Therapy Of Male Infertility Research Articles

Effect of hydrogen peroxide on Na+,K+-ATPase activity in spermatozoa of infertile men

Na+,K+-ATPase plays an essential role in sperm motility, hyperactivation, chemotaxis, acrosome reaction etc. Na+,K+-ATPase is sensitive to ROS insult. Apart from production of highly reactive molecules, H2O2 can exert a number of direct effects on cells, their metabolism and enzymes. In the present study, exposure to exogenous H2O2 was used to characterize the effects of H2O2 on Na+,K+-ATPase activity in spermatozoa of infertile men with different forms of pathospermia. It was shown that Na+,K+-ATPase activities in spermatozoa of infertile men with different forms of pathospermia were inhibited by exposure to H2O2 (50−500 μM). H2O2, one of the most toxic oxygen species, has the ability to depress Na+,K+-ATPase activity in a dose-dependent manner. Severe inhibition of the hydrolytic activity was observed when higher H2O2 were used. The time course of incubation with 100 μM H2O2 showed a sharp decrease in the enzyme activity during the first 5 min of incubation for both normozoospermic and pathozoospermic men. The enzymatic activity of Na+,K+-ATPase in the sperm was completely destroyed at 20 min for asthenozoospermic men and 30 min for normozoospermic men. We show that an administation of H2O2 inhibited Na+,K+-ATPase activity in normozoospermic samples with IC50 of 106.6 ± 7.9 μM. IC50 for patients with asthenozoospermia was two times less than for healthy men with preserved fertility. For other studied groups, the differences in IC50 were not significant. These observations suggest that Na+,K+-ATPase in pathozoospermic samples is more vulnerable to H2O2-induced damage than in normozoospermic men. The Hill coefficient was significantly increased only for patients with asthenozoospermia, indicating increased positively cooperative binding. The decreases in Na+,K+-ATPase hydrolase activity in H2O2-treated sperm cells in men with normozoospermia were largely attenuated by exogenous GSH at 5 mM. This suggests that GSH partially protects the Na+,K+-ATPase from inhibition under experimental oxidative stress. However, treatment of oligo-, astheno- and oligoasthenozoospermic samples with 100 μM H2O2 and 5 mM GSH did not result in protection of Na+,K+-ATPase against induced oxidation, suggesting that the impaired Na+,K+-ATPase in pathozoospermic samples appears to be an irreversible event. In contrast, presence of GSH only after H2O2 treatment does not reverse Na+,K+-ATPase inhibition. This study has provided a deeper insight into the role Na+,K+-ATPase plays in sperm cells,it also could offer clues to the clinical application of antioxidant therapy in male infertility therapy.

Laser therapy for male infertility. Part 1. Etiology and pathogenesis. experimental studies

Male infertility is associated with a wide range of pathological conditions affecting both the sexual and other body systems: endocrine, nervous, blood, and immune. Laser therapy is a form of modern physiotherapy that applies low-intensity laser radiation in various parts of the body. It is widely used in all areas of modern medicine due to its high efficiency, ease of use, the absence of contraindications and side effects. The authors reviewed the results of 171 studies investigating laser therapy, of which 93 were experimental (20 in Russian and 73 international publications), 4 clinical and experimental (all in Russian), 74 clinical (73 in Russian and 1in English). Along with domestic literature, publications in Russian from Belarus, Georgia, Kyrgyzstan, Uzbekistan and Ukraine were studied. The article analyzes the experimental studies, somehow related to the problem of infertility and its treatment using low-intensity lasers. The analysis findings suggest that there are all prerequisites to develop a laser therapy as an effective modality for treating infertile men.

Adjuvant antioxidant therapy in varicocele infertility

Infertility affects an estimated 15% of couples globally. Male factor infertility accounts for about a half of infertility cases. About 40% of infertile men have varicocele. The aim of this study was to investigate the efficacy and safety of a complex of acetyl-L-carnitine, L-carnitine fumarate and alpha-lipoic acid (SpermActin-forte) (SA) for adjuvant antioxidant therapy after microsurgical varicocelectomy (MVE) in men with varicocele and assess its impact on the level of DNA fragmentation in sperm cells. This is an open, prospective, randomized trial comprising 114 men aged 25-45 (mean 34.1+/-12.1) years who underwent MVE. The patients were allocated to receive either adjuvant SA (n=38), SA in combination with a vitamin complex (VC) (n=38) or no adjuvant therapy (n=38). The efficacy was assessed at 3 months by testing standard semen parameters and the level of sperm DNA fragmentation. MVE led to a 21.7% increase in the progressive sperm motility compared to the baseline level. In patients receiving SA, this was by 76.7% due to active sperm motility (category A) at 3 months. MVE with concurrent supplementation of SA resulted in a 22.3% decrease in the level of sperm DNA fragmentation at 3 months. When used in the postoperative period after MVE, SA and VC resulted in a 27% increase in the sperm concentration at 3 months. There were no side effects of pharmacotherapy. The statistical significance of the change in variables was calculated using the Wilcoxon test. Critical level =0.05 was established for all criteria. Using nutraceuticals (SA) after MVE is an effective and safe adjuvant antioxidant therapy of male infertility in patients with varicocele which leads to an improvement in the basic sperm parameters (sperm concentration and motility) and a decrease in the level of sperm DNA fragmentation in the short term. Adjuvant antioxidant therapy of varicocele infertility potentiates the effects of monotherapy (both medical and surgical), leads to an increase in its effectiveness and shortens the time to pregnancy. Further studies in this field are needed to assess long-term outcomes of the treatment.

The role of hyperbaric oxygen therapy and antioxidant administration in treatment of idiopathic male infertility

Introduction. In clinical andrology practice, the cause of infertility in 40–70 % cases is idiopathic male infertility. The treatment involves empirical therapy based on the knowledge of spermatogenesis. In recent years, dietary supplements are used as the first line therapy of male infertility. Among these multicomponent dietary supplements are combination supplement for increasing male fertility Speroton and antioxidant complex Sinergin (Acvion, Russia). The study objective is to optimize the results of hyperbaric oxygen therapy (HBOT) in combination treatment of patients with idiopathic abnormalities in semen analysis. Materials and methods. The study included 155 males aged 20–45 years. Among them, 120 males were diagnosed with idiopathic infertility, 35 were practically healthy (control group). Patients with infertility (n = 120) were divided into 3 groups, 40 patients each. In all 3 groups, patients received HBOT in combination with Speroton and/or Sinergin: the 1st group (n = 40) received Speroton (1 sachet/day); the 2 nd group (n = 40) received HBOT and Sinergin (2 capsules/day); patients in the 3 rd group received combination of HBOT + Speroton (one sachet/day) + Sinergin (2 capsules/day). Patients in the 3 groups continued to receive the supplements 3 months after the completion of HBOT. The HBOT course consisted of 10 60-minute daily sessions. In patients of the 4 groups, including the control group, the levels of oxidative stress products and endogenous antioxidants activity in plasma and ejaculate were evaluated, as well as the main semen characteristics. Results. The administered therapies normalized thiol-disulfide metabolism in plasma and semen, antioxidant protection, and the main semen characteristics. In patients of the 1st group (HBOT + Speroton), sperm count in the ejaculate increased 3-fold compared to the baseline (before treatment), motile sperm forms increased 2.4-fold, the number of degenerated sperm cells decreased 1.3-fold. In the 2 nd group (HBOT + Sinergin), sperm count in the ejaculate increased 2.2-fold in the course of treatment, motile sperm count increased 2.7-fold, the number of degenerated sperm cells decreased 1.4-fold. In the 3 rd group (HBOT + Speroton + Sinergin), sperm count in the ejaculate increased 4.7-fold, motile sperm count – 5.3-fold, the number of degenerated sperm cells decreased 1.8- fold. Conclusion. Combined administration of Speroton and Sinergin in combination with HBOT has shown high effectiveness in correction of the main ejaculate characteristics and can be recommended as a therapy for various forms of asthenozoospermia.

Histological, immunohistochemical, and morphometric study of peritubular cells and mast cells in the testicular interstitium of azoospermic men

Background Infertility continues to be a highly prevalent condition. Although assisted reproduction technology has been significantly effective, it is relatively costly and may yield disappointing clinical outcomes. There is still an unexplained correlation between the substantial contribution of the testicular somatic cells such as the peritubular cells and interstitial immune cells such as mast cells. Aim The aim of this work was to study the histological, immunohistochemical, and morphometric profiles of peritubular cells and mast cells in azoospermic men that might be a clue for male infertility therapy. Patients and methods This study was carried out on 39 infertile azoospermic men attending the Andrology Outpatient Clinic in Assiut University Hospital. Patients were divided into two groups. Group I (GI), which included patients with obstructive azoospermia (n=9), and group II (GII), which included patients with nonobstructive azoospermia. GII was subdivided as follows: subgroup A, which included patients with Sertoli cell-only syndrome (n=9); subgroup B, which included patients with germ cell arrest (n=10); and subgroup C, which included patients with hypospermatogenesis (n=11). Testicular biopsies were obtained and processed for light, immunohistochemical, morphometric analysis, and electron microscopic examinations. Results The testicular interstitium of GI showed an apparently normal pattern of organization as regards cellular and fibrous components. GIIA showed the highest mast cell number, which established cytoplasmic projections toward peritubular cells. The mean count of tumor necrosis factor α-positive cells showed a significant increase in GIIA compared with GI. The mean area% of collagen fibers increased significantly in GIIA, GIIB, and GIIC compared with GI. In GIIB, bundles of microfibrils were most evident between the basement membrane and the first layer of peritubular cells. In GIIC, they appeared flattened with slender interlacing processes forming bridges. Mast cells bore long cytoplasmic projections, which extended toward the neighboring fibroblasts. Conclusion The present data signify complex cell interactions in cases of testicular dysfunction within the interstitial compartment.

Asymptomatic Infection With Mycoplasma hominis Negatively Affects Semen Parameters and Leads to Male Infertility as Confirmed by Improved Semen Parameters After Antibiotic Treatment

To elucidate the association between asymptomatic infections caused by Mycoplasma hominis and male infertility and to evaluate the role of antibiotic therapy in the treatment of this failure. A total of 165 infertile men having abnormal semen parameters (study group) as well as 165 healthy fertile men (control group) were included in this study. Semen samples were taken from all participants and, after analyzing for semen parameters, real-time polymerase chain reaction, microbial culture, and reactive oxygen species (ROS) as well as total antioxidant capacity (TAC) assays were performed. Infected individuals of the study group were treated with antibiotic. One month after the treatment completion, second semen samples were taken and all the tests mentioned were performed. The data were analyzed using SPSS statistical software, version 22.0. The frequency of M. hominis was significantly higher in the infertile men compared with the fertile ones (14.5% vs 3.6%, P = .001). The mean cycle threshold (Ct) value was lower in infected infertile men than in infected fertile men (P < .001). All semen parameters, except volume, pH, and viscosity, were improved (P < .05), most of which reached their normal range; leukocytes in seminal fluid were eliminated (P = .04); the level of TAC elevated (P < .001); and the ROS level as well as the ROS-to-TAC ratio reduced after antibiotic treatment (P = .02). Moreover, wives of 14 infected infertile men (58.3%) became pregnant 4 months after the treatment completion. Our data suggest that asymptomatic infection caused by M. hominis is correlated with male infertility and antibiotic therapy can improve the semen quality and fairly treat the male infertility.

Stem Cell Therapy for Male Infertility: A Review

Infertility affects 15% of couples in reproductive age world-wide and male factor is solely responsible in about 50% of the cases and contributory in 30-40% of cases. Spermatogonial stem cells (SSCs) are small self-renewing cells found in the basal compartment of seminiferous tubules where they form the foundation of spermatogenesis and are required for the continuous production of sperm. Transplantation of spermatogonial stem cells results in a donor derived sperm production and fertility in rodent and non-rodent species like Goat. Transplantation of cryopreserved spermatogonial stem cells could help oncology patients undergoing radiotherapy or chemotherapy by prior isolation of the SSCs and transplantation after treatment. Induced pluripotent stem cells also have the ability to differentiate into late stage germ cells. The efficacy and safety of SSCs transplantation showed that offspring produced did not show any morphological differences or alteration of genetic material but is most effective through assisted reproduction and better on young/immature Animals. The transfer of germ cells through micro-injection of seminiferous tubules and cannulation of efferent ducts is most effective on rodent testis while injection into the rete testis guided by ultrasound was reported to be the best technique in higher Animals (Bulls and Apes). Researches are still ongoing to get a safe and successful method of SSCs transplantation with no or less side effects on humans.Keywords: Cryopreserved, Pluripotent, self-renewing, Spermatogonia, and Spermatogenesis

MOLECULAR AND METABOLIC ASPECTS OF MALE INFERTILITY

In a review on the basis of own and literary data the molecular and metabolic conditions of idiopathic infertility in men are considered. Key features of the metabolism and energy in the ejaculate are revealed. Cause-and-effect relationship between the parameters of seminal plasma metabolism and morphofunctional characteristics of spermatozoa, the violation of which is accompanied by a reduction in fertility potential are described. The necessity of increasing scientific research in this field in order to optimize the search of molecular targets and the development of targeted therapy of male infertility is substantiated.

The effect of R-(-)-deprenyl administration on antioxidant enzymes in rat testis

The aim of the study was to investigate the effect of R-(-)-deprenyl administration on the activity and localization of superoxide dismutases (SODs) and catalase (CAT) in rat testis. After 30 days of intraperitoneal administration of either saline (control) or R-(-)-deprenyl dissolved in saline at concentrations of 0.0025mg/kg (low dose of deprenyl, LDD) or 0.25mg/kg (high dose of deprenyl, HDD), males were killed by thiopental, and their testes were collected. We found that deprenyl administration significantly increased the activity of antioxidant enzymes, and this effect varied by dosage. LDD caused significant elevation of all monitored enzymes, but HDD did not increase the activity of SOD2. Employing immunohistochemistry, we detected enzymes predominantly in Leydig cells (SOD1, SOD2, CAT), in late spermatids and residual bodies (SOD1, SOD2), and in primary spermatocytes (SOD2). Histopathological examination did not reveal testicular damage in experimental groups compared to control. Deprenyl proved to be a potent stimulator of antioxidant enzymes in rat testes; therefore, it could be used in the therapy of male infertility. On the other hand, it is crucial to choose a proper dose, since lower dose was more competent compared to a dosage that was one hundred times higher.

Identification of New Epididymal Luminal Fluid Proteins Involved in Sperm Maturation in Infertile Rats Treated by Dutasteride Using iTRAQ.

Background: Spermatozoa become mature and acquire fertilizing capacity during their passage through the epididymal lumen. In this study, we identified new epididymal luminal fluid proteins involved in sperm maturation in infertile rats by dutasteride, a dual 5α-reductase inhibitor, in order to provide potential epididymal targets for new contraceptives and infertility treatment. Methods: Male rats were treated with dutasteride for 28 consecutive days. We observed the protein expression profiles in the epididymal luminal fluids in infertile and normal rats using isobaric tags for relative and absolute quantitation (iTRAQ) technique. The confidence of proteome data was validated by enzyme-linked immunosorbent assays. Results: 1045 proteins were tested, and 23 of them presented different expression profiling in the infertile and normal rats. The seven proteins were down-regulated, and 16 proteins were up-regulated. Among the seven proteins which were significantly down-regulated by dutasteride in the epididymal luminal fluids, there were three β-defensins (Defb2, Defb18 and Defb39), which maybe the key proteins involved in epididymal sperm maturation and male fertility. Conclusions: We report for the first time that dutasteride influences the protein expression profiling in the epididymal luminal fluids of rats, and this result provides some new epididymal targets for male contraception and infertility therapy.

Current medical management of endocrine-related male infertility.

Male factor contributes to 50%–60% of overall infertility but is solely responsible in only 20% of couples. Although most male factor infertility is ascertained from an abnormal semen analysis, other male factors can be contributory especially if the sample returns normal. Male infertility can be due to identifiable hormonal or anatomical etiologies that may be reversible or irreversible. This manuscript will highlight existing guidelines and our recommendations for hormone evaluation for male infertility and empiric therapies including multivitamins, estrogen receptor modulators (clomiphene), estrogen conversion blockers (anastrozole), and hormone replacement.

Successful Treatment of Oligoasthenozoospermia Using Traditional Korean Medicine Resulting in Spontaneous Pregnancy: Two Case Reports

Male factor subfertility accounts for at least half of all cases of subfertility. In 30-45% of cases, the cause of abnormal semen parameters is idiopathic; however, a standardized approach for the treatment of idiopathic male infertility remains elusive. In Korea, acupuncture and/or herbal remedies have been widely used to treat male infertility. Herein, we report two patients with oligoasthenozoospermia treated using traditional Korean medicine (TKM), who subsequently successfully impregnated their spouses. The sperm count and motility of patient 1 were 10 million/ml and 15%. The sperm count and motility of patient 2 were 8 million/ml and 3%. Both patients were treated using TKM therapy including acupuncture and/or herbal medicine for two months (patient 1) and five months (patient 2). After TKM therapy, both patients reported that their spouses were pregnant and subsequently gave birth to healthy, full-term infants. This study represents TKM therapy including acupuncture and/or herbal remedies might be an option for infertile men (oligoasthenozoospermia). Further large-scale randomized controlled trials investigating the effects of TKM therapy for male infertility are needed.

Deletion of the tyrosine phosphatase Shp2 in Sertoli cells causes infertility in mice.

The male’s ability to reproduce is completely dependent on Sertoli cells. However, the mechanisms governing the functional integrity of Sertoli cells have remained largely unexplored. Here, we demonstrate that deletion of Shp2 in Sertoli cells results in infertility in mice. In Shp2 knockout mice (SCSKO), a normal population of Sertoli cells was observed, but the blood-testis barrier (BTB) was not formed. Shp2 ablation initiated the untimely and excessive differentiation of spermatogonial stem cells (SSCs) by disturbing the expression of paracrine factors. As a consequence, the process of spermatogenesis was disrupted, and the germ cells were depleted. Furthermore, Shp2 deletion impaired the cell junctions of the primary Sertoli cells and failed to support the clonal formation of SSCs co-cultured with SCSKO Sertoli cells. As expected, Shp2 restoration largely restores the cell junctions of the primary Sertoli cells and the clonal formation of SSCs. To identify the underlying mechanism, we further demonstrated that the absence of Shp2 suppressed Erk phosphorylation, and thus, the expression of follicle-stimulating hormone (FSH)- and testosterone-induced target genes. These results collectively suggest that Shp2 is a critical signaling protein that is required to maintain Sertoli cell function and could serve as a novel target for male infertility therapies.

Editorial Commentary

No AccessUrology PracticePatient Care1 Jul 2015Editorial Commentary Ranjith Ramasamy, Jason M. Scovell, and Larry I. Lipshultz Ranjith RamasamyRanjith Ramasamy More articles by this author , Jason M. ScovellJason M. Scovell More articles by this author , and Larry I. LipshultzLarry I. Lipshultz More articles by this author View All Author Informationhttps://doi.org/10.1016/j.urpr.2014.10.021AboutFull TextPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail "Editorial Commentary." Urology Practice, 2(4), pp. 204–205 References 1 : The use of clomiphene citrate in male infertility. J Urol1984; 131: 425. Link, Google Scholar 2 : Drug therapy for idiopathic male infertility: rationale versus evidence. J Urol2006; 176: 1307. Link, Google Scholar 3 : WITHDRAWN: Clomiphene or tamoxifen for idiopathic oligo/asthenospermia. Cochrane Database Syst Rev1996; : CD000151. Google Scholar 4 : Enclomiphene citrate stimulates testosterone production while preventing oligospermia: a randomized phase II clinical trial comparing topical testosterone. Fertil Steril2014; 102: 720. Google Scholar © 2015 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 2Issue 4July 2015Page: 204-205 Advertisement Copyright & Permissions© 2015 by American Urological Association Education and Research, Inc.MetricsAuthor Information Ranjith Ramasamy More articles by this author Jason M. Scovell More articles by this author Larry I. Lipshultz More articles by this author Expand All Advertisement PDF downloadLoading ...

Sertoli cell-mediated differentiation of male germ cell-like cells from human umbilical cord Wharton's jelly-derived mesenchymal stem cells in an in vitro co-culture system.

BackgroundMicroenvironment signals play a critical role in directing the differentiation of stem cells. Sertoli cells (SCs) provide a unique microenvironment that is essential for germ cell differentiation.MethodsOur previous study has demonstrated that human umbilical cord Wharton’s jelly-derived mesenchymal stem cells (HUMSCs) could differentiate towards male germ cells in vitro, but HUMSC-derived germ-like cells expressed only few germ cell markers. The aim of this study was to investigate the effect of SCs on the differentiation of HUMSCs towards male germ cells using a co-culture system that mimicked the in vivo male germ cell microenvironment.ResultsHUMSCs formed clump-like features on SC monolayers after seeding for 3 weeks. Differentiated cells formed round colonies that share the morphological features of spermatogonial colonies. RT-PCR, immunofluorescence, confocal microscopy, and Western blot analyses revealed the expression of early germ cell markers STELLA and VASA and male germ cell-specific marker DAZL in differentiated HUMSCs, confirming the presence of cells with characteristics of male germ cells.ConclusionThe HUMSC-SC co-culture system mimics a native microenvironment for germ cell colonization without any in vitro artificial manipulation and can be used to explore the mechanisms controlling the differentiation of male germ cells from HUMSCs. Male germ cells derived from HUMSCs may be used in the therapy for male infertility.

Male Infertility: Present and Future

During the last decade, a sudden decline in human male fertility has become a global concern. Genetic, endocrine and life style related problems are generally considered to be the underlie causes. However, several cases have been reported where hormones fail to promote spermatogenesis. The reason of such reproductive malfunctioning leading to male infertility remains idiopathic. Therefore, a sincere attention with a coordinated multidisciplinary effort by the andrologists, urologists and basic scientists is needed to understand the regulation of the sperm production. Powerful tools like high through put gene expression analyses, genetically manipulated mouse models and tissue culture or transplantation based assays are now available, increasing the scope of a establishing a special line of therapy for idiopathic male infertility. In this review, we have briefly summarized various categories of male infertility with emphasis on current research advancement in this field.

A new model to study the effects of gonadotropins on an “in vitro” prepubertal artificial porcine mini-testis

At present, there is no reliable experimental model “in vitro” to analyze the complex interactions between gonadotropins on the pre-pubertal Sertoli cells (SC) and Leydig cells (LD). Considering that, in the pre-pubertal period, only the anti-mullerian hormone (AMH) is upregulated by FSH and down-regulated by androgens [1-2], AMH could be considered a potential marker of pre-pubertal testis function. The aim of our work was to study the effects of FSH, LH and HCG on an in-vitro model of “mini-testis”. SC and LD, obtained from 15-20 days old neonatal pigs, were isolated and evaluated in terms of purity by AMH (unique pre-pubertal SCs marker), INSL3 (LD marker), ASMI (peritubular cells marker) and PGP9.5 (gonocytes and spermatogonial cells marker). Finally, purified SC and LD were co-cultured to obtain the “mini-testis” and were stimulated with gonadotropins. We have then evaluated: a) AMH, inhibin B and testosterone levels released in the culture medium (by ELISA), both in basal conditions and after stimulations; b) analysis of the follicle-stimulating hormone receptor (FSHR), MAPkinasi (Erk1/2, AKT) by Real Time PCR. We show an increase in inhibin B levels after FSH and FSH/LH stimulation and a selectively increase in testosterone production after LH treatment. AMH secretion was downregulated by FSH treatment. These data seem to preliminarily suggest that ERK1/ ERK2 expression was up-regulated by FSH and FSH/LH stimulation while FSHreceptor expression was down-regulated by FSH and increased by FSH/LH treatment; AKT was up-regulated in all conditions. The proposed model, by creating an artificial mini- testis, could help better understanding the complex and still partially unknown interactions between human gonadotropins, SC and LD possibly creating a novel background to shed light inside a future therapy of male infertility

Androgenic effect of honeybee drone milk in castrated rats: Roles of methyl palmitate and methyl oleate

Ethnopharmacological relevanceNumerous honeybee (Apis mellifera) products have been used in traditional medicine to treat infertility and to increase vitality in both men and women. Drone milk (DM) is a relatively little-known honeybee product with a putative sexual hormone effect. The oestrogenic effect of a fraction of DM has recently been reported in rats. However, no information is available on the androgenic effects of DM. The purpose of the present study was to determine the androgen-like effect of DM in male rats and to identify effective compounds. Materials and methodsA modified Hershberger assay was used to investigate the androgenic effect of crude DM, and the plasma level of testosterone was measured. The prostatic mRNA and protein expression of Spot14-like androgen-inducible protein (SLAP) were also examined with real-time PCR and Western blot techniques. GC–MS and NMR spectroscopic investigations were performed to identify the active components gained by bioactivity-guided fractionation. ResultsThe crude DM increased the relative weights of the androgen-dependent organs and the plasma testosterone level in castrated rats and these actions were flutamide-sensitive. DM increased the tissue mRNA and protein level of SLAP, providing further evidence of its androgen-like character. After bioactivity-guided fractionation, two fatty acid esters, methyl palmitate (MP) and methyl oleate (MO), were identified as active compounds. MP alone showed an androgenic effect, whereas MO increased the weight of androgen-sensitive tissues and the plasma testosterone level only in combination. ConclusionThe experimental data of DM and its active compounds (MO and MP) show androgenic activity confirming the traditional usage of DM. DM or MP or/and MO treatments may project a natural mode for the therapy of male infertility.

Empirical medical therapy in idiopathic male infertility: Promise or panacea?

Male factors account for 20%-50% of cases of infertility and in 25% of cases, the etiology of male infertility is unknown. Effective treatments are well-established for hypogonadotropic hypogonadism, male accessory gland infection, retrograde ejaculation, and positive antisperm antibody. However, the appropriate treatment for idiopathic male infertility is unclear. Empirical medical treatment (EMT) has been used in men with idiopathic infertility and can be divided into two categories based on the mode of action: hormonal treatment and antioxidant supplementation. Hormonal medications consist of gonadotropins, androgens, estrogen receptor blockers, and aromatase inhibitors. Antioxidants such as vitamins, zinc, and carnitines have also been widely used to reduce oxidative stress-induced spermatozoa damage. Although scientifically acceptable evidence of EMT is limited because of the lack of large, randomized, controlled studies, recent systematic reviews with meta-analyses have shown that the administration of gonadotropins, anti-estrogens, and oral antioxidants results in a significant increase in the live birth rate compared with control treatments. Therefore, all physicians who treat infertility should bear in mind that EMT can improve semen parameters and subsequent fertility potential through natural intercourse.

A simple method for isolation, culture, and in vitro maintenance of chicken spermatogonial stem cells

Spermatogonial stem cells (SSCs) are expected to participate in male infertility therapy, endangered species preservation, and transgenic animal technology by their unique unipotency to differentiate into spermatozoa. The main challenges, however, remain to be addressed including the appropriate conditions to reach good number of these cells and how to derive, culture, and maintain them in vitro. In the present study, the testicular tissues were isolated from 1-d-old male chickens to establish primary cell cultures. This culture led to development of distinguished colonies which were further characterized by alkaline phosphatase (AP) activity assay and gene expression analysis. They were shown to be positive for AP activity and expressed two main transcription factors of OCT4 and STRA8 as indicated by reverse transcription-polymerase chain reaction. These were indications of carrying characteristics of SSCs by these colonies. The cultures were also exposed to different concentrations of glial cell line-derived neurotrophic factor (GDNF), basic fibroblast growth factor (bFGF), and leukemia inhibitory factor (LIF) growth factors to seek optimum colony-forming conditions. Colony-forming activity assay indicated that they were able to propagate in vitro with an increased self-renewal property when cultured in the presence of 15 ng/mL of GDNF, 20 ng/mL of bFGF, and 15 ng/mL of LIF. The present work provides an easy and practical method for isolation, culture, and in vitro maintenance of chicken spermatogonial stem cells and introduces appropriate cell culture conditions to improve and maintain their self-renewal property based on supplying the necessary growth factors.