Reproductive System Disorders Research Articles

Seminal plasma as a diagnostic fluid for male reproductive system disorders

Molecular biomarkers hold promise to advance the noninvasive diagnosis of male reproductive system disorders and facilitate the identification and management of these conditions through screening, early diagnosis and more accurate prognosis. Seminal plasma has great potential as a proximal fluid for protein biomarker discovery and as a clinical sample for noninvasive diagnostics. The seminal plasma proteome contains thousands of proteins and includes a large number of tissue-specific proteins that might accurately indicate a pathological process in the tissue of origin. Potential protein biomarkers for male reproductive system disorders are more abundant in seminal plasma than in blood serum or urine, and, therefore, are more easily identified and quantified in semen by mass spectrometry and other techniques. These methods have enabled elaboration of the composition of the seminal plasma proteome and the tissue specificity of seminal plasma proteins. Strategies have been developed to discover protein biomarkers in seminal plasma through integrated 'omics' approaches. Biomarkers of male infertility and prostate cancer are now emerging, and it is evident that seminal plasma has the potential to complement other diagnostic tools available in urology clinics.

The transcriptomic and proteomic effects of ectopic overexpression of miR-30d in human endometrial epithelial cells

miR-30d is known to be up-regulated during the acquisition of receptivity in the endometrium. In order to determine the transcriptomic and proteomic changes which occur after transient overexpression of miR-30d in primary endometrial epithelial cells, in vitro cultured human endometrial epithelial cells (hEECs) were studied experimentally. Two different miRNAs (scramble versus mimic; n = 15) were transiently transfected into primary hEECs from four different patients and were evaluated for mRNA and protein expression using Agilent's gene expression microarray and iTRAQ analysis techniques, respectively. A set of differentially expressed mRNAs were validated by qPCR and several differentially expressed proteins were validated by western blot. Finally, methylation differential immunoprecipitation (MeDIP) was used to validate the epigenetic changes in the H19 gene. The results showed that transient transfection with miR-30d miRNA induced the differential mRNA-expression of 176 genes (75 up-regulated and 101 down-regulated). Several of them have been associated with reproductive and endocrine system disorders, tissue development, and are implicated in epithelial cell proliferation. Also, the down-regulation of some genes such as H19 and N-methyltransferase (NNMT) may suggest that epigenetic alterations are induced. Furthermore, upstream effects of genes regulated by the estrogen receptor alpha 1 (ESR1) transcription factor have been predicted. Proteomic analysis identified 2290 proteins, of which 108 were differentially expressed (47 up-regulated and 61 down-regulated). Among these differentially expressed proteins DNA methyl transferase (DNMT)1 was found to be up-regulated; this protein participates in the maintenance of DNA methylation, supporting an epigenetic role for miR-30d. Finally MeDIP showed an increase in methylation in the H19 DMR region. In conclusion transient in vitro overexpression of the receptivity-up-regulated miRNA miR-30d in hEECs seems to activate genes which are associated with hormonal response and the epigenetic status of these cells.

Transcranial Physiotherapy in the Correction of Reproductive System Disorders in Adolescent Girls with Obesity

The study was undertaken to evaluate and optimize various modes of transcranial physiotherapy for reproductive system disorders in puberty girls with obesity. Combined use of transcranial magnetic therapy (TMT) and transcranial electrostimulation (TES) was substantiated by a study of the hormonal status, carbohydrate metabolism, anthropometric and clinical data, ultrasonography and electroencephalography. The application of AMO-ATOS-E apparatus for this purpose could normalize a menstrual cycle in 86,3% of the obese girls, by reducing body weight by an average of 9,26±4,28 kg for 3 months. This method was shown to ensure effective prevention of polycystic ovary syndrome. Analysis of electroencephalography indicates a baseline impairment of brain bioelectrogenesis in adolescent girls with reproductive system disorders and their correction with TMT and TES therapy.

Phytochemical, therapeutic, and ethnopharmacological overview for a traditionally important herb: Boerhavia diffusa Linn.

Boerhavia diffusa (BD) is a plant of rasayana category as per ayurvedic claims. It is reported to possess antiaging, disease prevention, and life strengthening activities which hold enormous influence in disease burden and affordability/availability of healthcare in the world. Objective. This paper has been compiled to comment on the studies reported for BD to highlight its chemical and therapeutic potential along with its ethnopharmacological considerations. Methods. In the present paper, a detailed account of chemical constituents and pharmacological activities has been presented. All the findings were correlated with modern pharmacological activities to appraise the value of BD. Results. Chemical analysis of BD gives a wide variety of chemical constituents, namely, rotenoids, flavonoids, xanthones, purine nucleoside, lignans, and steroids. Various ethnopharmacological reports emphasize its role in disorders of reproductive system, gastrointestinal system, respiratory system, urinary system, hepatic system/jaundice, cardiovascular system, and cancer. Conclusions. The studies on the therapeutic activities of BD range from studies on crude extracts to isolated compounds; however some of the studies require sophistication and validated results. BD is a plant of enormous importance in the purview of its chemical and therapeutic properties.

Natural and Preformed Physical Factors in Phase Medical Rehabilitation of Patients with Pubertal Menorrhagia

Relevance. The study presents the results of how the application of restorative treatment might be affective for people of puberty age with menstrual function. The appropriateness of the active use of resort related recourses of Krasnodar region during the medical rehabilitation of girls with reproductive system disorders has been shown in the study.

Studies on the Use of Extra-Pituitary Gonadotropins in Genital Disorders in Bitch

Normal 0 MicrosoftInternetExplorer4 Infertility in bitch is characterized by a variety of clinical manifestations depending on the disorders of reproductive system. Adjusting of ovarian hormonal activity is achieved by direct or indirect interconnection with other neuro - endocrine structures of hypothalamic- pituitary system . In the present study we aimed , based on the administration of pregnant mare's serum gonadotropin (PMSG) and human chorionic gonadotropin ( hCG ) combination , the induction of ovulatory oestrus in 10 bitches diagnosed with anoestrus . There were applied 2 therapeutic protocols . Therapeutic protocol no. 1 consisted of the administration for 9 consecutive days of 20 IU/Kg/day of PMSG, followed by the injection of an ovulatory dose of hCG - 500 IU/animal in the 10 th day . Therapeutic protocol no. 2 consisted of the administration for 5 consecutive days of 20 IU/Kg/day of PMSG, followed by the injection of an ovulatory dose of hCG - 500 IU/animal in the 6 th day. The results obtained in the two protocols were different . Rates of ovulation in bitches in the first protocol was higher compared to the ones of the second protocol .

Hypofertility factors. Role of exogenous impacts during lactation

Course of breastfeeding can cause disorders in somatic and sexual development of adult. This period is especially important for the reproductive system. Function of this system depends on the numerous endogenous and exogenous factors. Factors that have delayed manifestation actions include: stress, intake of hormone-like compounds, medications. Impact of these factors during breast feeding can lead to violations of physical and mental development, and reproductive system disorders in descendants.

Scientific journals' responsibility in shaping public health policy

The ultimate goal of a scholarly journal is to publish the highest quality scientific data, thus informing the research community and public at large about the latest findings. Because of the technical language and the required high degree of methodological detail, scientific publications are addressed first of all to other scientists. The public at large has to rely on intermediary messengers of scientific information, including scientists themselves and popular science writers, who ‘filter’ and ‘digest’ the technical data to make them ‘palatable’ for lay recipients. Among the most important and influential recipients of the scientific information are governmental and international policy makers, who are not trained to evaluate technical data, so the scientific information they use has to be first translated and summarized by advisory bodies (Brownson et al., 2009). While all areas of research are equally important, public health policy influences well-being of each of us and populations alike, and mistakes may have disastrous consequences. So it is crucially important that the health policy makers can rely on the best possible information provided by expert and impartial advisors, with rigorous adherence to the data quality. Naturally, the advisors base their opinion primarily on scientific reports published in scholarly bio-medical journals, which have to strive to maintain rigor and reproducibility, as discussed in another editorial in this issue of Andrology (Carrell & Rajpert-De Meyts, 2013). Once a while policy makers face a difficult and controversial issue, especially if the research area is novel, medical problem multifactorial and explanation of causes not straight-forward. Scientists, including those sitting in advisory bodies may have different views and interpret the data differently. An example of such a difficult issue in the public health is the recent debate on whether or not man-made endocrine disruptors (including pharmaceuticals) are responsible for changing trends in a number of diseases, such as cancer of endocrine organs, disorders of sex development, asthma, allergy, and behavioural disorders. This particular topic is of great interest for andrologists, because endocrine disruption has been implicated as one of the aetiologies of some disorders of the male reproductive system. According to recent reviews of the state of knowledge in the field prepared for international institutions and scientific societies (Zoeller et al., 2012; Bergman et al., 2013), there is growing evidence for such a causative link and there is a need for new and tightened regulation of endocrine disruptors. Where the evidence is not yet strong, one should err on the side of the health protection rather than risking exposing people. The main points of concern were recently formulated by a group of scientists in the so-called Berlaymont Declaration on Endocrine Disrupters (http://www.ipcp.ch/IPCP_Berlaymont.html), in which they appeal to the European Commission for action. The Declaration has provoked a public debate, including comments on the pages of scientific journals. One of the strongest opposing statements was signed by several expert toxicologists and has been since re-printed in a number of toxicology journals (Dietrich et al., 2013). Now a number of other scientists, mainly endocrinologists, came forward with their response which we support and have decided to publish also in this issue of Andrology (Gore et al., 2013). We believe that scientific journals and Andrology need to take part in debates that may potentially influence the health policy. We welcome our readers to comment and take part in this debate.

Clinical peptidomic analysis by a one-step direct transfer technology: Its potential utility for monitoring of pathophysiological status in female reproductive system disorders

To date, numerous studies have searched for candidate molecules or clinical examination methods as potential biomarkers for monitoring intractable diseases, such as carcinomas. Evidence accumulated over the past decade shows that many proteolytic peptides appear in human humoral fluids, including peripheral blood, in association with an individual's health condition. Although an analysis of the whole peptide (the 'peptidome') using mass spectrometry is thought to be one of the most powerful and promising experimental approaches, it has failed to identify biomarkers in the clinical blood samples, presumably due to the methodological limitations. In general, commonly used techniques for proteomic analysis of blood require the removal of large amounts of serum/plasma proteins prior to mass spectrometry analysis, and this step seems to have resulted in the overlooking of important biomarkers during the analytical process. Here, we provide a brief overview of a new quantitative peptidomic analysis by a one-step direct transfer technology without depletion of major blood proteins. Using this technology, we herein report experimental data on serum peptidomic analysis for patients with pregnancy-induced hypertension as a clinical model. In addition, we refer to the potential utility of this approach for the monitoring of pathophysiological status in female reproductive system disorders in general.

The opioid neuropeptides in uterine fibroid pseudocapsules: a putative association with cervical integrity in human reproduction

The myoma pseudocapsule (MP) is a fibro-vascular network rich of neurotransmitters, as a neurovascular bundle, surrounding fibroid and separating myoma from myometrium. We investigated the distribution of the opioid neuropeptides, as enkephalin (ENK) and oxytocin (OXT), in the nerve fibers within MP and their possible influence in human reproduction in 57 women. An histological and immunofluorescent staining of OXT and ENK was performed on nerve fibers of MP samples from the fundus, corpus and isthmian-cervical regions, with a successive morphometric quantification of OXT and ENK. None of the nerve fibers in the uterine fundus and corpus MPs contained ENK and the nerve fibers in the isthmian–cervical region demonstrated an ENK value of up to 94 ± 0.7 CU. A comparatively lower number of OXT-positive nerve fibers were found in the fundal MP (6.3 ± 0.8 CU). OXT-positive nerve fibers with OXT were marginally increased in corporal MP (15.0 ± 1.4 CU) and were substantially higher in the isthmian–cervical region MP (72.1 ± 5.1 CU) (p < 0.01). The distribution of OXY neurofibers showed a slight into the uterine corpus, while are highly present into the cervico-isthmic area, with influence on reproductive system and sexual disorders manifesting after surgical procedures on the cervix.

The influence of thyroid disorders upon the female reproductive function

The paper presents the aspects of functional disorders of the female reproductive system against a background of thyroid diseases. The frequency of reproductive disorders is found to depend on the course of endocrine diseases and hormonal disorders linked with them.

Iodine deficiency diseases and reproductive health of the women

A study of reproductive function at women with various iod deficiency diseases was held. High frequency of gynecological diseases in iodine deficiency disorders and interrelation between functional disorders of reproductive system and a thyroid gland are established. The necessity of a comprehensive study of the endocrine status, ultrasound and joint management of these patients by gynecologists and endocrinologists was shown.

Atividades antioxidante, antinociceptiva e anti-inflamatória das folhas de Mucuna pruriens (L.) DC

A espécie Mucuna pruriens (L.) DC. (Fabaceae) é popularmente conhecida no Brasil como "café berão" e "pó de mico". Suas sementes são bastante estudadas sendo empregadas na medicina popular para o tratamento de desordens nervosas e do aparelho reprodutor, porém poucas informações sobre a composição química e atividade farmacológica das folhas dessa espécie são descritas na literatura. Assim, os objetivos desse estudo foram: a determinação do perfil fitoquímico, a avaliação da atividade antioxidante in vitro, e a avaliação das atividades anti-inflamatória e antinociceptiva in vivo do extrato metanólico das folhas de M. pruriens nas concentrações de 100 e 300 mg/kg. O estudo fitoquímico foi realizado por meio da pesquisa qualitativa dos constituintes químicos e determinação quantitativa do teor de fenóis totais e flavonoides. O potencial antioxidante foi avaliado pelos métodos do DPPH e poder de redução. As atividades anti-inflamatória e antinociceptiva foram a ante foi promissor, com CI50 de 13,2∝g/mL pelo método do DPPH, e CE50 de 47,9∝g/mL pelo método que avalia o poder de redução. Em relação à atividade anti-inflamatória, o extrato foi capaz de inibir o edema de orelha em 63% (100mg/kg) e 28% (300mg/kg). Quanto à atividade antinociceptiva, houve redução do número de contorções abdominais de 64% (100 mg/kg) e 69% (300 mg/kg). Estes resultados indicam que M. pruriens apresenta um potencial farmacológico promissor e reforçam o conceito de que a pesquisa de plantas com usos etnofarmacológicos pode revelar um número substancial de respostas em ensaios in vitro e in vivo.

Human Seminal Plasma Proteome Study: a Search for Male Infertility Biomarkers

Seminal plasma is a potential source of biomarkers for many disorders of the male reproductive system including male infertility. Knowledge of the peptide and protein components of seminal fluid is accumulating especially with the appearance of high-throughput MS-based techniques. Of special interest in the field of male infertility biomarkers, is the identification and characterization of differentially expressed proteins in seminal plasma of men with normal and impaired spermatogenesis. However, the data obtained until now is still quite heterogeneous and with small percentage of overlap between independent studies. Extensive comparative analysis of seminal plasma proteome is still needed in order to establish a potential link between seminal plasma proteins and male infertility.

Ghrelin

Ghrelin is considered the endogenous ligand of growth hormone secretagogue receptor type 1a, and its modulatory actions have been demonstrated in a large array of endocrine and nonendocrine functions. According to recent studies, ghrelin seems to act at different levels of the reproductive system, exerting predominantly inhibitory effects on mammalian reproduction. It has been shown to influence the reproductive system by regulating hormone secretion from the brain and by acting directly on the gonads to affect tissue development and steroidogenesis. Thus, the endocrine network, which integrates energy balance and fertility, might involve ghrelin. Furthermore, ghrelin levels and actions have been assessed in various female reproductive system disorders. The findings could lead to a better understanding of the pathogenesis of these disorders and, possibly, to more beneficial therapeutic strategies.

An Encore for the Repeats: New Insights into an Old Genetic Variant

Throughout the human genome there are trinucleotide repeat sequences susceptible to either expansion or contraction during replication, giving rise to length polymorphisms in the general population. The polymorphic CAG repeat, which encodes an uninterrupted polyglutamine (polyQ) tract in the N-terminal transactivation domain of the androgen receptor (AR), is the most extensively studied genetic variant in individuals with disorders of the male reproductive system. Despite an impressive number of studies, the pathogenic role of this polymorphism and its clinical relevance are still a matter of debate. Although a recent meta-analysis of 33 publications (1) supports a pathogenetic role for longer polyQ length in male infertility, the authors conclude their work stating that there is a need for new, welldesigned studies (1). In fact, available data do not allow us to establish what range of AR-CAG repeat lengths predisposes impaired sperm production or to estimate the entity of the associated risk (1). Similar to other genetic variants, the literature related to CAG repeats suffers from an abundance in conflicting case-control association studies and a paucity of functional data (2). There are several plausible explanations for these apparent controversies, mostly related to: 1) poor study design (inappropriate selection of patients and controls, particularly with respect to their phenotype and their ethnic/geographic origin, and underpowered size of the study population); and 2) intrinsic complexity of the interaction between the AR and its endogenous/environmental ligands. An additional intricacy derives from the presence of another polymorphic trinucleotide repeat, (GGN)n, in the first exon of the AR gene, which may modulate the functional effect of the CAG repeat length, stressing the need for a combined analysis of the two AR polymorphisms (3, 4). It is commonly accepted that the length of the polyQ tract influences the transactivation capacity of the receptor in an inverse manner; that is, the longer the tract, the lower the activity. To support this hypothesis, a clear negative impact on AR activity is documented in relationship with pathological expansions of the repeat length (40 or more), known as the Kennedy syndrome (5). This syndrome is characterized by spinobulbar muscular atrophy and hypoandrogenism due to partial androgen insensitivity. On the other hand, controversies still exist about the effect of variations in polyQ within the normal polymorphic range. The normal distribution of the (CAG)n is reported as 6–39 repeats, with a median of 21–22 in White Caucasian, 19–20 in African-American, 22–23 in Asian, and 23 in Hispanic populations. Clinical observations showing a linear correlation between testosterone level and CAG repeat length support the notion of a functional effect of the polymorphism within the normal range. In fact, increased circulating testosterone and estradiol levels in men with a higher number of CAG repeats can be considered as a compensatory mechanism aimed to overcome the weaker AR activity (6, 7). However, such a linear correlation has not been clearly demonstrated by in vitro experiments. The first two functional studies reported that the longest tract (Q31) displayed lower activity when compared with the shortest one (Q15). However, no significant differences were observed by comparing these two types of alleles to an intermediate number of CAG repeats (20 or 24) (8, 9). Quite strikingly, two recent articles provided evidence for the lack of a stepwise reduction in activity with increasing CAG length across the polymorphic

Multi-residue analysis of free and conjugated hormones and endocrine disruptors in rat testis by QuEChERS-based extraction and LC-MS/MS

Endocrine disrupting compounds (EDCs) are suspected to be responsible for many disorders of the human reproductive system. To establish a causality relationship between exposure to endocrine disruptors and disease, experiments on animals must be performed with improved or new analytical tools. Therefore, a simple, rapid, and effective multi-residue method was developed for the determination of four steroid hormones (i.e., testosterone, androstenedione, estrone, and estradiol), glucuronide and sulfate conjugates of estrone and estradiol and four endocrine disruptors in rat testis (i.e., bisphenol A, atrazine, and active metabolites of methoxychlor and vinclozolin). The sample preparation procedure was based on the Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) approach. An analytical method was then developed to quantify these compounds at ultra-trace levels by liquid chromatography coupled to tandem mass spectrometry. The QuEChERS extraction was optimized with regard to the acetonitrile/water ratio used in the extraction step, the choice of the cleanup method and the acetonitrile/hexane ratio used in the cleanup step. The optimized extraction method exhibited recoveries between 89% and 108% for all tested compounds except the conjugates (31% to 58%). The detection limits of all compounds were below 20 ng g(-1) of wet weight of testis. The method was subsequently applied to determine the levels of hormones and EDCs in seven rat testis samples.

Proteomic analysis of seminal plasma in men with different spermatogenic impairment

Seminal plasma is a potential source of biomarkers for many disorders of the male reproductive system including male infertility. The identification and characterisation of differentially expressed proteins in seminal plasma of man with normal and impaired spermatogenesis can help in the elucidation of the molecular basis of male infertility. We compared the protein expression profiles of seminal plasma from four different groups of men as follows: normozoospermic, asthenozoospermic, oligozoospermic and azoospermic groups, using two-dimensional differential gel electrophoresis (2-D DIGE). We found eight proteins with statistically significant increased expression in azoospermia compared with at least one of the other studied groups. The differentially expressed spots were fibronectin, prostatic acid phosphatase (PAP), proteasome subunit alpha type-3, beta-2-microglobulin, galectin-3-binding protein, prolactin-inducible protein and cytosolic nonspecific dipeptidase. Notably, PAP was increased in patients with azoospermia compared with that of all other groups. We have observed no statistically significant differences in protein expression between three of the groups: normozoospermic, oligozoospermic and asthenozoospermic. We suggest that the identified panel of proteins in our study especially PAP have a strong potential to be used as azoospermia markers. However, further investigations will be necessary to validate these markers in samples of larger and independent patient cohorts and to clarify their role in the pathogenesis of male infertility.

DEPENDENCE OF THE REPRODUCTIVE FUNCTION OF SCHOOLGIRLS AGED 12–15 YEARS FROM THEIR LIFESTYLE PECULIARITIES

Judging upon the results of recent studies we can state that over the past years there is a negative tendency in reproductive health of schoolgirls aged 12–15. Study objective was to evaluate dependency of reproductive function of schoolgirls from their lifestyle. A hypothesis has been stated upon presence of causal relations in the biosocial system «Health–Lifestyle» and upon necessity of healthy lifestyle motivation to preserve girl’s reproductive health. 128 girls took part in the study from 5–7 forms from 2 Moscow schools. Results of the study stand for well-timed correction of the girl’s lifestyles in order to prevent disorders of reproductive system. m.

Purinergic signalling: Its unpopular beginning, its acceptance and its exciting future

Adenosine 5'-triphosphate (ATP) was identified in 1970 as the transmitter responsible for non-adrenergic, non-cholinergic neurotransmission in the gut and bladder and the term 'purinergic' was coined. Purinergic cotransmission was proposed in 1976 and ATP is now recognized as a cotransmitter in all nerves in the peripheral and central nervous systems. P1 (adenosine) and P2 (ATP) receptors were distinguished in 1978. Cloning of these receptors in the early 1990s was a turning point in the acceptance of the purinergic signalling hypothesis. There are both short-term purinergic signalling in neurotransmission, neuromodulation and secretion and long-term (trophic) purinergic signalling of cell proliferation, differentiation and death in development and regeneration. Much is known about the mechanisms of ATP release and its breakdown by ectonucleotidases. Recently, there has been emphasis on purinergic pathophysiology, including neurodegenerative and neuropsychiatric disorders. Purinergic therapeutic strategies are being developed for treatment of gut, kidney, bladder, lung, skeletal and reproductive system disorders, pain and cancer.