Showing 9 results for Dna.
Hamed Fanaei, Samira Khayat, Iman Halvaei, Vahid Ramezani, Yaser Azizi, Amir Kasaeian, Jalal Mardaneh, Mohammad Reza Parvizi, Maryam Akrami,
Volume 12, Issue 2 (2-2014)
Abstract
Background: Oxidative stress in teratozoospermic semen samples caused poor assisted reproductive techniques (ART) outcomes. Among antioxidants, ascorbic acid is a naturally occurring free radical scavenger and as such its presence assists various other mechanisms in decreasing numerous disruptive free radical processes.
Objective: The main goal of this study was to evaluate potential protective effects of ascorbic acid supplementation during in vitro culture of teratozoospermic specimens.
Materials and Methods: Teratozoospermic semen samples that collected from 15 volunteers were processed, centrifuged and incubated at 37PoPC until sperm swimmed-up. Supernatant was divided into four groups and incubated at 37PoPC for one hour under different experimental conditions: Control, 10 μm A23187, 600μm ascorbic acid and 10 μm A23187+600 μm ascorbic acid. After incubation sperm motility, viability, acrosome reaction, DNA damage and malondialdehyde levels were evaluated.
Results: Our results indicated that after one hour incubation, ascorbic acid significantly reduced malondialdehyde level in ascorbic acid group (1.4±0.11 nmol/ml) compared to control group (1.58±0.13 nmol/ml) (p<0.001). At the end of incubation, progressive motility and viability in ascorbic acid group (64.5±8.8% and 80.3±6.4%, respectively) were significantly (p<0.05 and p<0.001, respectively) higher than the control group (54.5±6.8% and 70.9±7.3%, respectively). A23187 significantly (p<0.0001) increased acrosome reaction in A23187 group (37.3±5.6%) compared to control group (8.5±3.2%) and this effect of A23187 attenuated by ascorbic acid in ascorbic acid+A23187 group (17.2±4.4%). DNA fragmentation in ascorbic acid group (20±4.1%) was significantly (p<0.001) lower than controls (28.9±4.6%).
Conclusion: In vitro ascorbic acid supplementation during teratozoospermic semen processing for ART could protect teratozoospermic specimens against oxidative stress, and it could improve ART outcome.
Viorica E Radoi, Camil L Bohiltea, Roxana E Bohiltea, Dragos N Albu,
Volume 13, Issue 10 (10-2015)
Abstract
Background: The discovery of circulating fetal DNA in maternal blood led to the discovery of new strategies to perform noninvasive testing for prenatal diagnosis.
Objective: The purpose of the study was to detect fetal aneuploidy at chromosomes 13, 18, 21, X, and Y by analysis of fetal cell-free DNA from maternal blood, without endangering pregnancy.
Materials and Methods: This retrospective study has been performed in Bucharest at Medlife Maternal and Fetal Medicine Department between 2013-2014. In total 201 women were offered noninvasive prenatal test. Maternal plasma samples were collected from women at greater than 9 weeks of gestation after informed consent and genetics counseling.
Results: From 201 patients; 28 (13.93%) had screening test with high risk for trisomy 21, 116 (57.71%) had advanced maternal age, 1 (0.49%) had second trimester ultrasound markers and the remaining 56 patients (27.86%) performed the test on request. Of those patients, 189 (94.02%) had a “low risk” result (<1/10,000). Of those who had a low risk result, 2 continued on to have amniocentesis with normal results.Five patients (2.48%) received “high risk” results (>99% risk) all for trisomy 21 (T21). T21 was confirmed by amniocentesis in 1 patient and the other 4 patients declined confirmation. The 7 remaining patients (3.48%) had a low fetal fraction of DNA.
Conclusion: It is probably that prenatal diagnosis using fetal DNA in maternal blood would play an increasingly role in the future practice of prenatal testing because of high accuracy.
Tayebeh Ghiasvand, Mohammad Taghi Goodarzi, Gholamreza Shafiee, Alireza Zamani, Jamshid Karimi, Marzieh Ghorbani, Iraj Amiri,
Volume 16, Issue 2 (2-2018)
Abstract
Background: Neopterin is a significant and sensitive marker in estimating the activity of cellular immune system. Oxidative stress plays a role in the etiology of male infertility. Increased reactive oxygen species is accompanied with increase in neopterin level. Hence neopterin may be involved in male infertility.
Objective: The objective of this case-control study was to determine neopterin level in idiopathic infertile and normospermic men; furthermore, to identify its relationship with oxidative stress markers including total oxidant, malondialdehyde, sperm DNA fragmentation, and total antioxidant capacity of seminal plasma.
Materials and Methods: Forty seven infertile and forty three normospermic males were selected according to WHO criteria. Their semen and blood samples were taken; subsequently, the levels of neopterin, total oxidant, total antioxidant, malondialdehyde, and sperm DNA fragmentation were measured.
Results: The levels of neopterin, total oxidant, and malondialdehyde in seminal plasma of infertile males were significantly higher than those of normospermic group (p=0.038, 0.018, and 0.028, respectively). Furthermore, sperm DNA fragmentation in infertile men was higher than that of control group (p<0.001). Moreover, total antioxidant capacity of seminal plasma in infertile males was significantly lower than that of normospermic subjects (p=0.002). No significant difference was observed in serum neopterin, total oxidant, and malondialdehyde between the infertile and normospermic groups.
Conclusion: The significant inverse correlation between seminal plasma neopterin and total antioxidant in the infertile males supports a possible role of neopterin in male infertility. Neopterin can be suggested as a marker in monitoring and diagnosis of idiopathic male infertility.
Ilyas Yusuf, Mathias Abiodun Emokpae,
Volume 19, Issue 2 (2-2021)
Abstract
Background: Studies have shown oxidative DNA damage is associated with male infertility.
Objective: This study determines the levels of 8-hydroxy-2′-deoxyguanosine (8-OHdG) and some markers of oxidative stress in seminal fluid of males investigated for infertility and men of proven fertility in Benin City, Nigeria.
Materials and Methods: Semen samples produced by self or assisted masturbation were analyzed by microscopic technique according to the World Health Organization guidelines. Thereafter, samples were centrifuged and seminal fluid plasma separated and stored at -20ºC prior to assay for 8-OHdG and oxidative stress biomarkers. Based on the sperm concentration/count, the overall samples were grouped into the following categories: normospermia (n = 20), oligozoospermia (n = 30), and azoospermia (n = 20). The control group comprised of 30 age-matched males of proven fertility. The seminal fluid 8-OHdG, total antioxidant status, superoxide dismutase and malondialdehyde (MDA) were assayed through ELISA and spectrophotometric methods, respectively.
Results: Seminal plasma level of 8-OHdG and MDA were significantly higher (p = 0.01) in infertile subjects than controls. The mean levels of 8-OHdG and MDA in infertile subjects were higher in azoospermia than oligospermia than normospermia and so, was least in the normospermia. Conversely, the mean levels of total antioxidant status and superoxide dismutase were significantly lower (p = 0.01) in infertile than fertile the control male subjects with levels higher in normospermia than oligospermia and least in azoospermia. Moreover, the seminal 8-OHdG correlated negatively with sperm count (r = -0.359, p = 0.01), percent motility (r = -0.388, p = 0.04), and percent morphology (r = -0.327, p = 0.02).
Conclusion: The assessment of sperm DNA damage in addition to routine seminal fluid analysis may play an important role in specific diagnosis and management of male infertility.
Mosavi A, Amjadi F, Barati M, Aflatoonian R, Amiri S, Mehdizadeh M, Mirsanei J,
Volume 19, Issue 5 (5-2021)
Abstract
Background: Preimplantation genetic diagnosis (PGD) is a useful clinical tool to identify embryos with or at risk of specific genetic malady before embryo implantation. Current procedures for embryo chromosomal screening require an invasive biopsy of the embryo. Blastomere biopsy has a potential lesion to the embryos may result in developmental defects or abortion. Thus, a non- invasive PGD is needed.This study hypothesized that embryonic DNA is present in the spent culture medium. We focused on X-linked disorders, these single-gene diseases due to the presence of defective genes on the X chromosome are dominant in males.
Objective: Therefore, the objective of this study was to discriminate between female (XX) and male (XY) embryos by detecting Y chromosome- specific genes in cell-free DNA and comparing to PGD results. It opens a new window for the development of a non-invasive PGD method.
Materials and Methods: Embryo´s spent media from day 3 and day 5 embryos development were collected. The modified phenol-chloroform solution was used for DNA extraction from spent media. DNA from spent media was evaluated using SRY, TSPY, and AMELOGENIN as targets using the qPCR method. IBM SPSS and Medcals were used for statistical analyses, to compare sex determination of embryos using spent medium with PGD results.
Results: Yield and purity of the extracted DNA as well as repeatability of the method were performed well using the modified phenol-chloroform solution. The amount of DNA at day 5 embryo culture medium was significantly higher than day 3. Results of sex determination using spent medium by Q-PCR were consistent with the results of PGD and 12th wk sonography. This invasive PGD method using a spent culture medium gave a sensitivity of 66.7%, specificity of 100%, positive predictive value of 100%, and negative predictive value of 67.6 (N = 56, Nxx = 23, Nxy = 33).
Conclusion: This investigation provides a potentially effective procedure that can help to avoid the invasive preimplantation genetic diagnosis, especially about X-linked diseases. Results of sex determination using spent medium by Q-PCR were consistent with the results of PGD. Improvements in DNA collection, amplification, and testing may allow for PGD without biopsy in the Future.
Asadollah Asadi, Rozita Ghahremani, Arash Abdolmaleki, Farzad Rajaei,
Volume 19, Issue 6 (6-2021)
Abstract
Activation of caspase, externalization of phosphatidyl serine, change in the mitochondrial membrane potential, and DNA fragmentation are apoptosis markers found in human ejaculated spermatozoa. Also, reactive oxygen species (ROS) play a vital role in the different types of male infertility. In this review, data sources including Google Scholar, Scopus, PubMed, and Science Direct were searched for publications with no particular time restriction to get a holistic and comprehensive view of the research. Apoptosis regulates the male germ cells, correct function and development from the early embryonic stages of gonadal differentiation to fertilization. In addition to maintaining a reasonable ratio between the Sertoli and germ cells, apoptosis is one of the well-known quality control mechanisms in the testis. Also, high ROS levels cause a heightened and dysregulated apoptotic response. Apoptosis is one of the well-known mechanisms of quality control in the testis. Nevertheless, increased apoptosis may have adverse effects on sperm production. Recent studies have shown that ROS and the consequent oxidative stress play a crucial role in apoptosis. This review aims to assimilate and summarize recent findings on the apoptosis in male reproduction and fertility. Also, this review discusses the update on the role of ROS in normal sperm function to guide future research in this area.
Serajoddin Vahidi, Nima Narimani, Laleh Dehghan Marvast, Esmat Mangoli, Ali Nabi, Mohammad Sadeghi,
Volume 20, Issue 5 (5-2022)
Abstract
Background: The sperm DNA fragmentation index (DFI) is one of the men's reproductive health criteria that affects assisted reproductive technique outcomes. Efforts in obtaining high-quality mature sperms seem to be necessary. Advanced sperm selection techniques (including physiological intracytoplasmic sperm injection [PICSI], zeta potential, microfluidic, etc.) have gained popularity in this regard.
Objective: The study aimed to compare the efficacy of zeta potential and PICSI sperm selection in obtaining sperms with better DNA integrity.
Materials and Methods: In this cross-sectional study, 48 couples were enrolled where the male partner had increased sperm DFI in his ejaculated sample and the female was in normal reproductive health. For each male partner, the semen sample was processed with zeta potential and PICSI techniques, then the sperm DFI of neat semen was compared to zeta and PICSI samples by the sperm chromatin dispersion test.
Results: Data showed that both the zeta potential and PICSI technique decreased sperm DFI in comparison with the neat semen sample (p < 0.001 for both). In addition, there was a statistically significant difference in sperm DFI between the PICSI and zeta potential samples (p < 0.01).
Conclusion: The current study showed that both zeta potential and PICSI could result in sperm with a lower DFI. However, PICSI seems to be superior to zeta potential in this regard.
Hassan Safari, Fatemeh Anbari, Saeed Ghasemi-Esmailabad, Behnam Maleki, Laleh Dehghan Marvast, Ali Reza Talebi,
Volume 20, Issue 5 (5-2022)
Abstract
Background: Total fertilization failure (TFF) is associated with essential mechanistic and cellular events.
Objective: The present study is a comprehensive examination of detrimental effects with well-known assays for predicting TFF in conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) cycles.
Materials and Methods: Semen parameters of 90 men, including 60 cases who had experienced IVF/ICSI failure and a control group of 30 individuals, were evaluated. Sperm chromatin/DNA quality assessments were done by aniline blue, toluidine blue, chromomycin A3, and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assays. A lipid hydroperoxide (LPO) kit was used to measure the LPO, and JC1 staining was used to evaluate mitochondrial membrane potential (MMP).
Results: There were statistically significant differences found between the IVF, ICSI and control groups by the toluidine blue (p = 0.01), TUNEL (p = 0.02), and chromomycin A3 (p < 0.001) tests, but not by the aniline blue staining. Furthermore, there was a significant difference regarding LPO concentration and high MMP in cases of IVF fertilization failure compared to the control group (p = 0.04, p = 0.02, respectively). The logistic regression model showed that sperm viability was predictive for fertilization failure in the ICSI group. Sperm chromatin and DNA quality assays were not predictors for TFF in either group.
Conclusion: Cellular events such as high DNA fragmentation damage, high levels of reactive oxygen species, and low MMP levels can cause TFF in IVF and ICSI programs. Diagnostic tests, especially in cases with previous fertilization failure, showed significant differences in sperm chromatin and DNA quality between groups but could not predict the risk of TFF.
Noorodin Karami, Farzaneh Iravani, Sareh Bakhshandeh Bavarsad, Samira Asadollahi, Seyed Mehdi Hoseini, Fateme Montazeri, Seyed Mehdi Kalantar,
Volume 22, Issue 3 (3-2024)
Abstract
To improve embryo transfer success and increase the chances of live birth in assisted reproductive methods, there is a growing demand for the use of pre-implantation genetic testing (PGT). However, the invasive approaches used in PGT have led to in vitro fertilization failure and abortions, increasing anxiety levels for parents. To address this, non-invasive PGT methods have been introduced, such as the detection of DNA in blastocoel fluid of blastocysts and spent culture media (SCM). These methods have proven to be minimally invasive and effective in detecting aneuploidy in the chromosomes of human embryos. This review aims to explore the different approaches to pre-implantation diagnosis, including invasive and non-invasive methods, with a particular focus on non-invasive PGT (niPGT). The search strategy involved gathering data from scientific databases such as PubMed, Google Scholar, and Science Direct using relevant keywords. The search was conducted until January 2023. In total, 22 studies have successfully reported the detection and amplification of cell-free DNA in the embryonic SCM. It is important to note that niPGT has some limitations, which include differences in indicators such as cell-free DNA amplification rate, concordance, level of maternal DNA contamination, sensitivity, and specificity between SCM samples and biopsied cells. Therefore, more extensive and detailed research is needed to fully understand niPGT's potential for clinical applications.
