Effects of cadmium chloride on testicular autophagy and blood-testis barrier integrity in prepubertal male rats / 中华劳动卫生职业病杂志

Objective: To study the effects of cadmium chloride (CdCl(2)) exposure on testicular autophagy levels and blood-testis barrier integrity in prepubertal male SD rats and testicular sertoli (TM4) cells. Methods: In July 2021, 9 4-week-old male SD rats were randomly divided into 3 groups: control group (normal saline), low dose group (1 mg/kg·bw CdCl(2)) and high dose group (2 mg/kg·bw CdCl(2)), and were exposed with CdCl(2) by intrabitoneal injection. 24 h later, HE staining was used to observe the morphological changes of testis of rats, biological tracer was used to observe the integrity of blood-testis barrier, and the expression levels of microtubule-associated protein light chain 3 (LC3) -Ⅰ and LC3-Ⅱ in testicular tissue were detected. TM4 cells were treated with 0, 2.5, 5.0 and 10.0 μmol/L CdCl(2) for 24 h to detect the toxic effect of cadmium. The cells were divided into blank group (no exposure), exposure group (10.0 μmol/L CdCl(2)), experimental group[10.0 μmol/L CdCl(2)+60.0 μmol/L 3-methyladenine (3-MA) ] and inhibitor group (60.0 μmol/L 3-MA). After 24 h of treatment, Western blot analysis was used to detect the expression levels of LC3-Ⅱ, ubiquitin binding protein p62, tight junction protein ZO-1 and adhesion junction protein N-cadherin. Results: The morphology and structure of testicular tissue in the high dose group were obvious changed, including uneven distribution of seminiferous tubules, irregular shape, thinning of seminiferous epithelium, loose structure, disordered arrangement of cells, abnormal deep staining of nuclei and vacuoles of Sertoli cells. The results of biological tracer method showed that the integrity of blood-testis barrier was damaged in the low and high dose group. Western blot results showed that compared with control group, the expression levels of LC3-Ⅱ in testicular tissue of rats in low and high dose groups were increased, the differences were statistically significant (P<0.05). Compared with the 0 μmol/L, after exposure to 5.0, 10.0 μmol/L CdCl(2), the expression levels of ZO-1 and N-cadherin in TM4 cells were significantly decreased, and the expression level of p62 and LC3-Ⅱ/LC3-Ⅰ were significantly increased, the differences were statistically significant (P<0.05). Compared with the exposure group, the relative expression level of p62 and LC3-Ⅱ/LC3-Ⅰ in TM4 cells of the experimental group were significantly decreased, while the relative expression levels of ZO-1 and N-cadherin were significantly increased, the differences were statistically significant (P<0.05) . Conclusion: The mechanism of the toxic effect of cadmium on the reproductive system of male SD rats may be related to the effect of the autophagy level of testicular tissue and the destruction of the blood-testis barrier integrity.

Perspectives of African American women about barriers to breast cancer prevention and screening practices: A qualitative study

Breast cancer is a severe illness that often has fatal consequences. Adherence to the recommendations for breast cancer surveillance is poorly practiced among African American women. The study aimed to identify barriers to preventative screening for breast cancer among African American women (AAW) using a qualitative research design. We explored the influence of personal barriers,stereotypes, socioeconomic status, culture, attitudes, and beliefs on African American women's behavior regarding breast cancer screening. Fourteen African American women were interviewed. Data analysis was completed with Interpretative Phenomenology Approach (IPA). This study's findings demonstrated that African American women perceived the barriers to breast cancer screening include lack of information about available resources, belief that screening cannot change genetic predisposition, embarrassment from exposing the breast for a mammogram, fear of mammograms, and fear of a positive result. These findings may be used to develop interventions to increase AAW's participation in breast cancer screening. (Afr J Reprod Health 2022; 26[7]: 22-28).

Torsión testicular, anticuerpos antiesperma y repercusión clínica / Testicular Torsion, Antisperm Antibodies, and Clinical Repercussions

La torsión testicular es una patología que afecta directamente la calidad del esperma de los pacientes debido a la ruptura de la barrera hematotesticular, lo cual hace que se formen anticuerpos antiesperma, que inclusive afectan al testículo contralateral. Múltiples estudios tanto en modelos animales como humanos, permiten hacer evidente la relación entre la torsión testicular y la formación de anticuerpos antiesperma y su repercusión clínica, principalmente, en la fertilidad.

Testicular torsion is a condition that directly affects the quality of the sperm of patients due to rupture of the blood-testis barrier; it leads to the formation of antisperm antibodies and can even affect the contralateral testis. This condition has been demonstrated previously, and multiple studies have been conducted in both animal and human models. In this review, the evidence from studies of testicular torsion, the formation of antisperm antibodies, and the clinical repercussions, mainly with respect to fertility, is presented.

Long-term exposure to PM2.5 from automobile exhaust results in reproductive dysfunction in male rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the effects of long-term exposure to particulate matter 2.5 (PM2.5) from automobile exhaust on the reproductive function of Sprague Dawley (SD) rats.</p><p><b>METHODS</b>Forty-five male SD rats, weighing 80 - 94 g and aged 28 days, were randomly assigned to receive intra-tracheal administration of 0.9% normal saline (control group, n = 15), PM2. 5 at 2 μg per 100 g body weight per day (low-dose PM2.5 group, n = 15), and PM2.5 at 16 μg per 100 g body weight per day (high-dose PM2.5 group, n = 15), qd, for 60 successive days. After the last 24-hour exposure, 10 rats were taken from each group for copulation with normal female ones, while the others were sacrificed, their testes removed for sperm count and deformity, pathological examination, and determination of the Connexin43 expression.</p><p><b>RESULTS</b>The conception rate was significantly decreased in the low- and high-dose PM2.5 groups as compared with that of the control (70% and 50% vs 100%), and so were the sperm count and quality. The rats in the PM2.5-exposed groups showed significantly disordered histological structure of the seminiferous tubules, reduced sperm count in the testicular lumen, some exfoliated secondary spermatocytes, downregulated Connexin43 expression in the testis, and damaged blood-testis barrier.</p><p><b>CONCLUSION</b>Long-term exposure to PM2.5 from automobile exhaust damages the reproductive function of male SD rats.</p>

Testicular CR16 and spermatogenesis / 中华男科学杂志

Spermatogenesis is a complex regulatory process depending on a variety of hormones (such as FSH, LH, T, and 17beta estradiol), cytokines, and genes. Research on gene regulation in spermatogenesis has become a hot spot and revealed some spermato-genesis-related genes, such as AYZ, DAZ, YRRM, NOSTRIN, and so on. Reports are rarely seen on the role of CR16 in male reproduction, and its action mechanism in spermatogenesis is not yet clear. This article updates the role of CR16 in spermatogenesis in the male reproductive system from the perspective of Sertoli cells forming a blood-testis barrier.

The actin filament network associated to Sertoli cell ectoplasmic specializations

Junctional devices in Sertoli cells conform the blood-testis barrier and play a key role in maturation and differentiation of germ cells. The spacial distribution of ectoplasmic specializations of Sertoli cells was studied by beta-actin immunolabelling, using laser confocal and transmission electron microscopy. For confocal microscopy, beta-actin immunolabelling of ectoplasmic specializations was studied over the background of either prosaposin or glutaredoxin immunolabelling of the Sertoli cytoplasm. Labelling was found near the basal lamina, surrounding early spermatocytes (presumably in leptotene-zygotene) or at one of two levels in the seminiferous epithelium: (1) around deep infoldings of the Sertoli cell cytoplasm, in tubular stages before spermiation, and (2) in the superficial part of the seminiferous epithelium, in tubular stages after or during spermiation. For transmission electron microscopy, beta-actin immunolabelling of ectoplasmic specializations was also used. Ectoplasmic specializations were found at two different levels of the seminiferous epithelium. We also used freeze fracture to analyze the characteristics of tubulo-bulbar complexes, a known component of apical ectoplasmic specializations. Also, these different approaches allowed us to study the complex arrangement of the actin cytoskeleton of Sertoli cells branches, which surround germ cells in different stages of the spermatogenic cycle. Our results show a consistent labelling for beta-actin before, during and after the release of spermatozoa in the tubular lumen (spermiation) suggesting a significant role of the actin network in spermatic cell differentiation. In conclusion, significant interrelations among the beta-actin network, the junctional complexes of the blood-testis barrier and the ectoplasmic specializations were detected at different stages of the seminiferous cycle.

Microwave radiation decreases the expressions of occludin and JAM-1 in rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the changes in the expressions of the tight junction related protein occludin and junctional adhesion molecule-1 (JAM-1) of the blood-testis barrier and their significance in rats after microwave radiation.</p><p><b>METHODS</b>Eighty male Wistar rats were exposed to microwave radiation with average power density of 0, 10, 30 and 100 mW/cm2 for five minutes, and dynamic changes in the expressions of testicular occludin and JAM-1 were observed by Western blot and image analysis at 6 h, 1 d, 3 d, 7 d and 14 d after the radiation.</p><p><b>RESULTS</b>There was a significant down-regulation in the expression of the occludin protein at 3 - 7 d, 6 h - 7 d and 6 h - 14 d (P < 0. 05), as well as in that of JAM-1 at 3 - 7 d, 1 - 7 d and 1-14 d (P < 0.05) after exposure to 10, 30 and 100 mW/cm2 microwave radiation.</p><p><b>CONCLUSION</b>The decreased protein expressions of occludin and JAM-1 may play an important role in the microwave radiation induced-damage to the blood-testis barrier.</p>

[ experimental model of hydatidosis in rabbit testis]

Although no part of human anatomy is invulnerable to hydatid disease, it has been reported to occur in most of vital organs. Hydatid disease of urinary tract is uncommon, accounting for only 2-3% of all such cases. Testes are extremely rare sites for echinococcosis. There are only three cases of testicular hydatid disease which were reported. In this laboratory animal model, we studied echinococcosis in rabbit testis. In this experimental trial study, 14 male Albino rabbits [body weight 2.5-3kg] were randomized into two groups: group A [study group], for testicular injection and group B [control group], for intraperitoneal injection of viable protoscoleces. All rabbits were infected, and then housed them under pathogen-free conditions for 10 weeks [70 days]. One rabbit from group A and three from group B died one day after injection, and the other rabbits survived during follow-up period. At 10th week they were all anaesthetized and then we studied all testes with ultrasonography. In group A all testes were excised, and in group B we removed liver, kidneys and took four biopsies from peritoneum, for histopathology investigation. There was demonstrable hydatid cyst [protoscoleces and germinative layer] in testes of five rabbits from group A, but in one rabbit both testes were normal. In group B, three out of four rabbits developed peritoneal hydatidosis. The mechanism of testicular resistance to echinococcosis could be blood-testis barrier because when we directly infected the testes of rabbits with protoscoleces, hydatid cyst developed

The Different Effects of Testicular Torsion on the Contralateral Testis between Pubertal and Adult Rats

PURPOSE: Our study aimed to determine whether the severity of damage to the contralateral testis by ipsilateral testicular torsion/detorsion in pubertal rats, which have an incomplete blood-testis barrier, is different from that in adult rats. MATERIALS AND METHODS: We divided pubertal (6 weeks, n=17) and adult (10 weeks, n=17) Sprague-Dawley (SD) rats into group S (sham; n=5), group O (orchiectomy; n=6), and group D (detorsion; n=6). After 4 hours' torsion of the ipsilateral testis, we applied orchiectomy (group O) and detorsion (group D) depending on the group and compared the histopathologic changes and germ cell apoptosis of the contralateral testis at the age of 13 weeks. RESULTS: In each age group, increased interstitial area, edema, and germ cell sloughing were observed in group D. The mean seminiferous tubule diameter decreased more in group D than in group S or O in each age group (p<0.05). The mean germ cell layer thickness and number of spermatids per tubule decreased more in group D than in group S or O in each age group; additionally, in group D, values decreased more in pubertal rats than in adult ones (p<0.05, respectively). The mean numbers of terminal deoxyuridine nick-end labeling (TUNEL)-positive cells were less than 1.0 in groups S and O, which was smaller than in group D (p<0.05); additionally, in group D, this value tended to be higher in pubertal rats than in adult ones (p=0.057). CONCLUSIONS: SD rats with a detorsioned testis had more severe damage to the contralateral testis than did those undergoing orchiectomy of the torsioned testis. Also, when comparing the severity of damage to the contralateral testis after ipsilateral torsion/detorsion between pubertal and adult rats, rats at a pubertal age, when most testicular torsions occur in clinical situations, had more severe damage than did those at an adult age.

High power microwave radiation damages blood-testis barrier in rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To determine the effect of high power microwave (HPM) radiation on the structure and function of blood-testis barrier (BTB) in rats.</p><p><b>METHODS</b>One hundred and sixty-six male Wistar rats were treated by heart perfusion of lanthanum-glutaraldehyde solution and tail vein injection of evans blue (EB) at 6 h, 1, 3, 7 and 14 d after exposed to 0, 10, 30 and 100 mW/cm2 HPM radiation for 5 minutes, the structural change of BTB and distribution of lanthanum or EB observed through the light microscope, electron microscope and laser scanning confocal microscopy (LSCM).</p><p><b>RESULTS</b>Testicular interstitial edema, vascular congestion or hyperemia with accumulation of plasma proteins and red blood cells in the inner compartment of seminiferous tubules were observed after exposure to HPM. The above-mentioned pathological changes were aggravated at 1-7 d and relieved at 14 d after radiation, obviously more severe in the 30 and 100 mW/cm2 exposure groups than in the 10 mW/cm2. Both lanthanum precipitation and EB were deposited in the inner compartment.</p><p><b>CONCLUSION</b>HPM radiation may damage the structure and increase the permeability of BTB.</p>

Estrogens affect Sertoli cells and the blood-testis barrier in pubertal rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the correlation of exogenous estrogens with the expression of FasL in Sertoli cells and the blood-testis barrier during the differentiation and maturation period of Sertoli cells, and to discuss the related factors that influence the blood-testis barrier of pubertal rats.</p><p><b>METHODS</b>Super-physiological doses of exogenous estrogenic compounds (diethylstilbestrol and estradiol) were administered to pubertal Sprague-Dawley rats in vitro and in vivo, the FasL expression in the Sertoli cells of the rats detected by immunohistochemistry and Western blot, and the changes in the blood-testis barrier observed with the electron microscope.</p><p><b>RESULTS</b>After the exposure to exogenous estrogens, the FasL expression was markedly up-regulated in the immature Sertoli cells (P < 0.05) as well as in the Sertoli cell membrane and the blood-testis barrier of the epithelium. The tracer lanthanum passed through the blood-testis barrier and reached the whole layer of the epithelium at 18 days.</p><p><b>CONCLUSION</b>Super-physiological dose of exogenous estrogens can change the expression and distribution of FasL in immature Sertoli cells and affect the structure of the blood-testis barrier.</p>

Effect of electromagnetic pulse exposure on permeability of blood-testicle barrier in mice / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To study the effect of electromagnetic pulse (EMP) exposure on the permeability of blood-testicle barrier (BTB) in mice.</p><p><b>METHODS</b>Adult male BALB/c mice were exposed to EMP at 200 kV/m for 200 pulses with 2 seconds interval. The mice were injected with 2% Evans Blue solution through caudal vein at different time points after exposure, and the permeability of BTB was monitored using a fluorescence microscope. The testis sample for the transmission electron microscopy was prepared at 2 h after EMP exposure. The permeability of BTB in mice was observed by using Evans Blue tracer and lanthanum nitrate tracer.</p><p><b>RESULTS</b>After exposure, cloudy Evans Blue was found in the testicle convoluted seminiferous tubule of mice. Lanthanum nitrate was observed not only between testicle spermatogonia near seminiferous tubule wall and sertoli cells, but also between sertoli cells and primary spermatocyte or secondary spermatocyte. In contrast, lanthanum nitrate in control group was only found in the testicle sertoli cells between seminiferous tubule and near seminiferous tubule wall.</p><p><b>CONCLUSION</b>EMP exposure could increase the permeability of BTB in the mice.</p>

Orchestration of occludins, claudins, catenins and cadherins as players involved in maintenance of the blood-epididymal barrier in animals and humans / 亚洲男科学杂志(英文版)

Although spermatozoa are formed during spermatogenesis in the testis, testicular spermatozoa are immature and cannot swim or fertilize. These critical spermatozoal functions are acquired in the epididymis where a specific luminal environment is created by the blood-epididymal barrier; proteins secreted by epididymal principal cells bind to maturing spermatozoa and regulate the maturational process of the spermatozoa. In the epididymis, epithelial cell-cell interactions are mediated by adhering junctions, necessary for cell adhesion, and by tight junctions, which form the blood-epididymal barrier. The regulation of these cellular junctions is thought to represent a key determinant in the process of sperm maturation within the epididymis. Tight junctions between adjacent principal cells permit the formation of a specific microenvironment in the lumen of the epididymis that is essential for sperm maturation. Although we have made significant progress in understanding epididymal function and the blood-epididymal barrier, using animal models, there is limited information on the human epididymis. If we are to understand the normal and pathological conditions attributable to human epididymal function, we must clearly establish the physiological, cellular and molecular regulation of the human epididymis, develop tools to characterize these functions and develop clinical strategies that will use epididymal functions to improve treatment of infertility.

Temporary Opening of the Testis-blood Barrier by Triolein Fat Emulsion / 대한비뇨기과학회지

PURPOSE: Although the purpose of the blood-testis barrier (BTB) is to protect germ cells from harmful influences, it also impedes the delivery of chemotherapeutic agents to the testis. This study was undertaken to determine whether a triolein emulsion could transiently alter the permeability of the BTB in cats. MATERIALS AND METHODS: An emulsion of 0.05ml triolein in 20ml of saline or just 20ml of normal saline, as the control, were infused into the testicular arteries in 18 and 15 cats, respectively (embolic and control group). Pre- and post-contrast magnetic resonance images (MRIs) were obtained 30 minutes and 2 hours after embolization. Qualitative and quantitative analyses of the MRIs were performed via the presence and degree of contrast enhancement and the contrast enhancement ratios (CERs), respectively. An electron microscopy (EM) study was subsequently performed, using a lanthanum tracer, to correlate with the MRI results. RESULTS: Contrast enhancement of the testis was observed in both groups and at both time points, but was more prominent in the embolic group. The CERs in the embolic group were significantly higher than those in the control group (p=0.0001). In each group, the CERs at 2 hours were significantly lower than those at 30 minutes (p=0.006). In the EM study, the entry of lanthanum was markedly increased at 30 mins, but recovered at 2 hours after embolization compared to the control. CONCLUSIONS: Intra-arterial infusion of triolein emulsion transiently increased the permeability of the BTB. This result may be useful in future studies for a chemotherapy delivery system to the testis.

Increase in Transepithelial Resistance Mouse Sertoli Cells by Leydig Cells Coculture / 대한남성과학회지

PURPOSE: To verify the regulation of transepithelial resistance (TER) of Sertoli cells by Leydig cells in mouse. MATERIALS AND METHODS: Primary culture of Sertoli cells was established on cell culture plate insert and monolayer culture was subjected to coculture in the Leydig cell culture. Changes in TER was monitored for 48 h using the conductivity meter equipped with two electrodes system. RESULTS: TER gradually increased according to the development of monolayer of Sertoli cells on the cell culture plate insert. Net changes in TER of Sertoli cells culture was significantly higher under the Leydig cells coculture compared to control after 48 h of coculture. CONCLUSIONS: It is the first report about the increase in TER of Sertoli cells by Leydig cells in vitro. Paracrine interaction between Leydig cells and Sertoli cells might be involved in the development of functional blood testis barrier which is made by tight junctions between Sertoli cells in mouse testis.

Aspectos imunológicos da torçäo testicular: trabalho experimental em ratos / Immunologic aspects of testicular torsion: experimental study in wistar rats

As atividades testiculares, hormonal e espermiogênica säo de extrema importância para o homem e para a manutençäo e o aprimoramento da humanidade. Trauma, tumores, varicocle, criptorquidia, anorquia, atrofias, torçäo de cordäo espermático, estäo entre as diversas causas da infertilidade masculina. A integridade da barreira testículo - sangüínea parece ter um papel fundamental na patôgenese da orquite auto-imune. A orquite auto-imune é um processo inflamatório testicular mediado por formaçäo de anticorpos, contra os próprios antígenos testiculares do indivíduo, causando lesöes no epitélio germinativo. Neste estudo foram utilizados oitenta (80) ratos Wistar, singênicos, machos, adultos, separados em dez grupos, submetidos à torçäo unilateral do testículo, sob anestesia, por um período de duas à doze horas, seguido, entäo, de destorçäo e exame anatomopatológico das gônadas contralateral e ipsilateral, em períodos que variaram de duas à dez semanas. Estes testículos foram analisados através das técnicas de histologia e de imunofluorescência com a utilizaçäo de anticorpos monoclonais para as imunoglobulinas IgA, IgG e IgM de coelho e cabra, contra os antígenos testiculares do rato, para que se estudasse as possíveis alteraçöes no tecido testicular do animal, objetivando a comprovaçäo da existência de complexos auto-imunes no epitélio germinal. O estudo demonstrou a presença de imunoglobulinas, principalmente das classes IgG e IgM na luza e no interstício dos túbulos semíferos. Alteraçöes estruturais do testículo contralateral, näo torcido, foram demonstradas. Os resultados deste experimento indicam que o sistema inume de animais sadios contém linfócitos com receptores para os próprios antígenos, capazes de, na torçäo unilateral do cordäo espermático, promoverem uma doença auto-imune. A utilizaçäo de substâncias imuno-supressoras, neste estudo, o corticóide, foi feita para tentar minimizar ou mesmo coibir o processo auto-agressivo. Existiu uma proteçäo testicular contralateral efetiva, com o uso precoce da referida droga. É sugerido que a permanência de um testículo necrótico na bolsa escrotal, após torçäo prolongada, pode levar à subfertilidade ou infertilidade.

La inmunología en la infertilidad / Infertility and inmunology

La influencia de los factores inmunológicos en la fertilidad humana ha cobrado interés en los últimos años, gracias a la facilidad de su estudio mediante nuevas técnicas de laboratorio y a la comprensión de las reacciones involucradas, lo que permite identificarlo como un proceso clínico real. Hasta hace unos pocos años sólo se contaba con métodos que evaluaban la presencia de aglutinación de los espermatozoides en torno a los anticuerpos, o su inmovilización gracias al complemento; dichos métodos tenían una gran variabilidad y sus resultados dificultaban más de lo que favorecían el diagnóstico. En la actualidad ya se dispone de métodos que detectan las inmunoglobinas específicas implicadas en el proceso, con una exactitud y confiabilidad más aceptable

Effect of sertoli cell changes on germ cells in experimentally produced varicocele in rats / 대한비뇨기과학회지

The relationship between Sertoli cells and germ cells in varicocele remains controversial. To study this relationship in varicocele, seminiferous tubular changes were observed in pubertal rats according to the length of time after induction of the varicocele and the interval between induction and repair of the varicocele. As the length of time of the varicocele increased, accumulation of lipid inclusions within the Sertoli cell cytoplasm appeared first and then premature sloughing of the early spermatids appeared. Lastly, decrease in testicular weight and mean seminiferous tubular diameter (MSTD) together with decrease in the number of late spermatids were observed. Inter-Sertoli cell junctions were preserved unrelated to the duration of the varicocele. When Sertoli cell changes were reversed after varicocele repair, premature sloughing of the early spermatids was not observed. The testicular weight, MSTD and number of late spermatids were significantly increased compared to controls. When Sertoli cell changes were not fully reversed after varicocele repair, premature sloughing of the early spermatids was still observed. The testicular weight, MSTD and number of late spermatids were not significantly increased compared to controls. These results suggest that the blood-testis barrier remains intact in varicocele. The Sertoli cell is the primary intratubular site of alteration leading secondarily to spermatogenic disruption in varicocele. Changes in the Sertoli cell cause premature sloughing of the early spermatids and affect maximally the spermatid Stage.

A study on the occurrence of circulating antisperm antibodies in patients with testicular rupture / 대한비뇨기과학회지

Immunologic factors including antisperm antibodies play a significant role in pathogenesis of 10-20% of unexplained infertility cases. and any event or circumstance that would breach the blood-testis barrier may result in the formation of antisperm antibodies. Using an immunobead test we studied the occurrence of circulating antisperm antibodies in twelve patients who had undergone surgical treatment for testicular rupture due to blunt scrotal trauma. On the follow-up ranging from 6 to 14 months after surgical treatment only one patient was found to have circulating antisperm antibodies but his semen remained normal in spite of the presence of circulating antisperm antibodies.