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

Survival assessment of mouse preimplantation embryos after exposure to cell phone radiation

Background: Using cellular phone has rapidly increased all over the world. Also, the concern on the possible health hazards of electromagnetic fields [EMF] induced from cell phones to reproduction has been growing in many countries. The aim of this study was to assess the consequences and effects of exposure to the cell phone radiation on the quality and survival rates of preimplantation embryos in mice

Methods: A total of 40 mice [20 females and 20 males], 6 weeks old and sexually mature BALB/c, were used for control and experimental groups. The ovary burses were removed and the zygotes were dissected in the morning after mating. Next, 2-cell embryos were divided into two groups of control [n=150] and experimental [n=150]. EMF [900-1800 MHz] was used for four days in experimental group for 30 min/day in culture at 37°C in a CO[2]incubator. The quality of embryos was recorded daily and the fluorescent staining was used for identification of viable blastocysts. All data were compared by Student's t-test and Mann-Whitney test [p<0.05]

Results: The rate of embryo survival to the blastocysts stage was similar in both groups. However, the percentage of dead embryos at the 2-cell stage was significantly higher in EMF-exposed group compared with controls [p=0.03]. Also, the loss of cell viability significantly increased in experimental blastocysts [p=0.002]

Conclusion: The normal embryonic development up to the blastocyst stage indicates that EMF-exposure commonly did not have adverse effect on embryo development in mice. But, it caused loss of blastocysts cell viability

Evaluating the effect of insulin sensitizers metformin and pioglitazone alone and in combination on women with polycystic ovary syndrome: an RCT

Background: Insulin resistance and hyperinsulinemia may play a role in pathogenesis of PCOS. One of the common therapeutic methods is using insulinsensitizing drugs such as metformin and thiazolidinediones

Objective: The purpose was to determine the effect of metformin and pioglitazone on clinical, hormonal and metabolic parameters in women with PCOS

Materials and Methods: Eighty four women randomly received one of the following for 3 months: metformin [n=28] [500 mg three times a day], pioglitazone [30 mg daily] [n=28] and combination of both metformin and pioglitazone [n=28] [30 mg/day pioglitazone plus 500 mg metformin three times a day]. Hormonal profile, fasting serum insulin, body weight, body mass index, menstrual status and waist to hip ratio were evaluated before and after treatment

Results: Metformin and pioglitazone and combination therapy induced favorable changes in fasting serum insulin, HOMA-IR index, QUICKI, fasting glucose to insulin ratio in women with PCOS. Body weight, BMI, and waist to hip ratio increased significantly after treatment with pioglitazone but the data were similar after administration of metformin or combination therapy. Total testosterone level decreased significantly only after treatment with metformin. After 3 months in patients who received pioglitazone or combination therapy, menstrual cycles became regular in 71.4% and 73.9% respectively. While menstrual improvement happened only in 36.4% of the patients treated with metformin

Conclusion: These findings suggest that insulin-sensitizing drugs induce beneficial effect in insulin resistance and menstrual cyclicity but only metformin ameliorated hyperandrogenemia in women with PCOS. Treatment with combination of metformin and pioglitazone did not show more benefit than monotherapy with each drug alone

role of ventilation mode using a laryngeal mask airway during gynecological laparoscopy on lung mechanics, hemodynamic response and blood gas analysis

Background: There are two methods for ventilation in gynecological laparoscopy: volume-controlled ventilation [VCV] and pressure-controlled ventilation [PCV]

Objective: To compare the lung mechanics, hemodynamic response and arterial blood gas analysis and gas exchange of two modes of VCV and PCV using laryngeal mask airway [LMA] at different time intervals

Materials and Methods: Sixty infertile women referred for diagnostic laparoscopy, based on ventilation mode, were randomly divided into two groups of VCV [tidal volume: 10 ml/kg] and PCV. In the PCV group, ventilation was initiated with a peak airway pressure [tidal volume: 10 ml/kg, upper limit: 35 cm H[2]O]. In both groups, the arterial blood samples were taken in several time intervals [5, 10 and 15 min after LMA insertion] for blood gas evaluation. Also the lung mechanics parameters were continuously monitored and were recorded at different time intervals

Results: There were no significant differences for patient's age, weight, height and BMI in two groups. The peak and plateau airway pressure were significantly higher in VCV group compared to PCV group 5 and 10 min after insertion of LMA. PaO[2] was significantly higher after 10 and 15 min in VCV group compared to PCV group [p=0.005 and p=0.03, respectively]. PaCO[2] showed significant increase after 5 min in PCV group, but the differences were not significant after 10 and 15 min in two groups. The end tidal CO2 showed significant increase after 10 and 15 min in VCV compared to PCV group

Conclusion: Both VCV and PCV seem to be suitable for gynecological laparoscopy. However, airway pressures are significantly lower in PCV compared to VCV

Effect of prenatal methamphetamine administration during gestational days on mice

Methamphetamine [MA] is one of most common illicit drugs which were reported that nearly half of MA consumers are women. MA can cross through placenta and affects pregnancy and fetus development. Our aim was to evaluate effects of injected MA on crown-rump length, head and placental circumference, body weight, histological changes and apoptosis in fetus. Twenty-four NMRI pregnant mice were randomly divided into five groups. First, second and third groups were injected intraperitoneally 10 mg/kg/day MA during gestational days [GD]: GD1-7, GD8-14, and GD1-14, respectively. Forth group, as sham, was injected saline from GD1-14, and finally control which was received neither MA nor saline. On GD15 cervical dislocated pregnant mice, fetus and placenta were weighed and fetus crown-rump length was measured. Hematoxylin and Eosin staining and TUNEL assay were applied to assess histological changes and apoptosis respectively. Fetus body weight and crown-rump length showed significant decrease in third compared to first and second groups [p

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

The effect of embryo catheter loading technique on the live birth rate / 대한생식의학회지

OBJECTIVE: Embryo loading (EL) is a major step in embryo transfer (ET) and affect on the success of in vitro fertilization (IVF). This study aimed to compare the effect of two different EL techniques on the rates of pregnancy and delivery in IVF/ET cycles. METHODS: 207 fresh ET and 194 Frozen-thawed ET (FET) cycles were included in this retrospective study. Two groups (A and B) were defined based on the EL technique used. In group A, the entire catheter was flushed with Ham's F-10 medium. The embryos were then drawn into the catheter using one air bracket. In group B, 70 microL of air was aspirated into the syringe and the catheter was flushed using Ham's F10 medium. The medium, air, embryos, air, and finally another layer of medium were then sequentially drawn into the catheter. The main outcome measures were the pregnancy and delivery rates. RESULTS: The groups did not differ with respect to the etiology of infertility, the source of spermatozoa, the quality of the embryos, the type of EL catheter, and the ease of transfer. The pregnancy rate was similar between two groups. In fresh ET cycles, a higher delivery rate was observed in group B than it group A (78.1% vs. 60%, p=0.1). In FET cycles, the rate of delivery was significantly higher in group B than in group A to a nonsignificant extent (88.9% vs. 58.8%, p=0.06). CONCLUSION: EL techniques did not have a significant impact on the delivery rate in either fresh or FET cycles.

[ investigation of bacterial infection of seminal fluid in men with infertility with unknown etiology]

One of the most important factors that cause infertility in men is urogenital infections. In most cases, these infections cause impairment in parameters, [such as sperm count, motility, viability, and morphology] and sperm function, and also induction of inflammation in epididymis and prostate gland is occurred. Therefore, the identification of bacterial species causing infection and administration of appropriate antibiotic therapy can result in improvement of sperm parameters and consequently fertility. This study was performed with the purpose of determining the frequency of bacterial infection of seminal fluid in infertile men with unknown etiology. In this descriptive-analytical study, seminal fluid of 65 infertile men with unknown cause referred to Yazd Institute for Reproductive Sciences, were studied. At first, seminal fluid analysis was performed according to WHO guideline. Then, blood agar and EMB [eosin methylene blue] culture media were used for detection of bacterial infection. In semen. Supplementary microbial tests were used to detect bacterial species. Pearson correlation test was used to determine the correlation between sperm parameters, between the two groups with and without infection. Significance level was considered less than 0.05. In this study, 40 specimens had bacterial infection. Seven different bacterial species were detected in these specimens, of which the staphylococcus aureus had the highest incidence, 16.9%, and pseudomonas and enterococcus had the lowest incidence, 1.5%. According to the high incidence of seminal infection in infertile men, use of microbial tests for infertile men with unknown etiology and attempting to treat urogenital infection are suggested

Effects of ascorbic acid on sperm motility, viability, acrosome reaction and DNA integrity in teratozoospermic samples

Oxidative stress in teratozoospermic semen samples caused poor assisted reproductive techniques [ART] outcomes. Among antioxidants, ascorbic acid is a naturally occurring free radical scavenger and as such its presence assists various other mechanisms in decreasing numerous disruptive free radical processes. The main goal of this study was to evaluate potential protective effects of ascorbic acid supplementation during in vitro culture of teratozoospermic specimens. Teratozoospermic semen samples that collected from 15 volunteers were processed, centrifuged and incubated at 37°C until sperm swimmed-up. Supernatant was divided into four groups and incubated at 37°C for one hour under different experimental conditions: Control, 10 micro m A23187, 600 micro m ascorbic acid and 10 micro m A23187+600 micro m ascorbic acid. After incubation sperm motility, viability, acrosome reaction, DNA damage and malondialdehyde levels were evaluated. Our results indicated that after one hour incubation, ascorbic acid significantly reduced malondialdehyde level in ascorbic acid group [1.4 +/- 0.11 nmol/ml] compared to control group [1.58 +/- 0.13 nmol/ml] [p<0.001]. At the end of incubation, progressive motility and viability in ascorbic acid group [64.5 +/- 8.8% and 80.3 +/- 6.4%, respectively] were significantly [p<0.05 and p<0.001, respectively] higher than the control group [54.5 +/- 6.8% and 70.9 +/- 7.3%, respectively]. A23187 significantly [p<0.0001] increased acrosome reaction in A23187 group [37.3 +/- 5.6%] compared to control group [8.5 +/- 3.2%] and this effect of A23187 attenuated by ascorbic acid in ascorbic acid+A23187 group [17.2 +/- 4.4%]. DNA fragmentation in ascorbic acid group [20 +/- 4.1%] was significantly [p<0.001] lower than controls [28.9 +/- 4.6%]. In vitro ascorbic acid supplementation during teratozoospermic semen processing for ART could protect teratozoospermic specimens against oxidative stress, and it could improve ART outcome

comparison of anxiety and depression rate between medical staff of infertility centers and obstetrics and gynecology centers of Yazd, Shiraz, Isfahan and Kerman hospitals

Regarding the close and continuous interaction of infertility staff with hopeless infertile couples and in the contrary the atmosphere of happiness especially in obstetric wards make a sense that considering anxiety and depression it would be a difference between these two wards. The objective of this study is the comparison of the rate of depression and anxiety between the two wards of infertility and obstetrics and gynecology. This study is a descriptive-correlation study based on cross-sectional method. 199 individuals who were the staff of infertility and obstetrics and gynecology wards in four provinces enrolled in this study through stratified sampling. Data collection was done by demographic questionnaire, Spiel Berger and Beck depression inventory tests. Data were analyzed by SPSS software using ANOVA test. The result showed the rate of anxiety in obstetrics and gynecology staff of Isfahan center [54.69 +/- 13.58] and depression rate had increased level in infertility staff of Shiraz center [14.94 +/- 10.87]. Overall, there was significant correlation between anxiety, depression and work place [p=0.047, 0.008 respectively]. According to ANOVA test, the mean value of anxiety level was higher in the staff of four obstetrics and gynecology centers and one infertility center. As long as we know that infertile couples have little chance for success rate and obstetrics and gynecology wards patients have little risk of failure in treatment, it could be mentioned that the anxiety and depression in the staff are not correlated with the client illness

Zoroastrians support oocyte and embryo donation program for infertile couples

The main goal was to evaluate the attitudes and knowledge of Zoroastrians living in Iran towards oocyte donation [OD] and embryo donation [ED] program. This cross sectional study consisted of 318 Zoroastrians [n=175 for OD and n=143 for ED] of both sexes. The questionnaire form comprised two parts of general demographic characteristics of the participants and twenty multiple-choice questions about attitude and knowledge of participants towards OD and ED. For statistical analysis, the chi-square test was applied for comparison of data generated from ED and OD groups. Majority of the participants supported OD [69.7%] and ED [71.3%] for infertile patients. In addition, 40% and 42% preferred donation program [OD and ED, respectively], compared to adoption. About 60% of the respondents believed that the donors have no right to find the child and claim it as their own. In addition, more than half of the respondents thought that the recipients of oocyte/embryo should never know the name and address of the donors. More than half of the participants did not know whether their religion accepts donation program or not. Approximately, 80% of respondents supported psychological counseling for both donors and recipients. Moreover, about 56% of the participants necessitated the advertisement on OD/ED program in the mass media. Our preliminary data showed that Zoroastrians supported both OD and ED program equally for infertile couples

Evaluating the role of silver nanoparticles on acrosomal reaction and spermatogenic cells in rat

Nanoparticles have wide range of application while there are some reports regarding their probable effects on male reproductive system and spermatozoa. The aim of this study was to evaluate the effect of different doses of silver nanoparticles [AgNPs] [70nm] on acrosome of rat spermatozoa and number of spermatogenic cells. In this experimental study, in experimental group, 32 male wistar rats [8 rats/group] received oral feeding AgNPs every 12 hr in one spermatogenesis period [48 days] by means of gavages in 25, 50, 100 and 200 mg/kg concentration [experimental groups 1-4, respectively]. The control group [8 rats] was treated on schedule with distilled water. Spermatozoa were stained by triple staining protocol for acrosome reaction. Histological evaluation on testis sections was performed using tissue processing and hematoxylin-eosin [H and E] staining. There was significant difference between the control group and the experimental group 1 for acrosome reaction [11.00 +/- 0.00 and 24.25 +/- 3.68, respectively, p=0.01]. There was only significant reduction in spermatogonia cells in experimental group 4. Experimental groups 2, 3 and 4 showed a significant reduction in the number of primary spermatocytes and spermatids as well as spermatozoa. But there were no significant differences between different groups for Sertoli cell number and seminiferous tubule diameter. It seems that Ag NPs have acute and significant effects on spermatogenesis and number of spermatogenic cells and also on acrosome reaction in sperm cells. More experimental investigations are necessary to elucidate better conclusion regarding the safety of nanoparticles on male reproduction system.

Seminal bacterial contaminations: probable factor in unexplained recurrent pregnancy loss

It is estimated that about 50% of causes of recurrent pregnancy loss [RPL] cases remain unknown. Sperm factors are suggested to have probable role in cases with RPL. The goal was to determine the possible relationship between semen bacterial contaminations with unexplained RPL. Also, the correlation between number of bacterial colony and sperm chromatin condensation was examined This study consisted of 30 fertile men [group A] and 30 infertile [group B] men with unknown RPL. Semen collection and analysis were done according to WHO manuals. Sperm count and motility were evaluated by Makler chamber. Eosin-Nigrosin and Papanicolaou staining methods were applied for viability and morphology assessment, respectively. The semen samples from both groups were cultured for aerobic bacteria. Aniline blue [AB] and toluidine blue [TB] staining methods were applied for evaluating sperm chromatin condensation. The numbers of colonies were significantly higher in group B when compared to group A. Also, S. aureus and E. coli contaminations showed significant differences between two groups. Both AB+ and TB+ sperm cells showed significant increase in group B compared to group A. There was a significant negative correlation between colony number and progressive motility [p=0.01], and sperm viability [p=0.007]. In addition, positive correlations were found between colony number and AB+ [p=0.001] and TB+ [p=0.004] as well. Bacterial contaminations in semen of men from RPL couples had significantly higher levels when compared to fertile controls. Presence of microorganisms in semen may be correlated with irregular sperm parameters and quality

Laser assisted zona hatching does not improve live birth rate in patients undergoing their first ICSI cycles

Routine use of assisted hatching [AH] following ICSI is a controversial issue in the literature. There are rare studies regarding the effect of laser assisted hatching [LAH] on live birth rate. Our main goal was to evaluate the effect of LAH on delivery rate as well as congenital anomaly in patients undergoing their first ICSI cycle. A total of 182 patients subjected to ICSI were randomly aliquot into two groups of experiment and control. In experiment group, the embryos were subjected to LAH to open a hole in ZP [about 10-12 Micro m] while in control group, the transferred embryos were intact with no AH. The patients were followed for clinical pregnancy and delivery rate as well as congenital anomaly. All the patients were infertile due to male factor infertility and LAH and embryo transfer were done on day 2. Laboratory and clinical characteristics of two groups of experiment and control were the same. There were insignificant differences between two groups of experiment and control for clinical pregnancy rate [20% vs. 23.9%, respectively, p=0.3] and live birth rate [11.11% vs. 8.6%, respectively, p=0.6]. Also no significant differences were observed between two groups of experiment and control for multiple pregnancy as well as congenital anomaly. Routine use of LAH in first ICSI cycle for male factor patients may have no beneficial effects on clinical pregnancy and live birth rate

Acute effects of ruta graveolens L. on sperm parameters and DNA integrity in rats

Increase in world population is one of the serious and threatening issues in this century. Therefore, it is vitally important to find safe and effective contraceptive methods, especially for men which already have few choices in this regard. Medicinal plants that were used for contraception in ancient times could be good sources of investigation in this filed. Ruta graveolens L. is one the plants introduced in the Iranian traditional medicine as an oral male contraception to be used before intercourse. In this study we tried to investigate the probable effects of the plant on the spermatozoa of male rats. Ruta graveolens L. aqueous extract [5 g/kg] was administered orally to five groups of male rats and sperm motility was checked after half, one, two, four and six hours later. Moreover, one group of rats served as the control group. Subsequently, viability of cells [Eosin-Nigrosin staining], morphological changes [Diff-Quick staining], DNA status [acridine orange dye] and serum testosterone levels were assessed in the treated groups which had significant immotile spermatozoa. For statistical analysis, Student's t-test and one-way ANOVA with Tukey's post-hoc test were employed for comparison between groups. A significant reduction in sperm motility was seen one hour after administration of the extract in the case groups compared to the controls [36% vs. 68.15%, respectively, p<0.01]. The motility gradually increased afterwards, and by 6 hours, it was the same as the control group [65.43% and 68.15%, respectively]. No significant changes were seen in viability, morphology or DNA structure of spermatozoa in each group. Testosterone levels did not show any significant changes in the treated groups when compared with the controls. Since a significant temporary immobility of spermatozoa without any adverse effects on other sperm characteristics occurred upon the administration of Ruta graveolens L. aqueous extract, it seems that this plant might have the potential to be used for the suggested male contraception

Evaluating the role of first polar body morphology on rates of fertilization and embryo development in ICSI cycles

Recent studies have demonstrated that morphology of the first polar body [1st PB] is related to oocyte viability, which can be used as a prognostic tool to predict oocyte performance and pregnancy outcomes in an intracytoplasmic sperm injection [ICSI] program. According to some studies, there is a correlation between oocyte performance and 1st PB morphology, while others have not reported any correlation. The objective of this study is to evaluate the role of 1st PB morphology on rates of fertilization and embryo development in ICSI cases. In this prospective study morphological characteristics of 470 metaphase II [MII] oocytes were assessed in 80 ICSI cycles. The women were ages 21-42 years [mean 32.6 +/- 0.2]. Their oocytes were retrieved after a hyperstimulation protocol. After denudation, all oocytes were evaluated for 1st PB morphology. The oocytes were divided into two groups of A [normal 1st PB] and B [abnormal fragmented 1st PB]. In addition, other abnormalities, such as refractile bodies [RF], wide previtelline space [wPVS], central and general granulation, bull's eye, vacuole, smooth endoplasmic reticulum cluster [SERc], debris in PVS, shape and dark oocyte were checked. For verifying of fertilization, about 18-19 hours post-ICSI, oocytes were checked for two-pronuclear. Assessments of embryo quality, development and embryo transfer were done at day two. Chisquare, Fisher's exact and independent sample t tests were chosen for statistical analysis. Twenty-seven percent of oocytes had fragmented 1st PB, while the remainder was associated with other morphological abnormalities. A total of 46.1% and 26.9% of oocytes showed double and multiple defects, respectively. RF was the most common abnormality observed in group B. No significant differences in women's' ages between groups A and B were noted [p=0.3]. A total of 179 and 107 oocytes [61.5% vs. 59.8%] were fertilized in groups A and B, respectively [p=0.7]. The rates of good embryo formation for A and B groups were 66. 5% and 55. 6% [p=0.07], and cleavage rates were 77. 7% and 68.5%, respectively [p=0.09]. The data demonstrated that 1st PB morphology does not appear to be a prognostic factor for rates of fertilization and embryo development in ICSI cycles