bluuebunny/crossref_metadata_2025 huggingface dataset:
Variation in Semen Parameters Derived From Computer‐Aided Semen Analysis, Within Donors and Between Donors
Authors: WIJCHMAN JANNY G. , DE WOLF BEN T. H. M. , GRAAFF REINDERT , ARTS EUGENE G. J. M.
Published: November 2023

ABSTRACT: The development of computer‐aided semen analysis (CASA) has made it possible to study sperm motility characteristics objectively and longitudinally. In this 2‐year study of 8 sperm donors, we used CASA to measure 7 semen parameters (concentration, percentage of motile spermatozoa, curvilinear velocity, average path velocity, straight‐line velocity, amplitude of lateral head displacement, and beat/cross frequency). The frequency distributions of the 7 parameters in the semen samples of each donor were investigated. All parameters but one were normally distributed; concentration was distributed log‐normally. Variation within individual donors and between donors was studied. Analysis of variance demonstrated that variation between donors was not explained by the longitudinal variation within individual donors. Variations in motility characteristics between donors were substantial, which may make motility characteristics of limited value as a tool for establishing fertility. Strong correlations were found between the 7 parameters, partly because by definition, motility characteristics are interdependent. Fisher's discriminant analysis demonstrated that each donor appeared to have his own set of semen characteristics and, more specifically, his own motility signature. From this data set it can be predicted that in order to find population means among sperm, it may be more efficient to measure more subjects than to increase the number of samples per subject.
Testicular Pathology in 46,XY Dysgenetic Male Pseudohermaphroditism: An Approach to Pathogenesis of Testis Cancer
Authors: SłOWIKOWSKA‐HILCZER JOLANTA , SZARRAS‐CZAPNIK MARIA , KULA KRZYSZTOF
Published: November 2023

ABSTRACT: Eleven children with dysgenic male pseudohermaphroditism (DMP) and 18 boys with isolated penile hypospadias, all with 46,XY karyotype, were studied. Testicular dysgenesis was associated with significantly lower testosterone response to human chorionic gonadotropin (0.9 ± 0.2 ng/mL) than it was in hypospadias (3.3 ± 0.1 ng/mL), and with significantly higher mean serum follicle‐stimulating hormone (FSH) levels (8.4 ± 2.3 IU/L vs 1.5 ± 0.3 IU/ L). Gonadoblastoma, a tumor that arises from the sex cords, was found in more than ¼ of patients with DMP, whereas testicular carcinoma in situ (CIS) cells were present in all of these patients. Forty‐two percent to 98% of CIS cells revealed an aneuploid pattern of nuclear DNA, indicating that most of them are neoplastic cells. In patients with hypospadias, CIS was not seen, and no other abnormalities were detected. In children with DMP, the percentage of tubules populated with germ cells was significantly lower than it was in those with hypospadias (48.3% ± 10.6% vs 92.4% ± 4.0%). The total number of germ cells (CIS cells + spermatogonia) did not differ significantly between the 2 groups, but the number of spermatogonia was significantly reduced in children with DMP (0.08 ± 0.05 vs 3.65 ± 0.2), suggesting impaired differentiation of gonocytes to spermatogonia. The following significant correlations were present with DMP: 1) the higher the seminiferous tubule cross‐section area, the higher the number of CIS cells (r = 0.78); and 2) the higher the serum gonadotropin levels, the higher were tubular diameter (r = 0.93 for FSH and r = 0.75 for luteinizing hormone [LH]), area (r = 0.79 for FSH and r = 0.82 for LH), percentage of tubules populated with germ cells (r = 0.86 for FSH and r = 0.81 for LH), and number of CIS cells (r = 0.87 for FSH and r = 0.79 for LH). The results indicate that in intersex children with 46,XY karyotype, CIS occurs in dysgenetic testes in all cases and is frequently associated with gonadoblastoma. Impaired organogenesis of sex cords, relative inhibition of testosterone secretion, and the associated increased secretion of gonadotropins may create a milieu that induces or is favorable for the formation or maintenance of neoplastic lesions in dysgenetic testes early in childhood.
Impaired Seminal Antioxidant Capacity in Human Semen With Hyperviscosity or Oligoasthenozoospermia
Authors: SICILIANO L. , TARANTINO P. , LONGOBARDI F. , RAGO V. , DE STEFANO C. , CARPINO A.
Published: November 2023

ABSTRACT: Antioxidant capacity of seminal plasma was evaluated in 120 semen samples subdivided into asthenozoospermic and oligoasthenozoospermic specimens with normal consistency and into asthenozoospermic and oligoasthenozoospermic specimens with hyperviscosity. Semen samples (n = 25) from normozoospermic donors were used as a control group. Scavenger antioxidant capacity of reactive oxygen species was evaluated by superoxide dismutase and catalase activity measurements, whereas the chain‐breaking antioxidant efficiency was detected by total antioxidant status assessment. In semen with normal viscosity, unaltered enzymatic and nonenzymatic antioxidant capacity was revealed in the asthenozoospermic specimens, whereas low superoxide dismutase activity was detected in oligoasthenozoospermic samples. On the contrary, impairment of both the scavenger and chain‐breaking antioxidative systems was revealed in asthenozoospermic and oligoasthenozoospermic hyperviscous ejaculates, regardless of sperm count. Catalase activity and total antioxidant status values were also reduced in the 2 subgroups of hyperviscous ejaculates compared with their respective matched controls, whereas similar superoxide dismutase activities were detected in oligoasthenozoospermic samples with normal and high consistencies. These results suggest that asthenozoospermia could be related to an antioxidant deficiency only in combined ejaculate pathologies, and that a severe impairment of the low and high molecular weight seminal antioxidative capacities could be associated with semen hyperviscosity.
CAG Repeat Length Analysis and Mutation Screening of the Androgen Receptor Gene in Japanese Men With Idiopathic Azoospermia
Authors: SASAGAWA ISOJI , SUZUKI YASUHIRO , ASHIDA JUNKO , NAKADA TERUHIRO , MUROYA KOJI , OGATA TSUTOMU
Published: November 2023

ABSTRACT: Because androgens are required for normal spermatogenesis, we are investigating abnormalities in the androgen receptor as a possible cause of impaired spermatogenesis in patients with idiopathic male infertility. The CAG repeat length in exon 1 and mutations of the androgen receptor gene were studied in 30 men with idiopathic azoospermia and in 51 fertile men. In men with azoospermia, plasma luteinizing hormone (LH), follicle‐stimulating hormone (FSH), and testosterone levels were measured and testicular biopsies were performed. The CAG repeat length ranged from 19 to 30 (mean 23.4 ± 2.9) and from 17 to 28 (mean 23.7 ± 3.2) in men with azoospermia and in controls, respectively. There was no significant difference between the 2 groups. In men with azoospermia, the Johnsen testicular biopsy score negatively correlated with plasma FSH (P < .01). However, the Johnsen testicular biopsy score did not correlate with plasma LH and testosterone levels. The CAG repeat length did not correlate with the Johnsen testicular biopsy score, or with plasma concentrations of LH, FSH, and testosterone. No abnormalities in the androgen receptor gene were detected. These facts suggest that the CAG repeat length and alterations in the androgen receptor gene are not associated with the etiology of idiopathic azoospermia.
GnRH Agonists and Antagonists Stimulate Recovery of Fertility in Irradiated LBNF1 Rats
Authors: MEISTRICH MARVIN L. , WILSON GENE , SHUTTLESWORTH GLADIS , HUHTANIEMI ILPO , REISSMANN THOMAS
Published: November 2023

ABSTRACT: The goal of this study was to determine whether both gonadotropin‐releasing hormone (GnRH) agonists and antagonists could enhance fertility in rats given sterilizing doses of irradiation, to quantify the levels of fertility, and to measure their relative effectiveness in stimulating recovery of spermatogenesis. Irradiated rats were treated with either the GnRH agonist Lupron or the GnRH antagonist Cetrorelix, which have different mechanisms of action. The antagonist suppressed luteinizing hormone (LH), reducing intratesticular testosterone from 75 ng/g‐testis to about 5 ng/g‐testis, whereas the agonist reduced intratesticular testosterone only moderately to about 20 ng/g‐testis, presumably by direct action on the Leydig cell since LH was elevated. These differences were reflected in Leydig cell morphology. When hormone treatment was started immediately after 3.7‐Gy irradiation, fertility was normal at week 20 in the agonist‐treated rats and was near normal in antagonist‐treated rats, whereas irradiated‐only rats were sterile. At week 22 in the GnRH antagonist‐treated rats, testicular weights and sperm counts were maintained at greater than 80% of control values; in GnRH agonist‐treated rats, they were slightly but significantly lower than in GnRH antagonist‐treated rats, and in irradiated‐only rats, they were very low. When the treatment was initiated 10 weeks after 5‐Gy irradiation, after spermatogenesis had ceased, fertility was restored at week 30 to subnormal levels in 83% of GnRH agonist‐ and 50% of GnRH antagonist‐treated rats. Testis weights and sperm counts were restored to about 50% and 20% of control levels, respectively. The percentages of tubules with differentiated germ cells were higher in all groups of antagonist‐treated rats than in those of agonist‐treated rats. Thus, both GnRH agonists and antagonists produced dramatic recovery of spermatogenesis and fertility in irradiated rats, although there were differences in mechanism and perhaps also in effectiveness.
Sonographic Testicular Microlithiasis as an Indicator of Premalignant Conditions in Normal and Infertile Men
Authors: VON ECKARDSTEIN SIGRID , TSAKMAKIDIS GEORGIOS , KAMISCHKE AXEL , ROLF CLAUS , NIESCHLAG EBERHARD
Published: November 2023

ABSTRACT: Sonographic detection of multiple, small hyperechogenic lesions in the testis (testicular microlithiasis; TM) can indicate germ cell tumors. However, it has not been well established whether this finding signifies a risk factor for development of testicular neoplasm in all cases or whether it indicates premalignant changes only in those men with additional risk factors for germ cell cancer, such as infertility, a history of testicular maldescent, orthe presence of an atrophic testis. In a retrospective analysis of 1701 consecutively performed scrotal sonographies of patients with (n = 1399) and without (n = 219) infertility or with contralateral testicular tumors (n = 83), the prevalence of TM was compared with that in 198 healthy men who volunteered for different clinical trials. TM was equally frequent in all groups (2.3% [32/1399] of infertile patients, 2.3% [5/219] of other patients without infertility, and 1.5% [3/198] of healthy men). Results of testicular biopsies were available for a subgroup of infertile men. Carcinoma in situ (CIS) was present only in cases with TM (2/11). In addition, sonographic follow‐up examinations were performed in another 14 men with TM. Testicular tumors had developed in 2 patients, one whom was infertile and one in the control group. None of these patients had a history of testicular maldescent but all testes affected either by CIS or tumors were reduced in volume. We conclude that diagnosis of TM, especially if it is present in an atrophic testis, demands a diagnostic biopsy or at least sonographic follow‐up examinations.
Comparative Evaluation of Fresh and Washed Human Sperm Cryopreserved in Vapor and Liquid Phases of Liquid Nitrogen
Authors: SARITHA K.R. , BONGSO ARIFF
Published: November 2023

ABSTRACT: Infectious organisms such as hepatitis B were recently shown to survive in liquid nitrogen. To prevent cross‐contamination of semen samples via liquid nitrogen, studies were undertaken to evaluate human sperm survival in the vapor phase of liquid nitrogen at −189°C. The study was conducted in 2 separate experiments. In the first experiment, a total of 30 unwashed, fresh semen samples (15 normozoospermic and 15 oligozoospermic) were evaluated for motility, vitality, and morphology after freeze‐thaw survival in vaporous (‐189°C) and liquid nitrogen (‐196°C; control) phases. Similar evaluations were carried out in a second experiment on 27 samples (15 normozoospermic and 12 oligozoospermic) that were previously washed by the swim‐up method. Motile sperm recovery rates were significantly different between liquid and vapor phases (unwashed, normozoospermic: 42.76% ± 3.23% vs 45.52% ± 4.44%, P < .05; washed, normozoospermic: 34.44% ± 4.41% vs 37.58% ± 3.90%, P < .05; unwashed, oligozoospermic: 16.53% ± 3.34% vs 18.25% ± 4.36%, P < .05; washed, oligozoospermic: 10.32% ± 2.54% vs 12.25% ± 2.81%, P < .05). Recovery rates for motility were much higher for unwashed samples compared with washed semen samples. In all experiments the recovery of normal and live forms showed no significant differences between the vapor and liquid nitrogen storage phases (P > .05). The results demonstrate that both washed and unwashed human sperm survive satisfactorily with good recovery in the vapor of liquid nitrogen and can be recommended for future storage in medically assisted conception programs.
Effect of Scrotal Insulation on Clusterin‐Positive Cells in Ram Semen and Their Relationship to Semen Quality
Authors: IBRAHIM NASRELDIN M. , ROMANO JUAN E. , TROEDSSON MATS H.T. , CRABO BO G.
Published: November 2023

ABSTRACT: Scrotal contents of 2 rams were insulated for 96 hours and the fraction (as a percentage) of clusterin‐positive cells (CPCs) and its relationship to semen quality was investigated. Semen collection was started 18 days before insulation and was terminated on day 78 and day 63 after insulation in animals 1 and 2, respectively. Sperm clusterin was localized by immunostaining with anti‐bovine clusterin antibody (anti‐bCAb) and fluorescein isothiocyanate‐conjugated immunoglobulin G. Scrotal insulation led to deterioration of semen quality and increased the percentage of CPCs in both rams. Two types of sperm reactivity were observed: an extensive, intensive staining pattern (ESP); and a localized, less‐intensive staining pattern (LSP). The percentage of ESP‐CPCs began to increase from day 6 and reached 88.8% and 100% on day 15 after insulation in animals 1 and 2, respectively. The increase in CPCs coincided with the presence of a high percentage of teratoid forms (88.3%) in semen from animal 1, and detached heads (81.4%) in semen from animal 2. After normal semen production was restored on day 60 in animal 1, the percentages of ESP‐CPCs and LSP‐CPCs returned to preinsulation rates, whereas only the ESP‐CPCs returned to normal in animal 2. A negative relationship was observed between ESP‐CPCs and total sperm/ ejaculate (r = ‐.62), motility (r = ‐.78), viability (r = ‐.68), and filtration rate (r = ‐.71) in semen from animal 1. Conversely, a positive relationship was seen between ESP‐CPCs and total abnormal spermatozoa (r = .82). Similar results were obtained in semen from animal 2. CPCs were nearly completely absent in glass wool‐Sephadex (GWS)‐filtered semen, suggesting a role for clusterin in the process of trapping abnormal spermatozoa in the GWS filters. We conclude that aberrant spermatogenesis induced by scrotal insulation increases the percentage of CPCs in ram semen. We suggest that the percentage of CPCs in ram semen could be a useful marker in poor‐quality ejaculates.
Altered Growth Factor Expression in the Aging Penis: The Brown‐Norway Rat Model
Authors: RAJASEKARAN MAHADEVAN , KASYAN ARMEN , JAIN AMITA , KIM SAE‐WOONG , MONGA MANOJ
Published: November 2023

The objective of the present study was to evaluate age‐related changes in the protein and gene expression of modulators of erectile function (nitric oxide [NO] and endothelin‐1 [ET‐1]) and growth factors such as transforming growth factor (TGF‐β1) and vascular endothelial growth factor (VEGF) in the penile tissue of Brown‐Norway (BN) rats. Young and old BN male rats were euthanized, and the penile tissue was processed for immunohistochemical and molecular analyses. Total RNA was extracted, and an Access reverse transcription‐polymerase chain reaction (RT‐PCR) system was used for messenger RNA (mRNA) expression analysis. Immunohistochemical studies showed a decreased expression of endothelial nitric oxide synthase (eNOS) protein and an increased staining for ET‐1. Quantitative analysis of PCR products revealed decreased levels of VEGF mRNA expression in the old population of rats. The most significant decrease was detected between bands corresponding to splice forms 164 (21%) and 120 (18%). The observed alterations in the gene expression of growth factors such as VEGF may contribute to the abnormal age‐related morphological and physiological alterations in the erectile tissue.
Peptide and Nonpeptide Reactive Oxygen Scavengers Provide Partial Rescue of the Testis After Torsion
Authors: LYSIAK JEFFREY J. , NGUYEN QUOC AN T. , TURNER TERRY T.
Published: November 2023

Ischemia‐reperfusion (IR) of the testis results in germ cell‐specific apoptosis, followed by a reduction in testis weight and daily sperm production (DSP). This has been associated with an increase in the adhesion of neutrophils to testicular subtunical venules and an increase in reactive oxygen species (ROS). The present study investigated: 1) the effects of a direct, non‐IR‐related ROS insult to the testis and 2) the effects of catalase, superoxide dismutase (SOD), and a novel nonpeptide mimic of SOD, M40403, on neutrophil recruitment, ROS production, testis weight, and DSP following IR of the rat testis. Results revealed that the infusion of H2O2 increased testicular lipid peroxidation 1 hour after administration and increased germ cell apoptosis within 24 hours of administration. Four hours after the repair of torsion plus vehicle infusion, there was a significant increase in myeloperoxidase (MPO) activity, an indicator of neutrophil accumulation, and thiobarbituric acid reactive substances (TBARS), a measure of ROS production, compared to equivalent data in sham‐treated testes. Animals sacrificed 30 days after the torsion plus vehicle infusion revealed a significant decrease in testis weight and DSP compared to the same parameters in sham‐operated animals. The treatment of animals with catalase plus SOD or M40403 showed a significant decrease in MPO activity and TBARS 4 hours after IR of the testis. Animals treated with SOD, SOD plus catalase, and M40403 provided a partial rescue of DSP 30 days after IR of the testis. These results demonstrate that oxidative stress can directly cause germ cell apoptosis, even outside the IR model, and confirm the importance of oxidative stress in testicular IR injury. Also, following testicular IR, there is a recruitment of neutrophils and an increase in ROS production in the testis. The administration of ROS scavengers significantly reduced the IR‐induced responses. Interestingly, the administration of all ROS scavengers also blocked neutrophil recruitment to the testis. The mechanism by which ROS modulates neutrophil adhesion to venules is presently under investigation, as are additional therapeutic regimens to block oxidative stress.
Treatment of Male Hypogonadism With Testosterone Undecanoate Injected at Extended Intervals of 12 Weeks: A Phase II Study
Authors: VON ECKARDSTEIN SIGRID , NIESCHLAG EBERHARD
Published: November 2023

ABSTRACT: This paper reports the result of an open‐label, nonrandomized clinical trial investigating the efficacy and safety of an injectable preparation of testosterone undecanoate (TU) dissolved in castor oil and given over a 3.2‐year period. In a previous study we demonstrated that injections of TU every 6 weeks resulted in satisfactory substitution but a tendency toward testosterone accumulation. Here we investigate prolonged TU treatment at extended injection intervals in 7 hypogonadal men. Injections were given at gradually increasing intervals between the fifth and 10th injection, and from then on every 12 weeks. Steady state kinetics were obtained after the 13th injection. Well‐being, sexual activity, clinical chemistry, prostate volume, and prostate‐specific antigen (PSA) and serum hormone levels were monitored. Patients were clinically well‐adjusted throughout the study. Before the next injection, testosterone, dihydrotestosterone, and estradiol levels were mostly within the normal range and showed a tendency to decrease with increasing injection intervals. Body weight, hemoglobin, serum lipids, PSA, and prostate volume did not change significantly during the 3.2 years of treatment. PSA levels were always within the normal limit. Maximal testosterone levels during steady state kinetics were measured after 1 week with 32.0 ± 11.7 nmol/L (mean ± SD). Before the last injection, mean testosterone concentrations were 12.6 ± 3.7 nmol/L. Compared with conventional testosterone enanthate or cypionate treatment requiring injection intervals of 2–3 weeks and resulting in supraphysiological serum testosterone levels, injections of TU at intervals of up to 3 months offer an excellent alternative for substitution therapy of male hypogonadism.
Lack of Glutamate Transporter EAAC1 in the Epididymis of Infertile c‐ros Receptor Tyrosine‐Kinase Deficient Mice
Authors: WAGENFELD ANDREA , YEUNG CHING‐HEI , LEHNERT WILLI , NIESCHLAG EBERHARD , COOPER TREVOR G.
Published: November 2023

ABSTRACT: Transgenic male mice carrying inactive mutations of the receptor tyrosine kinase c‐ros lack the caput epididymidis initial segment and are infertile because sperm volume regulation is compromised. Complementary DNA arrays were used to detect differences in gene expression in the caput epididymidis of heterozygous fertile and homozygous infertile males. The glutamate transporter excitatory amino acid carrier 1 (EAAC1) was expressed in all epididymal regions with high expression in the initial segment and cauda epididymidis. Homozygous knockout mice did not express EAAC1 messenger RNA (mRNA) in the caput but they did express the gene in the corpus and cauda. Immunohistochemical staining for EAAC1 confirmed regional mRNA expression and demonstrated an adluminal location on stereocilia/microvilli of principal cells. The glutamate transporter‐associated protein (GTRAP) 3–18 was detected in all epididymal regions independent of genotype, but a highly abundant novel transcript of 4.2 kilobases was found only in the initial segment of heterozygous c‐ros mice. High‐performance liquid chromatography measurement of glutamate revealed a significantly higher content in the proximal caput of infertile mice than fertile mice, and tissue glutamate content decreased distally in both genotypes. Because glutamate is used as an osmolyte in somatic cells, the lack of EAAC1 reported here may disturb normal osmolyte balance in the proximal epididymal lumen and compromise sperm maturation, in particular the development of sperm volume regulatory mechanisms.
Molecular Cytogenetic Detection of Meiotic Segregation Patterns in Sperm Nuclei of Carriers of 46,XY,t(15;17)(q21; q25)
Authors: ĈORA TULIN , ACAR HASAN , KAYNAK MURAT
Published: November 2023

ABSTRACT: Structural chromosomal abnormalities in gonadal tissue represent an important category of parentally transmittable unbalanced chromosomal abnormalities to the offspring. A child with multiple anomalies was sent for cytogenetic analysis, and his karyotype was 46, XY, der(17)t(15; 17)(q21; q25). This abnormality was transferred from his grandfather to his father and to the proband. In this family, 5 persons (1 female and 4 male) are the carriers of this abnormality. In this study, fluorescence in situ hybridization (FISH) on sperm nuclei of 4 male carriers was studied to determine the distribution of segregation patterns of the balanced translocation 15q; 17q. The segregation results showed that the segregation products in the third carrier (the grandfather) were different, but they were not statistically significant. The segregation patterns in the other carriers were similar. Overall, 50.3% of the sperm nuclei (mean value for 4 carriers) analyzed were the result of alternate segregation; 36.9%, of adjacent I segregation; 9.0%, of adjacent II segregation; and 2.4%, of 3:1 segregation; the remaining 1.3% could be diploid sperm nuclei or of 4:0 segregation. Multicolor FISH analysis appears to be a rapid and reliable method for the direct analysis of segregation patterns in sperm nuclei of carriers of balanced reciprocal translocation, and it also provides interesting information for determining the possible risks for the offspring.
CAG Repeat Expansion in the Androgen Receptor Gene Is Not Associated With Male Infertility in Indian Populations
Authors: THANGARAJ KUMARASAMY , JOSHI MANJUNATH B. , REDDY ALLA G. , GUPTA NALINI J. , CHAKRAVARTY BAIDYANATH , SINGH LALJI
Published: November 2023

ABSTRACT: CAG repeat expansion in exon 1 of the androgen receptor (AR) gene has been reported to be associated with male infertility in some but not all populations. Until now, studies have not been carried out to examine this among Indian populations. For the first time, we have analyzed the CAG repeat motif in the AR gene in 280 men with azoospermia and in 201 men with normal fertility. The mean number of CAG repeats in the AR gene of men with azoospermia was 21.7 ± 0.18, with a high incidence of repeat number 22. Among fertile‐control men, the mean number of CAG repeats was 22.4 ± 0.19, with a predominance of repeat number 23. The highest number of CAG repeats (32) was found with low frequency in both fertile and azoospermic groups. Comparison of fertile men and those with azoospermia on the basis of CAG repeats revealed that the number of CAG repeats in both groups were similar, as revealed with a paired t test (t = 0.04; P = .967). Expansion of the CAG repeat in the AR gene is therefore not associated with male infertility in Indian populations. This suggests that what is true for one population may not be true for other populations.
Immunohistochemical Localization of the Retinoic Acid Receptors in Human Prostate
Authors: RICHTER FRANK , JOYCE ALINA , FROMOWITZ FRANK , WANG SHULUN , WATSON JOHN , WATSON RICHARD , JR ROBERT J. IRWIN , HUANG HOSEA F. S.
Published: November 2023

ABSTRACT: Retinoic acid receptors (RARs) are nuclear transcription factors that mediate the effects of retinoids. Aberrant expression and regulation of RARs have been linked to various malignancies, including steroid‐related breast and cervical cancers. Our previous results also suggest that prostate cancer is associated with altered RAR signaling. To understand the relationship between RAR signaling and prostate cancer, the current study examined the cellular distribution of RAR‐α,‐β, and ‐γ in human prostate tissues exhibiting different pathologic conditions. In histologically normal epithelium, both RAR‐α and ‐γ were present throughout the epithelium with minimal nuclear accumulation. RAR‐β was present only in basal epithelial nuclei. On the contrary, RAR‐α was significantly increased in the nuclei of luminal epithelial cells, and both RAR‐β and ‐γ were increased in basal and luminal epithelial nuclei in glands exhibiting benign prostatic hyperplasia (BPH). RAR‐α was also increased in luminal epithelial nuclei in glands exhibiting prostatic intra‐epithelial neoplasia (PIN). In these glands, RAR‐β was persisting in basal epithelial nuclei that were also RAR‐γ positive. In low‐ and intermediate‐grade cancerous glands, RAR‐α was also significantly increased in luminal epithelial nuclei, and a strong RAR‐γ signal was seen in some cells. RAR‐β was absent in these glands. Both RAR‐α and ‐γ were also increased in high‐grade cancer cells. In conclusion, current results demonstrated changes in cellular distribution of RAR‐α and ‐γ in human prostate tissues exhibiting different pathologies. These results suggest links between altered RAR signaling and deregulated cell growth and/or tumorigenic transformation of prostate epithelial cells.
Inhibin B Regulating Follicle‐Stimulating Hormone Secretion During Testicular Recrudescence in the Male Golden Hamster
Authors: JIN WANZHU , HERATH CHANDANA B. , YOSHIDA MIDORI , ARAL KOJI Y , SAITA ERINA , ZHANQUAN SHI , REN LONGQUAN , WATANABE GEN , GROOME NIGEL P. , TAYA KAZUYOSHI
Published: November 2023

ABSTRACT: In the present study, to clarify whether inhibin affects follicle‐stimulating hormone (FSH) secretion in the recrudescence of the male golden hamster, we used a recently developed specific enzyme‐linked immunosorbent assay (ELISA) in order to measure 2 forms of inhibin molecules: inhibin B and inhibin pro‐αC. In addition, we used the radioimmunoassay (RIA) to measure immunoreactive (ir‐)inhibin, FSH, luteinizing hormone (LH), and testosterone. And finally, we used the proliferating cell nuclear antigen (PCNA) and computer‐assisted sperm motion analysis (CASA) methods to ascertain how well spermatogenesis and sperm motility recover from the photoinhibition caused by exposure to a short‐day (SD; 10‐hour light:14‐hour dark) photoperiod. Animals were exposed to SD for 15 weeks, and then their testes were checked carefully and found to be completely regressed. Thereafter, those animals were transported to a long‐day (LD; 14‐hour light:10‐hour dark) photoperiod. Sampling was carried out at weeks 0 (exposed SD 15 weeks), 1, 2, 4, 6, 8, and 10. Plasma FSH rapidly increased and reached peak levels 2 weeks after transferral to the LD photoperiod and then declined to normal LD levels at week 6. Circulating ir‐inhibin, inhibin B, and inhibin pro‐αC rose to normal LD levels by week 4. A highly significant inverse correlation was observed between plasma FSH and inhibin B but not between FSH and either ir‐inhibin or inhibin pro‐αC. Plasma testosterone recovered to normal LD levels within 1 week. Sperm motility parameters were low until week 2 and recovered to normal LD levels from weeks 4 to 10. PCNA‐labeled cells were confined to the spermatogenic cells of the seminiferous tubules, though Leydig and Sertoli cell nuclei were never stained for PCNA during the period studied. The number of pachytene spermatocytes and the diameter of seminiferous tubules increased in a time‐dependent manner after transferral from SD to LD. In conclusion, these results suggest that 1) secretion of inhibin B may be stimulated by an early rise in FSH; 2) inhibin B suppresses FSH secretion from weeks 2 to 10, after transferral to the LD photoperiod; and 3) testes recrudescence is based on the increase in the number of sperm cells instead of the increase in the number of Sertoli and Leydig cells of the male golden hamster.
Immortalized Epididymal Cell Lines From Transgenic Mice Overexpressing Temperature‐Sensitive Simian Virus 40 Large T‐Antigen Gene
Authors: ARAKI YOSHIHIKO , SUZUKI KICHIYA , MATUSIK ROBERT J. , OBINATA MASUO , ORGEBIN‐CRIST MARIE‐CLAIRE
Published: November 2023

ABSTRACT: Epididymal epithelium is well known as a site of secretion of various proteins present in epididymal luminal fluid. Although there have been many reports of primary cultures of epididymal epithelial cells, their growth is limited over time. We have established immortalized epididymal epithelial cell lines from primary cultures of epididymal cells from transgenic mice harboring temperature‐sensitive simian virus 40 large T‐antigen gene in order to study the regulatory mechanisms of epididymal function, including specific factor secretion. These cell lines (PC1 from proximal caput; and DC1, DC2, and DC3 from distal caput) have been maintained for more than 1 year and show temperature‐dependent growth and expression of cytokeratin, a marker of epithelial cells. These cells express the androgen receptor as well as markers of the murine epididymal epithelium, PEB‐like protein (ie, phosphatidye ethanolamine binding protein), E‐RABP (ie, epididymal retinoic acid‐binding protein), and EP17 (ie, epididymal protein of 17 kd). The androgen‐regulated 5‐kilobase mE‐RABP promoter DNA fragment ligated to the neomycin‐resistant gene was used for stable transfection of DC1 cells. Because the mE‐RABP gene is specifically expressed in the distal caput, neomycin selection provides a pure population of epithelial cells from that segment. This neomycin‐resistant immortalized cell line from the distal caput was cultured for more than 6 months. Such immortalized cell lines should be valuable tools for studying the regulation of tissue‐specific gene expression, and may be used to identify one or more epididymal specific transcription factors involved in the expression of epididymal specific proteins.
Serum Androgen Bioactivity During 5α‐Dihydrotestosterone Treatment in Elderly Men
Authors: RAIVIO TANELI , TAPANAINEN JUHA S. , KIMELIUS PEKKA , JÄNNE OLLI A.
Published: November 2023

ABSTRACT: The androgens used in the treatment of age‐related androgen decline have different bioactivities that cannot be evaluated with conventional detection methods for serum steroids. We have recently developed a recombinant cell bioassay for the determination of androgen bioactivity in human serum that is based on androgen‐specific interaction between the ligand‐binding domain (LBD) and the N‐terminal region of the androgen receptor (AR). In this work, we examined the effect of topically applied 5α‐dihydrotestosterone (DHT; 7.5–10 g of 2.5% DHT gel daily for 6 months) on circulating androgen bioactivity in 14 men (age range, 51–63 years) with symptoms of andropause and pretreatment serum testosterone less than 15 nM, or serum sex hormone‐binding globulin concentration greater than 30 nM, or both. The mean (±SEM) pretreatment androgen bioactivity was 3.3±0.3 nM testosterone equivalents, and the levels correlated with serum testosterone concentration (r = .55, P #lT .05). DHT gel treatment induced a sixfold increase (from 1.5±0.1 nM to 9.0±0.7 nM) in mean serum DHT level, whereas endogenous testosterone and estradiol levels measured with radioimmunoassays were suppressed by approximately 70% and approximately 50%, respectively (P < .0001). Concomitantly, serum androgen bioactivity increased by sevenfold (from 3.3±0.3 to 23.6±2.8 nM testosterone equivalents; P < .0001). We conclude that DHT gel therapy in elderly men significantly increases their circulating androgen bioactivity as measured with a mammalian cell bioassay. An androgen‐specific bioassay such as ours may enable investigation of other androgens with different bioactivities, such as selective AR modulators.
Nocturnal Penile Tumescence and Rigidity Testing in Bicycling Patrol Officers
Authors: SCHRADER STEVEN M. , BREITENSTEIN MICHAEL J. , CLARK JOHN C. , LOWE BRIAN D. , TURNER TERRY W.
Published: November 2023

ABSTRACT: A health assessment study was conducted in response to complaints of groin numbness in a bicycling police unit. Seventeen male cyclists were compared with 5 nonbiking men. The cyclists rode an average of 5.4 hours per day, and 91% indicated they experienced groin numbness on occasion. Each man wore the RigiScan Plus Rigidity Assessment System for one normal sleep session. Pressure measurements were also taken between the cyclist and the bicycle saddle. The percentage of sleep sessions that recorded an erectile event was significantly lower in the cyclists than it was in noncyclists (cyclists 27.1%; noncyclists 42.8%; P = .008). This duration percentage is negatively correlated with average hours a day that cyclists rode their bikes (r = ‐.41; P = .05), the number of days a week they rode (r = ‐.55; P = .009), and the average pressure exerted on the nose of the bike saddle (r = ‐.39; P = .08). The other measures of erectile quality (tumescence activity units [TAUs] and rigidity activity units [RAUs] of both the base and tip of the penis) were lower in the cyclists, but did not reach statistical significance. The number of hours cyclists rode during the day of RigiScan Plus assessment was negatively correlated with penis tip RAU (r = ‐.41; P = .04), and tip TAU (r = ‐.45; P = .04). These data suggest that prolonged bicycle riding may have negative effects on nocturnal erectile function and indicate a need for innovative bicycle saddle designs.
Effect of Different Incubation Conditions on Phosphatidylserine Externalization and Motion Parameters of Purified Fractions of Highly Motile Human Spermatozoa
Authors: SCHUFFNER ALESSANDRO , MORSHEDI MAHMOOD , VAAMONDE DIANA , DURAN E. HAKAN , OEHNINGER SERGIO
Published: November 2023

ABSTRACT: The objective of this study was to assess the temporal effects of sperm incubation at body temperature with various amounts of human serum albumin (HSA) on motion parameters and phosphatidylserine externalization, an expression of membrane integrity. Purified sperm populations were prepared by discontinuous gradient separation, incubated at 37°C in 3 different culture conditions (human tubal fluid [HTF] alone, HTF plus 0.3% HSA, and HTF plus 3% HSA) and evaluated at 0, 1, 3, 6, and 24 hours. Annexin V binding was used to monitor membrane translocation of phosphatidylserine and a computer‐assisted semen analyzer was used to evaluate motion parameters. All incubation conditions led to a time‐dependent, significant decline in sperm motion parameters and an increase in exposure of phosphatidylserine (annexin V+, live cells) to the outer leaflet of the plasma membrane in both patients and donors. Patients had a higher degree of motility loss and externalization of phosphatidylserine than donors. The decline in the percentage of normal cells (annexin V−, live) was greater in HTF alone up to 6 hours, and the decline in the percentages of motile and rapid sperm were greater in HTF alone throughout 24 hours when compared with HSA supplementation. We conclude that prolonged incubation of purified populations of highly motile human spermatozoa at body temperature was associated with significant motility loss and membrane changes as revealed by phosphatidylserine translocation. A higher concentration of HSA resulted in a relative protective effect against such impairments, particularly during the first 6 hours of incubation. Under the experimental conditions tested, significant differences were observed between infertile men and fertile controls.
Enolase Isoforms Activities in Spermatozoa From Men With Normospermia and Abnormospermia
Authors: FORCE ANDRE , VIALLARD JEAN‐L. , GRIZARD GENEVIEVE , BOUCHER DANIEL
Published: November 2023

ABSTRACT: Total enolase as well as enolase‐αα (ENO‐aa, ubiquitous) and enolase‐S (ENO‐S, sperm‐specific) activities were measured in total and Percoll‐selected sperm from 30 normospermic fertile men and 20 abnormospermic infertile patients. The total enolase activity was significantly higher in total sperm from patients with abnormospermia compared with normospermic patients (11.1 ± 1.9 vs. 4.8 ± 0.5 mIU/107 sperm P < .05). ENO‐αα activity was significantly higher in total sperm from abnormospermic men than from normospermic men (P < .05). ENO‐αα activity in Percoll‐selected sperm was significantly lower compared with total sperm in both group of patients; however, for the same sperm fraction ENO‐αα activity did not differ between normospermic and abnormospermic men. ENO‐αα activity was related to the cell contamination ratio and to the percentage of spermatozoa with abnormal morphology. Furthermore, ENO‐αα was positively correlated with the percentage of immature sperm showing an excess of residual cytoplasm. ENO‐S activity was significantly higher in total sperm from normospermic patients than from abnormospermic patients (P < .05). ENO‐S activity in Percoll‐selected sperm was not significantly different compared with total sperm in both group of patients. However, this activity was significantly lower in Percoll‐selected sperm from abnormospermic men compared with normospermic men (P < .05). ENO‐S activity was not related to the cell contamination ratio but was significantly correlated with the percentage of spermatozoa with normal morphology. The 2 enolase isoforms seem to reflect 2 opposite aspects of sperm cells quality: ENO‐αα is associated with abnormal spermatozoa, immature spermatozoa, or both; and ENO‐S is associated with normal spermatozoa. As an additional index to distinguish normal from abnormal semen, the ENO‐S:ENO‐αα ratio was evaluated for total and Percoll‐selected sperm in both groups. This ratio seems to be a new, valuable marker of the global sperm quality in a given semen sample, and may represent a predictive index of sperm fertilizing potential.
Importance of Glycosylation and Disulfide Bonds in Hyaluronidase Activity of Macaque Sperm Surface PH‐20
Authors: LI MING‐WEN , YUDIN ASHLEY I. , ROBERTSON KATHRYN R. , CHERR GARY N. , OVERSTREET JAMES W.
Published: November 2023

ABSTRACT: PH‐20 is a glycoprotein located on the surface of the sperm plasma membrane and on the inner acrosomal membrane. The best understood function of sperm surface PH‐20 is its hyaluronidase activity, which results in hydrolysis of the hyaluronic acid‐rich cumulus matrix during sperm penetration of this extracellular oocyte investment. In this study, we investigated whether alterations in the secondary and tertiary structures of sperm surface PH‐20 would affect its enzyme activity. Proteins were isolated from the sperm plasma membrane by treatment of living cells with phosphatidylinositol‐specific phospholipase C (PI‐PLC). PH‐20 was purified from the PI‐PLC released proteins by immunoaffinity chromatography. Two‐dimensional electrophoresis of purified PH‐20 revealed 6 isoforms with isoelectric points ranging from 5.1 to 6.0. Removal of the N‐linked glycans from PH‐20 with N‐glycosidase F shifted the molecular weight from 64 kd to approximately 54 kd, its deduced molecular weight based on sequence analysis, suggesting that most if not all, of the potential N‐glycosylation sites are linked to oligosaccharides. The lectins Con A and PSA recognized purified sperm surface PH‐20 after Western blotting, suggesting that mannose is a major sugar within or at the terminal end of the linked glycan. The lectins UEA and LPA did not recognize PH‐20 Western blot, suggesting that fucose and sialic acid are not terminal sugars of sperm surface PH‐20. Deglycosylation of sperm surface PH‐20 resulted in a complete loss of its hyaluronidase activity. The reduction of disulfide bonds with β‐mercaptoethanol or dithiothreitol also resulted in loss of enzyme activity. We conclude that the hyaluronidase activity of sperm surface PH‐20 is dependent on structural features established by sulfhydryl linkages, as well as glycosylation.
Differences in Sperm Motion Between High‐ and Low‐Shuttlebox Avoidance Rats (Hatano Strains)
Authors: SATO MASAKO , OHTA RYO , KOJIMA KOHICHI , SHIROTA MARIKO
Published: November 2023

ABSTRACT: Sperm from the caudal epididymis of 2 inbred strains of Sprague‐Dawley (SD) rats, selected on the basis of their high‐or low‐shuttlebox avoidance responses, were analyzed for motion characteristics by a computer‐assisted sperm motion analysis (CASA) system. Sperm motion in high‐avoidance animals (HAA) was characterized by high velocities, high amplitude of lateral head displacement (ALH), and low beat cross frequency (BCF). Conversely, sperm from low‐avoidance animals (LAA) displayed low velocities, low ALH, and high BCF. These characteristics in sperm motion were not changed by washing. Furthermore, after treatment with alpha‐chlorohydrin (aCH) as a male antifertility agent affecting rat epididymal sperm motion, sperm velocities in HAA rats were significantly reduced to levels similar to those in untreated LAA rats. However, ALH and BCF in HAA rats treated with aCH were different from those in untreated LAA rats. Sperm adenosine triphosphate (ATP) content was higher in HAA than in LAA rats, correlating with values of their sperm velocities. These data suggest there are apparent strain differences in sperm motion between HAA and LAA rats and that these differences are dependent on factors, including sperm energy production.
Aneuploidy and Chromosome Breakage in Swim‐Up Versus Unprocessed Semen From Twenty Healthy Men
Authors: ONG THUAN D. , XUN LIN , PERREAULT SALLY D. , ROBBINS WENDIE A.
Published: November 2023

ABSTRACT: Toxicological and epidemiological studies have investigated several factors that are believed to induce cytogenetic damage in human sperm cells in an effort to estimate heritable risk to future generations. Most of these studies have not differentiated damage based on cell fertility or motility. In the clinical setting, intracytoplasmic sperm injection (ICSI) bypasses the natural process of sperm selection. Although practitioners attempt to select motile sperm for ICSI, the sperm may not always demonstrate motility, maturity, or even viability. Knowing whether cytogenetic damage differs in motile versus unselected sperm would improve our ability to estimate heritable risk and lead to improved ICSI procedures, and would expand the body of toxicology and epidemiology research. We divided semen samples from 20 healthy donors and compared aneuploidy and chromosome breakage in sperm cells gathered directly from the ejaculate (unprocessed semen) with cells enriched for motility using the swim‐up assay. Sperm fluorescence in situ hybridization was used to detect aneuploidy for chromosomes 13, 18, 21, X, and Y. Tandem labeling probes were used to detect breakage in the 1cen‐1q12 region of chromosome 1. The occurrence of disomy 1818 and XY18 was significantly lower in specimens enriched for motility (P = .004 and P = .001, respectively). Sperm that carried duplication errors and diploid sperm were also seen less frequently in semen analyzed by the swim‐up assay (P < .008). Chromosome 1 breakage did not differ between swim up‐assayed and unprocessed specimens. Findings suggest that unprocessed semen may overestimate heritable aneuploidy risk in sperm biomarker studies, and may be biologically relevant to ICSI in disomy categories 18–18 and XY18, demonstrating 1.4‐fold to 1.8‐fold differences.
The effect of tyrosol on diclofenac sodium‐induced acute nephrotoxicity in rats
Authors: Çömez Mehmet , Cellat Mustafa , Kuzu Müslüm , Uyar Ahmet , Türk Erdinç , Kaya Yusuf Selim , Etyemez Muhammed , Gökçek İshak , Güvenç Mehmet
Published: November 2023

Although diclofenac (DCF) is a nonsteroidal anti‐inflammatory drug that is considered safe, its chronic use and overdose may show some toxic effects. The protective effect of tyrosol (Tyr) pretreatment against DCF‐induced renal damage was investigated in this study. The 32 rats used in the study were randomly divided into four groups of eight rats each. According to the data obtained, it was determined that creatinine, urea, and blood urea nitrogen (BUN) levels increased in serum samples of the DCF group. Besides, the levels of reduced glutathione (GSH) and glutathione peroxidase (GPx) activity decreased and the malondialdehyde (MDA) level increased in the kidney tissue. However, no change was observed in catalase (CAT) activity. Cyclooxygenase‐2 (COX‐2), nuclear factor kappa B (NF‐κB), and tumor necrosis factor‐alpha (Tnf‐α) levels increased and nuclear factor erythroid 2‐related factor 2 (Nrf‐2) levels decreased. No change was detected in the level of interleukin 1 beta (IL‐1β). When the DCF+Tyr group and the DCF group were compared, it was assessed that Tyr had a curative effect on all biochemical parameters. Also, kidney damages, such as degeneration and necrosis of tubular epithelium and congestion of veins, were obviated by treatment with tyrosol in histopathological examinations. It was determined that Tyr pretreatment provided a protective effect against nephrotoxicity induced by DCF with its anti‐inflammatory and antioxidant properties.
Protecting yourself: Due diligence in mergers and acquisitions
Authors: Conway A. Carr , Rouse Robert W.
Published: November 2023

PublicComp spent a lot of money to acquire another company. It seemed well worth it, based on financial
reports. But a few weeks later, the CEO learned about some unusual journal entries on the acquired company's
books—and some troubling letters from customers. How could this have been prevented? ©
2002 John Wiley & Sons, Inc.