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Showing 19 results for Chromatin

Tahereh Esmaeilpour, Leila Elyasi, Soghra Bahmanpour, Alireza Ghannadi, Ahmad Monabbati, Farzaneh Dehghani, Marjaneh Kazerooni,
Volume 10, Issue 5 (10-2012)
Abstract

Background: It has been claimed that by using different washing methods, the sperms can be separated according to size, motility, density, chromosomal content and surface markings and charge. These methods also reduce sperm chromatin deficiencies and screen the sperms before applying in assisted reproduction techniques.
Objective: This study compared simple density gradient methods and a combined method with albumin density gradient and PureSperm separation (alb/PureSperm) for sex preselection by double fluorescence in situ hybridization (FISH) versus chromomycin A3 staining to determine chromatin integrity. Materials and Methods: 30 normal semen samples were prepared with PureSperm, albumin gradients and alb/PureSperm. All samples were then stained by FISH and chromomycin A3. The results were compared with SPSS 11.5 and the Kruskal-Wallis test.
Results: The proportion of X-bearing spermatozoa by PureSperm separation (47.58±5.67) and Y-bearing spermatozoa by albumin gradient (46.13±3.83) methods were slightly higher than in putative normal sperm samples (1:1), but there were no significant differences in the X- or Y- bearing spermatozoa counts among the three methods. Albumin gradient separation tended to underestimate abnormal spermatozoa compared to PureSperm and combined alb/PureSperm.
Conclusion: Routine separation methods slightly enriched X- or Y- bearing spermatozoa, but the differences were not significant for clinical purposes. The combined alb/PureSperm method had no advantages for assessing sex ratio or chromatin integrity compared to simpler gradient methods.
Esmat Mangoli, Ali Reza Talebi, Morteza Anvari, Majid Pourentezari,
Volume 11, Issue 1 (4-2013)
Abstract

Background: Diabetes mellitus (DM), primary or idiopathic is a chronic disorder of the carbohydrate, lipid and protein metabolism. DM may impact male reproductive function at several levels. It is shown that DM has detrimental effects on sperm parameters in human and experimental animals.
Objective: The aim of this study was to observe the effects of diabetes on sperm parameters (viability, count, morphology and motility) and evaluation of sperm chromatin quality in mice.
Materials and Methods: Totally twenty adult male Syrian mice were divided randomly into 2 groups (n=10). The animals of group A were considered as controls while group B mice were diabetic that received a single dose (200 mg/kg) streptozotocin (STZ) intra peritoneally. After 35 days, the cauda epididymis of each diabetic mouse was dissected and placed in culture medium for 30 min. The swim-out spermatozoa were analyzed for count, motility, morphology and viability. The sperm chromatin quality and DNA integrity, was evaluated with Aniline Blue (AB), Toluidine blue (TB), Acridine orange (AO) and Chromomycin A3 (CMA3) staining.
Results: In sperm analysis, the diabetic mice had poor parameters in comparison with control animals (p=0.000). Regarding sperm chromatin quality, the results of TB and AO tests showed statically significant differences between two groups, but in AB and CMA3 staining, we didn’t see any differences between them.
Conclusion: The results showed that STZ-induced diabetes mellitus may influence the male fertility potential via affecting sperm parameters and DNA integrity in mice. However, according to our data, the diabetes doesn’t have any detrimental effects on histone-protamines replacement during the testicular phase of sperm chromatin packaging.
Mohammad Mardani, Ahmad Vaez, Shahnaz Razavi,
Volume 12, Issue 5 (6-2014)
Abstract

Background: Currently, relation between reactive oxygen species (ROS) ROS concentration and semen quality was indicated. Saffron has traditionally been not only considered as a food additive but also as a medicinal herb, which has a good antioxidant properties.
Objective: The aim of this study was to evaluate the protection potency of saffron and vitamin E on sperm chromatin integrity.
Materials and Methods: Thirty adult male Wistar rats divided equally into saffron (100 mg/kg), vitamin E (100 mg/kg) and control (0.5cc distilled water /day) groups. After 60 days, cauda epididymis dissected and sperm cells were used for analysis of sperm chromatin packaging by chromomycin A3 (CMA3) staining, and sperm chromatin susceptibility to acid denaturation by acridine orange (AO) staining.
Results: The mean percentage of CMA3 positive sperm was significantly decreased in saffron and vitamin E groups relative to control group (p<0.001). Moreover, the AO staining results showed that the mean percentage of sperm with DNA damage was significantly decreased in saffron and vitamin E groups as compared with control group (p<0.001).
Conclusion: Our results purposed that saffron can protect sperm against DNA damage and chromatin anomalies.
Majid Pourentezari, Alireza Talebi, Abulghasem Abbasi, Mohammad Ali Khalili, Esmat Mangoli, Morteza Anvari,
Volume 12, Issue 5 (6-2014)
Abstract

Background: Acrylamide (AA) is an important industrial chemical primarily. AA is also found in carbohydrate-rich foods that are prepared at high temperatures, such as French fries and potato chips. It is demonstrated that AA is a carcinogen and reproductive toxin and has ability to induce sperm damage.
Objective: The aim of this study was to observe the effects of AA on sperm parameters and evaluation of sperm chromatin quality and testosterone hormone in mice.
Materials and Methods: Totally, 16 adult male mice were divided into two groups. Mice of group A fed on basal diet; group B received basal diet and AA (10 mg/kg, water solution) for 35 days. The right cauda epididymis was incised and then placed in Ham’s F10 culture media at 37oC for 15 min. Released spermatozoa were used to analyze count, motility, morphology and viability. To determine the sperm DNA integrity and chromatin condensation, the cytochemical techniques including Aniline blue, Acridine orange and Chromomycin A3 staining were used.
Results: AA-treated mice had poor parameters in comparison with control animals. In sperm chromatin assessments, except TB (p=0.16), significant differences were found in all of the tests between two groups. It was also seen a significant decrease in concentration of blood testosterone in AA-treated animals when compared to controls (p<0.001).
Conclusion: According to our results, AA can affect sperm parameters as well as sperm chromatin condensation and DNA integrity in mice. These abnormalities may be related to the reduction in blood testosterone.
Fatemeh Roodbari, Nahid Abedi, Ali Reza Talebi,
Volume 13, Issue 11 (11-2015)
Abstract

Background: There are few studies indicating the detrimental effects of ibuprofen on sperm fertility potential and DNA integrity. Objective: To determine the effects of Ibuprofen on sperm parameters, chromatin condensation and DNA integrity of mice. Materials and Methods: In this experimental study, 36 adult male mice with average weight 37 gr were divided into three groups, including control (group I, n=12), normal dosage of ibuprofen (group II, n=12) and high dosage (group III, n=12). Ibuprofen with different doses was dissolved in daily water of animals. After 35, 70 and 105 days, the cauda epididymis of mice were cut and incubated in Ham’s F10 media. Sperm samples were analyzed for parameters (motility, morphology and count), DNA integrity (SCD test) and chromatin condensation (chromomycin A3 and Aniline blue staining). Results: After 35 days, in addition to above mentioned sperm parameters, all of the treated mice showed statistically significant increase in spermatozoa with immature chromatin (P<0.05). However, after 70 days, the rate of sperm DNA fragmentation assessed by SCD was increased in group II (66.5±0.7) and the percentage of immature spermatozoa (AB+ and CMA3+) was higher in group III (77.5±0.7 and 49.5±6.3 respectively) than other groups. After 105 days, the AB+ spermatozoa were increased in both normal dose and high dose groups. Conclusion: Ibuprofen may cause a significant reduction in sperm parameters and sperm chromatin/DNA integrity in mice. It should be noted that these deleterious effects are dose-dependent and can be seen in early and late stage of drug treatments.
Neda Taghizabet, Esmat Mangoli, Fatemeh Anbari, Seyed Ali Masoodi, Ali Reza Talebi, Malihe Mazrooei,
Volume 14, Issue 6 (6-2016)
Abstract

Background: Evaluating the significance and the effects of plant-derived drugs on laboratory animal’s fertility was recognized. There was antioxidant activity reported from Heracleum persicum (Golpar).
Objective: Current study aims to study the antioxidant effect of Golpar extracts on sperm parameters and chromatin quality in mice.
Materials and Methods: Eighteen adult male mice were divided to 3 groups (10 wk old, 35 gr weight): group1 received hydro alcoholic extract (1000 mg/kg, ip), group 2 received oil extract (200 mg/kg, ip) and group 3 serving as the sham control group that received sterile water. Finally, left cauda epididymis of each animal was dissected and sperm analysis was done accordingly. To asses sperm chromatin and DNA quality, we used aniline blue (AB), toluidine blue (TB), chromomycin A3 (CMA3) and acridine orange (AO) staining.
Results: Progressive and non-progressive sperm motility were significantly increased in group 1 in comparison with group 3 (p=0.032). There was an increasing trend in progressive sperm motility and decreasing trend in non-progressive sperm motility in group 2 in comparison with group 3, but the differences were not significant (p=0.221 and p=0.144, respectively). According to the sperm chromatin quality, the results of TB and AO tests revealed significant differences (p=0.004, p=0.000, respectively) between those groups and showed that the extracts of Golpar cause DNA damage, but no differences can be observed between them in AB and CMA3 staining (p>0.05).
Conclusion: The results showed that Heracleum persicum extracts may improve sperm motility. Also, it has harmful effects on sperm chromatin condensation and DNA integrity in mice
Mahsa Nazari, Ali Reza Talebi, Mohammad Hosseini Sharifabad, Abolghasem Abbasi, Arezoo Khoradmehr, Amir Hossein Danafar,
Volume 14, Issue 10 (10-2016)
Abstract

Background: The particles in the range of 1-100 nm are called nanoparticles. Gold nanoparticle is one of the most important metal nanoparticles with wide usage.
Objective: This study investigated the effects of gold nanoparticles on sperm parameters and chromatin structure in mice.
Materials and Methods: In this experimental study, 72 male bulb-c mice were divided into 9 groups including: 4 Sham groups (Sc 1-4), 4 experimental groups (Au 1-4), and 1 control group (C). Experimental groups received 40 and 200 µg/kg/day soluble gold (Au) nano-particles for 7 and 35 days, by intra peritoneal injection, respectively. Sham groups were treated with 1.2 mM sodium citrate solution with 40 and 200 µg/kg/day doses for same days and control group did not receive any materials. Motility and Morphology of spermatozoa were analyzed. Chromatin quality was also evaluated using AB (Aniline blue), TB (Toluidine blue) and CMA3 (Chromomycin A3) staining methods.
Results: The sperm analysis results showed that motility and morphology of sperm in experimental groups (especially in groups that have been treated for 35 days with nano-particles) had significant decrease in comparison with control group. TB, AB and CMA3 results showed a significant increase in abnormal spermatozoa from all Au-treated groups.
Conclusion: Gold nano-particles firstly can reduce the sperm parameters such as motility and normal morphology and secondly affect sperm chromatin remodeling and cause the increase instability of chromatin and also increase the rate of sperm DNA damage. These deleterious effects were more obvious in maximum dose and chronic phase.

Mojdeh Sabour, Arezoo Khoradmehr, Seyyed Mehdi Kalantar, Amir Hossein Danafar, Marjan Omidi, Iman Halvaei, Ali Nabi, Saeed Ghasemi- Esmailabad, Ali Reza Talebi,
Volume 15, Issue 3 (5-2017)
Abstract

Background: Methamphetamine (MA) was shown to have harmful effects on malereproductive system.
Objective: To investigate probable effects of daily administration of MA on spermparameters and chromatin/DNA integrity in mouse.
Material and Methods: Thirty-five NMRI male mice were divided into five groupsincluding low, medium, and high dosage groups which were injectedintraperitoneally with 4, 8 and 15 mg/kg/day for 35 days, respectively. Normalsaline was injected in sham group and no medications were used in control group.Then, the mice were killed and caudal epididymis of each animal was cut and placedin Ham’s F10 medium for sperm retrieval. To evaluate sperm chromatinabnormalities, the aniline blue, toluidine blue and chromomycine A3 were used. Forsperm DNA integrity and apoptosis, the acridine orange, sperm chromatindispersion, and TUNEL assay were applied. For sperm morphology, Papanicolaoustaining was done
Results: Normal morphology and progressive motility of spermatozoa decreased inmedium and high dosage groups in comparison with the control group (p=0.035).There was a significant increase in rate of aniline blue, toluidine blue, andchromomycine A3 positive spermatozoa in high dosage group. In a similar manner,there was an increase in rates of acridine orange, TUNEL and sperm chromatindispersion positive sperm cells in high dosage group with respect to others.
Conclusion: MA abuse in a dose-dependent manner could have detrimental effectson male reproductive indices including sperm parameters and spermchromatin/DNA integrity in mice
Parvin Sabeti, Fardin Amidi, Seyed Mahdi Kalantar, Mohammad Ali Sedighi Gilani, Soheila Pourmasumi, Atefeh Najafi, Ali Reza Talebi,
Volume 15, Issue 5 (6-2017)
Abstract

Background: Teratoasthenozoospermia (TA) is a severe form of male infertilitywith no clear etiology.
Objective: To compare the level of intracellular anion superoxide (O2–), heat shockprotein A2 (HSPA2) and protamine deficiencies in ejaculated spermatozoa betweenteratoasthenozoospermic and normozoospermic men.
Materials and Methods: In this case- control study, semen samples of 20 infertilemen, with TA (with normal morphology lower than 4%_ and total motility lowerthan 40% ) as the case group and 20 normozoospermic fertile men as the controlgroup were evaluated for intracellular O2– and HSPA2 by flow cytometry andprotamine deficiency by Chromomycin A3 (CMA3) test.
Results: The rate of CMA3+ spermatozoa in the case group was higher thancontrols (p=0.001). The percentages of HSPA2+ spermatozoa in the cases weresignificantly lower than controls (p=0.001). Also, intracellular O2– levels in the casegroup were significantly higher than controls (p=0.001) and had positivecorrelations with sperm apoptosis (r=0.79, p=0.01) and CMA3 positive sperm(r=0.76, p=0.01), but negative correlations with normal morphology (r=-0.81,p=0.01) and motility (r=-0.81, p=0.01). There was no significant correlation betweenintracellular O2– and HSPA2 in the case group (r=0.041, p=0.79).
Conclusion: We suggest that the increase in intracellular O2–, decrease inspermatozoa HSPA2+, and high percentages of spermatozoa with immaturechromatin might be considered as etiologies of infertility in TA patients.
Soheila Pourmasumi, Parvin Sabeti, Tahereh Rahiminia, Esmat Mangoli, Nasim Tabibnejad, Ali Reza Talebi,
Volume 15, Issue 6 (7-2017)
Abstract

The sperm DNA damage may occur in testis, genital ducts, and also after ejaculation. Mechanisms altering chromatin remodeling are abortive apoptosis and oxidative stress resulting from reactive oxygen species. Three classifications of intratesticular, post-testicular, and external factors have been correlated with increased levels of sperm DNA damage which can affect the potential of fertility. Alcohol consumption may not increase the rate of sperm residual histones and protamine deficiency; however, it causes an increase in the percentage of spermatozoa with DNA fragmentation and apoptosis. In a medical problem as spinal cord injury, poor semen parameters and sperm DNA damage were reported. Infection induces reactive oxygen species production, decreases the total antioxidant capacity and sperm DNA fragmentation or antigen production that lead to sperm dysfunctions and DNA fragmentation. While reactive oxygen species generation increases with age, oxidative stress may be responsible for the age-dependent sperm DNA damage. The exposing of reproductive organs in older men to oxidative stress for a long time may produce more DNA-damaged spermatozoa than youngers. Examining the sperm chromatin quality in testicular cancer and Hodgkin’s lymphoma patients prior to chemotherapy demonstrated the high incidence of DNA damage and low compaction in spermatozoa at the time of diagnosis. In chemotherapy cycles with genotoxic agents in cancer patients, an increase in sperm DNA damage was shown after treatment. In overall, those factors occurring during the prenatal or the adult life alter the distribution of proteins associated with sperm chromatin induce changes in germ cells which can be detected in infertile patients.
Ladan Bandegi, Morteza Anvari, Mahmood Vakili, Arezoo Khoradmehr, Aghdas Mirjalili, Ali Reza Talebi,
Volume 16, Issue 6 (6-2018)
Abstract

Background: Prescribing antidepressant drugs is becoming common. These drugs are known to affect sexual functions.
Objective: The study is aimed to assess the effects of amitriptyline and venlafaxine on sperm parameters and evaluate Malondialdehyde (MDA) and 1, 1-Diphenyl-2-picryl-hydrazyl values in BALB/ mice spermatozoa.
Materials and Methods: Forty adult male BALB/c mice were separated into five groups. Group Ι (control) received distilled water; group ΙΙ amitriptyline (4 mg/kg); group ΙΙΙ amitriptyline (4 mg/kg) +vitamin C (10 mg/kg); group ΙV venlafaxine (2 mg/kg); and group V received vitamin C (10 mg/kg) + venlafaxine (2 mg/kg). All drugs were administered by oral gavage for 35 days. After excision of caudal epididymis, it was located in 1 mL Ham's F10 medium at 37oC for 15 min and then analysis of sperm parameters was performed. To examine lipid peroxidation and total antioxidant capacity, the MDA and 1, 1-Diphenyl-2-picryl-hydrazyl were measured, respectively.
Results: The mean sperm parameters in the group treated with amitriptyline were significantly lower than in the other groups. MDA tests showed a significant difference between amitriptyline and control groups (p=0.007).
Conclusion: The results of this study showed that amitriptyline consumption can weaken sperm parameters, which can be attributed to the increased production of ROS and toxicity resulting from amitriptyline consumption. Moreover, venlafaxine improved sperm parameters in mice and the lipid peroxidation in this group did not change compared to the control group.
Mahnaz Heidari, Niknam Lakpour, Mahsa Darbandi, Sara Darbandi, Saeideh Shani, Leila Goharbakhsh, Ghazaleh Cheshmi, Mohammad Mehdi Akhondi, Mohammad Reza Sadeghi,
Volume 16, Issue 7 (7-2018)
Abstract

Background: Sperm processing methods separate motile sperms with good morphology from dead and abnormal forms of sperms, immature germ cells, and non-sperm cells.
Objective: The propose of this study was to compare the efficacy of upstream and swim-up processing techniques to separate sperms with the high quality especially in relation to sperm chromatin integrity.
Materials and Methods: This experimental study used semen samples from 60 normozoospermic men. Specimens were divided into equal aliquots for processing by swim up (group A), and upstream (group B) methods and compare with control by raw semen (group C). Sperm concentration, morphology, motility, DNA fragmentation and chromatin maturation were measured in these three groups.
Results: The results revealed that sperm concentration in the swim up samples was significantly greater than upstream samples (p≤0.04). as addition, motile sperm recovery including the percentage of progressive motility and a total number of motile sperm was better in the swim-up compared to an upstream method and raw semen (p≤0.001). The cell debris and seminal fluid were equally removed by both methods and the percentage of normal forms was also similar in both procedures (p≥0.4). In addition, sperm DNA fragmentation and chromatin maturation were not significantly different between the three groups (p≥0.1).
Conclusion: According to results, apparently the upstream method had no significant efficiency to separate good quality sperms compare to swim up. Therefore, swim up seems to be a simple, inexpensive, reliable and widely available method with an efficient yield to separate motile sperm with good morphology and better chromatin integrity for insemination in the infertility clinics.
Sepideh Sadeghi, Ali Reza Talebi, Abbas Shahedi, Mohammad Reza Moein, Abolghasem Abbasi-Sarcheshmeh,
Volume 17, Issue 4 (4-2019)
Abstract

Background: Tamoxifen (TX) is widely used for the treatment of male factor and idiopathic infertility. It has been shown that TX induces sperm production and so improves male fertility.
Objective: This study evaluated the effects of different doses of TX on the sperm parameters and chromatin quality in mice.
Materials and Methods: In this research, 24 male NMRI mice were divided into three groups including group A: control animal receiving vehicle; group B: the group receiving basal diet and TX 0.4 mg/kg/day; and group C: the group receiving basal diet and TX 0.6 mg/kg/day for 35 days. Thereafter, epididymal spermatozoa were analyzed for standard parameters and nuclear chromatin quality using Aniline Blue (AB) and Toluidine Blue (TB) staining.
Results: The results indicated that although the TX did not affect the sperm count, motility, and viability parameters, it could elevate the percentage of sperm cells with abnormal morphology and abnormal chromatin at both doses. In addition, in comparison with the control mice, a significant elevation was observed in spermatozoa with residual histones (assessed by AB staining) at high doses of TX.
Conclusion: Our experimental data in mice suggested that the use of TX for treating male infertility might increase the rates of spermatozoa with abnormal chromatin in a dose-dependent manner.
Masoomeh Mohammadzadeh, Mohammad Ali Khalili, Vahid Ramezani, Hamed Hamishehkar, Laleh Dehghan Marvast, Esmat Mangoli, Mahya Rajabi, Zhima Akhavan Sales, Prof Ali Reza Talebi,
Volume 18, Issue 9 (9-2020)
Abstract

Background: Previous studies have examined the effect of resveratrol as a potent antioxidant for free radicals in semen. While, the prepared spermatozoa are more affected by ROS factors due to centrifugation and incubation.
Objective: To evaluate the RSV's effects on the prepared sperm parameters and chromatin quality in both normozoospermic and asthenozoospermic cases before and after freezing.
Materials and Methods: The sample of 10 normozoospermic and asthenozoospermic men was prepared through the swim-up method. The groups were then divided into two samples of control and experimental (exposure to 30 µmol/l of RSV) to evaluate and compare the sperm parameters and chromatin quality before and after freezing.
Results: The motility and viability of spermatozoa were seen to be significantly different before and after freezing separately in the control and treatment samples of the groups (p ≤ 0.001 and p = 0.001, respectively). However, the stated difference between the control and treatment samples of normozoospermic and asthenozoospermic patients were not significant (p > 0.05). In addition, the sperm morphology and chromatin quality were not significantly different between the two samples of each group; nonetheless, chromatin quality of the treated sample was better than that of the control before and after freezing.
Conclusion: Despite the protective effects of RSV on the semen samples, RSV cannot affect significantly the prepared sperms parameters and chromatin quality in normozoospermic and asthenozoospermic patients.
Daniyal Ezati, Reyhane Vardiyan, Alireza Talebi, Morteza Anvari, Majid Pourentezari,
Volume 18, Issue 10 (10-2020)
Abstract

Background: Formalin is commonly applied as an antiseptic and tissue fixative. It has reactive molecules that lead to its cytotoxic effects. According to recent studies, formalin causes a change in the testicular and sperm structure and L-carnitine (LC) acts as an antioxidant to counteract its effects.
Objective: This study aimed to investigate the protective effects of LC on the parameters, chromatin condensation and apoptosis of mice sperm exposed to formalin.
Materials and Methods: In this experimental study, 24 balb/c mice (25-40 gr ,10-12 wk) were divided into three groups (n = 8/each): group I without any injections or gavage; group II, received 10 mg/ kg formalin intraperitoneally (I.P); and group III was exposed to formalin and LC, where a dose of 10 mg/kg formalin was injected I.P daily and LC the dose of 100 mg/kg was kept in a solvent solution. After 31 days, the sperm examination was performed as follows: to evaluate chromatin and DNA quality of the sperm, we applied aniline blue (AB), toluidine blue (TB), chromomycin A3 (CMA3), and terminal transferase-mediated deoxy uridine triphosphate biotin end labeling (TUNEL) tests.
Results: Sperm parameters such as count, motility, morphology, and viability displayed a significant decrease in the formalin group. While the data exhibited a considerable augment in sperm parameters in the formalin + LC than the formalin and control groups (p < 0.001), significant differences were detected between groups with respect to TB staining, TUNEL test, CMA3 test and AB staining in the formalin and formalin + LC groups.
Conclusion: LC can reduce the negative effects of formalin on sperm parameters, chromatin stability, and percentage of apoptosis in an animal model.
Ghasemi-Esmailabad S, Talebi Ah, Talebi Ar, Amiri S, Moshrefi M, Pourentezari M,
Volume 19, Issue 5 (5-2021)
Abstract

Background: Morphine is one of the major psychoactive chemicals in opium that can increase the production of free radicals and thus can negatively affect spermatogenesis.
Objective: The purpose of this survey was to demonstrate the effect of morphine consumption on sperm parameters, DNA integrity and apoptosis in men taking morphine.
Materials and Methods: In this case-control study, 30 man abusing morphine (cases) and 30 healthy men (controls) were compared for sperm parameters (count, motility and morphology) and sperm chromatin quality, with aniline blue, toluidine blue and Chromomycin A3 stainings. The participants were matched for age, weight, amount and duration of cigarette smoking.
Results: In men with morphine dependency, sperm progressive and total motility (p = 0.038 and p < 0.001 respectively) showed significant decreasing compared to control group. Regard to morphine abusing, although morphine can decrease the sperm chromatin condensation and increases the rate of sperm apoptosis, but these differences were not statistically significant.
Conclusion: According to our result morphine dependence can reduce male fertility by affecting sperm parameters and also it may affect sperm chromatin/DNA integrity.

Ghasemi-Esmailabad S, Moshrefi M, Pourentezari M, Talebi Ar, Talebi Ah, Niknahad S, Mirjalili A, Eskandarian Borojeni S,
Volume 19, Issue 5 (5-2021)
Abstract

Background: Oxidative stress affects male fertility by defecting spermatozoa. Asthenozoospermia is referred to reduced or complete sperm motility. Reactive oxygen species is one of the major reasons of higher sperm DNA fragmentation. Sperms with high DNA fragmentation are higher in asthenozoospermia, teratozoospermia and oligozoospermia. Also, sperm DNA fragmentation level is higher in men with sperm motility defects. The imbalance between the production of Reactive oxygen species and physiological status leads to damage which is known as oxidative stress. So, antioxidants supplements and Pentoxifylline (PTX) probably improve sperm quality by reducing oxidative damage.
Objective: The present retrospective study aimed to investigate the possible effect of oral co-administration of PTX + folic acid (FA) + vitamin E (Vit E) on sperm parameters, apoptosis, and sperm chromatin in asthenospermic men.
Materials and Methods: Semen samples of 30 infertile asthenospermic men, who referred to Yazd Reproductive Sciences Institute ere collected. Sperm parameters (count, motility, morphology, and viability), apoptosis, and DNA and chromatin quality were evaluated before and three months after consumption of PTX + FA + Vit E. DNA integrity and chromatin quality were assessed by Aniline blue (AB), Toluidine blue (TB), and Chromomycin A3 (CMA3) staining. Also, apoptosis was assessed by Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). Sperm morphology was assessed by Papanicolaou staining.
Results: Our results showed that after co-administration of PTX and antioxidants, sperm motility and morphology increased significantly (p < 0.0001). Semen volume and sperm count were also increased, but not significantly (p < 0.05). Following the intervention, AB, TB, and CMA3 staining showed that the number of sperms with good chromatin quality and DNA integrity was increased, although it was not significant (p < 0.05). The mean ± S.D. of chromatin condensation, which were measured by AB, TB, and CMA3 before taking the drugs were 37.48 ± 8.84, 47.10 ± 15.43. and 37.72 ± 9.52, respectively and after taking the drug were 34.51 ± 7.27, 44.51 ± 14.99, and 35.68 ± 9.37, respectively. Also, the mean ± S.D. of sperm apoptosis by TUNEL test before and after taking the drug was 14.89 ±3.49, and 14.06 ± 3.74, respectively.
Conclusion: Based on these data, the cocktail of PTX + FA + Vit. E significantly increased the normal motility and morphology of sperm in asthenospermic men. But still more studyies with larger sample size is needed.

Amirhossein Danafar, Arezoo Khoradmehr, Mahya Hosseini Bondarabadi, Fahime Mazaheri, Amin Tamadon, Soheila Pourmasoumi, Lida Gholizadeh, Mojgan Moshrefi, Iman Halvaei, Akram Hosseini, Jalal Golzadeh, Tahereh Rahiminia, Morteza Anvari,
Volume 19, Issue 12 (12-2021)
Abstract

Background: Titanium dioxide nanoparticles (TiO2NPs) are widely used in many compounds. Recent evidence has displayed some cytotoxic effects of TiO2NPs on male reproduction.
Objective: The effects of TiO2NP administration on sperm parameters and chromatin and seminiferous histopathology of male mice were investigated.
Materials and Methods: In this experimental study, 32 NMRI male mice (35 ± 3 gr, 8-12-week-old) were divided into four groups (n = 8/each): treated groups were fed orally with 2.5 (group I), 5 (group II) and 10 (group III) mg/kg/day TiO2NPs for 40 days and the control group received phosphate buffered saline. Sperm parameters, DNA integrity and chromatin quality were assessed using chromomycin A3, aniline blue, toluidine blue staining and TUNEL. Hematoxylin eosin staining was performed to measure spermatogenic cells and the total diameter of seminiferous tubules. Also, sex hormone and malondyaldehyde levels were measured.
Results: Abnormal sperm tails rose in group III (28.87 ± 4.91) in comparison with the control group (12.75 ± 3.95). However, chromomycin A3 staining and TUNEL showed higher levels in group III in comparison with the control group, whereas aniline blue and toluidine blue staining showed no differences. A significantly lower spermatogenesis index and lumen parameters were observed in group III. Leydig cell numbers, cellular diameters and the area of the seminiferous tubules were lower in the treated groups. The testosterone level was also lower in these groups and the percentage of malondyaldehyde in the seminal fluid was higher.
Conclusion: Exact mechanisms of TiO2NPs are not clear; however, cytotoxic and genotoxic effects of TiO2NPs may relate to oxidative stress. Given their widespread use, TiO2NPs should be a public health focus of attention.

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.


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