 for T cell activation. The aim of this study was to characterize CAR T cells generated using this aAPC-mediated approach in terms of expansion efficiency, immunophenotype, and cytotoxicity.

In this experimental study, we generated an aAPC line by engineering K562 cells to express a membrane-bound anti-CD3 (mOKT3). T cell activation was performed by co-culturing PBMCs with either mitomycin C-treated aAPCs or surface-immobilized anti-CD3 and anti-CD28 antibodies. Untransduced and CD19-CARtransduced T cells were characterized in terms of expansion, activation markers, interferon gamma (IFN-γ) secretion, CD4/CD8 ratio, memory phenotype, and exhaustion markers. Cytotoxicity of CD19-CAR T cells generated by aAPCs and antibodies were also investigated using a bioluminescence-based co-culture assay.

Our findings showed that the engineered aAPC line has the potential to expand CAR T cells similar to that using the antibody-based method. Although activation with aAPCs leads to a higher ratio of CD8+ and effector memory T cells in the final product, we did not observe a significant difference in IFN-γ secretion, cytotoxic activity or exhaustion between CAR T cells generated with aAPC or antibodies.

Our results show that despite the differences in the immunophenotypes of aAPC and antibody-based CAR T cells, both methods can be used to manufacture potent CAR T cells. These findings are instrumental for the improvement of the CAR T cell manufacturing process and future applications of aAPC-mediated expansion of CAR T cells.

There are several methods for genotyping single nucleotide polymorphisms (SNPs). Polymerase chain reaction with confronting two-pair primers (PCR-CTPP) is an inexpensive, quick, reliable, time saving and cost effective method that is applicable for SNPs genotyping compared to other related methods. In the PCR-CTPP technique, allele-specific products of the genes are selectively amplified by adding two pairs of four primers and ordinarily prepared PCR mixture in a single PCR tube. Four allele-specific primers are F1 (sense) and R1 (antisense) primers for one allele that R1 has an anti-sense nucleotide at the 3′ end and F2 (sense) and R2 (antisense) for the other allele that F2 has a sense nucleotide at the 3′ end. PCR-CTPP produces three products with different sizes which can make possible genotyping of SNP by gel electrophoresis. To increase the reproducibility and accuracy of the PCR-CTPP results, it is better to optimize protocol before sample genotyping. The aim of this study was to review the currently used strategies for optimum application and determination of advantages and disadvantages of the PCR-CTPP method in SNPs genotyping.

The articles related to the PCR-CTPP method were searched from databases such as ISI Web of science, Science direct, Pubmed, Google Scholar, SID, and Scopus.

For more efficiency and reproducibility of the PCR-CTPP method, we should consider the specificity and similarity of melting temperatures of the designed primers by using algorithms  and specific primer design softwares.

Strategies for optimizing the concentration ratio of internal and external primers, adding amplification additives to the PCR reaction mix, and use of the touchdown program are also recommended.

The association of 1,25-dihydroxy vitamin D3 (1,25(OH)2D3) and its receptor, vitamin D receptor (VDR), with cancer types have been studied. However, there are controversial findings regarding the association of specific VDR polymorphisms with different kinds of cancers. In the current study, we investigated the association of VDR polymorphisms (Fok1 (rs2228570), ApaI (rs7975232), BsmI (rs1544410), and TaqI (rs731236)) with the risk of gastric cancer in a Kurdish population of Kermanshah in Iran for the first time.

In this case-control study, the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used in 99 gastric cancer patients and 100 healthy subjects as controls.

The frequencies of f (FokI), b (BsmI), t (TaqI), and a (ApaI) alleles were: 55.6%, 27.3%, 62.1%, and 44.95% in the patient group, respectively and 42%, 29.5%, 54.5%, and 46.0% in the control group, respectively. Analysis of the results indicated that there was a positive association between the frequency of FokI genotypes with gastric cancer risk (p= 0.021). However, no statistically significant association of BsmI, Taq1, and ApaI polymorphisms of VDR was detected in gastric patients when compared with healthy individuals.

VDR-FokI polymorphism could increase the risk of GC development and predispose to the disease by mechanisms.

In recent years, several studies have shown the association between exercise and decreased risk of mortality in patients with breast cancer. However, the effects of combined resistance and endurance training on salivary Interleukin-12 (IL-12), tumor necrosis factor (TNF-α), Cortisol, and Testosterone levels in patients with breast cancer have not been investigated.

This study aimed at determining the effect of 8 weeks of combined resistance and endurance training on salivary IL-12, TNF-α, Cortisol, and Testosterone levels in women with breast cancer.

Forty-two postmenopausal women with breast cancer were randomly selected and divided into training (intervention) and control groups. The training group performed resistance training with 2 to 3 sets, 10 to 18 repetitions, 50 to 70% 1 repetition maximum (1RM), and aerobic exercise with 50 to 70% maximum heart rate (maxHR) (12-14 degrees borg scale) for 20 to 40 minutes for 8 weeks. The salivary IL-12, TNF-α, cortisol, and testosterone levels were measured, using the enzyme-linked immunosorbent assay (ELISA) method. Two-way analysis of variance repeated measure was also used to analyze variance with the confidence interval of 95%.

In the training group, there was a significant decrease in salivary TNF-α levels, cortisol, TNF-α/IL-12 ratio, and variables of weight, fat percentage, body mass index (BMI), and waist circumference (P < 0.05). Also, the results showed a significant increase in salivary testosterone and testosterone/cortisol ratio in the intervention group (P < 0.05). However, no significant changes were observed in the interaction between-group and time in IL-12 and waist–hip ratio (WHR) values (P > 0.05).

The results indicate that resistance and endurance training could be used as a useful method to improve salivary pro-inflammatory factors and hormonal levels in patients with breast cancer. Medical oncologists can underline a resistance and endurance training program for patients with breast cancer under their care.

Chronic myeloid leukemia (CML) is a common type of cancer. Leukemia is associated with diverse molecular and genetic changes, including loss and gain of chromosomes, gene deletions, duplications, point mutations, and gene fusions derived from chromosomal translocations. Advanced genetic tests are done to diagnose leukemia; however, many patients are not diagnosed because many of the symptoms are vague, unspecific, and referable to other diseases.

Following the extraction of genomic DNA from CML patient, we performed exome sequencing. Variants were detected via GATK software. Novel alterations in CML cases were then visualized in the integrated genome browser. To enrich our findings, we included exome sequencing data pertaining to 11 individuals. The data was deposited in the ENA database in our analysis. Afterwards, we verified insertion in the ATXN3 gene through performing PCR reactions for both healthy and CML cases.

We identified an alteration in the genomic sequence of the ATXN3 gene in the CML cases.

A correlation existed between insertion in the ATXN3 gene and positive CML cases. These findings might be conducive to the detection of CML at the early stages of the disease.

Gastric cancer is one of the most common malignancies in the world. It may result from a defect in the genes involved in DNA repair. One of the essential genes in the repair pathway is the XRCC1 gene that its polymorphisms in the human population play a role in gastric cancer susceptibility. The main purpose of this study was to investigate the association of 194C/T and 399G/A polymorphisms of the XRCC1 gene with gastric cancer in an Iranian population.

A total of 66 patients with gastric cancer and 67 control individuals were enrolled in our study. Following DNA extraction from blood samples, polymorphisms were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay.

The allele frequencies of C/T of XRCC1-194C/T in the control and patients groups were 83.17% and 71.29%, respectively. Moreover, The allele frequencies of G/A of XRCC1-399G/A in control and patient groups were 66.34% and 62.38%, respectively. Our results indicated a significant positive association between the distribution T/C alleles and the risk of gastric cancer (χ2: 5.37 and P=0.02), but no significant association was found in the distribution G/A alleles (χ2: 0.47 and P=0.48).

Altogether, these findings indicate a positive association between the distribution of 194T/C alleles of XRCC1 and the risk of gastric cancer and the presence of the C allele may increase the risk of gastric cancer.

Concurrent training is more effective in developing fitness indicators than doing endurance and resistance training separately. However, there has been limited research to evaluate the effects of this type of exercise training on improvement of body composition and quality of life indicators in postmenopausal women with cancer.

The present study aimed to determine the effects of eight-week of concurrent training on body composition, quality of life, and sleep quality in postmenopausal women with breast cancer.

This study was conducted on 42 women with breast cancer who were selected randomly and divided into exercise training and control groups. The training group followed eight-week of resistance training (2 - 3 sets, 10 - 18 repetitions, and 50% - 70% 1RM) and aerobic training (50% - 70% maximum heart rate, 12 - 14 Borg scale, and 20 - 40 minutes). Anthropometric characteristics were measured based on body composition (ZEUS 9.9), the sleep quality was measured by the Pittsburgh sleep quality index (PSQI), and the quality of life was measured by the McGill quality of life (MQOL) questionnaire. Two-way repeated measures ANOVA has been used for McGill’s analysis of variance (P < 0.05).

The results showed a significant decrease in sleep quality score, weight, fat percentage, BMI, and waist circumference in the training group (P < 0.05), as well as an increase in quality of life index in the training group (P < 0.05). However, no significant changes were observed in the Waist-hip ratio (WHR) values of the training group compared with the control group (P > 0.05).

Although the changes in WHR index were not significant after eight weeks of concurrent training, this type of training program could be considered as a beneficial way for improving body composition, quality of life, and sleep quality in patients with breast cancer.

The key enzyme of methylenetetrahydrofolate reductase (MTHFR) is involved in DNA biosynthesis and repair. The role of MTHFR C677T polymorphism in susceptibility to breast cancer is controversial. In the present case-control investigation, 297 individuals consisted of 100 patients with breast cancer and 197 healthy women were studied for MTHFR C677T genotypes, using PCR-RFLP method. The frequency of MTHFR TT genotype was 10% in patients and 3% in controls (P = 0.008). The presence of TT genotype was associated with susceptibility to breast cancer [OR = 1.97, 95%CI: 1.16-3.36, P = 0.012]. The T allele of MTHFR was found in 30% of the patients compared to 27.6% healthy controls (P = 0.024) that enhanced the risk of breast cancer by 1.56 times (95% CI: 1.06 - 2.3, P = 0.024). There were 71 individuals (71%) with the age of breast cancer diagnosis ≤ 51 years old. Comparing patients with the age of cancer diagnosis ≤ 50 years old with those > 51 years old group indicated a higher frequency of MTHFR TT genotype in the latter (20.7%) compared to the first group (5.6%, P = 0.05). Our study demonstrated an association between the MTHFR C677T polymorphism with the risk of breast cancer among population of Western Iran. Also, our study suggests that the MTHFR TT genotype could be a risk factor for breast cancer in postmenopausal women.

Semaphorins are large family of secretory and membrane-bound proteins that first were recognized in the nervous system as axon guidance molecules. Semaphorins family has more than 30 members and has been classified into eight subclasses. Different classes of these molecules involved in various phases of immune responses are considered as immune semaphorins. Main receptors for semaphorins are plexins and neuropilins. Moreover, other types of molecules can act as receptor for semaphorins, such as TIM-2 (T cell, immunoglobulin, and mucin domain protein 2), CD72 that bind to Sema 4A (Semaphorin 4A), and Sema 4D. Both forms of semaphorins, namely secretory and membrane-bound semaphorins, play important roles in the immune system. Multiple sclerosis (MS), a chronic inflammatory autoimmune disease, is characterized by infiltration of lymphocytes into the central nervous system and demyelination. Recent investigations have shown that increased serum level or increased expression of some immune semaphorins is associated with severity of MS disease. Moreover, immune semaphorins-deficient mice are resistant to experimental autoimmune encephalomyelitis, which is attributed to impaired production of myelin basic protein-specific T cells. Identification of specific expression patterns of semaphorins and their receptors in the nervous system and a comprehensive understanding of their function in autoimmune brain disorders could provide a novel biomarker and therapeutic target for these disorders. The present study reviews the role of semaphorins and their receptors in the development and differentiation of immune cells and their relation to MS

Formation of secondary structure such as DNA hairpins or loops may influence molecular genetics methods and PCR based approaches necessary for genetic engineering, in addition to gene regulation.
A polymerase chain reaction with splice overlap extension (SOE-PCR) was used to create fully synthetic 1F5 chimeric anti-CD20 heavy- and light-chain genes. The chimeric genes were cloned into the pCR-Blunt II-TOPO vector following by cloning into the pBudCE4.1 expression vector. Prediction of secondary structure was performed with the Vienna RNAfold webserver. PCR and sequencing across the predicted secondary structure of chimeric 1F5 heavy-chain gene was performed with multiple protocols for standard and GC-rich templates.
In our attempt to design vectors aimed to generate mouse-human chimeric antibody against CD20 (1F5), we found that the coding sequence of 1F5 chimeric heavy-chain gene constructed by SOE-PCR was resistant to polymerase during both PCR and sequencing reactions. Furthermore, we were also unable to analysis some positive transformants by restriction enzyme digestion. Encountering such difficulties to identify the cloned anti-CD20 chimeric heavy-chain gene, we found that the chimeric heavy-chain sequence is highly GC-rich and predicted to form a stable secondary structure.
In conclusion, for the first time, we reported several difficulties with production of therapeutic chimeric 1F5 anti-CD20 antibody due to a predicted hairpin cluster correlates with barriers to PCR, sequencing and possibly restriction analysis. Our findings provide a probable note for researchers experiencing technical difficulties with construction of chimeric anti-CD20 antibody 1F5 gene vectors and also with other genes and molecular biology techniques requiring PCR-based method or restriction enzyme analysis.

Stevia is a natural and non-caloric sweetener which most used in food and drug industries. In the present study, we focused on optimization of cell dedifferentiation and callus induction in stevia. In order to evaluation of growth regulators and explant types effects on callus induction, a factorial experiment was carried out in two factors and based on completely randomized design in six replications. The factor A included different levels of benzene adenin in three levels (0.0, 0.5 and 1.0 mg/l), Naphthalene acetic acid in four levels (0.0, 0.5, 1 and 2 mg/l), 2-4-D in three levels (0, 1 and 2 mg/l). Factor B comprised two levels of explant (leaf and stem) that were evaluated. The experiment was performed on Tissue Culture Lab in Kermanshah Industrial University. Analysis of variance for data results showed that there were significant differences among levels of plant explants for callus induction (P

The DNA molecule has been known to be the cellular target for many cytotoxic anticancer agents for several decades. Understanding how drug molecules interact with DNA has become an active research area in the interface between chemistry, molecular biology and medicine. DNA extraction has been suggested as a main step affecting molecular DNA technology such as PCR and PCR-based methods. Therefore, researchers have used several modified protocols for efficient DNA extraction from whole blood. In this study, we focused on a fast and reliable protocol with inexpensive and non-poisonous reagents for DNA extraction from whole blood. Current method was optimized based on a combination of conventional salting-out and boiling methods. Also the quality and quantity of the extracted DNA were surveyed by gel electrophoresis and Nanodrop spectrophotometry methods, respectively. Results showed that high quantity and quality of isolated DNA by this method is enough to do hundreds of PCR-based reactions and also to be utilized in other DNA manipulation assay such as restriction digestion, drug- DNA interaction and methylation detection survey. In conclusion, we described a fast, low-cost, non-toxic and enzyme free protocol for high yield genomic DNA extraction from whole blood.

Aim and The Wdhn13 gene codes the LEA 2 protein family. LEA proteins were identified and discussed in wheat and cotton as a proteins cumulative first time in the late embryo period. The primary structure of LEA proteins، including early charged domains and non-charged of polar amino acid residues non hydrophilic is that LEA proteins are being implicated on hydrophilic. in this research، the Phylogenetic and bioinformatics relationship of Wdhn13 in 8 wheat species was investigated. The HindIII (AAGCTT)، AluBI (AGCT) and TaqI (TCGA) restriction enzyme were used for molecular analysis and by biological CLC main workbench 6 software the Insilco and out of vitro were applied in sequence Phylogenetic analysis result based on UPGMA algorithm showed that studied wheat species were divided into 3 clusters. The first cluster includes 4 wheat cultivar NCBI database’s sequence، T. aestivum CV Sardari، T. durum CV shosh، T. durum CV borojerd. The T. urartu was located in the second cluster alone. The third cluster includes T. aestivum CV gonbad and T. dicocoides. The results indicated a relationship among some wheat species.

Background and The 50 KDa protein (50 µg) in carboxylic domain of the neurotoxin heavy chain (BoNT/A-Hc) recognizes surface receptors on target neurons and this fragment contains the principle protective antigenic determinants. Recently, this fragment has been used as a recombinant vaccine candidate for botulism. The study aimed to compare the evaluation of BoNT/A-Hc production in fed-batch (high cell density cultivation) and batch cultivation of recombinant Escherichia coli (E. coli.). In this research, growth of recombinant E. coli in batch culture was studied. In order to maximize protein expression, induction time and Isopropyl β-D-1-thiogalactopyranoside (IPTG) inducer concentration were optimized by the Taguchi statistical method. Then, fed-batch culture was applied to achieve high cell density cultivation. Finally, the recombinant protein expression level was determined. In optimized conditions, 62 and 486 mg/l of soluble recombinant BoNT/A-Hc were produced in batch and fed-batch cultivation. According to the results, high cell density in fed-batch cultivation is a very effective for improve of recombinant proteins productivity.

Primary culture takes place following the cell isolation from tissues. Isolation and culture of melanocytes based on their roll in the protection of body against hazardous sun rays، production of skin، cornea and hair color is really important. This study was done to set isolation، culture and proliferation of melanocytes from children foreskin and adult eyelashes، and also comparison of two types of melanocyte culture medium.

Human foreskin and eyelash samples were used for melanocyte isolation and culture. After isolation of epidermis from dermis، epidermis cell suspensions were prepared by enzymatic digestion. The isolated cells were cultured in two melanocyte selective culture media. Immunocytochemistary and reverse transcription-polymerase chain reaction (RT-PCR) assays were used for confirmation of isolated and cultured melanocytes.

Our results indicated that isolated melanocyte cultured in the selective medium without phorbol esters is better than the melanocytes cultured in selective medium cont-aining phorbol esters not only morphologically but also physiologically and from the aspect of cell adhesion. In addition، the results showed that isolated melanocyte from adult eyelashes are more dendritic than melanocytes isolated from children foreskin. Conversely، our results indicated that the number of cell passages in melanocyte isolat-ed from foreskin is more than melanocytes isolated from adult eyelashes.

Melanocytes cultured in selective medium containing convenient growth factors in absence of phorbol esters show more native physiological and adhesive properties. In addition، melanocyte isolated from younger tissues such as foreskin have better proliferative and sub-culturing properties so we suggest isolation and culture of younger tissues.

Acute lymphoblastic leukemia (ALL) is the most common pediatric malignancy. The risk of thrombophilia increases in patients with ALL during chemotherapy. The present study aimed to investigate the frequency of factor V Leiden (FVL) mutation in children with acute lymphoblastic leukemia and its possible association with ALL.Patients and We studied 92 patients with ALL and 249 healthy individuals from Kermanshah Province of Iran. Detection of FVL mutation was performed by PCR-RFLP using restriction enzymes of Mnl I. The frequency of FVL G1691A polymorphism was 7.8% in patients compared to 3.2% in controls (p=0.052). There was a trend towards increased risk of ALL in the presence of FVL mutation [OR=2.54, 95% CI 0.9-7.2, p=0.08]. Our results indicated that the frequency of both thrombophilic mutation of FVL was higher in ALL patients from Kermanshah province compared to healthy individuals and FVL mutation tended to be associated with the increased risk of ALL. Further studies needed to evaluate the association between FVL mutation and the occurrence of thromboembolism in ALL patients.

In 30 to 40% of patients with type 2 diabetes mellitus (T2DM) nephropathy is developed. The aim of the present study was to find the synergistic effect of two polymorphisms of methylenetetrahydrofolate reductase (MTHFR) C677T and angiotensin converting enzyme insertion/ deletion (ACE I/D) polymorphism on the risk of diabetic nephropathy and its progression.

In a case-control study, the MTHFR C677T and ACE I/D were detected using PCR and PCR-RFLP, respectively in 72 patients with macroalbuminuria, 68 patients with microalbuminuria and 72 normoalbuminuric patients. The data were analyzed using SPSS.

In the presence of T allele of MTHFR the risk of microalbuminuria increased 1.54-fold (p=0.58). Also, non significant increased risk of macroalbuminuria was observed in the presence of ACE D allele (1.44-fold). However, in the presence of both MTHR 677T and ACE D alleles the risk of macroalbuminuria increased 4-fold (95%CI=1.1-14.5, p=0.035). Also, in the presence of both alleles the risk of progression from micro- to macro-albuminuria increased 2.07-fold (p=0.27).

The results of present study indicate the synergistic effect of MTHFR 677T and ACE D alleles on the increased risk of diabetic nephropathy and its progression.