HLA-DQB1 as a potential prognostic biomarker of hormonal therapy in patients with non-obstructive azoospermia.

The gonadotropin treatment of infertile men may improve spermatogenesis and lead to sperm cell production, however, only a small fraction of treated patients positively responds to such therapy. To identify individual treatment prognostic biomarkers associated with responsiveness to gonadotropins, we compared the gene expression profiles of testicular oligobiopsies from 3 patients with non-obstructive azoospermia (NOA) who positively responded to therapy with a combination of human chorionic gonadotropin and recombinant follicle-stimulating hormone (hCG/rFSH) to those of 3 non-responders. We used Affymetrix Human Gene 1.

Effects of knockout on energy chain transportation and spermatogenesis: implications for male infertility.

Study Question: Is the mutation causative for male infertility?

Summary Answer: Our collected data underline the complex and devastating effect of the single-gene mutation on the testicular molecular network, leading to male reproductive failure.

What Is Known Already: Recent data have revealed mutations in genes related to axonemal dynein arms as causative for morphology and motility abnormalities in spermatozoa of infertile males, including dysplasia of fibrous sheath (DFS) and multiple morphological abnormalities in the sperm flagella (MMAF). The nexin-dynein regulatory complex (N-DRC) coordinates the dynein arm activity and is built from the DRC1-DRC7 proteins.

The role of the intrinsic pathway of apoptosis in human ejaculated sperm damage under a state of scrotal heat stress.

Purpose: The study aimed to determine the associations among standard sperm characteristics and oxidative/apoptotic markers in ejaculated sperm of men exposed to prolonged scrotal hyperthermia of either environmental or clinical origin.

Methods: The original study design included four research groups: professional drivers (n = 54), infertile men with varicocele (n = 78), infertile men not exposed to prolonged genital heat stress (n = 37), and fertile individuals serving as the control group (n = 29). Standard semen analysis was performed according to the 5th WHO laboratory manual.

Proteomic approach towards identification of seminal fluid biomarkers from individuals with severe oligozoospermia, cryptozoospermia and non-obstructive azoospermia: a pilot study.

Background: Infertility becomes a global problem that affects to the same extent females and males. As reasons of male infertility can differ among individuals, the accurate diagnostics is essential for effective treatment. The most problematic both in diagnostics and in treatment are disturbances of spermatogenesis.

Global 5mC and 5hmC DNA Levels in Human Sperm Subpopulations with Differentially Protaminated Chromatin in Normo- and Oligoasthenozoospermic Males.

Epigenetic modifications play a special role in the male infertility aetiology. Published data indicate the link between sperm quality and sperm chromatin protamination. This study aimed to determine the relationship between methylation (5mC) and hydroxymethylation (5hmC) in sperm DNA, with respect to sperm chromatin protamination in three subpopulations of fertile normozoospermic controls and infertile patients with oligo-/oligoasthenozoospermia.

Human live spermatozoa morphology assessment using digital holographic microscopy.

Digital holographic microscopy (DHM) was applied for the morphological assessment of live intact spermatozoa from fertile and infertile men directly after semen liquefaction. This method allowed us to study the sperm population directly from the sample droplet and not only from the focal plane of the microscope as in classical optical microscopy. The newly implemented 3-dimensional sperm morphological parameters (head height, acrosome/nucleus height, head/midpiece height) were included in morphological assessment of semen samples from fertile and infertile individuals.

The Role of Seminal Oxidative Stress Scavenging System in the Pathogenesis of Sperm DNA Damage in Men Exposed and Not Exposed to Genital Heat Stress.

Responding to the need for the verification of some experimental animal studies showing the involvement of oxidative stress in germ cell damage in the heat-induced testis, we investigated the possibility of a direct relationship between seminal oxidative stress markers (total antioxidant capacity, catalase activity, superoxide dismutase activity, and malondialdehyde concentration) and ejaculated sperm chromatin/DNA integrity (DNA fragmentation and chromatin condensation abnormalities) in distinct groups of men exposed and not exposed to prolonged scrotal hyperthermia. A statistical increase in the proportion of sperm with DNA fragmentation was observed in all the studied subgroups compared to the fertile men. In turn, the groups subjected to heat stress as professional drivers or infertile men with varicocele presented greater disturbances in the oxidative stress scavenging system than men not exposed to genital heat stress.

Evaluation of seminal plasma HSPA2 protein as a biomarker of human spermatogenesis status.

In mammals, testicular Heat shock-related 70 kDa protein 2 (HSPA2) is a chaperon strictly linked to spermatogenesis status, whereas its presence in spermatozoa ensures successful oocyte fertilization. However, there is little information on this protein in seminal plasma in infertile males. Based on our previous two independent studies, we have selected HSPA2 to evaluate this seminal plasma protein is a potential biomarker of correct spermatogenesis.

Seminal Plasma Analysis of Oxidative Stress in Different Genitourinary Topographical Regions Involved in Reproductive Tract Disorders Associated with Genital Heat Stress.

The pathophysiological mechanisms responsible for male subfertility/infertility caused by or complicated by genital heat stress remains unclear in many respects. Because seminal plasma creates the environment for the proper functioning of spermatozoa, in this study, we verified the associations among standard spermiograms, seminal biochemical parameters (neutral alpha-glucosidase, fructose, and citric acid) and oxidative stress markers (total antioxidant capacity, catalase activity, superoxide dismutase activity, and malondialdehyde concentration) in distinct entities associated with male infertility with and without long-time exposure to local hyperthermia. We demonstrated that men exposed to prolonged environmental or clinically recognized local heat stress in adulthood may suffer from dysregulation of seminal antioxidant components, which can be directly associated with epididymal and prostate function.

Human sperm proteins identified by 2-dimensional electrophoresis and mass spectrometry and their relevance to a transcriptomic analysis.

The aim of this study was to identify and analyse human sperm proteins from normozoospermic men using 2-dimensional electrophoresis (2-DE) and mass spectrometry (MS). We identified 73 different sperm proteins, including two less characterized human sperm proteins, Annexin A7 (ANXA7) and c14orf105. Bioinformatic analysis of detected sperm proteins revealed new carbohydrate and lipid metabolic pathways, which supply energy to motile sperm.

Killer cell immunoglobulin-like receptor gene association with cryptorchidism.

Cryptorchidism is a condition where a testis persists in the abdominal cavity. Thus, due to elevated temperature we may expect induction of aberrant immune reactions depending on genetic constitution of individual. This may be reflected by development of anti-sperm antibodies (ASA) in cryptorchid males.

Semen Quality, Hormonal Levels, and Androgen Receptor Gene Polymorphisms in a Population of Young Male Volunteers from Two Different Regions of Poland.

Background: The population of healthy Polish men has not been frequently and systematically investigated for fertility status. The aim of this study was to assess the quality of semen in a randomly recruited population of young males. The most important task was to find a relationship between semen parameters, sex hormones, and AR gene polymorphism.

Weak association of anti-sperm antibodies and strong association of familial cryptorchidism/infertility with HLA-DRB1 polymorphisms in prepubertal Ukrainian boys.

Background: Cryptorchidism is a frequent syndrome occurring in 1-2% of males within the first year of age. Autoimmune reactions, particularly directed to testicular elements and/or spermatozoa have been found to be often associated with cryptorchidism. Therefore we investigated in this study the frequency of HLA class II alleles in order to recognize possible genetic predisposition for antisperm antibodies development in prepubertal boys with diagnosed cryptorchidism in Caucasoid population.

An isoimmune response to human sperm clathrin in an infertile woman with systemic lupus erythematosus.

We have employed a proteomic approach to study the immune response to human sperm in an infertile female patient suffering from systemic lupus erythematosus (SLE). Human sperm antigenic extracts were resolved by means of two-dimensional electrophoresis and electroblotted onto nitrocellulose membranes. The membranes were incubated with serum from the SLE patient.

[Flow cytometry application in the evaluation of antisperm antibodies in sera samples of infertile people and prepubertal boys with gonadal disorders].

Objectives: The aim of the following study was to assess antisperm antibodies in sera samples of infertile men and women, as well as from prepubertal boys by means of flow cytometry.

Material And Methods: We tested sera samples of infertile and fertile adult populations, prepubertal boys with gonadal disorders and healthy prepubertal boys. The indirect immunobead test and flow cytometry were used to detect antisperm antibodies.

Cryptorchidism and long-term consequences.

Cryptorchidism has been on the rise for several decades and can be observed with frequency of 1-2% of males within the first year of age. It may appear as an isolated disorder or can be a consequence of genetic and endocrine abnormalities connected with somatic anomalies. Its genetic background still seems to be unclear although a range of genes can be responsible for the development of this syndrome.

In situ reconstruction of humoral immune response against sperm: comparison of SCID and NOD/SCID mouse models.

Problem: Comparison of two types of immunocompromised mouse strains (SCID and NOD/SCID) for production of human antisperm antibodies (AsA).

Method Of Study: Human peripheral blood lymphocytes (PBL) were grafted to mouse peritoneal cavity and sensitized with natively glycosylated and N-deglycosylated sperm extracts.

Results And Conclusion: NOD/SCID mice inoculated with hu-PBLs exhibited higher AsA titres with a tendency for greater sperm agglutination than human AsA raised in SCID mouse model.

Specific Fab fragments recovered by phage display technique recognizing human spermatozoa.

Human hybridoma cell lines are often unstable and loose ability for antibody production. Sometimes, they show low and varying levels of heavy and light chains synthesis. Therefore it is reasonable to preserve generated specificities of light and heavy chains by cloning them to phagemid vector and creating phage display library.

Proinflammatory cytokines as an intermediate factor enhancing lipid sperm membrane peroxidation in in vitro conditions.

We have examined the effect of white blood cells (WBCs), various proinflammatory cytokines, or a combination of the two on the peroxidation of human sperm membrane lipids in in vitro conditions. Six recombinant cytokines, such as interleukin-1beta (IL-1beta), IL-6, IL-8, IL-12, IL-18, and tumor necrosis factor alpha (TNF-alpha), used singly or in combinations, were analyzed. WBCs were isolated from the whole heparinized blood using a density gradient technique (Histopaque 1.

Bacteria trigger oxygen radical release and sperm lipid peroxidation in in vitro model of semen inflammation.

Objective: To assess the in vitro effect of five bacterial strains isolated from semen samples (Escherichia coli, Staphylococcus haemolyticus, Streptococcus oralis, Bacteroides ureolyticus, and Ureaplasma urealyticum) on reactive oxygen intermediate (ROI) release and lipid sperm membrane peroxidation in the coincubated suspensions of white blood cells (WBC) with spermatozoa.

Design: An in vitro model of semen infection.

Setting: Basic research laboratory.

Antisperm antibodies in prepubertal boys with cryptorchidism.

The presence of antisperm antibodies in male individuals before puberty is controversial due to the lack of finally differentiated male germ cells. It was questioned whether the pathologic conditions of the male gonad may influence antisperm antibody formation in individuals before puberty. Sera samples of 76 individuals and 10 healthy boys with testicular failure (mainly uni- or bilateral cryptorchidism) were examined by means of indirect immunobead-binding test (IDIBT).

Peritoneal fluid cytokines and sICAM-1 in minimal endometriosis: search for discriminating factors between infertility and/or endometriosis.

Objective: To evaluate cytokine levels (IL-1beta, TNF-alpha, IL-6, IL-8), soluble intercellular adhesion molecule-1 (sICAM-1) and number of macrophages in peritoneal fluid (PF) of women with no minimal endometriosis and associated (or not) infertility in order to discriminate between infertility and/or endometriosis.

Study Design: Cytokines and sICAM-1 were measured by using quantitative enzyme-linked immunosorbent assay (ELISA) while the macrophages were identified by May-Grunwald-Giemsa and non-specific esterase staining and presented as medians. The measurements were performed in 78 women belonging to four selected subgroups according to their endometriosis and/or infertility status.

The frequency of antisperm antibodies in infertile couples--a Polish pilot study.

Background: One of every four Polish married couples is infertile. Numerous factors may contribute to the lack of conception in these couples, one of which may be the presence of antisperm antibodies in semen, cervical mucus and serum in infertile individuals. Thus immunological examinations of these individuals should be standard diagnostic practice.

Infertility status of male individuals with abnormal spermiogram evaluated by cytogenetic analysis and in vitro sperm penetration assay.

Background: The possibility of a link between altered sperm morphology and functional ability, possibly reflected on the genetic level, is still a matter of controversy.

Material/methods: 60 infertile males with pathological spermiogram and 14 healthy individuals with confirmed in vivo fertility were selected for our study. Ejaculated sperm was assessed according to the standard criteria set by the World Health Organization, which constituted a basis for subgroup classification.