A new cryotop vial device system provides an aseptic cryoprotectant-free and centrifuge-free cryopreservation of human spermatozoa (a closed system).

Single sperm cryopreservation is a new method to preserve small numbers of spermatozoa in small droplets. So far, several devices have been introduced for this technique, but more studies are needed for its optimization. The aim of this study was optimizing the previous device for low number of spermatozoa and low volume semen, which led to design of Cryotop Vial device. Normal semen samples from 25 patients were prepared by swim-up method and divided into four groups: Fresh (F), Rapid freezing (R) and ultra-rapid freezing with Cryotop Device (CD) and Cryotop Vial Device (CVD). In R group, the diluted sperm suspension with sperm freezing medium was cooled in the vapor phase and then immersed in liquid nitrogen. Ultra rapid freezing was performed with sucrose in small volume using the Cryotop Device (CD) or Cryotop Vial Device (CVD). Sperm viability, motility, fine morphology, mitochondrial activity and DNA fragmentation were assessed in all samples. All sperm parameters decreased significantly in all the cryo groups compared to the fresh group. The comparison between the cryo groups showed that progressive motility (69.28 ± 6.82 vs. 55.68 ± 9.04, and 54.76 ± 5.34, p < 0.001) and viability (77.36 ± 5.48 vs.68.84 ± 8.51, p < 0.001, and 70.04 ± 7.44, P = 0.002) were significantly higher in the CVD compared to the CD and R groups respectively. DNA fragmentation was also significantly lower in both ultra-rapid freezing groups (CD and CVD) compared to the R group. Fine morphology and mitochondrial activity were not different between the cryo groups. The CVD as a cryoprotectant and centrifuge-free technique preserved sperm motility, viability and DNA integrity after cryopreservation better than other groups.

The effect of pentoxifylline and calcium ionophore treatment on sperm cell biology in oligoasthenoteratozoospermia samples.

The objective of this study was to assess the effects of pentoxifylline (PTX) and Ca2+ ionophore (CI) A12387 treatment on some biological characteristics of sperm cells in oligoasthenoteratozoospermia (OAT) patients. After processing, each sample was divided into four groups: 1, control; 2, exposed to 3.6 mM PTX; 3, exposed to 5 µm calcium ionophore (CI); and 4, exposed to both PTX and CI; 30 min at 37°C. Sperm motility was measured before and after preparation. Acrosome reaction (AR), status of sperm vacuoles, mitochondrial membrane potential (MMP) and DNA fragmentation were assessed using PSA-FITC staining, motile sperm organelle morphology examination (MSOME), JC-1 staining and sperm chromatin dispersion (CSD) test, respectively. Treatment with PTX and CI led to increased and decreased sperm motility, respectively (P < 0.05). Furthermore, vacuole status and rates of sperm DNA fragmentation were not significantly different among groups (P > 0.05). Moreover, the data showed that the rates of AR and disrupted MMP were significantly different between groups (P < 0.05). In conclusion, in vitro application of PTX not only did not have any adverse effects on sperm cell biology characteristics, but also can rectify the harmful effect of CI.

Improvement of human sperm properties with platelet-rich plasma as a cryoprotectant supplement.

PURPOSE: Platelet-rich plasma (PRP) is a remarkable substance, which involves the growth and proliferation of all cell types. As a source of growth factors, we evaluated whether sperm cryopreservation supplemented with PRP improves the rates of sperm motility, viability, and DNA integrity after vitrification compared with conventional cryo-medium. MATERIALS AND METHODS: 20 normal semen specimens were collected from healthy men. After swim-up preparation, each sample was divided into four aliquots. One, as control, received no treatment, and the other three experimental samples were treated with three different concentrations of PRP as cryoprotectant. Sperm parameters were examined before and after freezing procedure. RESULTS: PRP had no significant effect on sperm count. Meanwhile, the percentage of sperm progressive motility and viability in the PRP treated samples with 1×105 /µL concentration was significantly higher than control group. Besides, the rate of immotile sperms in these samples was significantly lower than the control. Sperm viability was significantly higher in the PRP samples at 1×105/µL concentration. In the case of DNA integrity, CMA3 staining showed that the lower PRP concentration was correlated with the higher rate of abnormal spermatozoa. SCD showed that the rate of abnormal sperms in the PRP samples with 1×105 /µL concentration was significantly lower than control group. CONCLUSIONS: This study showed a protective effect of PRP on human sperm quality at an optimized concentration after vitrification. Besides, the effects of PRP supplementation of sperms on successful fertility following sperm preservation will be of interest.

Prolonged exposure of human spermatozoa in polyvinylpyrrolidone has detrimental effects on sperm biological characteristics.

Polyvinylpyrrolidone (PVP) has been utilized in intracytoplasmic sperm injection (ICSI) for immobilization and manipulation of spermatozoa. This study aims to determine the suitable time that sperm cells could be safely exposed to PVP during ICSI procedure. Twenty-five normal semen samples were prepared using the swim-up method and then were exposed to 10% PVP at different time intervals (15, 30 and 60 min). The effect of PVP on sperm parameters (viability and morphology), DNA fragmentation index (sperm chromatin dispersion test), chromatin quality (aniline blue, toluidine blue and chromomycin A3 staining), acrosome reaction, mitochondrial membrane potential and sperm ultrastructure was assessed at different time intervals. Our results showed that prolonged sperm exposure in PVP for 15, 30 and 60 min significantly affects viability and morphology with a concomitant increase in DNA fragmentation and abnormal chromatin structure, while the percentage of acrosome-reacted spermatozoa was additionally increased. In addition, the spermatozoa with high mitochondrial membrane potential were significantly decreased compared to unexposed spermatozoa to PVP. In conclusion, the detrimental effects of PVP were increased significantly following sperm exposure in PVP after 15 min. Therefore, the sperm exposure to PVP should be limited to less than 15 min during ICSI procedure.

Proliferating cell nuclear antigen presentation, as a marker of folliculogenesis, in the transplanted ovarian tissue.

AIM: One of the most important ways to understand the ovarian biology is studding the initiation of primordial follicle development and subsequent folliculogenesis control. In this study, proliferating cell nuclear antigen (PCNA) presentation was used as a marker of follicular development in the thawed ovarian tissue (OT) following transplantation onto chick embryo chorioallantoic membrane (CAM) using two methods of freezing of slow freezing and vitrification. METHODS: Samples of OT from 10 patients were subjected to slow freezing and vitrification. After warming, CAM transplantation was done and PCNA proliferation index (PI; percent of PCNA-positive granulosa cells) was calculated for each follicle stage. Image J software was used to determine the mean staining intensity. RESULTS: PCNA was positive for granulosa cells and oocytes nuclei, but negative for ooplasm. There were no remarkable PCNA staining in the granulosa cells of primordial follicles, but increased significantly as follicle progression (p < 0.05). Proliferation rate was also insignificantly higher in the vitrified than slow freezing group, before and after transplantation (p < 0.05). Lower PCNA presentation index was observed after CAM transplantation (p < 0.05). The earliest stage of follicular recruitment took place in the transitional follicles, before squamous cells transform to cuboidal cells. CONCLUSION: PCNA showed that follicles had proliferation power after cryopreservation. Higher presentation after vitrification may indicate accelerated folliculogenesis in the thawed OT.

The effect of testosterone and antioxidants nanoliposomes on gene expressions and sperm parameters in asthenospermic individuals.

BACKGROUND: Antioxidants that used for the infertility treatment cannot have their complete effectiveness, because of their instability in the culture medium. SIGNIFICANCE: One of the most advances, in the drug delivery systems, is nanoliposomes-loaded, as biodegradable and bioavailable carriers. Hormonal and antioxidant agents encapsulating inside the nanoliposomes were used, to increase the effectiveness of antioxidants in the sperm culture medium. MATERIALS: Semen sample from 15 asthenospermia were divided into 10 equal parts. After preparation, the sperms were incubated with free form of drugs and nanocarriers contained resveratrol, catalase, resveratrol-catalase and testosterone for 45 min. All sperm parameters, sperm DNA and gene expressions were evaluated before and after freezing. RESULTS: Before freezing, all nanocarriers and free testosterone showed higher sperm motility compared to free drugs (p=.000). Free Testosterone and free resveratrol-catalase had higher DNA damage compared to nanocarriers (p=.000). Before freezing, the blank nanoliposome and testosterone nanoliposomes had the lowest HSP70 gene expression respectively (p = 0.005) (p = 0.001). After freezing, a significant reduction in sperm motility was observed in the free resveratrol-catalase group (p=.003). Also, a significant increase in sperm viability was observed in the free testosterone and nanoliposomes of blank and testosterone (p > 0.05). The least DNA fragmentation was related to catalase nanoliposomes (p=.000). All nanoliposomes, especially catalase, had the highest percentage of class I morphology compared to the control group (p=.000). CONCLUSIONS: Nanoliposomes could improve the sperm parameters and DNA integrity before and after freezing, by increasing the effectiveness of antioxidants. So, it can be recommended in the ART lab.

Does sperm DNA fragmentation have negative impact on embryo morphology and morphokinetics in IVF programme?

Evaluation of sperm integrity may predict the in vitro fertilisation (IVF) outcomes. The aim was to evaluate the relationship between the sperm DNA fragmentation (sDNAf) with embryo morphology and morphokinetic using time-laps monitoring (TLM) and to select the best time points for normalisation in IVF setting. After evaluating the fertilisation and pronuclei (Z) scoring, 328 normally fertilised oocytes were assessed to time of pronuclei fading, time of 2 to 8 discrete cells (t2-t8) and abnormal cleavage patterns, such as multinucleation, direct cleavage, reverse cleavage and fragmentation. Sperm chromatin dispersion (SCD) assay was used for assessment of prepared sperm chromatin status. SCD was categorised into 4 groups of <6.5, 6.5-10.7, 10.7-20.1 and >20.1. The finding showed significant differences in t6 (p = .012), t7 (p = .045), t8 (p = .013) and s1 (p = .001) between 4 SCD groups. When morphokinetic variables were normalised to tPNf, this difference was observed in t2 (p = .003) and t6 (p = .017). Subsequently, the percentage of top quality embryos and Z1 scoring were dependent to the sDNAf rate. In conclusion, tPNf was the best reference time point in IVF cycles. Also, we found high sDNAf rate had no negative impact on embryo morphology and morphokinetics in conventional IVF.

Effects of microsurgical varicocelectomy on semen analysis and sperm function tests in patients with different grades of varicocele: Role of sperm functional tests in evaluation of treatments outcome.

It seems that varicocele play a role in male infertility, as such, their prevalence increases from 15% in the normal population to 80% in secondary infertility subjects. Varicoceles may have negative effects on semen quality. Our goal was to assess the effects of microsurgical varicocelectomy on semen analysis and sperm functional tests in men with different grades of varicoceles. Thirty infertile men with different grades of varicoceles (grades 1 to 3) were enrolled in our study. Semen quality was assessed by semen analysis according to the WHO guideline (WHO, 1999) and four different sperm functional tests (aniline blue, toluidine blue, chromomycin A3 and TUNEL test) were carried out before and 3 months after microsurgical varicocelectomy (M-varicocelectomy). When considered all three grades together, we showed that M-varicocelectomy had statistically significant effects on all four types of sperm functional tests (p value<0.05). It also had positive effects on conventional semen parameters, although the effects were not statistically significant for some parameters (for example sperm count). When analysed separately (based on varicocele grades) the surgery, although caused improvements in semen quality, but may have more statistically significant effects on patients with varicocele of higher grade. In addition, in varicocele of lower grade (for example grade 2), sperm function test may be a better predictor of surgical success than the conventional semen analysis. Thus, we show that not only M-varicocelectomy has significant positive effect on semen quality but also if sperm functional tests become more affordable in the future, because they yield more precise results, their use in daily practice may increase significantly in patients with varicoceles.

Pentoxifylline increase sperm motility in devitrified spermatozoa from asthenozoospermic patient without damage chromatin and DNA integrity.

The freeze-thaw process results in reduced motility, viability and fertilization potential of human spermatozoa. So, a variety of substances were evaluated in order to enhance human sperm resistance to the stress of cryopreservation, such as Pentoxifylline (PTX) for improving the Intracytoplasmic sperm injection (ICSI) outcomes. The aim was to investigate the effect of PTX on sperm parameters and chromatin/DNA integrity of asthenozoospermic semen post vitrification. A total of 30 semen specimens were obtained from infertile men with asthenozoospermia. The cryoprotectant-free vitrification was performed for the samples after assessment of sperm parameters. After warming, each sample was exposed for 30 min to 3.6 mmol/l PTX in experimental group and the control group without any treatment apposing at 37 °C for 30 min in regard, to repeat all in vitro analysis (sperm parameters and DNA integrity assay). Regardless of the vitrification devastating impacts on sperm parameters, incubation of post vitrified samples with PTX increased the rate of progressive motility (P < 0.01). Moreover, PTX addition did not significantly damage DNA integrity of asthenozoospermic sperm samples. The data showed that PTX was able to improve sperm movement without any adverse effects on sperm chromatin/DNA integrity in vitrification program.

Vitrification is not superior to rapid freezing of normozoospermic spermatozoa: effects on sperm parameters, DNA fragmentation and hyaluronan binding.

Human sperm vitrification is a new cryopreservation method. This study compared the effects of rapid freezing and vitrification on various sperm parameters, hyaluronan-binding assay and DNA fragmentation and assessed the impact of cryoprotectant agents (CPA) with vitrification. A total of 30 normo-ejaculates were prepared by swim up and the motile sperm fraction was divided into four: fresh (control), rapid freezing, and two vitrification groups (a, lacking CPA; b, with CPA). For rapid freezing, a cryovial of sperm suspension was held just above the liquid nitrogen surface, and for vitrification, 30µl suspension was dropped directly into liquid nitrogen. Sperm parameters, including motility, viability and morphology, declined after cryopreservation in both groups. DNA fragmentation was not significantly higher in the vitrification (15.7±4.4%) or rapid freezing (16.6±5.6%) groups when compared with controls (11.6±4.5%). The rates of hyaluronan binding were similar between the control and cryopreserved groups. Moreover, addition of CPA for vitrification had a neutral effect on rates of sperm recovery. In conclusion, vitrification has great potential for human sperm cryopreservation and does not require CPA, with its possible toxicity. However, it is not superior to rapid cryopreservation regarding sperm recovery rate in normozoospermia. Human sperm vitrification is a new cryopreservation method that has been introduced recently. This study compared the effects of rapid freezing with vitrification on rates of sperm parameters, hyaluronan-binding assay and DNA fragmentation after thawing/warming and assessed the impact of cryoprotectant agent (CPA) on vitrification. The study was performed on 30 ejaculates prepared using the swim-up technique. Each motile sperm suspension was divided into four: control (fresh); rapid freezing; and two vitrification groups (a, lacking CPA; b, with CPA). For rapid freezing, a cryovial of sperm suspension was held above the surface of liquid nitrogen. For vitrification, 30µl sperm suspension was dropped into liquid nitrogen directly. The rates of progressive motility (86.6±5.9%) and viability (95.8±3.9%) in controls declined significantly, to 40.0±13.0% and 63.2±7.7% for rapid freezing and 41.9±10.3% and 64.4±10.0% for vitrification, respectively. Normal sperm morphology was also significantly decreased after cryopreservation in all groups. DNA fragmentation was higher with rapid freezing compared with fresh controls (16.6±5.6% vs. 11.6±4.5%, P=0.01), but DNA fragmentation did not increase significantly in vitrified samples (15.7±4.4%). The rates of hyaluronan binding were similar between the control and cryopreserved groups. Moreover, addition of CPA for vitrification had a neutral effect on rates of sperm recovery. In conclusion, vitrification has great potential for human sperm cryopreservation and does not require CPA, with its possible toxicity. However, it is not superior to rapid cryopreservation regarding sperm recovery rate in normozoospermia.

Application of Autologous Platelet-rich Plasma Exerts Cryoprotective Effects on Biological Characteristics of Human Oligoasthenoteratospermia Samples after Freezing and Thawing Procedures.

OBJECTIVE: Platelet-rich plasma (PRP) is enriched with active biological components which showed proliferative and cytoprotective properties in healing different injuries in medicinal fields. This study was designed to assess cryoprotective effects of autologous PRP on quality of oligoasthenoteratospermia (OAT) samples during freezing and thawing procedure. MATERIALS AND METHODS: The present study is an experimental research. Twenty OAT semen samples were obtained from individuals and prepared by discontinuous density - gradients technique. Control group is sperm samples after DGC. After the procedure, the specimen divided into four groups. Freeze group which has no additive and other three groups were cryopreserved with different concentrations of PRP (1×105/µL, 0.5×105/µL and 0.25×105/µL). Autologous PRP was provided by each participant. After thawing, sperm parameters, DNA fragmentation by sperm chromatin dispersion test (SCD), protamine deficiency by (Chromomycin A3) CMA3 staining, acrosome integrity and malondialdehyde (MDA) level were evaluated. RESULTS: Cryopreservation resulted in significant decreased in all factors compared to the control group. There were no significant changes on sperm count, morphology, non-progressive motility and acrosome reaction by adding PRP as cryoprotectant in comparison with freeze group. PRP at all three concentrations showed significant increase in progressive motility (3.05±2.01 vs. 14.05±4.13, 12.35±4.90 and 12.15±9.65, P<0.001) and viability (36.85±10.25 vs. 47.85±5.86, 51.30±5.54 and 50.05±5.67, P<0.001) compared to the sperm samples without PRP. The percentage of immotile sperms decreased at all PRP concentrations compared to the freeze group. Moreover, PRP at 1×105/µL concentration showed cryoprotective effects on DNA fragmentation, protamine deficiency and MDA level compared to the other three concenterations. CONCLUSION: Cryopreservation and thawing procedures may exert adverse effects on biological factors of sperm samples. Therefore, adding PRP as cryoprotectant at all three concentrations especially 1×105/µL can be promising strategy to reduce adverse effects of cryopreservation on OAT samples.

Minimum energy desynchronizing control for coupled neurons.

We employ optimal control theory to design an event-based, minimum energy, desynchronizing control stimulus for a network of pathologically synchronized, heterogeneously coupled neurons. This works by optimally driving the neurons to their phaseless sets, switching the control off, and letting the phases of the neurons randomize under intrinsic background noise. An event-based minimum energy input may be clinically desirable for deep brain stimulation treatment of neurological diseases, like Parkinson's disease. The event-based nature of the input results in its administration only when it is necessary, which, in general, amounts to fewer applications, and hence, less charge transfer to and from the tissue. The minimum energy nature of the input may also help prolong battery life for implanted stimulus generators. For the example considered, it is shown that the proposed control causes a considerable amount of randomization in the timing of each neuron's next spike, leading to desynchronization for the network.

Possible Harmful Effects of Smoking Hookah on Sperm DNA Fragmentation Index and Protamine Genes Expression in Normozoospermic Men.

Objectives: In recent years, smoking water pipes or hookah has increased among adolescents in most countries. Although there is evidence in support of the negative effects of this type of smoking on human health, such as the increased risk of lung disease, little is known about the potential effects of hookah smoking on the male reproductive system, especially on the molecular aspects of sperm. Patients and methods: This cross-sectional study examined sperm DNA fragmentation index, protamine 1 and 2 (PRM1 and PRM2) genes expression, and oxidant status in normozoospermic hookah smokers in comparison with non-smoker controls. Results: Our results showed significantly higher rates of DNA fragmentation, protamine deficiency, and abnormal chromatin condensation in the spermatozoa of hookah smokers (P < .0001). Also, protamine gene expression showed a remarkable decrease in hookah smokers (1.55 ± 2.54 and 0.33 ± 0.54) compared to the controls (3.49 ± 5.41 and 1.22 ± 1.96), although the reduction was not statistically significant (P = .155 and P = .066, respectively). Moreover, a significantly higher level of semen MDA was observed in the case group compared to the controls (0.39 ± 1.04 vs 0.15 ± 0.21; P = .013). Conclusion: According to our study, although hookah smoking does not have a significant effect on sperm parameters, it may have deleterious effects on DNA integrity, oxidative status, and nuclear protein levels of spermatozoa.

Time optimal control of spiking neurons.

By injecting an electrical current control stimulus into a neuron, one can change its inter-spike intervals. In this paper, we investigate the time optimal control problem for periodically firing neurons, represented by different one-dimensional phase models, and find analytical expressions for the minimum and maximum values of inter-spike intervals achievable with small bounded control stimuli. We consider two cases: with a charge-balance constraint on the input, and without it. The analytical calculations are supported with numerical results for examples of qualitatively different neuron models.

Comparison of zeta potential and physiological intracytoplasmic sperm injection in obtaining sperms with a lower DNA fragmentation index: A cross-sectional study.

Background: The sperm DNA fragmentation index (DFI) is one of the men's reproductive health criteria that affects assisted reproductive technique outcomes. Efforts in obtaining high-quality mature sperms seem to be necessary. Advanced sperm selection techniques (including physiological intracytoplasmic sperm injection [PICSI], zeta potential, microfluidic, etc.) have gained popularity in this regard. Objective: The study aimed to compare the efficacy of zeta potential and PICSI sperm selection in obtaining sperms with better DNA integrity. Materials and Methods: In this cross-sectional study, 48 couples were enrolled where the male partner had increased sperm DFI in his ejaculated sample and the female was in normal reproductive health. For each male partner, the semen sample was processed with zeta potential and PICSI techniques, then the sperm DFI of neat semen was compared to zeta and PICSI samples by the sperm chromatin dispersion test. Results: Data showed that both the zeta potential and PICSI technique decreased sperm DFI in comparison with the neat semen sample (p < 0.001 for both). In addition, there was a statistically significant difference in sperm DFI between the PICSI and zeta potential samples (p < 0.01). Conclusion: The current study showed that both zeta potential and PICSI could result in sperm with a lower DFI. However, PICSI seems to be superior to zeta potential in this regard.

Prolonged semen incubation alters the biological characteristics of human spermatozoa.

OBJECTIVE: The present study assessed the biological characteristics of human spermatozoa at different time intervals (0, 1, 1.5, and 2 hours) after incubation at 37°C. METHODS: Twenty-five normozoospermic semen samples were incubated at 37°C. Incubation was performed at four time intervals of 0 (after liquefaction), 1, 1.5, and 2 hours. The samples were evaluated for sperm parameters at each time interval. RESULTS: The rate of sperm progressive motility decreased at 1.5 hours compared to 0 hours as well as 2 hours compared to 1 hour and 0 hours. The rate of non-motile spermatozoa also decreased after 2 hours compared to after 0 hours. No significant changes were observed in sperm viability (p=0.98) and non- progressive motility (p=0.48) at any time intervals. Abnormal sperm morphology increased at 1.5 hours of incubation time (p<0.001). No significant changes were observed in DNA fragmentation at 1 hour compared to 0 hours (median [interquartile range]: 19.5 [4] vs. 19 [4]), as well as at 1.5 hours compared to 1 hour (20 [5]). However, a significant increase in DNA fragmentation was observed at 1.5 hours compared to 0 hours. The mitochondrial membrane potential decreased remarkably after 1 hour of incubation time. No significant differences were observed in the acrosome reaction or malonaldehyde levels at any time point (p=0.34 and p=0.98, respectively). CONCLUSION: The incubation of normozoospermic samples before use in assisted reproductive technology should be less than 1.5 hours to minimize the destructive effects of prolonged incubation time on general and specific sperm parameters.

Comparison of the Efficiency of Magnetic-Activated Cell Sorting (MACS) and Physiological Intracytoplasmic Sperm Injection (PICSI) for Sperm Selection in Cases with Unexplained Infertility.

Background: The cases with unexplained infertility may have an abnormality in their sperm chromatin structure. Sperm selection methods can be used to separate sperm with low DNA fragmentation. The purpose of this study was to compare the efficacy of physiological intracytoplasmic sperm injection (PICSI) with magnetic-activated cell sorting (MACS) in assisted reproductive techniques in cases with unexplained infertility. Methods: The semen samples were collected from couples with unexplained infertility. After semen analysis and sperm DNA fragmentation (SDF) evaluations, samples were prepared with swim-up method. The rates of SDF in different fractions including raw semen (n=20), swim-up (n=20), only motile sperm after swim-up (swim-up selection) (n=20), MACS sperm selection (n=20), only motile sperm after MACS (MACS selection) (n=20), and PICSI sperm selection (n=16) were evaluated. Also, the main sperm characteristics and fine morphology of sperm suspension after MACS were assessed. Statistical analysis was performed using GraphPad Prism. The p<0.05 was considered statistically significant. Results: DNA fragmentation index (DFI) values in PICSI and MACS groups were significantly reduced as compared to the swim-up group. The rate of this reduction was more pronounced in MACS (58.20±13.02) than PICSI (36.57±15.52) group. Also, our results showed that MACS resulted in decreased sperm motility, with no alteration in their fine morphology. Conclusion: MACS was found to be more efficient in reduction of SDF rates than PICSI. However, none of the sperm selection techniques can not totally eliminated the spermatozoa with DNA fragmentation in the final sperm sample.

A comparison of different O-antigen serogroups of Escherichia coli in semen samples of fertile and infertile men.

OBJECTIVE: Male genital tract infections have been associated with infertility, and Escherichia coli has drawn increasing attention as an important bacterium in this context. This investigation aimed to characterize and compare the distributions of O-antigen serogroups of E. coli in the semen samples of fertile and infertile men. METHODS: In this case-control study, semen samples were collected from 618 fertile and 1,535 infertile men. The E. coli-positive samples were evaluated in terms of concentration, morphology, viability, and motility parameters according to the World Health Organization 2010 guidelines. Finally, different serogroups of E. coli were identified by multiplex polymerase chain reaction targeting the O-antigen variations of the bacterium. RESULTS: The prevalence of E. coli among fertile men was significantly higher than among infertile men (p<0.001). The sperm morphology, viability, and motility in the E. coli-positive fertile group were significantly higher than in the E. coli-positive infertile group (p<0.001). E. coli O6 was the most prevalent serogroup found in both groups. However, there was no significant difference in the frequency of different serogroups of E. coil between the two groups (p=0.55). CONCLUSION: Despite the higher prevalence of E. coli among fertile men, E. coli had more detrimental effects on semen parameters in infertile men. There was no significant difference in E. coli serogroups between the fertile and infertile groups.

Effect of Coronavirus Disease (COVID-19) on Human Semen: No Evidence of Coronavirus in Semen of Patients.

Background: In December 2019, a severe acute respiratory syndrome (SARS-COV-2) was found in China. The coronavirus can impact different organs, as shown by the virus having been detected in urine, blood, oropharyngeal, and feces. This study was done to assess the impact of COVID-19 on semen analysis and to evaluate the existence of the virus in the semen of infected men. Methodology. Forty fertile men with COVID-19 were confirmed by an oropharyngeal sample. The men were divided into two groups. The semen of twenty men in the acute stage of COVID-19 and twenty men in the clinical recovery stage was analyzed, and the parameters of semen were compared between two groups. In addition, a PCR test of patients' semen was done. Result: The analysis showed that all patients' semen specimens tested negative. Semen analysis revealed no significant difference in sperm count, concentration, or motility, and the sperm of both groups was found to be normal. However, viability and morphology parameters were significantly lower in men with the acute disease. Conclusion: Coronavirus (COVID-19) was not secreted in the semen of infected men but had a negative effect on the morphology and viability of the sperm of men in the acute stage.

Predictive accuracy of CNN for cortical oscillatory activity in an acute rat model of parkinsonism.

In neurological and neuropsychiatric disorders neuronal oscillatory activity between basal ganglia and cortical circuits are altered, which may be useful as biomarker for adaptive deep brain stimulation. We investigated whether changes in the spectral power of oscillatory activity in the motor cortex (MCtx) and the sensorimotor cortex (SMCtx) of rats after injection of the dopamine (DA) receptor antagonist haloperidol (HALO) would be similar to those observed in Parkinson disease. Thereafter, we tested whether a convolutional neural network (CNN) model would identify brain signal alterations in this acute model of parkinsonism. A sixteen channel surface micro-electrocorticogram (ECoG) recording array was placed under the dura above the MCtx and SMCtx areas of one hemisphere under general anaesthesia in rats. Seven days after surgery, micro ECoG was recorded in individual free moving rats in three conditions: (1) basal activity, (2) after injection of HALO (0.5 mg/kg), and (3) with additional injection of apomorphine (APO) (1 mg/kg). Furthermore, a CNN-based classification consisting of 23,530 parameters was applied on the raw data. HALO injection decreased oscillatory theta band activity (4-8 Hz) and enhanced beta (12-30 Hz) and gamma (30-100 Hz) in MCtx and SMCtx, which was compensated after APO injection (P ¡ 0.001). Evaluation of classification performance of the CNN model provided accuracy of 92%, sensitivity of 90% and specificity of 93% on one-dimensional signals. The CNN proposed model requires a minimum of sensory hardware and may be integrated into future research on therapeutic devices for Parkinson disease, such as adaptive closed loop stimulation, thus contributing to more efficient way of treatment.