The effect of lipopolysaccharide from uropathogenic Escherichia coli on the immune system, testis tissue, and spermatozoa of BALB/c mice / 대한생식의학회지

Objective@#Uropathogenic Escherichia coli is known to cause urinary tract infections, and the endotoxin (lipopolysaccharide [LPS]) of this bacterium may cause deficiencies of sperm quality and morphology. In the present study, the effects of LPS on mouse sperm were studied, and the levels of interleukin (IL)-17A and possible changes in testis tissue were evaluated. @*Methods@#LPS of uropathogenic E. coli was extracted using the methanol-chloroform method, followed confirmation using sodium dodecyl sulfate-polyacrylamide electrophoresis. Purified LPS (100 µg/kg) or phosphate-buffered saline was injected intraperitoneally into BALB/c mice for 7 days consecutively in the test and control groups, Mice were sacrificed on days 3, 7, and 42 after the first injection. Blood was tested for levels of IL-17A using the enzyme-linked immunosorbent assay method. Testis tissue and sperm were collected from each mouse and were studied according to standard protocols. @*Results@#The mean sperm count and motility significantly decreased (p=0.03) at 3, 7, and 42 days after the injections. The level of IL-17A in the test groups increased, but not significantly (p=0.8, p=0.11, and p=0.15, respectively). Microscopic studies showed no obvious changes in the morphology of the testis tissue; however, significant changes were observed in the cellular parenchyma on day 42. @*Conclusion@#LPS can stimulate the immune system to produce pro-inflammatory cytokines, resulting in an immune response in the testis and ultimately leading to deficiency in sperm parameters and testis tissue damage. In addition, the presence of LPS could significantly impair sperm parameters, as shown by the finding of decreased motility.

The effect of lipopolysaccharide from uropathogenic Escherichia coli on the immune system, testis tissue, and spermatozoa of BALB/c mice / 대한생식의학회지

Objective@#Uropathogenic Escherichia coli is known to cause urinary tract infections, and the endotoxin (lipopolysaccharide [LPS]) of this bacterium may cause deficiencies of sperm quality and morphology. In the present study, the effects of LPS on mouse sperm were studied, and the levels of interleukin (IL)-17A and possible changes in testis tissue were evaluated. @*Methods@#LPS of uropathogenic E. coli was extracted using the methanol-chloroform method, followed confirmation using sodium dodecyl sulfate-polyacrylamide electrophoresis. Purified LPS (100 µg/kg) or phosphate-buffered saline was injected intraperitoneally into BALB/c mice for 7 days consecutively in the test and control groups, Mice were sacrificed on days 3, 7, and 42 after the first injection. Blood was tested for levels of IL-17A using the enzyme-linked immunosorbent assay method. Testis tissue and sperm were collected from each mouse and were studied according to standard protocols. @*Results@#The mean sperm count and motility significantly decreased (p=0.03) at 3, 7, and 42 days after the injections. The level of IL-17A in the test groups increased, but not significantly (p=0.8, p=0.11, and p=0.15, respectively). Microscopic studies showed no obvious changes in the morphology of the testis tissue; however, significant changes were observed in the cellular parenchyma on day 42. @*Conclusion@#LPS can stimulate the immune system to produce pro-inflammatory cytokines, resulting in an immune response in the testis and ultimately leading to deficiency in sperm parameters and testis tissue damage. In addition, the presence of LPS could significantly impair sperm parameters, as shown by the finding of decreased motility.

Protective effects of curcumin on chromatin quality, sperm parameters, and apoptosis following testicular torsion-detorsion in mice / 대한생식의학회지

Objective@#The chief outcome of testicular torsion in clinical and experimental contexts is testicular ischemia. Curcumin, a compound with anti-inflammatory and antioxidant properties, has fascinated researchers and clinicians for its promise in the treatment of fertility diseases. @*Methods@#Thirty-five fully grown male mice were randomly classified into five groups: control, sham, testicular torsion, treatment group 1 (testicular torsion+short-term curcumin), and treatment group 2 (testicular torsion+long-term curcumin). Thirty-five days later, spermatozoa from the right cauda epididymis were analyzed with regard to count and motility. Toluidine blue (TB), aniline blue (AB), and chromomycin A3 (CMA3) staining assays were used to evaluate the sperm chromatin integrity. In addition, the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) test was used to assess apoptosis.Result: Treatment group 1 exhibited a remarkably elevated sperm count compared to the testicular torsion group. Additionally, notably lower sperm motility was found in the testicular torsion group compared to the control, treatment 1, and treatment 2 groups. Staining (CMA3, AB, and TB) and the TUNEL test indicated significantly greater testicular torsion in the torsion group compared to the control group (p<0.05). The data also revealed notably lower results of all sperm chromatin assays and lower apoptosis in both treatment groups relative to the testicular torsion group (p<0.05). Significantly elevated (p<0.05) AB and TB results were noted in treatment group 1 compared to treatment group 2. @*Conclusion@#Curcumin can compensate for the harmful effects of testicular ischemia and improve sperm chromatin quality in mice.

The effects of sesame oil and different doses of estradiol on testicular structure, sperm parameters, and chromatin integrity in old mice / 대한생식의학회지

Objective@#Studies of the effects of estrogens on the male reproductive system have emphasized the role of these hormones in male fertility. Sesame oil has many phytoestrogenic compounds and may improve male fertility. This study investigated the effects of sesame oil and different concentrations of estrogen on sperm parameters and DNA integrity in male mice. @*Methods@#Twenty old NMRI (The Naval Medical Research Institute) male mice (40 weeks; weight, 30–35 g) were treated with sesame oil or different concentrations of estrogen (estradiol, 1 and 10 μL/kg/ day) or received no treatment (controls). After 35 days, sperm parameters and DNA integrity were assessed and analyzed. @*Results@#Sperm count, progressive motility, and morphology were decreased in the group that received 10 μL/kg of estradiol. A remarkably lower percentage of DNA fragmentation and protamine deficiency were detected in the group that received 1 μL/kg of estradiol. In the groups that received sesame oil and 1 μL/kg of estradiol, the numbers of spermatogonia and Leydig cells were higher than in controls. The combination of sesame oil and 1 μL/kg of estradiol led to improved sperm parameters and chromatin and testicular structure. @*Conclusion@#Based on this study, consumption of sesame oil and a low concentration of estradiol may improve testicular function in older mice.

The effect of vitamin C on the gene expression profile of sperm protamines in the male partners of couples with recurrent pregnancy loss: A randomized clinical trial / 대한생식의학회지

Objective@#Since sperm abnormalities are known to be a major reason for recurrent pregnancy loss (RPL), any defects in DNA structure and chromatin condensation can place embryos at risk in the early stage of development and implantation. As antioxidants such as vitamin C may play a protective role against the destruction of protamine genes in sperm chromatin, this study was conducted to evaluate the effects of vitamin C on chromatin and the expression of protamine genes in the male partners of couples with RPL. @*Methods@#Twenty male partners of couples with RPL were selected as the intervention group and received vitamin C supplementation (250 mg daily for 3 months). Healthy fertile men (n=20) were included as controls. Sperm chromatin, DNA integrity, and the expression levels of protamine genes were evaluated before and after treatment. @*Results@#Significant differences were found in sperm morphology, protamine deficiency, and apoptosis between the two groups and before and after vitamin C administration. A significant change was found in mRNA levels of PRM1, PRM2, and the PRM1/PRM2 ratio after treatment. @*Conclusion@#Daily oral administration of vitamin C may improve human sperm parameters and DNA integrity by increasing protamine gene expression levels in the male partners of couples with RPL. The beneficial effects of vitamin C supplementation as an antioxidant for the male partners of couples with RPL could lead to improved pregnancy outcomes in these cases.

Effects of acrylamide in the presence of vitamin E on sperm parameters, chromatin quality, and testosterone levels in mice / 대한생식의학회지

Objective@#The present study investigated sperm chromatin quality and testosterone levels in acrylamide-treated mice and the possible protective effects of vitamin E on the fertility potential of spermatozoa. @*Methods@#Thirty-two adult male mice were divided equally into four groups. Group 1 was the control, group 2 received acrylamide (10 mg/kg, water solution), group 3 received vitamin E (100 mg/kg, intraperitoneal), and group 4 received both acrylamide and vitamin E. After 35 days, spermatozoa from the right cauda epididymis were analyzed in terms of count, motility, morphology, and viability. Sperm DNA integrity and chromatin condensation were assessed by acridine orange (AO), aniline blue (AB), toluidine blue (TB), and chromomycin A3 (CMA3) staining. @*Results@#In acrylamide-treated mice, significantly lower sperm concentration, viability, motility, and testosterone levels were found in comparison with the control and acrylamide+vitamin E groups (p<0.05). In the vitamin E group, significantly more favorable sperm parameters and testosterone levels were found than in the other groups (p<0.05). There were also significantly more spermatozoa with less condensed chromatin in the acrylamide-treated mice than in the other groups. Moreover, significantly more spermatozoa with mature nuclei (assessed by AB, CMA3, AO, and TB staining) were present in the vitamin E group than in the control and acrylamide+vitamin E groups. @*Conclusion@#This study revealed the deleterious effects of acrylamide on sperm parameters and sperm chromatin quality. Vitamin E can not only compensate for the toxic effects of acrylamide, but also improve sperm chromatin quality in mice.

Sperm chromatin and DNA integrity, methyltransferase mRNA levels, and global DNA methylation in oligoasthenoteratozoospermia / 대한생식의학회지

OBJECTIVE: To investigate sperm chromatin/DNA integrity, global DNA methylation, and DNMT mRNA transcription in men with oligoasthenoteratozoospermia (OAT) compared with normozoospermic men. METHODS: Semen samples from 32 OAT patients who comprised the case group and 32 normozoospermic men who comprised the control group were isolated and purified using a standard gradient isolation procedure according to World Health Organization criteria. DNMT1, DNMT3A, and DNMT3B transcripts were then compared between groups using real-time quantitative reverse-transcription polymerase chain reaction. Global DNA methylation in sperm was determined by an enzyme-linked immunosorbent assay. Protamine deficiency and the proportion of apoptotic spermatozoa were evaluated using chromomycin A3 (CMA3), aniline blue (AB), and toluidine blue (TB) staining, as well as the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. The p-values < 0.05 were considered to indicate statistical significance. RESULTS: Significantly higher proportions of AB+, TB+, CMA3+, and TUNEL+ spermatozoa, as well as DNMT3A and DNMT3B transcription, were found in the OAT group. Positive correlations were detected between sperm parameters, DNA/chromatin damage, and DNMT3A and DNMT3B transcripts. Global DNA methylation was significantly higher in the OAT patients and had a significant correlation with abnormal results of all sperm chromatin integrity tests, but was not associated with DNMT1, DNMT3A, or DNMT3B expression. CONCLUSION: Oligoasthenoteratozoospermic men showed abnormal sperm parameters, abnormal chromatin/DNA integrity, and a higher global DNA methylation rate, as well as overexpression of DNMT mRNA.

Evaluation of sperm protamine deficiency and apoptosis in infertile men with idiopathic teratozoospermia / 대한생식의학회지

OBJECTIVE: Sperm morphology plays an important role in infertility, especially in cases of defects in the heads of spermatozoa. Tapered-head or elongated-head spermatozoa are examples of morphological abnormalities. The aim of this study was to compare the semen parameters, levels of protamine deficiency, and frequency of apoptosis between patients with normozoospermia and those with teratozoospermia with tapered-head spermatozoa. METHODS: Fifty-two semen samples (27 patients with tapered-head sperm and 25 fertile men) were collected and semen analysis was performed according to the World Health Organization criteria for each sample. Protamine deficiency and the percentage of apoptotic spermatozoa were evaluated using chromomycin A3 (CMA3) staining and terminal deoxynucleotidyl transferase dUTP nick-end labelling (TUNEL) assays, respectively. RESULTS: Sperm concentration, motility, and normal morphology in the tapered-head spermatozoa (cases) were significantly lower than in the normozoospermic samples (controls). CMA3-reactive spermatozoa (CMA3+) in the case group were more common than in the controls. Apoptotic spermatozoa (TUNEL-positive) were significantly more common in the cases than in the controls. CONCLUSION: This analysis showed that tapered-head spermatozoa contained abnormal chromatin packaging and exhibited a high rate of apoptosis, which can be considered to be an important reason for the impaired fertility potential in teratozoospermic patients with tapered-head spermatozoa.

Administration of high dose of methamphetamine has detrimental effects on sperm parameters and DNA integrity in mice

Background: methamphetamine [MA] was shown to have harmful effects on male reproductive system

Objective: to investigate probable effects of daily administration of MA on sperm parameters and chromatin/DNA integrity in mouse

Material and Methods: thirty-five NMRI male mice were divided into five groups including low, medium, and high dosage groups which were injected intraperitoneally with 4, 8 and 15 mg/kg/day for 35 days, respectively. Normal saline 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 placed in Ham's F10 medium for sperm retrieval. To evaluate sperm chromatin abnormalities, the aniline blue, toluidine blue and chromomycine A3 were used. For sperm DNA integrity and apoptosis, the acridine orange, sperm chromatin dispersion, and TUNEL assay were applied. For sperm morphology, Papanicolaou staining was done

Results: normal morphology and progressive motility of spermatozoa decreased in medium 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, and chromomycine A3 positive spermatozoa in high dosage group. In a similar manner, there was an increase in rates of acridine orange, TUNEL and sperm chromatin dispersion positive sperm cells in high dosage group with respect to others

Conclusion: MA abuse in a dose-dependent manner could have detrimental effects on male reproductive indices including sperm parameters and sperm chromatin/DNA integrity in mice

Relationship between sperm protamine deficiency and apoptosis in couples with unexplained repeated spontaneous abortions

Background: Etiology of more than half of Recurrent Spontaneous Abortion. The etiology of more than 50 percent of Recurrent Spontaneous Abortions [RSA] cases has been remained unexplained. It is supposed that RSA may have "paternal effect" due to supply 50% of embryonic genomic content by male gamete

Objective: The aim of present study was to evaluate the role of sperm apoptosis and protamine deficiency at same time in RSA cases

Materials and Methods: Forty fertile [control] and 40 unfertile men with RSA [case] were enrolled in this case-control study. Semen analysis was performed in accordance with WHO criteria and sperm apoptosis and protamine deficiency were evaluated by cell apoptosis detection kit and chromomycin A3, respectively

Results: Results showed significant different between normal morphology and total motility in two groups. Case group had higher percentage of spermatozoa with protamine deficiency and apoptosis compared to controls significantly

Conclusion: Our results showed that in cases of RSA, in addition to abnormal sperm parameters, we have a high percentage of spermatozoa with protamine deficiency and apoptosis and these two anomalies may consider as important causes of idiopathic recurrent abortions. It should be advised that sperm chromatin and DNA examinations are useful tools in the process of RSA treatments

Etiologies of sperm oxidative stress

Sperm is particularly susceptible to reactive oxygen species [ROS] during critical phases of spermiogenesis. However, the level of seminal ROS is restricted by seminal antioxidants which have beneficial effects on sperm parameters and developmental potentials. Mitochondria and sperm plasma membrane are two major sites of ROS generation in sperm cells. Besides, leukocytes including polymer phonuclear [PMN] leukocytes and macrophages produce broad category of molecules including oxygen free radicals, non-radical species and reactive nitrogen species. Physiological role of ROS increase the intracellular cAMP which then activate protein kinase in male reproductive system. This indicates that spermatozoa need small amounts of ROS to acquire the ability of nuclear maturation regulation and condensation to fertilize the oocyte. There is a long list of intrinsic and extrinsic factors which can induce oxidative stress to interact with lipids, proteins and DNA molecules. As a result, we have lipid peroxidation, DNA fragmentation, axonemal damage, denaturation of the enzymes, over generation of superoxide in the mitochondria, lower antioxidant activity and finally abnormal spermatogenesis. If oxidative stress is considered as one of the main cause of DNA damage in the germ cells, then there should be good reason for antioxidant therapy in these conditions

effect of heracleum persicum [Golpar] oil and alcoholic extracts on sperm parameters and chromatin quality in mice

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

Mitochondrial genetic variation in Iranian infertile men with varicocele

Background: Several recent studies have shown that mitochondrial DNA mutations lead to major disabilities and premature death in carriers. More than 150 mutations in human mitochondrial DNA [mtDNA] genes have been associated with a wide spectrum of disorders. Varicocele, one of the causes of infertility in men wherein abnormal inflexion and distension of veins of the pampiniform plexus is observed within spermatic cord, can increase reactive oxygen species [ROS] production in semen and cause oxidative stress and sperm dysfunction in patients. Given that mitochondria are the source of ROS production in cells, the aim of this study was to scan nine mitochondrial genes [MT-COX2, MT-tRNA[Lys], MT-ATP8, MT-ATP6, MT-COX3, MT-tRNA[Gly], MT-ND3, MT-tRNA[Arg]and MT-ND4L] for mutations in infertile patients with varicocele

Materials and Methods: In this cross-sectional study, polymerase chain reaction-single strand conformation polymorphism [PCR-SSCP] and DNA sequencing were used to detect and identify point mutations respectively in 9 mitochondrial genes in 72 infertile men with varicocele and 159 fertile men. In brief, the samples showing altered electrophoretic patterns of DNA in the SSCP gel were sent for DNA sequencing to identify the exact nucleotide variation

Results: Ten type nucleotide variants were detected exclusively in mitochondrial DNA of infertile men. These include six novel nucleotide changes and four variants previously reported for other disorders

Conclusion: Mutations in mitochondrial genes may affect respiratory complexes in combination with environmental risk factors. Therefore these nucleotide variants probably lead to impaired ATP synthesis and mitochondrial function ultimately interfering with sperm motility and infertility

Large nuclear vacuoles in spermatozoa negatively affect pregnancy rate in IVF cycles

Recently, motile sperm organelle morphology examination [MSOME] criteria as a new real time tool for evaluation of spermatozoa in intracytoplasmic sperm injection [ICSI] cycles has been considered. The aim was to investigate the predictive value of MSOME in in vitro fertilization [IVF] in comparison to ICSI cycles and evaluation of the association between MSOME parameters and traditional sperm parameters in both groups. This is a cross sectional prospective analysis of MSOME parameters in IVF [n=31] and ICSI cycles [n=35]. MSOME parameters were also evaluated as the presence of vacuole [none, small, medium, large or mix]; head size [normal, small or large]; cytoplasmic droplet; head shape and acrosome normality. In sub-analysis, MSOME parameters were compared between two groups with successful or failed clinical pregnancy in each group. In IVF group, the rate of large nuclear vacuole showed significant increase in failed as compared to successful pregnancies [13.81 +/- 9.7vs7.38 +/- 4.4, respectively, p=0.045] while MSOME parameters were the same between successful and failed pregnancies in ICSI group. Moreover, a negative correlation was noticed between LNV and sperm shape normalcy. In ICSI group, a negative correlation was established between cytoplasmic droplet and sperm shape normalcy. In addition, there was a positive correlation between sperm shape normalcy and non-vacuolated spermatozoa. The high rate of large nuclear vacuoles in sperm used in IVF cycles with failed pregnancies confirms that MSOME, is a helpful tool for fine sperm morphology assessment, and its application may enhance the assisted reproduction technology success rates

Sperm parameters, protamine deficiency, and apoptosis in total globozoospermia

Globozoospermia is a severe form of teratozoospermia [incidence < 0.1%] in infertile men that is characterized by round headed sperm and acrosomeless in semen. To compare the semen parameters, protamine deficiency, and apoptosis in ejaculated spermatozoa between globozoospermic and normozoospermic men. Thirty six semen samples were divided into two groups including 15 infertile men with total globozoospermic [> 90% round-headed sperm] and 21 healthy donors with normal spermograms as controls. Semen analysis was performed according to World Health Organization criteria [2010]. Sperm protamine deficiency was assessed using Chromomycin A3 [CMA3] staining and the rate of apoptotic spermatozoa was evaluated with TUNEL assay. Sperm concentration, motility, and normal morphology in globozoospermic men were significantly decreased compared with controls [p<0.05]. The rate of CMA3-reacted spermatozoa [CMA3+] in globozoospermic men was higher than controls [65.93 +/- 11.77 vs. 21.24 +/- 7.37, respectively, p<0.0001]. The rate of apoptotic spermatozoa [TUNEL positive] were significantly increased in globozoospermic cases with respect to the controls [17.60 +/- 10.72 and 5.95 +/- 3.02, respectively, p<0.0001]. There was no significant correlation between sperm protamine deficiency and apoptosis in globozoospermic men. Globozoospermic samples contain a higher proportion of spermatozoa with abnormal chromatin packaging and DNA fragmentation than normozoospermic samples. Therefore, in addition to absence of acrosome in the spermatozoa of globozoospermic patients, the high percentage of spermatozoa with immature chromatin and apoptotic marker may be considered as the other etiologies of infertility in these patients

Early and late effects of Ibuprofen on mouse sperm parameters, chromatin condensation, and DNA integrity in mice

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

Effects of silver nano-particles on sperm parameters, number of Leydig cells and sex hormones in rats

Nano-particles are extensively employed in most industries. Several studies have been started to explore the probable detrimental effects of nano-particles on human reproduction. However, there is insufficient and controversially evident of effects of silver nano-particles on sperm parameters and other reproductive indices. Investigation of the effects of silver nano-particles on sperm parameters, sex hormones and Leydig cells in rat as an experimental model. In this experimental study, 75 male prepubertal Wistar rats were categorized in five groups including control group and 4 experimental groups [n=15 in each group]. The rats in the experimental groups were fed silver nano-particles [60 nm in dimension] with concentrations of 25, 50, 100, and 200 mg/kg/day. After 45 days [about one duration of spermatogenesis in rat], samples of blood were taken from the rats for testosterone, leuteinizing hormone [LH], and follicle stimulating hormone [FSH] assessments. Afterwards, the epididymis and the testis of each rat were dissected for analyzing sperm parameters and Leydig cells. The results demonstrated a statistically significant reduction in number of Leydig cells in experimental groups compared to control one. In addition, the data showed a reduction in testosterone and a rise in LH level which was more obvious in high doses [p<0.05]; however, FSH level showed a reduction but it was not statistically significant. A significant decrease was also found in sperm motility and normal sperm morphology in the experimental groups compared to the control one. Our results demonstrated that silver nano-particles, in addition to interruption in functions of sex hormones, can diminish the number of Leydig cells and sperm parameter indices. It should be noted that the effects of nano-particles on reproductive indices are dose-dependent

Mutation analysis of TNP1 gene in infertile men with varicocele

Varicocele is associated with the failure of ipsilateral testicular growth and development, and the symptoms of pain and reduced fertility. The highly condensed structure of the sperm nuclear chromatin is provided by proper expression of Transition Nuclear Protein [TNP] genes, so any dysregulational expression of these genes results in abnormal spermatogenesis and infertility. The aim of present study was to assess the association between TNP1 mutations and varicocele in Iranian infertile men. Analysis of association between TNP1 gene mutation and varicocele phenotype was performed using PCR and Single-Stranded Conformational Polymorphism technique and DNA sequencing in 82 varicocele infertile men and 80 control subjects. Sequence analysis was identified one variant in this gene that found in 15 infertile men and was absent in control group. This variant was a single nucleotide polymorphism that were identified in the intron region of this gene at position g.IVS1+75T>C. The effect of this nucleotide substitution in intronic region of the TNP1 gene and their role on expression remains to be determined

Saccharin consumption increases sperm DNA fragmentation and apoptosis in mice

Saccharin is an artificial non-caloric sweetener that used to sweeten products such as drinks, candies, medicines, and toothpaste, but our bodies cannot metabolize it. Sodium saccharin is considered as an important factor in tumor promotion in male rats but not in humans. The objective of this study was to investigate the effect of saccharin consumption on sperm parameters and apoptosis in adult mice. Totally 14 adult male mice were divided into 2 groups. Group 1 served as control fed on basal diet and group 2 or experimental animals received distilled water containing saccharin [0.2% w/v] for 35 days. After that, the left cauda epididymis of each mouse was cut and placed in Ham's F10. Swimmed-out spermatozoa were used to analyze count, motility, morphology [Pap-staining] and viability [eosin-Y staining]. Sperm DNA integrity, as an indicator of apoptosis, was assessed by SCD [sperm chromatin dispersion] and terminal deoxynucleotidyl transferase [TUNEL] assay. Following saccharin consumption, we had a reduction in sperm motility with respect to control animals [p=0.000]. In addition, the sperm count diminished [17.70 +/- 1.11 in controls vs. 12.80 +/- 2.79 in case group, p=0.003] and the rate of sperm normal morphology decreased from 77.00 +/- 6.40 in control animals into 63.85 +/- 6.81 in saccharin-treated mice [p=0.001]. Also, we saw a statistically significant increase in rates of sperm DNA damage and apoptosis in experimental group when compared to control one [p=0.001, p=0.002 respectively]. Saccharin consumption may have negative effects on sperm parameters, and increases the rate of sperm DNA fragmentation and apoptosis in mice

Sperm nuclear DNA in ejaculates of fertile and infertile men

Our aim was to compare the nuclear DNA integrity of the spermatozoa from infertile men with abnormal semen parameters with that from normospermic fertile men, and to evaluate the relationship between the sperm DNA integrity and semen parameters. Thirty ejaculate samples with abnormal semen analysis and 30 ejaculates with normal semen parameters were randomly collected from infertile and fertile men, respectively. The acridine orange test was used to assess the integrity of sperm DNA. The number of ejaculates with a DNA Fragmentation Index [DFI] above 30% were 16 [53.3%] and 22 [73.3%] in fertile and infertile subjects, respectively [P = .10]. The mean DFI was 37.7 +/- 19.6% and 46.1 +/- 16.7% in the fertile and infertile subjects, respectively [P = .24]. The DFI of the fertile men with normal sperm morphology [12 patients] ranged from 1% to 80%. In the samples with oligoasthenospermia, mean DFI was 52.7 +/- 17.9%. There were 2 samples with severe teratospermia [normal morphology less than 4%] and DFIs of 70% and 87%. There were no significant correlations between the DNA integrity and the 3 parameters of semen quality in our 60 subjects. Our results failed to show any significant difference in the DNA integrity of the spermatozoa between infertile and fertile men. Also, no correlation was noticed between the DNA abnormality and the semen parameters in the studied samples