The role of staining techniques in seminological analysis of mammalian semen

The most important semen parameters are the concentration, motility and morphology of sperm cells. Sperm morphology is regarded as the most reliable parameter for predicting fertility in males. A problem in evaluating sperm morphology and morphometry is the lack of standardization of staining techniques. The staining procedure and reagents used can significantly affect the morphometric parameters of the sperm cell. The use of stains with different pH or osmotic pressure, as well as the duration of the procedure, may influence the shape and size of the sperm, and thus the result of the morphological evaluation of the semen. It is necessary to develop an evaluation procedure for sperm morphology and morphometry that will minimize the changes in the structure of the evaluated semen in relation to the native semen

Use of silver nitrate for the assessment of sperm measurements in selected farm and free-living animal species

The study was conducted on spermatozoa of selected farm and free-living animal species, isolated post mortem from the tail of the epididymis, and stained with silver nitrate – AgNO3. The material was collected from pigs, goats, wild boar, and European roe deer. Twenty morphologically normal spermatozoa randomly selected from each animal and well visible under the microscope, were analysed. The following measurements were considered: head length, width, perimeter and area, acrosome area, mid-piece length, tail length, and overall sperm length. AgNO3 staining differentiated the acrosomal (light hue) and distal (dark hue) part of the sperm head, and a light-hued mid-piece was visible within the sperm tail. Silver nitrate staining revealed species and variety-related differences, particularly in reference to the sperm head. Clear-cut differentiation within the head and tail area made it possible to perform detailed morphometric measurements of the spermatozoa

The risk of mental disorders in patients with disorders/differences of sex differentiation/development (DSD) and Y chromosome

Introduction: Patients with disorders/differences of sex differentiation/development (DSD) are exposed to physical and mental suffering. The aim of the study was to assess the following: the mental health status and the risk of mental problems in adult DSD patients, their dependence on therapeutic procedures, and to identify groups of disorders that require particular psychological support. Material and methods: The study involved 59 patients with DSD (gonadal dysgenesis — GD, androgen insensitivity syndrome — AIS, 5-alpha reductase deficiency, ovotestis), and with the Y chromosome in the karyotype, aged 16–65 years. All completed the General Health Questionnaire (GHQ-28) for the assessment of their mental health status. Raw results were converted into sten scores using norms for the Polish adult population to assess the risk of mental problems. Results: A high risk of mental problems was identified in 24% of individuals (26% men, 21% women). Women, when compared with men, displayed a significantly higher mean level of anxiety and insomnia (7.3 vs. 4.6 scores) and somatic symptoms (7.4 vs. 5.5), and worse general mental health status (25.6 vs. 18.8). The most disturbing symptoms were observed among patients with complete and partial AIS, and complete GD (general mental health status: 39.5, 24.3, and 24.2, respectively), women lacking a vagina (27.2), and without an enlarged clitoris (27.5). Patients after genital surgery had significantly fewer somatic symptoms (5.4 vs. 7.8; p < 0.05) and better general mental health status in comparison to those without surgery (20.1 vs. 24.9; p < 0.05). No significant differences were observed between patients using hormone replacement therapy and those who were not. Conclusions: The individuals with DSD and Y chromosome in the karyotype have increased risk of developing mental problems in comparison to the general Polish population. The risk factors seem to be as follows: female gender, the lack of a vagina, the lack of virilisation (no enlarged clitoris), and no genital operations performed. In some cases, sex hormone replacement therapy may be also the risk of mental problems. Particularly vulnerable groups are CAIS, PAIS, and CGD. The psychological support and an individual approach to particular needs of these patients is necessary.

Di(n-butyl) phthalate has no effect on the rat prepubertal testis despite its estrogenic activity in vitro

The aim of this study was to assess the impact of di(n-butyl) phthalate (DBP) on the rat’s prepubertal testis. Male Wistar rats were given daily subcutaneous injections with DBP (20 or 200 μg) or a vehicle from the 5th to the 15th postnatal day (pd). On the 16th pd, the rats were euthanized, and the testes were dissected, weighed, and paraffin embedded. The blood was collected to determine the serum levels of testosterone (T), estradiol (E) and FSH. The following parameters were assessed in the testis sections: diameter and length of seminiferous tubules (st), numbers of spermatogonia A + intermediate + B (A/In/B), preleptotene spermatocytes (PL), leptotene + zygotene + pachytene spermatocytes (L/Z/PA) and Sertoli cells per testis, percentage of st containing gonocytes or pachytene spermatocytes or lumen. An estrogenicity in vitro test was performed by means of a transgenic yeast strain expressing human estrogen receptor alpha. At both doses, DBP had no influence on testis and seminal vesicle weight, st diameter and length, number of germ and Sertoli cells per testis, percentage of st containing gonocytes or pachytene spermatocytes or lumen. DBP did not change E, T or FSH serum levels. The in vitro yeast screen showed that DBP was a weak estrogenic compound, approximately six to seven orders of magnitude less potent than 17β-estradiol. In conclusion, exposure of a rat to DBP in doses 100 or 1,000-fold higher than a Tolerable Daily Intake for humans had no effect on its testicular development. (Folia Histochemica et Cytobiologica 2011; Vol. 49, No. 4, pp. 685–689

Use of silver nitrate for the assessment of sperm measurements in selected farm and free-living animal species

Abstract The study was conducted on spermatozoa of selected farm and free-living animal species, isolated post mortem from the tail of the epididymis, and stained with silver nitrate -AgNO3. The material was collected from pigs, goats, wild boar, and European roe deer. Twenty morphologically normal spermatozoa randomly selected from each animal and well visible under the microscope, were analysed. The following measurements were considered: head length, width, perimeter and area, acrosome area, mid-piece length, tail length, and overall sperm length. AgNO3 staining differentiated the acrosomal (light hue) and distal (dark hue) part of the sperm head, and a light-hued mid-piece was visible within the sperm tail. Silver nitrate staining revealed species and variety-related differences, particularly in reference to the sperm head. Clear-cut differentiation within the head and tail area made it possible to perform detailed morphometric measurements of the spermatozoa

Zależność pomiędzy funkcją seksualną a wskaźnikiem masy ciała i stężeniem hormonów steroidowych u młodych mężczyzn

Introduction: In older men, sexual disorders may be the result of a decrease in testosterone and an increase in sex hormone binding globulin (SHBG) serum levels. Although obesity may enhance the decline of testosterone, it is also the cause of metabolic disorders, which are additional risk factors of erectile dysfunction. The purpose of this study was to investigate whether elevated body weight is associated with decreased serum testosterone concentrations and reduced sexual function in young men.Material and methods: Data on general health, medication, depressive symptomatology and sexual life was obtained from 136 men aged 20–49 years. Blood levels of LH, total testosterone (TT), dehydroepiandrosterone sulfate (DHEA-S), oestradiol, SHBG, total cholesterol, LDL- and HDL-cholesterol, and triglycerides were determined. Body mass index (BMI), waist to hip ratio (WHR) and free testosterone index (FTI) were calculated.Results: A significantly reduced occurrence of sexual fantasies, morning erections and erectile function scores was observed in the oldest group compared to the youngest men with normal BMI, although orgasmic function was unchanged. A significant decrease in TT serum levels was observed in obese 30-year-olds compared to men with normal BMI, while in obese 40-year-olds decreased LH and SHBG levelswere also found. No differences in the levels of lipids and sexual achievements were found among men with different BMI. However, erectile function and morning erections significantly negatively correlated with age, BMI and WHR, and positively with FTI, but not with other studied hormones and lipids.Conclusions: In young men, obesity can lead to a deterioration of erectile function as a result of lower testosterone levels as the only reason.Wstęp: U starszych mężczyzn zaburzenia seksualne mogą być następstwem zmniejszania się stężenia testosteronu i wzrostu stężeniaglobuliny wiążącej steroidy płciowe (SHBG) we krwi. Otyłość może nasilać obniżanie stężenia testosteronu, ale jest także przyczyną zaburzeńmetabolicznych, które stanowią dodatkowe ryzyko pojawienia się zaburzeń erekcji. Celem pracy było zbadanie, czy nadmiernamasa ciała jest związana z obniżonym stężeniem testosteronu i pogorszeniem funkcji seksualnych już u młodych mężczyzn.Materiał i metody: Dane na temat stanu zdrowia, stosowanych leków, objawów depresji i życia seksualnego uzyskano od 136 mężczyzn w wieku 20–49 lat. We krwi oznaczono stężenia LH, testosteronu całkowitego (TT), siarczanu dehydroepiandrosteronu (DHEA-S), estradiolu, SHBG, cholesterolu całkowitego, cholesterolu LDL i HDL oraz triglicerydów. Wyliczono wskaźniki: masy ciała (BMI), talia/biodra (WHR) i wolnego testosteronu (FTI).Wyniki: Znamienne zmniejszenie częstości fantazji seksualnych i porannych erekcji oraz pogorszenie jakości erekcji, ale nie zdolności do osiągania orgazmu, obserwowano w najstarszej grupie badanych w porównaniu z grupą najmłodszych mężczyzn z prawidłowym BMI. Znamienne zmniejszenie stężenia TT stwierdzono u otyłych 30-latków w porównaniu z mężczyznami z prawidłowym BMI, a u 40-latków dodatkowo stwierdzono zmniejszenie stężenia LH i SHBG. Nie stwierdzono znamiennych różnic w stężeniu lipidów, a także w jakości reakcji seksualnych u mężczyzn z różnym BMI. Jednakże, jakość erekcji oraz częstość występowania porannych erekcji znamiennie negatywnie korelowały z wiekiem, BMI i WHR, a pozytywnie z FTI, ale nie z innymi badanymi hormonami i lipidami.Wnioski: U młodych mężczyzn otyłość może prowadzić do pogorszenia erekcji na skutek obniżenia stężenia testosteronu jako jedynej przyczyny

Cardiovascular and metabolic effects of estrogen in men

A b s t r a c t Although in men with an inherited mutation of gene encoding ERα (estrogen resistance) the occurrence of premature coronary artery disease was documented, and in men with inherited lack of aromatase (estrogen deficiency) an unfavorable lipid milieu was reported, the predominant number of both epidemiological and interventional studies suggests that in men estrogens may either not influence or may promote the develpoment of coronary artery disease. It is possible therefore the beneficial effect of estrogen administration on the lipid milieu in patients with estrogen deficiency is limited only to this unique situation. There may exist a sex-specific difference in the response to estrogen action. In contrast to women where estrogens generate nitric oxide (NO) production in the vascular endothelium, they do not do so in men. NO is responsible for vascular dilation and inhibits lipoprotein oxydation and monocyte adhesion to the endothelium. There may exist also a difference between short-term, non-genomic effect of estrogen and the effects of a long-term exposition to the hormone. Several reports are available indicating that estrogen administartion may have an unfavourable effect not only on blood lipid profile but also on venous thromboembolism in both sexes. In this context the role of estrogens in the regulation of the cardiovascular system gains a special importance and needs further studies. K Ke ey y w wo or rd ds s: : estardiol, coronary artery disease, human male

Physiological significance of estrogens in men - breakthrough in endocrinology

Estradiol (E2) jest tradycyjnie uznanym żeńskim hormonem płciowym. Przez 40 lat wierzono, że u mężczyzn E2 nie wywiera żadnego wpływu lub wywołuje uszkodzenie czynności gonady. Głównym źródłem E2 u mężczyzn jest tkanka tłuszczowa i mózg. E2 jest również wytwarzany w nadnerczach, wątrobie, gruczołach sutkowych i włosach oraz w gonadzie męskiej. Dzienne wytwarzanie i stężenie E2 we krwi u mężczyzn są wyższe niż u kobiet w okresie pomenopauzalnym. W końcu lat 80. wykazaliśmy po raz pierwszy, że w okresie dojrzewania płciowego E2 jest istotnym sygnałem hormonalnym dla zapoczątkowania spermatogenezy. Tradycyjny pogląd o braku lub hamującej roli E2 u płci męskiej został ostatecznie obalony dzięki odkryciu receptorów estrogenowych u samców zwierząt. W latach 90. wytworzono też samce myszy transgenicznych pozbawione receptora estrogenowego (ER), a także genu kodującego enzym aromatazę, umożliwiającego konwersję testosteronu do E2. Obserwacje dorosłych mężczyzn z wrodzonymi mutacjami tych genów, znacznie poszerzyły naszą wiedzę na temat roli E2 u mężczyzn przy tworzeniu zrębu kości, hamowaniu ich wzrostu, metabolizmie lipidów i dojrzewaniu płciowym, efektom przypisywanym dotychczas działaniu testosteronu. Nowe dane wskazują także na ważną rolę estrogenów i ER w czynności układu sercowo-naczyniowego i w przeciwdziałaniu chorobie wieńcowej u mężczyzn.Estradiol (E2) is traditionally recognised as the female sex hormone. It has been believed for 40 years, that E2 didn’t exert any influence or caused impairment of the gonadal function in men. The main source of E2 in men is adipose tissue and the brain. E2 is also produced in adrenals, liver, mammary glands, hair and in male gonad. Daily production and the level of E2 in the blood in men are higher than those in postmenopausal women. At the end of the 80-ties we were first reporting that during sexual maturation E2 can be the important hormonal signal for the initiation of spermatogenesis. The traditional view about unimportant or inhibitory role of E2 in male physiology was finally refuted thanks to discovering estrogen receptors in males. In the 90-ties, transgenic mice with the lack of estrogen receptor (ER) or gene encoding enzyme aromatase, that enable the conversion of testosterone into E2, were also produced. Observations of men with inherited mutations of these genes, considerably extended our knowledge about E2 in men in stroma bones formation, inhibition of their growing, lipids metabolism and sexual maturation, the effects that were attributed to testosterone action until today. New data also points at important role of estrogens and ER in the function of the cardio-vascular system and in counteracting coronary disease in men

Wolne i biodostępne frakcje steroidów płciowych mogą wpływać na kości u młodych mężczyzn w zależności od wieku i stężenia estradiolu

Introduction: Longitudinal bone growth ceases by the end of puberty, and it is thought to be a result, in both sexes, of increased pubertal oestrogen serum concentrations. Since peak bone mass is achieved by the third decade of life or later, the aim of this study was to relate sex steroid hormones and sex hormone binding globulin (SHBG) levels to bone quality in men during their third and fourth decades of life.Material and methods: Eighty men, healthy volunteers aged between 18 and 39 years, were subjected to an interviewer-administered questionnaire, body mass index (BMI) measurement, blood sample and calcaneal quantitative ultrasound (QUS) (Hologic-SAHARA). Blood was assessed for testosterone (T), oestradiol (E2), dehydroepiandrosterone sulfate (DHEAS), SHBG, luteinising hormone (LH) and follicle stimulating hormone (FSH). Free and bioavailable T and E2 levels were calculated knowing SHBG and albumin levels.Results: While T, E2, DHEAS, LH and FSH levels were not related, free and bioavailable fractions of T and E2 were positively associated with QUS readings. SHBG level was associated negatively. After dichotomisation for age, the associations remained significant only for younger subjects (18–30 years, n = 47). After adjustment for other co-variants, only SHBG in younger subjects retained its negative association with QUS. Older subjects (31–39 years, n = 33) revealed higher BMI and lower serum concentrations of total (–17 %), free (–18.5%) and bioavailable (–22.5%) levels of E2 than younger subjects.Conclusion: Free and bioavailable fractions of sex steroids may influence bones in young men, depending on age and E2 level. (Endokrynol Pol 2014; 65 (5): 357–364)Wstęp: Wzrost kości na długość ustaje wraz z końcem dojrzewania płciowego i wykazano, że u obu płci jest to wynik wzrostu stężenia estrogenów we krwi. Skoro przyjęto, że szczytowa masa kostna jest osiągana dopiero w trzeciej dekadzie życia lub po trzydziestce, badano związki pomiędzy stężeniami steroidów płciowych i białka wiążącego steroidy płciowe (SHBG) a jakością kości u mężczyzn w trzeciej i czwartej dekadzie życia.Materiał i metody: Osiemdziesięciu mężczyzn, zdrowych ochotników w wieku 18–39 lat wypełniło kwestionariusz z wywiadem, zmierzono u nich wskaźnik masy ciała (BMI) i wykonano ilościową analizę ultrasonograficzną kości piętowej (QUS) (Hologic-SAHARA). We krwi oznaczono stężenia testosteronu (T), estradiolu (E2), siarczanu dehydroepiandrosteronu (DHEAS), SHBG, hormonu luteinizującego (LH) i hormonu folikulotropowego (FSH). Znając stężenia SHBG i albumin, wyliczano stężenia wolnego i biodostępnego T i E2.Wyniki: Podczas gdy stężenia T, E2, DHEAS, LH i FSH nie wykazywały powiązań, stężenia wolnych i biodostępnych frakcji T i E2 były dodatnio związane z parametrami QUS. Stężenie SHBG wykazywało związek ujemny. Relacje te zależały od wieku. Mianowicie, po podziale na dwie grupy wiekowe, relacje pozostały znamienne tylko wśród młodszych mężczyzn (18–30 lat, n = 47). Analiza wieloczynnikowa wykazała, że tylko stężenie SHBG u młodszych mężczyzn zachowało znamiennie ujemny związek ze stanem kości. Starsi (31–39 lat, n = 33) wykazali wyższy BMI, a niższe stężenie całkowitego (–17%), wolnego (–18,5%) i biodostępnego (–22,55%) E2 w porównaniu z młodszymi badanymi.Wnioski: Wolne i biodostępne frakcje steroidów płciowych mogą wpływać na kości u młodych mężczyzn, w zależności od wieku i stężenia E2. (Endokrynol Pol 2014; 65 (5): 357–364