جستجوی مقالات مرتبط با کلیدواژه « DNA fragmentation » در نشریات گروه « پزشکی »

Vitrification is a recently introduced yet widely applied assisted reproduction technique. So far, the effects of the chemicals and devices in vitrification on sperm motility and DNA integrity are still unclear.

This study aimed to examine sperm quality, as determined by semen analysis and sperm DNA integrity when vitrified with or without cryoprotectant agents (CPAs) using pulled-glass capillaries.

Between February and June 2020, 50 infertile men from the Hue Center for Reproductive Endocrinology and Infertility, Hue University of Medicine and Pharmacy, Vietnam, were enrolled. Sperm samples, prepared using the swim-up technique, were divided into 2 groups: vitrification with CPAs (group 1) and without CPAs (group 2). Vitrified sperm samples were preserved in 10 µL pulled-glass capillaries. Motility, sperm membrane integrity, and the DNA fragmentation index were tested.

Sperm motility in vitrified media with CPAs (54.4 ± 11%) was statistically higher than in media without CPAs (51.14 ± 10.6%, p < 0.05). CPAs did not affect sperm membrane integrity or large halo ratio (71.34 ± 8.47 vs. 70.38 ± 8.11 and 50.84 ± 18.92 vs. 51.98 ± 19.44, respectively). Group 2 exhibited a lower DNA fragmentation index than group 1 after vitrification (14.2 ± 8.47 vs. 12.60 ± 9.03, p = 0.021).

Using a pulled-glass capillary for sperm vitrification, the presence of CPAs in the vitrification medium resulted in higher progressive motility and lower DNA fragmentation index than the medium without CPAs.

Oral cancer’s aggressive nature poses a significant health risk, demanding timely diagnosis and effective treatment. Despite progress in conventional therapies like chemotherapy and surgery, their limitations drive the exploration of alternative strategies. This study assessed Solanum trilobatum-derived silver nanoparticles (AgNPs) in impacting cancer cell cycles, inducing apoptosis, and modulating key pathways, including the phosphoinositide 3-kinase (PI3K)/Protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway, leveraging phytotherapy and nanotechnology—a promising frontier in cancer treatment.

AgNPs were synthesized through the reduction of ions and stabilized using aqueous leaf extracts of S. trilobatum. After the characterization of AgNPs, the mRNA Gene Expression and mitochondrial membrane potentials (MMPs) were assessed. DNA fragmentation was done and DNA pattern by gel documentation system was observed. The study also assessed the modulation of the PI3K/Akt/mTOR cascade, impacting tumor growth and proliferation.

Biosynthesized AgNPs were characterized using UV-visible spectrophotometry (UVvis), energy-dispersive X-ray spectroscopy (EDX), scanning electron microscopy (SEM), and Fourier-transform infrared (FTIR) spectroscopy. DNA fragmentation exhibited a typical ladder pattern. Dose-dependent changes in MMP were observed in the treated oral cancer cells. The effect of S. trilobatum-derived AgNPs in targeting the cell signaling pathway correlated significantly with their anticancer potency (P<0.001).

This study reveals S. trilobatum leaf extract-based AgNPs as a natural cytostatic agent against oral squamous carcinoma. Utilizing nature’s resources and nanoscale science, they hold promise for enhancing oral cancer treatment outcomes and survival rates.

Glioblastoma (GBM) stands out as the most prevalent primary brain tumor characterized by its high aggressiveness. Numerous therapeutic approaches have been employed, and the utility of combination therapies has been substantiated, particularly in GBM treatment. Cisplatin, an anticancer chemotherapeutic agent, is employed for the management of various malignancies, including GBM; however, it is associated with significant systemic toxicity. In the realm of combination therapy, metformin, a biguanide drug conventionally used as a first-line treatment for type 2 diabetes, has recently emerged as a valuable adjunct in the treatment of a diverse spectrum of tumors. This study aimed to elucidate the impact of metformin on sensitizing the human cerebral GBM cancer cell line (AMGM) to cisplatin chemotherapy by employing the comet assay as a means to assess DNA damage, thereby advocating the potential of metformin as an adjuvant for cisplatin-based therapy. In this experimental study, the AMGM cell line was cultured and subsequently treated with either single-agent cisplatin, metformin, or a combination of both drugs. Cell viability was assessed through growth inhibition calculations. The Chou–Talalay analysis was used to assess the cooperative effect of this drug combination. Furthermore, DNA fragmentation was quantified using the alkaline comet assay technique. The findings demonstrate that metformin significantly potentiates the therapeutic efficacy of cisplatin by synergistically inhibiting the growth of AMGM cells and reducing DNA damage. These results underscore the potential utility of metformin as a valuable adjunct in enhancing the clinical effectiveness of chemotherapy regimens.

DNA fragmentation index (DFI) enhances routine semen analysis by providing valuable insights into male reproductive potential. Utilizing Halosperm test, a sperm chromatin dispersion (SCD) assay based on induced condensation. The purpose of this study was to assess sperm DNA damage both before and after freezing. By following the specified kit instructions, an attempt was made to validate the SCD test protocol, with a particular emphasis on the implications of sperm freezing on its DNA integrity.

In total, 380 fresh human semen samples from normozoospermic patients were frozen at -20°C for 10 days, using SCD cryopreservation reagent. Routine semen analysis and DNA fragmentation index (DFI) were determined for each sample before freezing and after thawing. Semen morphology and sperm DFI were compared before and after freezing/thawing process.

There was a significant decrease in sperm normal morphology after thawing (9.31±2.42% vs. 7.1±1.53%, p<0.05, respectively). The sperm head, midpiece, and tail defect rate increased after freezing at -20°C. Moreover, DFI was significantly higher after thawing compared to before freezing (20.71±1.61% before freezing vs. 29.1±0.21% after thawing with p<0.001).

Cryoconservation of semen samples at -20C for 10 days using SCD cryopreservation reagent seems to damage sperm morphology, resulting in a reduction in sperm DNA integrity. The measurement of DFI on a fresh sample remains the most reliable technique for obtaining accurate results.

Sperm DNA fragmentation (SDF) can affect fertilization rate and embryo development, making it a useful measure for assessing male fertility. Available evidence supports the association between high sperm DNA fragmentation and poor outcomes, with regard to natural conception. Several treatment options are being adopted with varying degrees of success. Some of the commonly used treatment options are the intake of oral antioxidants, varicocele repair, and techniques like micromanipulation- based sperm selection and use of testicular sperm for intracytoplasmic sperm injection.

Studies have shown that around 29% of couples depend on complementary and alternative medicine (CAM) modality for the treatment of infertility. However, there is a lack of substantial evidence regarding its efficacy in treating various aspects of infertility in couples. The current case report is about a 44 year-old male patient with infertility, who has a known diagnosis of sex chromosome abnormalities. Meanwhile, the SDF study reports indicated the presence of chromosomal abnormalities. This patient was treated exclusively with Ayurveda therapy aimed towards qualitative improvement in reproductive tissues (Shukra Dhatu as per Ayurveda). Patient was assessed periodically for changes in chromosomal abnormality. After four months of treatment, the evaluations demonstrated the presence of completely normal chromosomes.

This case study indicates the potential of Ayurveda therapy in treating cases of male infertility caused by DNA fragmentation. Furthermore, observations and systematically designed clinical trials are warranted to establish a stronger level of evidence before making further clinical recommendations.

Varicocele is one of the most common treatable causes of male infertility, and its treatment may bebeneficial for fertility. This study aimed to evaluate fertility rate and DNA fragmentation index (DFI) following varicocelectomyin primary infertile men with clinical varicocele. This prospective longitudinal study was conducted on primary infertility men, in a tertiarycenter from December 2018 to December 2019 with one-year follow-up. Data of the semen parameters, DFI (%), andfertility rate were gathered before, as well as 4 and 12 months after undergoing varicocelectomy. For data analysis,SPSS software and analytical test were used. Out of 76 patients who were analyzed, 22 (29%) became fertile and 54 (71%) remained infertile. Semenparameters and DFI (%) were improved significantly following varicocelectomy (P<0.001). Smoking history, occupationalheated exposure, body mass index (BMI), and infertility duration were determined as predictors associatedwith fertility status (P<0.05). Although varicocele repair improved the DFI, the fertility rate was achieved in less than one-third ofpatients; it seems that the other parameters, such as the history of smoking, occupational heated exposure, overweight,and duration of infertility should be considered as predictors of fertility status, in primary infertile men who are a candidatefor varicocelectomy.

Sperm chromatin abnormalities are defects in nuclear maturation and DNA integrity. These defectsoriginated from defective spermatogenesis due to a lack of DNA repair during chromatin remodeling. Changes insemen elements can cause damage to chromatin. There is little information about the relationship between changesin trace metal elements and total antioxidant capacity (TAC) with sperm chromatin damage. The present study wasconducted to determine the relationship between Selenium (Se), Iron (Fe), Zinc (Zn), Copper (Cu) and the TAC ofsemen with the status of human sperm chromatin.

In this experimental study, semen samples (n=30) were collected from healthy men referredto Kermanshah Motazadi Hospital and stored in liquid nitrogen; after thawing and centrifugation, sperm were separated.The atomic absorption method was used to measure the concentration of metal elements. The TAC was evaluatedusing the ferric-reducing antioxidant capacity of the plasma method. Furthermore, the integrity of sperm chromatinwas measured using the sperm DNA fragmentation (SDF) method.

The status of sperm chromatin had a non-significant correlation with body mass index (BMI, P=0.25, r=0.21)and a non-significant negative correlation with sperm count (P=0.71, r=-0.71) and motility (P=0.75, r=0.61). In addition,there was no significant relationship between sperm chromatin and the TAC of semen (P=0.92, r=0.01). Additionally,there was no significant correlation between Se, Zn, or Cu concentration (P>0.05) and Fe concentration,which had a partially positive relationship with the chromatin state of sperm (P=0.24, r=0.20).

The trace metal elements in the seminal fluid did not play a significant role in the status of sperm chromatin.

Sperm DNA fragmentation can affect reproductive outcomes in assisted reproductive techniques (ART), and it is a concern in density gradient centrifugation (DGC). By contrast, microfluidic approaches allow the selection of highly motile sperm with low DNA fragmentation index (DFI). The purpose of current study, was to compare the efficacy of a microfluidic device designed in-house in comparison with DGC.

Nineteen healthy men with normal semen profiles were included in the study. Semen samples were individually aliquoted for three sperm preparation analyses (crude and processed with to either DGC or the microfluidic method). Sperm parameters of the samples were evaluated along with DNA fragmentation using the terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) method.

Sperm processed using the microfluidic method showed a significantly lower DFI than those obtained using DGC and in crude semen, with DFI of 1.1%, 3.5%, and 4.9%, respectively. Although the microfluidic method yielded significantly lower sperm concentrations than DGC, no significant differences were observed in total motility, progressive motility, curvilinear velocity, straight-line velocity, or normal morphology.

Using the in-house microfluidic device, sperm with lower DFI was effectively isolated when compared with DGC. The motility and normal morphology rates were comparable among the samples.

To study the alterations on the lenticules extracted after femtosecond (Femto) small incision lenticule extraction (SMILE) versus the corneal free cap removed using a microkeratome.

The visuMax (500 kHz; laser energy: 180 nJ) was used for small-incision lenticule extraction. Free caps from human cadaveric corneas were excised by microkeratome. The collected lenticules were examined with the light and transmission electron microscope (TEM) for histological analysis, DNA fragmentation was assessed by agarose gel electrophoresis, DNA damage was evaluated using comet assay, and corneal proteins secondary structure was assessed by Fourier transform infrared spectroscopy (FTIR).

Light microscopic examination showed the presence of more edematous stroma under Femto SMILE than under free cap with a percentage change of 101.6%. In the Femto SMILE group, TEM examination showed pyknotic keratocytes, disruption, and cavitation of the collagen arrays stromal area under Femto SMILE. The DNA fragmentation for the Femto SMILE group revealed one undefined band with a size of 1.1 Kbp. The comet assay analysis indicated the presence of 3% and 8.0% tailed cells for the free cap and Femto SMILE groups, respectively. The tail lengths were 1.33 ± 0.16 and 1.67 ± 0.13 µm (P < 0.01), the percentage of tail DNA was 1.41 ± 0.18% (P < 0.01) and 1.72 ± 0.15%, and the tail moments were 1.88 ± 0.12 AU and 2.87 ± 0.14 AU (P < 0.001) for the free cap and Femto SMILE groups, respectively. FTIR spectroscopy of the Femto smile group revealed disorders in the secondary and tertiary structure of the proteins.

Femto SMILE technique induced more structural changes, DNA fragmentation, DNA damage, and corneal proteins secondary structure alteration than those induced by a microkeratome cutting. These changes may be attributed to the deep penetration of high energy levels to the corneal layer. These findings may highlight the potential impact of the Femto SMILE on the cornea and the necessity for managing the laser parameters used.

Hormonal imbalance is one of the important etiological factors for Oligoasthenoteratospermias (OAT).

This study aimed to evaluate the effects of hormonal changes including prolactin, TSH, testosterone, luteinizing hormone, follicle-stimulating hormone and anti-Mullerian hormone on sperm DNA fragmentation in normal men compared with OAT to design a clinical algorithm for the comprehensive study of male factor infertilities.

We consecutively selected 60 candidates referred to the infertility clinic to collect the semen and blood samples. Then, a terminal deoxynucleotidyl transferase dUTP nick end labeling test was performed to evaluate the sperm DNA fragmentation index (DFI). After semen analysis and DFI checking, they were classified into 4 groups consisting of normospermia and OAT men each with or without increased DFI. Hormone parameters were analyzed using enzyme-linked immunoassay.

Follicle-stimulating hormone and luteinizing hormone levels showed positive correlations with DFI in a significant way (p ≤ 0.01), while testosterone and thyroid-stimulating hormone were associated with sperm concentration. Prolactin and anti-Mullerian hormone levels significantly correlated (p ≤ 0.01) with sperm concentration and DFI value simultaneously.

Decreased and increased levels of serum hormones could adversely affect semen profile and sperm DNA integrity which lead to severe male infertility. Although we investigated the effects of the main hormones related to male infertility on DNA damage, the role of these hormones on the fertilization rate and embryo quality needs to be evaluated in further studies.

Recurrent pregnancy loss is a distinct disorder defined as the loss of at least 2 pregnancies before the 20th wk of gestation. With one half of the genome of the embryo belonging to the father, the integrity of the sperm genome is crucial for a successful pregnancy. Semen analysis is recommended for men in such cases to evaluate sperm concentration, morphology, vitality and motility. However, other important sperm parameters such as sperm epigenetics, aneuploidy, Y chromosome microdeletion and chromatin integrity also correlate with successful pregnancy and delivery rate. This article examines the use of different sperm tests and their importance in male partners of women suffering from recurrent pregnancy loss.

Polyvinylpyrrolidone (PVP) is a chemical material used in intracytoplasmic sperm injection (ICSI) pro-gram. The aim of this study was to investigate the ideal time that sperm can be safely incubated in PVP with less structure and DNA damage.

Thirty-one Oligoasthenoteratospermia (OAT) samples were used. Sperm samples were prepared by dis-continuous density-gradients method and incubated in 10% PVP at different time intervals (0, 5, 10, 15, 20, and 30 min). The effect of PVP was assessed on sperm DNA fragmentation and viability via SCD assay and Eosin–ni-grosin staining respectively.

Data showed there was a significant increase in sperm DNA fragmentation at 10 min compared to 0 min. The viability rate also significantly reduced at 10 min compared to 0 min.

As a result, sperm samples could be incubated with PVP for less than 10 min. While prolonged in-cubation may significantly damage the sperm DNA integrity and viability.

Total fertilization failure (TFF) is associated with essential mechanistic and cellular events.

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.

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).

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.

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.

Today, vitamin D deficiency (VDD) is one of the major health issues around the world and VDD is associated with several diseases. This study was conducted to find the relationship between vitamin D status in male’s serum with sperm function and clinical outcomes in infertile men candidate for intracytoplasmic sperm injection (ICSI).

In this cohort study, different parameters of male fertility such as sperm parameters, oxidative stress, and sperm chromatin status were evaluated in sperm samples of 30 infertile couples candidate for ICSI. Clinical outcomes like fertilization, embryo quality, and implantation were also assessed. Data were analyzed using SPSS Statistics 25.0 software. Besides, assessment of the correlation between aforementioned parameters with the level of serum vitamin D, in this study, ICSI candidates were divided into three groups [individuals with sufficient vitamin D levels (>30 ng/ml), insufficient vitamin D levels (between 20-29 ng/ml), and VDD (<20 ng/ml)]. The aforementioned parametesr were also compared between these study groups.

Analysis of all the data revealed a significant correlation between the level of vitamin D with sperm concentration (P=0.000, r=0.5), sperm count (P=0.03, r=0.31) and sperm reactive oxygen species (ROS) level (P=0.000, r=-0.77). Moreover, comparing clinical outcomes within study groups showed a significant difference in implantation rate between sufficient and other groups (insufficient and deficient) (P=0.02).

Considering the association between sperm concentration and level of ROS with vitamin D and, higher implantation rate in individuals with vitamin D sufficient group compared to other two groups, our data call for vitamin D supplementation as part of male infertility treatment. But considering our sample size, further research is needed to verify these findings.

The effect of storage time and temperature on the prepared semen sample was evaluated, but the optimal condition is unclear. The aim of this study was to assess the effect of long-term incubation of prepared sperm at testicular temperature versus room temperature on semen parameters and DNA fragmentation index (DFI).

Sperm samples were collected from 40 patients between 2019 and 2020. Each sample was separated into two parts and underwent a non-direct swim-up method. One group was placed in a 35°C incubator, and the other group was kept at room temperature (26°C) in the dark. Both groups were evaluated at intervals of 45 minutes, 24 hours and 48 hours after sampling in terms of sperm concentration, motility, morphology, and DFI. Student t-test and repeated measures analysis of variance were used.

Sperm count (P=0.007) and motility (P<0.001) at 26°C in three-time intervals of 45 minutes, 24 hours and 48 hours were significantly higher than 35°C. The proportion of normal morphology spermatozoa at 26 and 35°C at 45 min, 24 h, and 48 h did not show a significant difference (P=0.08). DFI at 26°C in three-time intervals was significantly lower than 35°C (P=0.008).

 The results of this study indicated that when the prepared sperm samples are incubated for 24 h at 26°C compared to 35°C, they show significantly better quality and good quality of sperm can be retained for several hours if stored at room temperature.

Testicular dysfunction is one of the common side effects that results from the treatment with oxaliplatin® as a chemotherapy drug, and pharmaceutical search for agents to relieve the side effects are underway. The current study explored the possible ameliorative and regenerative effects of Costus ethanolic extract against testicular degeneration in male rats induced by oxaliplatin. 

Male Wistar albino rats weighing 150-200g were divided into four groups of 10 rats each as follows: group-1 control rats; group-2 rats treated orally with the extract at 50 mg/kg/day for six weeks; group-3 rats injected oxaliplatin intraperitoneally at 10 mg/kg/week for six successive weeks; group-4 rats were injected intraperitoneally with oxaliplatin at 10 mg/kg/week combined with a daily oral dose of the Costus extract for six weeks. 

Data from the current study revealed that the extract lowered the toxic effect of oxaliplatin on the testicular tissue samples. This was evident by the significant rise in the serum of total & free testosterone and CD4 cells, and the levels of GSH, SOD and CAT activities in the testis coupled with a marked reduction of serum TNF-α and IL-1β and testis MDA, nitric oxide levels and DNA fragmentation. Also, the extract promoted the regeneration of the histopathological structure of the testis. 

This study proposes a novel therapeutic application for the Costus extract as a therapeutic agent against testicular toxicity induced by oxaliplatin treatment through its promising anti-inflammatory and antioxidant properties.

Emamectin benzoate (EMB) is a biopesticide which used in agriculture as an insecticide. It is easier to reach ecologically and affects human health. This study aims to evaluate the protective effect of chitosan and chitosan nanoparticles against EMB-induced hepatotoxicity.

Male mice were distributed into four groups: G1: the negative control, G2: EMB group (5 mg/kg diet), G3: EMB with Chitosan, (600 mg/kg diet), and G4: EMB with Chitosan nanoparticles (600 mg/kg diet). The experiment continues for 8 weeks, and the animals were sacrificed, and their organs were removed and immediately weighed after sacrifice. The liver was quickly removed and processed for histopathological and genetic studies.

Emamectin benzoate (EMB) treatment induced oxidative stress by increased levels of Malondialdehyde (MDA), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) with inhibition of acetylcholinesterase (AChE), Superoxide dismutase (SOD) and Catalase (CAT) levels. EMB produced several histopathological changes in the liver. Relative expressions of studied genes elevated in the liver with increase in DNA damage. Co-treatment with chitosan and chitosan nanoparticles reduced EMB related liver toxicity that belong to biochemical, histopathological, gene expression, and DNA damage by increasing antioxidant capacity.

This study offers insight into the potential for Chitosan and chitosan nanoparticles as a novel natural material against the oxidative stress induced by EMB.