Exosomes from SHED-MSC regulate polarization and stress oxidative indexes in THP-1 derived M1 macrophages.

OBJECTIVE: The inhibition of M1 macrophages may be interesting for targeted therapy with mesenchymal stem cell-derived Exosomes (MSC-EXOs). This study aimed to investigate the stem cells of human exfoliated deciduous teeth-derived EXOs (SHED-MSC-EXOs) effect on regulating the pro- and anti-oxidant indexes and inhibiting M1 macrophage polarization. Besides, an in-silico analysis of SHED-MSC-EXO miRNAs as the highest frequency of small RNAs in the exosomes was performed to discover the possible mechanism. METHODS: The flow cytometry analysis of CD80 and CD86 as M1-specific markers confirmed the polarization of macrophages derived from THP-1 cells. After exosome isolation, characterization, and internalization, THP-1-derived M1 macrophages were treated with SHED-MSC-EXOs. M1-specific markers and pro- and anti-oxidant indexes were evaluated. For in-silico analysis of SHED-MSC-EXOs miRNAs, initial miRNA array data of SHED-EXOs is collected from GEO, and the interaction of the miRNAs in M1 macrophage polarization (M1P), mitochondrial oxidative stress (MOS) and LPS-induced oxidative stress (LOS) were analyzed by miRWalk 3.0 server. Outcomes were filtered by 75th percentile signal intensity, score cut-off ≥0.95, minimum free energy (MEF)≤ -20 kcal/mol, and seed = 1. RESULTS: It shows a decrease in the expression of CD80 and CD81, a reduction in pro-oxidant indicators, and an increase in the anti-oxidant indexes (P < 0.05). Computational analysis showed that eight microRNAs of SHED-MSC-EXO miRNAs can bind to and interfere with the expression of candidate genes in the M1P, MOS, and LOS pathways simultaneously. CONCLUSION: SHED-MSCs-EXOs can be utilized to treat conditions related to M1 macrophage-induced diseases (M1IDs) due to their unique physical properties and ability to penetrate target cells easily.

Association between PON1-rs662 Gene Polymorphism and Diabetic Retinopathy in Population of the Qom, Iran.

Background: Diabetic retinopathy is the most severe diabetic microvascular complication that causes changes in the vessel wall. One of the genes involved in this disease is PON1, which encodes paraoxanase1 protein in liver and kidney. It might regulate inflammatory and microvascular responses to the disease. The rs662 T>C is one of the single nucleotide polymorphisms of this gene that changes glutamine to arginine at position 192. Methods: In this study, 300 samples were collected, including 100 healthy and 100 diabetics without retinopathy, and 100 diabetics retinopathies were studied and their age range was from 30 to 80 years. Then 2.5 ml of blood was collected from all relevant individuals in tubes containing EDTANa2. This polymorphism was examined by tetra-ARMS PCR. Results: Results showed that there is no significant correlation between genotypes and alleles related to PON1 and Diabetes (CC genotype: p=0.609; C allele: p=0.228). On the other hand, an association was observed between PON1 and diabetic retinopathy (CT+CC genotype: p<0.001; CT allele: p<0.001). Considering that the Polyphen database examined the changes caused by replacing the amino acid arginine instead of glutamine at position 129 on the protein, it does not consider these changes dangerous and has introduced this polymorphism as benign. Conclusion: Based on the findings of this study, the rs662 locus could be considered as one of the molecular markers in future research.

Myopic regression after photorefractive keratectomy: a retrospective cohort study.

Background: Myopic regression is a major complication of photorefractive keratectomy (PRK). The rates and causes vary considerably among different studies. This study aimed to investigate myopic regression at six months after myopic PRK. Methods: In this retrospective cohort study, we included all eligible patients with myopia ranging from - 0.75 to - 9 D, aged 18 to 50 years, who underwent PRK by a single surgeon with the availability of preoperative and postoperative data at six months after the initial procedure. All participants underwent comprehensive ophthalmic examinations preoperatively and at six months post-PRK. Overcorrection was planned based on the participant's age range to achieve the desired refractive result after PRK. All patients received the same postoperative antibiotic and steroid eye drops in a similar dosage regimen, and the contact lenses were removed after complete corneal epithelial healing. Based on the spherical equivalent of refraction six months after PRK, eyes without and with myopic regression were allocated into groups 1 and 2, respectively. Results: We included 254 eyes of 132 patients who underwent myopic PRK with a mean (standard deviation) age of 30.12 (7.48) years; 82 (62.12%) were women and 50 (37.88%) were men. The frequency of myopic regression was significantly lower in patients with younger age, lower preoperative cylindrical refraction, and lower ablation depth (all P < 0.05). Overcorrection was more successful in eyes with low myopia than in eyes with high myopia (P < 0.05). The highest frequency of myopic regression occurred in eyes with moderate myopia (25.68%), followed by eyes with high myopia (20.0%) and low myopia (6.54%). Among different age groups, patients aged ≤ 30 years had a lower frequency of myopic regression. The frequency of myopic regression in the different age groups was 5.0% at 18 - 20 years, 7.46% at 26 - 30 years, 12.28% at 21 - 25 years, 21.31% at 31 - 35 years, and 26.53% at 36 - 50 years. Conclusions: Overcorrection was more successful in eyes with low myopia than in eyes with high myopia. The success rate was higher in younger patients with lower astigmatism and ablation depths. Myopic regression was most frequent in eyes with moderate myopia, followed by those with high and low myopia. Further studies should replicate our findings over a longer follow-up period with a larger sample size before generalization is warranted.

Missense polymorphisms potentially involved in mandibular prognathism.

Objective: The current study aimed to identify and analyze missense single nucleotide polymorphisms (SNPs) that can potentially cause mandibular prognathism. Methods: After reviewing the articles, 56 genes associated with mandibular prognathism were identified and their missense SNPs were retrieved from the NCBI website. Several web-based tools including CADD, PolyPhen-2, PROVEAN, SNAP2, PANTHER, FATHMM, and PON-P2 were used to filter out harmful SNPs. Additionally, ConSurf determined the level of evolutionary conservation at positions where SNPs occur. I-Mutant2 and MUpro predicted the effect of SNPs on protein stability. Furthermore, to investigate the structural and functional changes of proteins, HOPE and LOMETS tools were utilized. Results: Based on predictions in at least four web-based tools, the results indicated that PLXNA2-rs4844658, DUSP6-rs2279574, and FBN3-rs33967815 are harmful. These SNPs are located at positions with variable or average conservation and have the potential to reduce the stability of their respective proteins. Moreover, they may impair protein activity by causing structural and functional changes. Conclusions: In this study, we identified PLXNA2-rs4844658, DUSP6-rs2279574, and FBN3-rs33967815 as potential risk factors for mandibular prognathism using several web-based tools. According to the possible roles of PLXNA2, DUSP6, and FBN3 proteins in ossification pathways, we recommend that these SNPs be investigated further in experimental research. Through such studies, we hope to gain a better understanding of the molecular mechanisms involved in mandible formation.

Optimization of Affinity Chromatography Based on Sepharose 4B- chitin for Rapid Purification of Urtica dioica Agglutinin.

Background: Today, numerous antimicrobial and anticancer properties have been reported for plant lectins due to their ability to bind to carbohydrates. The Urtica dioica agglutinin (UDA lectin) is a monomeric, small, and low molecular weight glycoprotein. It has attracted the attention of many researchers for identification, treatment, and other clinical purposes. Objectives: The aim of this study is the optimization of the chitin affinity chromatography based on Sepharose 4B (CNBr-activated Sepharose 4B) for the rapid purification of UDA lectin from Urtica dioica rhizome. Materials and Methods: The chitin ligands were dissolved in 40% Trichloroacetic acid and attached to Sepharose 4B according to the Amersham-Biosciences instructions. The attachment of the ligand to the Sepharose 4B beads was investigated by Fourier transform infrared (FTIR) spectroscopy. An acidic crude extract of nettle rhizome passes from chromatographic columns in two sizes with dimensions: 24 x 0.51 cm and 8.44 x 0.86 cm. Quantity and quality of purified lectin were calculated by the Bradford microplate method: SDS-PAGE gel electrophoresis and human erythrocyte cell (RBC) hemagglutination, respectively. Results: The analysis of FTIR spectrograms showed that major changes were observed in the fingerprint regions. Besides, due to the dissolution of Sepharose 4B and chitin in the aqueous phase, this difference was not significant in the Imine and Nitrile regions. On the other hand, the comparative results of purification chromatograms showed that increasing the column length causes a smaller half-width and increases the length of the purified peak. Also, it leads to high-quality purified UDA lectin, with a molecular weight of almost 12.5 kDa in gel electrophoresis. Hemagglutination activity on trypsinized red blood cells was displayed, and agglutination of purified UDA lectin started at least at 300 µg.mL-1 concentration. Conclusion: According to our findings, we suggested that dissolving chitin in the polar solvent of Trichloroacetic acid, using Sepharose 4B as the beads of a matrix, and increasing the column length might lead to a decrease in the half-width of the peak. These can increase the purity and concentration of purified UDA lectin, and speed up the purification process. These findings could be used by researchers to accelerate the purification of UDA lectin in other studies, dealing with drug delivery systems, ELISA techniques, and cell growth.

Mutant Allele of CD44 (rs8193C>T) and Pum2 Regulatory Element as A Prognosis Factor of Prostate Neoplasms: A Case-Control and In Silico Studies.

OBJECTIVE: Expression of CD44 variant 6 (CD44v6) as a homing-associated cell adhesion molecule (HCAM), has proved to change most cancer cells. Aim of the study is the effect of mutant allele of CD44 (rs8193C>T) and Pum2 regulatory element as a prognosis factor of prostate neoplasms: a case-control and in silico studies in the Mazandaran province-Iran. MATERIALS AND METHODS: In a case-control study, CD44-rs8193C>T genotyping of the 420 prostate neoplasms (210 benign prostatic hyperplasia (BPH) patients and 210 prostate cancer patients) and 150 healthy samples are performed by the touchdown polymerase chain reaction with confronting two-pair primers (PCR-CTPP) method. The T mutant allele effects on the mRNA structure and cell pathways were also investigated in silico methods. RESULTS: Our results showed that the increase of T mutant allele frequency was significantly associated with BPH compared with prostate cancer. Furthermore, results showed TT genotype was significantly associated with BPH [odds ratio (OR)=0.572 and P=0.015], and also influenced the CD44v6 transcript secondary structure, miRNA binding, and regulatory element-binding site for Pum2 protein. Attachment of Pum2 to standard CD44 transcript may lead to transcript isoform-switching and shift-expression to a variety of CD44 isoforms, which can trigger some of the cell signaling pathways, such as Nanog-Stat, PKC-Nanog, and PKC-Twist. CONCLUSION: Based on this, the presence of the T mutant allele of CD44 (rs8193C>T) in the populations may create a regulatory element-binding site for Pum2. So, it could be known as a prognosis factor and prediction of prostate neoplasms. However, more comprehensive studies in different populations (with various ethnicities and large population sizes), and also CD44v6 gene expression studies in protein and transcript levels are required to confirm our data.

Effects of Urtica dioica agglutinin on glycotargeting of the vasculature: an in ovo study on chicken embryo.

The angiogenesis process is a pivotal cellular process involved in both developmental and pathological circumstances. In this study we investigated effect of Urtica dioica agglutinin (UDA), as an unusual phyto-lectin from the chitin-binding protein family, on the angiogenesis of chicken embryos. The UDA was extracted from plant rhizomes and purified by affinity chromatography column. The activity of this lectin was assayed by hemagglutination test on the human RBCs. Anti-angiogenic effect of UDA on the extra-embryonic layer of the chick egg was studied in the different concentrations. Our results showed that the minimum concentration of UDA for agglutination were 48.00 and 15.00 µg mL-1 in macro- and microscopic studies, respectively. Because the number and length of the vessels were dramatically decreased at 100 µg kg-1 of UDA, the lectin had an inhibitory effect on angiogenesis of the embryonic vasculature of the chick. We concluded that UDA might target the vascularization events through binding to GlcNAc-conjugates. More investigations are needed to clarify the angiogenesis-related therapeutic roles of this interesting biomolecule.

In Silico Analysis of Collagens Missense SNPs and Human Abnormalities.

Collagens are the most abundant proteins in the extra cellular matrix/ECM of human tissues that are encoded by different genes. There are single nucleotide polymorphisms/SNPs which are considered as the most useful biomarkers for some disease diagnosis or prognosis. The aim of this study is screening and identifying the functional missense SNPs of human ECM-collagens and investigating their correlation with human abnormalities. All of the missense SNPs were retrieved from the NCBI SNP database and screened for a global frequency of more than 0.1. Seventy missense SNPs that met the screening criteria were characterized for functional and stability impact using six and three protein analysis tools, respectively. Next, HOPE and geneMANIA analysis tools were used to show the effect of SNPs on three-dimensional structure (3D) and physical interaction of proteins. Results showed that 13 missense SNPs (rs2070739, rs28381984, rs13424243, rs1800517, rs73868680, rs12488457, rs1353613, rs59021909, rs9830253, rs2228547, rs3753841, rs2855430, and rs970547), which are in nine different collagen genes, affect the structure and function of different collagen proteins. Among these polymorphisms, COL4A3-rs13424243 and COL6A6-rs59021909 were predicted as the most effective ones. On the other hand, designed mutated and native 3D of rs13424243 variant illustrated that it can disturb the protein motifs. Also, geneMANIA predicted that COL4A3 and COL6A6 are interacting with some proteins including: DDR1, COL6A1, COL11A2 and so on. Based on our findings, ECM-collagens functional SNPs are important and may be considered as a risk factor or molecular marker for human disorders in the future studies.

Features of Pathobiology and Clinical Translation of Approved Treatments for Coronavirus Disease 2019.

BACKGROUND: Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is currently the most important etiological agent of acute respiratory distress syndrome (ARDS) with millions of infections and deaths in the last 2 years worldwide. Several reasons and parameters are responsible for the difficult management of coronavirus disease-2019 (COVID-19) patients; the first is virus behavioral factors such as high transmission rate, and the different molecular and cellular mechanisms of pathogenesis remain a matter of controversy, which is another factor. SUMMARY: In the present review, we attempted to explain about features of SARS-COV-2, particularly focusing on the various aspects of pathogenesis and treatment strategies. KEY MESSAGES: We note evidence for the understanding of the precise molecular and cellular mechanisms of SARS-CoV-2 pathogenesis, which can help design the appropriate drug or vaccine. Additionally, and importantly, we reported the updated issues associated with the history and development of treatment strategies such as, drugs, vaccines, and other medications that have been approved or under consideration in clinics and markets worldwide.

The Functional Roles of Curcumin on Astrocytes in Neurodegenerative Diseases.

Progressive abnormality and loss of axons and neurons in the central nervous system (CNS) cause neurodegenerative diseases (NDs). Protein misfolding and its collection are the most important pathological features of NDs. Astrocytes are the most plentiful cells in the mammalian CNS (about 20-40% of the human brain) and have several central functions in the maintenance of the health and correct function of the CNS. Astrocytes have an essential role in the preservation of brain homeostasis, and it is not surprising that these multifunctional cells have been implicated in the onset and progression of several NDs. Thus, they become an exciting target for the study of NDs. Over almost 15 years, it was revealed that curcumin has several therapeutic effects in a wide variety of diseases' treatment. Curcumin is a valuable ingredient present in turmeric spice and has several essential roles, including those which are anticarcinogenic, hepatoprotective, thrombosuppressive, cardioprotective, anti-arthritic, anti-inflammatory, antioxidant, chemopreventive, chemotherapeutic, and anti-infectious. Furthermore, curcumin can suppress inflammation; promote angiogenesis; and treat diabetes, pulmonary problems, and neurological dysfunction. Here, we review the effects of curcumin on astrocytes in NDs, with a focus on Alzheimer's disease, Parkinson's disease, multiple scleroses, Huntington's disease, and amyotrophic lateral sclerosis.

Human DT-diaphorase expression in Escherichia coli: optimization, purification and structural stability.

Expression and purification of human DT-diaphorase, also referred to as NAD(P)H quinone oxidoreductase 1 (NQO1; EC. 1.6.99.2), which is a flavoprotein belongs to the family of oxidoreductases are optimized. The DT-diaphorase plays an important role in biosensor design for laboratory analysis and also developing biosensor for measurement of glucose level in blood. The aim of this study was to investigate various parameters regarding the expression of DT-diaphorase in Escherichia coli BL21 (DE3) and thermal stability of DT-diaphorase activity at different temperatures in the presence of sucrose. Expression conditions of DT-diaphorase in E. coli were optimized with an induction time (22.00 hr), induction temperature (18.00 ˚C) and also lactose (5.00 mM) and isopropyl ß-D-1-thiogalactopyranoside (1.00 mM) concentrations as inducers. The Km, Vmax and kcat values for NADH as a substrate were 25.50 µM, 357 µM per min and 446.40 µM mg-1 per min, respectively. Results of our research revealed that different concentrations of sucrose at 40.00 ˚C did not have any significant effect on enzyme structure; while, relatively significant changes, especially in the presence of sucrose (0.75 M) at 50.00 ˚C were observed. The results presented show that sucrose causes DT-diaphorase inactivation rate reduction and relatively little increases in thermal stability and thus, sustains its conformation against thermal unfolding.

The decellularized ovary as a potential scaffold for maturation of preantral ovarian follicles of prepubertal mice.

ABBREVIATIONS: GAG: glycosaminoglycan; ECM: extracellular matrix; 2D: two-dimensional; E2: estradiol; P4: progesterone; BMP15: bone morphogenetic protein 15; GDF9: growth differentiation factor 9; ZP2: zona pellucida 2; Gdf9: growth/differentiation factor-9; Bmp6: bone morphogenetic protein 6; Bmp15: bone morphogenetic protein 15.

PTGS2 Over-Expression: A Colorectal Carcinoma Initiator not an Invasive Factor.

BACKGROUND: Cyclooxygenase-2 (COX-2) main product is Prostaglandin E2 (PGE2) which cause mitogenesis and inflammation. COX-2 is the product of prostaglandin-endoperoxide synthase 2 (PTGS2) gene expression. COX-2 dysregulation can cause angiogenesis, differentiation, and promotion of cancer and its suppression related to control of the tumor's size, number, and cell shape. This study focused on the association of COX-2 expression with colorectal carcinoma (CRC) among Iranian patients on mRNA level and in the Cancer Genome Atlas Program (TCGA) colon and rectum RNAseq dataset, and its relation with pathological features. METHODS: PTGS2 expression was assayed by quantitative-PCR method from 90 tissue samples collected from 45 participants. The control samples come from the non-tumor area of the same patients. The data analyzed based on ΔΔCq. The PTGS2-RNAseq data extracted and analyzed by UCSC Xena browser, and its association assessed the occurrence of CRC and invasive-features. RESULTS: PTGS2 showed very significant over-expression in tumor tissues (p< 0.0001) with an N-fold expression of 2.25. But, there was not any significant association between PTGS2 and CRC invasive-pathological features such as Lymphatic, vascular and perineural invasion, the Grades of cancer, and Pathologic-M in both parts of this study. CONCLUSION: The increase in PTGS2 is related to the occurrence of CRC among patient samples. But in both part of this study, PTGS2 is not an invasive factor, and it does not affect the cell differentiation of tumors and metastasis. Based on the high N-fold for patient samples, it can be a strong candidate as a CRC initiator biomarker.

Subcellular partitioning of cadmium and lead in Eisenia fetida and their effects to sperm count, morphology and apoptosis.

Earthworms and their biomarkers are considered good indicators for assessing the effects of toxic chemicals. Therefore, in this study, we exposed Eisenia fetida to lethal and sub-lethal concentrations of Cd and Pb nitrate in artificial soil for 14 and 28 days to evaluate the impact on subcellular partitioning, lethal toxicity (LC50), growth, sperm count, morphology and apoptosis (using TUNEL assay). The soluble internal pools of both metals were good predictors of the responses of biomarkers. We found sperm deformation, TUNEL positive sperms and weight loss positively and sperm count negatively correlated with the concentrations of Cd and Pb in the total internal and cytosolic fraction (p < 0.01) and to a lesser extent with Pb concentrations in the granular fraction (p < 0.05). Fourteen days LC50 for Cd and Pb were 2169 ±â€¯322 and 6387 ±â€¯904 µg/g, respectively. Cadmium and Pb caused a significant depression in sperm count after 14 (Cd: up to 46.9%; Pb: up to 36.24%) and 28 (Cd: up to 72.47%; Pb: up to 43.12%) days of exposure relative to the control (p < 0.05). Cadmium induced higher abnormality in sperm heads than Pb. For both metals, TUNEL positive sperms significantly increased after 14 (Cd: up to 14.17%; Pb: up to 16.33%) and 28 (Cd: up to 16.33%; Pb: up to 11.67%) days of exposure compared with the control (p < 0.05). The findings of this study, illustrate the importance of considering sperm parameters as a rapid, easy and sensitive biomarker for the evaluation of metal toxicity.

Interspecific differences in toxicological response and subcellular partitioning of cadmium and lead in three earthworm species.

Earthworms are often used as test subjects in toxicological studies, due to their ubiquitousness and sensitivity to contaminant exposure. Such testing is typically conducted using Eisenia fetida as the test subject, but continued use of E. fetida (eco) toxicology is questionable. Therefore, in this study three earthworm species, Aporrectodea rosea, Aporrectodea trapezoides and E. fetida, were exposed to lethal and sublethal concentrations of cadmium (Cd) and lead (Pb) nitrate in artificial soil for 7, 14 and 28 days. A biomarker of genotoxicity (TUNEL assay), biochemical markers [malondialdehyde (MDA) and total antioxidant capacity (TAC)], weight loss, lethal toxicity (LC50) and subcellular partitioning were assessed. Cadmium and Pb caused significant inhibition in TAC and growth and significant increases in DNA damage and lipid peroxidation in the earthworms. Acute toxicity rank (14 days) for both Cd and Pb were E. fetida > A. trapezoides > A. rosea. Subcellular partitioning of Cd and Pb in the earthworms were cytosol > debris > granules and debris > granules > cytosol, respectively. Comparison of biomarker responses between study species showed that E. fetida proved to be less susceptible to Cd and Pb exposure than A. rosea and A. trapezoides. Therefore, this study confirms that A. rosea and A. trapezoides are more suitable as subjects than E. fetida for the soil toxicity tests, because of both their greater susceptibility to toxicants and in their abundance in the field.

Comparison of large-scale deletions of the sperm mitochondrial DNA in normozoospermic and asthenoteratozoospermic men.

BACKGROUND AND OBJECTIVE: Mitochondria play a crucial role in energy metabolism for the survival and motility of sperm during fertilization. The aim of this study was to determine the association of large-scale mitochondrial DNA deletions with abnormal sperm motility and morphology in asthenoteratozoospermic patients. MATERIALS AND METHODS: In this case-control study, 41 semen samples were collected from 18 normozoospermic healthy men and 23 asthenoteratozoospermic patients, according to the WHO guidelines. The swim-up technique was used for separation of spermatozoa on the basis of their motility. Long-range polymerase chain reaction (PCR) was used for screening of mitochondrial DNA (mtDNA) large-scale deletions, and primer shift PCR was used for confirmation of deletions. RESULTS: The mean sperm motility, normal morphology, and progressive motility in asthenoteratozoospermic patients were significantly lower than in the normozoospermic group (P < 0.0001). There was a positive significant correlation between motility and normal sperm morphology ( P < 0.0001, r = 0.741). The results of long-range PCR revealed the existence of 4866-bp deletion along with the two common 4977-bp and 7436-bp deleted mtDNA in both groups. However, the frequency of multiple mtDNA deletions in the asthenoteratozoospermic group (15/23, 65.22%) was significantly higher than that in the normozoospermic group (7/18, 38.89%). Direct sequencing of the 534-bp PCR product revealed that it was amplified from the mtDNA with a 4866-bp deletion flanked by a seven-nucleotide direct repeat (5'-ACCCCCT-3'). CONCLUSIONS: Our findings suggested that these large-scale deletions of mtDNA may be genetic risk factors for poor sperm quality in asthenoteratozoospermia-induced male infertility. Thus, it is necessary to understand the mechanisms behind the generation of these deletions.

Association of the human aryl hydrocarbon receptor repressor (AhRR)-c.565C>G transversion with male infertility: A case-control study from Iran.

Dioxins (eg, 2,3,7,8-tetrachlorodibenzo-p-dioxin/TCDD), as environmental endocrine disruptors and toxic carcinogens, can affect male reproductive health. The influence of dioxins is mediated via the aryl hydrocarbon receptor (AhR) pathway and its repressor (AhRR). In this study, we investigated the association of AhRR-c.565C>G transversion polymorphism with male infertility. In a hospital-based case-control study, 221 semen samples (111 infertile and 110 healthy controls) based on World Health Organization guidelines were collected from in vitro fertilization centers of Babol, Iran. The AhRR-c.565C>G (rs2292596) polymorphism was genotyped using a polymerase chain reaction-restriction fragment length polymorphism analysis. The difference in the allele frequency of AhRR-c.565C>G transversion polymorphism did not reach a significant level. The genotype frequency was statistically significantly different between fertile and infertile men. We found that polymorphism rs2292596 (Pro185Ala) was statistically significantly associated with the risk of male infertility. In addition, the statistical difference became more significant when the frequency was compared between the Pro/Pro genotype and the Pro/Ala plus Ala/Ala genotype. The 185 Pro wild-type alleles of AhRR may be associated with the risk of male infertility. The proallele also may diminish inhibition of AhR-mediated signaling of exposure to environmental pollutants.

Magnetically Separable Fe3 O4 @CdS Type-II Nanohybrids with Excellent Photocatalytic Activity and Antibacterial Properties.

A magnetically separable Fe3 O4 @CdS type-II core-shell nanohybrid (NH) photocatalyst, with excellent antibacterial properties and photocatalytic activity, was synthesized and characterized by means of structural, elemental, and morphological analyses. Steady-state and time-resolved emission and absorption spectroscopy were also exploited to realize the location of charge-carrier wave functions in the NHs. The antibacterial activity of NHs was then evaluated against Escherichia coli (1.4×108  CFU mL-1 ; CFU=colony forming units) and Staphylococcus saprophyticus (1.2×108  CFU mL-1 ) as model strains of Gram-negative and -positive microbes. Compared with CdS and Fe3 O4 , the as-synthesized Fe3 O4 @CdS composite exhibited highly improved bactericidal activity and recyclability. Concentration values of 5.0 and 4.0 mg mL-1 were required for Fe3 O4 @CdS NHs to inhibit the growth of E. coli and S. saprophyticus, respectively. The contribution of OH. radicals to photocatalysis at the Fe3 O4 @CdS surface was also considered. Moreover, thiobarbituric acid reactive substances and protein carbonyl assay protocols have been exploited to monitor levels of oxidative damage to lipids and proteins in cells. Additionally, the photocatalytic activity of Fe3 O4 @CdS NHs was monitored for the degradation of xylenol orange dye. Compared with CdS and Fe3 O4 , as-synthesized Fe3 O4 @CdS displayed superior performances in the photodegradation of xylenol orange. Possible mechanisms involved in the degradation of xylenol orange are also discussed.

Human MTHFR-G1793A transition may be a protective mutation against male infertility: a genetic association study and in silico analysis.

In this paper, we evaluate the association of the human methylenetetrahydrofolate reductase (MTHFR)-G1793A transition with male infertility using a case-control study, a meta-analysis and an in silico analysis. In the case-control study, 308 blood samples (169 infertile and 139 fertile men) were collected. MTHFR-G1793A genotyping was performed by PCR-RFLP. The study revealed a significant protective association between the GA genotype (OR: 0.3737, 95%CI: 0.1874-0.7452, p = 0.0052) and A allele (OR: 0.4266, 95%CI: 0.2267-0.8030, p = 0.0083) with male infertility. Meta-analysis showed that the G1793A transition might be a protective mutation against male infertility in both A vs. G (OR: 0.608, 95%CI: 0.466-0.792, p < 0.001), and GA vs. GG (OR: 0.534, 95%CI: 0.394-0.724, p < 0.001) genetic models. In silico-analysis revealed that although G1793A could not make fundamental changes in the function and structure of MTHFR, it could modify the structure of the mRNA (Distance =0.1809, p = 0.1095; p < 0.2 is significant). The results suggest that G1793A substitution might be a protective genetic factor against male infertility. However, further case-control studies are required to provide a more robust conclusion.

Genetic polymorphisms and haplotypes of the DJ-1 gene promoter associated with the susceptibility to male infertility.

PURPOSE: In this study, we evaluate the relationship between genetic polymorphisms of the DJ-1 gene, g.-6_+10del, and g.168_185del with male infertility susceptibility. METHODS: Four hundred and twenty-two male infertile patients and 285 fertile male controls were recruited. Genotyping was performed by polymerase chain reaction. In silico analysis was performed by EPD, ElemeNT, SNPnexus, and PROMO to predict the potential functions of rs901561484 and rs373653682 polymorphisms. RESULTS: The Del (D) allele carriers of DJ-1 g.-6_+10del polymorphism were significantly associated with the risk of male infertility in total infertile, asthenozoospermia, and oligoasthenozoospermia patients. Moreover, the Del (D) allele of DJ-1 g.-6_+10del polymorphism significantly increased in total male infertile, asthenozoospermia, and oligoasthenozoospermia groups. In addition, the frequencies of different genotypes and the Del allele and Dup allele carriers of DJ-1 g.168_185del gene polymorphisms were associated with male infertility in total infertile and four different sub-group patients. Furthermore, haplotype analysis of DJ-1 g.-6_+10del and g.168_185del polymorphisms revealed that the D-Dup and I-Del haplotype frequencies significantly increased the risk of male infertility, while I-Ins haplotypes were associated with a decreased risk of male infertility in total and sub-group patients. The in silico analysis showed that the presence of Ins and/or Dup alleles of the DJ-1 g.-6_+10del and g.168_185del polymorphisms could provide additional binding sites of more nuclear factors and probably affect transcriptional activity. CONCLUSION(S): Our study presents evidence of a strong association between functional polymorphisms of the DJ-1 promoter, g.-6_+10del, and g.168_185del with the risk of male infertility.