bluuebunny/crossref_metadata_2025 huggingface dataset:
A Study of Phospholipase in Albumin and its Role in Inducing the Acrosome Reaction of Guinea Pig Spermatozoa in Vitro
Authors: SINGLETON CRYSTAL L. , KILLIAN GARY J.
Published: December 2015

Phospholipase activity associated with guinea pig spermatozoa during capacitation and the acrosome reaction (AR) was determined by measuring the production of [14C] glycerylphosphorylcholine (GPC) from [14C phosphatidylcholine (PTC) over a 2‐hour period at the beginning and at the end of a 6‐hour incubation. Spermatozoa converted 13 × more [14C] PTC to [14CJ GPC during the last 2 hours of incubation than during the first 2 hours which corresponded to an increase in AR from 7% at 2 hours to 55% at 6 hours. In vitro studies also were performed to assess the role of phospholipase activity in bovine serum albumin (BSA) in inducing the AR of guinea pig spermatozoa. Spermatozoa, obtained by backflushing the cauda epididymides, were incubated for up to 6 hours in Biggers, Whitten, and Whittinghams' (BWW) medium alone or containing various additives. These additives were: 0.1% BSA, phospholipase A2, p‐bromophenacyl bromide, DMSO, 0.1% BSA plus p‐bromophenacyl bromide, or 0.1% BSA plus DMSO. Each hour of incubation, samples were assessed for the percentage of AR sperm and sperm motility. The percentage of AR sperm after 6 hours in BWW supplemented with phospholipase A2, at the level of PTC hydrolyzing ability detected in BWW with 0.1% BSA, was similar to that of BWW and BSA. However, the percentage of AR sperm in BWW with BSA and the phospholipase inhibitor p‐bromophenacyl bromide was significantly lower than that in BWW with BSA or phospholipase A2 and similar to that in BWW without BSA. Sperm motility was significantly less in incubations containing phospholipase A2, but lacking BSA. It was concluded that phospholipase activity in BSA may contribute to capacitation and the AR of guinea pig sperm in vitro by mimicking the action of native sperm phospholipase.
Synthesis and Secretion of Glycoprotein by the Epididymal Epithelium
Authors: FLICKINGER CHARLES J.
Published: December 2015

The secretory activity of the middle segment of the mouse caput epididymidis was studied using 3H‐fucose as a precursor to glycoprotein. Young adult male mice were injected with a concentrated solution of 3H‐fucose interstitially, ie beneath the connective tissue capsule of the epididymis. Two animals were killed and prepared for light microscopic radioautography at each of six intervals between 10 minutes and 24 hours after injection. Silver grains were concentrated over the supranuclear Golgi region at 10 minutes and over the apical ends of the cells 30 minutes and 1 hour after injection. Quantitative analysis showed that luminal radioactivity increased greatly beginning with the 2‐hour samples. The results indicate that the epididymal epithelium synthesizes and secretes glycoproteins, and that 1 to 2 hours are required for terminal glycosylation, intracellular transport, and release of the secretory product.
Estrogens and Cytosolic Estrogen Receptors in Aged Male Rats
Authors: WINTERS STEPHEN J. , TAKAHASHI JUN
Published: December 2015

To determine whether estrogens may play a role in the decline in Leydig cell function in aged rats, plasma levels of testosterone, estradiol, and estrone; intratesticular concentrations of testosterone and estradiol; as well as cytosolic estrogen receptors were studied. Plasma and intratesticular testosterone levels were reduced in aged Sprague‐Dawley rats, but circulating levels of estrone and estradiol, as well as intratesticular estradiol concentrations, were similar in aged and young‐adult animals. Furthermore, in both testicular and kidney cytosol, the dissociation constants for estradiol and the concentrations of estradiol binding sites remained unchanged during aging. These data suggest that estrogens are unlikely to be important mediators of the reduced Leydig cell function in old rats.
Human Reproductive Tissues Contain Thyrotropin Releasing Hormone (TRH) and TRH‐homologous Peptides
Authors: PEKARY A. EUGENE , ROSS RICHARD , DEKERNION JEAN , HERSHMAN JEROME M.
Published: December 2015

Thyrotropin releasing hormone (TRH) immunoreactive peptides occur in high concentration within rat ventral prostate and human semen. To extend these observations to human reproductive organs, tissue fragments from human prostates undergoing benign hypertrophy were obtained by transurethral resection or open surgery. Human prostate, seminal vesicles, testes, and epididymis were also obtained from cadavers within 24 hours postmortem. After tissue extraction, the total TRH immunoreactivity (TRH IR) was measured by TRH radioimmunoassay. The total TRH IR in autopsy seminal vesicles was significantly greater (P < 0.01) than in all other autopsy reproductive tissues. The mean autopsy TRH IR of prostate was not significantly different from that measured in prostatic tissue obtained at surgery. High pressure liquid chromatography of extracts of autopsy seminal vesicles, prostate, and testis revealed multiple peaks of TRH IR. The two major peaks corresponded to the two TRH‐homologous peptides of human semen and one of the minor peaks cochromatographed with synthetic TRH. The distribution of TRH IR in human reproductive tissues appears to be very different from that in the rat, where ventral prostatic TRH IR levels exceed that of any other reproductive tissue by one to two orders of magnitude.
Acute Hormonal Changes Following Chemotherapy for Hodgkin's Disease in Man
Authors: NADER SHAHLA , SCHULTZ PAMELA , DEINZER DOROTHY , SAMAAN NAGUIB A.
Published: December 2015

Eleven men, aged 21 to 60 years, with Hodgkin's disease have been studied before, during, and after chemotherapy. Blood samples were obtained for hormone analyses. In all 11 patients FSH levels rose following chemotherapy, in ten LH and testosterone rose, and in seven estradiol rose above pretreatment values. For each hormone the mean peak post‐treatment value was significantly higher than pretreatment values. While there are mechanisms to explain the rise in testosterone and estradiol, the rise in LH in the face of increasing testosterone and estradiol values is unexplained. We propose that feedback regulation of LH by the testis includes a mechanism that is independent of testosterone and estradiol and that this mechanism is damaged during chemotherapy.
Morphology of Spermatozoa in Infertile Men with and without Varicocele
Authors: PORTUONDO JOSÉ A. , CALABOZO MARCELO , ECHANOJAUREGUI ABEL D.
Published: December 2015

This study was carried out to evaluate the morphology of spermatozoa in infertile men with and without varicocele. A series of 285 ejaculates were fully evaluated for seminal volume, sperm count, motility, and morphology, and classified into fertile (165 subjects), infertile without varicocele (93 subjects) and infertile with varicocele (27 subjects). Sperm morphology was classified by multiple entry criteria and recorded as normal, abnormal with head, midpiece, or tail single anomaly or abnormal with simultaneous multiple abnormalities.

Semen volume was almost identical in the three groups. Among the infertile men, sperm count was lower in those having a varicocele, but conversely those with varicocele had a higher percentage of motile spermatozoa, higher percentage of spermatozoa with forward movement and higher sperm velocity. There were higher proportions of spermatozoa with abnormal morphology, total number of anomalies, and multiple anomalies in infertile men, both with and without varicocele, than in fertile men. The percentage of abnormal spermatozoa was higher in infertile men with varicocele than in those without varicocele. The pattern of sperm morphology differed between the infertile and the fertile group, and with the presence or absence of varicocele. In the presence of varicocele, only the incidence of elongated (tapered) forms was significantly increased.
Reproductive History and Semen Analysis in Prevasectomy Fertile Men with and without Varicocele
Authors: DE CASTRO MARCOS PAULO P. , MASTROROCCO DIOGO A. M.
Published: December 2015

A group of 598 allegedly fertile men requesting vasectomy were investigated; varicocele was found in 97 (16.2%) of these men. The mean ages and age distributions of men with and without varicocele were not significantly different. Reproductive histories (number of pregnancies, living children and spontaneous abortions, as well as incidence of present pregnancy) were similar in both groups. The average seminal characteristics (semen volume, sperm count, total sperm count, percentage of motile spermatozoa, quality of motility, morphology) were not different for men with and without varicocele, except for a slight, but significantly higher incidence of oval‐headed sperm in men without varicocele. However, the incidence of varicocele was significantly higher in men with sperm counts below 40 million/ml. Three important observations may be made from this study: 1) the incidence of varicocele in this prevasectomy population was similar to that reported for the general population, but lower than the incidence reported for male partners of infertile couples; 2) in this population of allegedly fertile men, the presence of a varicocele did not significantly affect reproductive performance; 3) even though the incidence of varicocele was higher in men with sperm counts below 40 million/ml, the average seminal characteristics were not different in men with and without varicocele.
Specific Binding of the Glycosaminoglycan 3H‐Heparin to Bull, Monkey, and Rabbit Spermatozoa In Vitro
Authors: HANDROW RICHARD R. , BOEHM SHELLEY K. , LENZ RICHARD W. , ROBINSON JERRY A. , AX ROY L.
Published: December 2015

In Vitro binding and some binding parameters of the glycosaminoglycan heparin to viable epididymal or ejaculated bull spermatozoa, ejaculated rabbit spermatozoa, and frozen‐thawed rhesus monkey spermatozoa were investigated. Nonspecific binding was affected only by the concentration of 3H‐heparin, whereas specific binding was saturable, reversible, and dependent on thepH, temperature, and calcium concentration of the incubation medium. Magnesium concentration dependence was observed in the presence of calcium but could not be detected in the absence of calcium. Bound 3H‐heparin was displaced by several orders of magnitude greater concentrations of chondroitin sulfate. Scatchard plot analysis suggested multiple binding affinities for 3H‐heparin to spermatozoa. 3H‐heparin was shown to bind to sperm heads and flagella. Fluorescein‐labeled heparin bound to acrosomal, postacrosomal, and flagellar membranes. It was concluded that the specific binding of heparin involved a protein‐aceous component on, or intercalated with, spermatozoal membranes. Thus, glycosaminoglycans present in the female reproductive tract may contribute to sperm capacitation and enhance the likelihood of successful fertilization in mammals.
Seasonal Changes in Steroidogenesis in the Testis of the Rhesus Monkey (Macaca mulatto)
Authors: HIGASHI YOTSUO , TAKAHASHI JUN , YOSHIDA KEN‐ICHIRO , WINTERS STEPHEN J. , OSHIMA HIROYUKI , TROEN PHILIP
Published: December 2015

To investigate the influence of seasonal changes from breeding to nonbreeding season on testicular steroidogenesis in the adult rhesus monkey, two animals were hemiorchidectomized in November and the second testis of each was removed the following April. Intratesticular steroid concentrations related to the formation of testosterone from pregnenolone were measured by radioimmunoassay. Related steroidogenic enzyme activities were also assayed and dynamic studies of the bioconversion of radiolabeled precursor steroids by testicular microsomes were performed.

Testosterone was synthesized from pregnenolone through both dehydroepiandrosterone and 17α‐hydrox‐yprogesterone. No distinct differences were found in the testosterone biosynthetic pathway between testes in breeding and nonbreeding seasons. In the breeding season, intratesticular testosterone concentrations, the sum of intratesticular steroids measured, and the ratio of testosterone to the steroid sum were increased when compared to those in the nonbreeding season. No major seasonal changes in microsome enzyme activities were found, although enzyme activities, except for Δ5‐3β‐hydroxysteroid dehydrogenase for C21‐steroids, were slightly higher in the breeding season. We conclude that increased testosterone production in the testis during the breeding season is due primarily to increased pregnenolone supply from Leydig cell mitochondria and, to a lesser extent, to an enhanced pregnenolone bioconversion to testosterone in the microsomes.
Fibroblast Androgen Receptors in Patients with Genitourinary Anomalies
Authors: EIL CHARLES , CRAWFORD JOHN D. , DONAHOE PATRICIA K. , JOHNSONBAUGH ROGER E. , LORIAUX D. LYNN
Published: December 2015

The etiology of certain disorders of sexual differentiation is unclear. The authors have examined the hypothesis that hypospadias and other disorders compatible with a defect in androgen action, such as cryptorchidism, micropenis, chordee/penile torsion, and ectopic testis, might be explained by androgen receptor abnormalities. Therefore, 25 subjects were studied who were selected only because they had one of these developmental defects, together with a predominantly male phenotype, and no readily ascertainable explanation for the defect. Four of these subjects had mixed gonadal dysgenesis with multiple genito‐urinary anomalies. They were included for comparative purposes, since there is no evidence for androgen resistance in this disorder. Patients with testicular regression syndrome (gross testosterone deficiency), impaired testosterone biosynthesis (relative testosterone deficiency), 5α‐reductase deficiency (altered T/DHT ratio), and a family history or endocrine profile suggestive of androgen resistance, were all excluded from evaluation. Androgen receptor content (Ro) and binding affinity (Kd) were measured in 26 genital or pubic skin fibroblast strains cultured from 25 affected patients using a dispersed, whole cell assay at 22 C. There was no difference in the mean androgen receptor content (approximately 10,000 sites/cell) or binding affinity (approximately 1 nM) between the patients' fibroblasts and those from 26 fibroblast strains established from 26 normal males. Moreover, there were no differences in the nuclear uptake of [3H]dihydrotestosterone into dispersed, intact fibroblasts incubated at 37 C when 11 patient and seven normal male fibroblast strains were compared. In conclusion: these disorders of sexual differentiation cannot be attributed to androgen resistance on the basis of decreased androgen receptor binding affinity or receptor number, as reflected in whole cells or nuclei of cultured skin fibroblasts.
Inhibition of 5a‐Reductase Activity and Alteration of Nuclear Testosterone
Authors: BERKOVITZ GARY D. , BROWN TERRY R. , MIGEON AND CLAUDE J.
Published: December 2015

17β‐N, N‐diethylcarbamoyl‐4‐methyl‐4‐aza‐5α‐an‐drostan‐3‐one (4‐MA) inhibits 5α‐reductase activity in cultured human genital skin fibroblasts. Enzyme kinetic studies analyzed by Eadie‐Hofstee plots demonstrated that 4‐MA is a competitive inhibitor, with an apparent Ki of 15 nM. 4‐MA had a very low binding affinity for the androgen receptor. When fibroblasts were incubated in the presence of testosterone (T) and 4‐MA, nuclear uptake of 5α‐dihydrotestosterone (DHT) decreased in parallel with the inhibition of 5?‐reduc‐tase activity. While the overall sum for the nuclear uptake of T and DHT diminished, the nuclear uptake of T increased. Biological androgen inactivity cannot be precluded on the basis of nuclear T plus DHT uptake.
Scrotal Hypothermia and Testicular Blood Flow in the Dog
Authors: GLODE L. MICHAEL , ROBINSON JULIE , HORWITZ LAWRENCE D.
Published: December 2015

Two clinically tolerable methods of producing testicular hypothermia have been studied in dogs. Evaporative cooling produced a 3.7 degree fall and ice water a 9.7 degree fall in intratesticular temperatures. Scrotal skin blood flow (mls/100 gm/min) fell from 4.9 ± 4.6 to 2.3 ± 1.2 and 1.3 ± .05 with the two techniques and was associated with increased vascular resistance. In contrast, no significant change was noted in testicular blood flow. These data show that testicular blood flow cannot be altered by clinically tolerable hypothermia.
A New Microsurgical Technique for Varicocele Correction
Authors: BELGRANO E. , PUPPO P. , GABOARDI F. , TROMBETTA C.
Published: December 2015

Varicocele is the consequence of a hemodynamic abnormality causing venous hypertension and consequent dilatation of the spermatic veins. In many cases, the correction of varicocele has led to improvement in semen quality. Conventional techniques of varicocele correction have the sole aim of occluding a nonfunctioning venous pathway, but do not attempt restoration of a hemodynamically correct venous drainage of the testis. We propose a new microsurgical technique for varicocele correction that achieves anastomosis between the spermatic vein and the epigastric vein. Either stump of the epigastric vein can be used, depending on the type of varicocele. In this way, a hemodynamically correct surgical solution can be achieved.
Seminiferous Tubule Degeneration in Human Cryptorchid Testes
Authors: AMAT PEDRO , PANIAGUA RICARDO , MONTERO JUAN
Published: December 2015

Two types of degenerating seminiferous tubules were found in cryptorchid testes with Sertoli cell hyperplasia of children and adults: 1) tubules with central degeneration, and 2) tubules with total degeneration. Central degeneration begins with degenerative changes in germ cells that accumulate in the lumen of the seminiferous tubule. Some Sertoli cells may also be affected. Degenerated cells finally disappear, and the remaining tubule is composed of only a cuboidal epithelium, which consists mainly of Sertoli cells and occasional germ cells surrounding a wide lumen. Total degeneration is principally seen in tubules with severe germinal hypoplasia. All the seminiferous epithelium cells degenerate and lose their characteristic distribution, forming a disorganized Sertoli cell nodule surrounded by a thickened basement membrane. Lastly, Sertoli cells disintegrate, and the seminiferous epithelium disappears. Tubular degeneration might be related to the thickening of the basement membrane, which hinders metabolic interchange between the seminiferous epithelium and the interstitium.
Effect of Gossypol Acetate on Guinea Pig Epididymal Spermatozoa In Vivo and Their Susceptibility to Capacitation In Vitro
Authors: SHI QI‐XIAN , FRIEND DANIEL S.
Published: December 2015

To determine the effects of gossypol acetate on guinea pig epididymal and vas deferens sperm maturity and in vivo susceptibility to in vitro capacitation and the acrosome reaction, we examined spermatozoa removed from 37 animals fed gossypol acetate (10–15 mg/kg/day) for 5 to 9 weeks, and 15 vegetable oil‐fed, age‐paired control animals. In gossypol‐treated, reproductively immature guinea pigs, the number of spermatozoa in the epididymis was markedly reduced (P < 0.01) compared to controls, whereas the presence of spermatids and spermatocytes increased in the epididymis with the duration of gossypol administration. In sexually mature guinea pigs (given 15 mg/kg/day for 5 weeks), the epididymal sperm survival and forward motility were decreased significantly (P < 0.025 and P < 0.01, respectively), although the density of mature spermatozoa was the same as in control animals. The percentage of induced acrosome reactions (26.4 ± 12%) was almost three‐fold lower than that of control animals (72.8 ± 4.6%). Also, in 31.5 ± 3.8% of spermatozoa from gossypol‐treated animals, as compared to only 2.4 ± 0.7% of controls, the cytoplasmic droplet failed to migrate to its proper position in the midpiece and was retained in the neck region. With a few exceptions, spermatozoa from both experimental and control groups had comparable patterns of freeze‐fractured membrane differentiations. Susceptibility to the induced acrosome reactions and the position of the retained cytoplasmic droplet reversed within 3 weeks after the end of gossypol feeding. This study helps establish the suitability of the guinea pig for studies on gossypol‐induced infertility.
Dose–Response Studies on Male Reproductive Parameters in Dogs with Nafarelin Acetate, a Potent LHRH Agonist
Authors: VICKERY BRIAN H. , McRAE GEORGIA I. , BRIONES WILLIAM V. , ROBERTS BRYAN B. , WORDEN ANNE C. , SCHANBACHER BRUCE D. , FALVO RICHARD E.
Published: December 2015

Adult male beagle dogs were administered daily subcutaneous injections of either 0.5 or 2.0 μg/kg of a potent LHRH agonist, nafarelin acetate, for 44 days. Although there was a rise in the circulating levels of the gonadotropins and of testosterone following the early injections of agonist, continued treatment caused a marked decline in acute response and basal levels of both LH and testosterone and smaller decreases in the acute FSH response. The decline in LH and testosterone was accompanied by decreases in testicular volume, ejaculated sperm count, sperm motility, ejaculate volume, and duration of ejaculation. The decline in these parameters was more rapid at 2.0 μg/kg than at 0.5 μg/kg. The profile of responses to 2.0 μg/kg could be superimposed on that previously shown for the injection of 10.0 μg/kg. At the end of treatment, prostate weights were 36% and 68% of vehicle‐treated controls for high‐ and low‐dose animals, respectively. Spermatogenesis was absent in the testes of all agonist‐treated animals. Over the dose range tested, the dose–response on all parameters was characterized by a slower evolution to the same maximal effect, rather than by a partial effect. If these data can be extrapolated to man, they would suggest that administration of higher dose levels of LHRH agonists than presently reported should be explored.
Testosterone Levels after Bromocriptine Treatment in Patients Undergoing Long‐term Hemodialysis
Authors: VIRCBURGER MIROSLAV I. , PRELEVIĆ GORDANA M. , PERIĆ LJILJANA A. , KNEŽEVIĆ JASMINA , DJUKANOVIĆ LJUBICA
Published: December 2015

Seventeen out of 34 male patients undergoing long‐term hemodialysis had increased basal plasma prolactin levels (mean = 1344 ± 1158.76 mU/L). Seven of these 17 patients having the greatest degree of erectile impotence were treated with 3.5 to 7.5 mg/day of bromocriptine. After a 4‐week treatment period, basal plasma prolactin levels in all seven patients were within normal limits (mean = 210.2 ± 66.97 mU/L). The treated patients reported an improvement in both libido and potency. At the same time, an increase in plasma testosterone levels was observed, while plasma LH and FSH levels were essentially unchanged.
Comparison of the Penetration Ability of Human Spermatozoa into Bovine Cervical Mucus and Zona‐free Hamster Eggs
Authors: TAKEMOTO FERN S. , ROGERS B. JANE , WILTBANK MILO C. , SODERDAHL DOUGLAS W. , VAUGHN WILLIAM K. , HALE RALPH W.
Published: December 2015

In vitro bovine cervical mucus (BCM) penetration tests, sperm penetration assays (SPA) using zona‐free hamster eggs, and routine semen analyses were performed on a total of 136 freshly collected semen samples from men who were seen at an infertility clinic. The correlations between bovine cervical mucus penetration and other semen parameters were the percent motile spermatozoa (r = 0.48), progressive motility grade (r = 0.44), sperm count (× 106/ml) (r = 0.47), the percent normal morphology (r = 0.32) and the percent eggs penetrated (r = 0.46) (P < 0.0001 for each correlation coefficient). When known fertile (n = 32) and infertile (n = 18) groups were tested, positive mucus penetration was associated 75% correctly and positive egg penetration was associated 90% correctly to clinical status. The mucus test had no false‐negative results and the SPA had no false‐positive results in these groups. It appears, then, that the mucus test and sperm penetration assay, although contributing different elements of data to an infertility evaluation, are both useful adjuncts to a semen analysis.
Rat Cauda Epididymal Fluid is a Mucus
Authors: USSELMAN MARION C. , CONE RICHARD A. , ROSSIGNOL DANIEL P.
Published: December 2015

Immobilin, the highly viscoelastic glycoprotein isolated from rat cauda epididymal fluid, exhibits all of the key biochemical characteristics of a mucin: 1) it has a very high molecular weight (will not pass through a 106 dalton cut‐off filter; 2) it contains 56% carbohydrate, with low or undetectable levels of mannose, xylose and uronic acid; 3) the carbohydrates (primarily galactose, N‐acetylglucosamine and N‐acetylgalactosamine) are arranged in short, oligosaccharide chains (4–20 monosaccharides per chain); 4) these oligosaccharide chains can be cleaved by NaOH in the presence of NaBH4, suggesting O‐glycosidic linkages; and 5) the protein core is pronase‐resistant. Immobilin, however, contains no detectable sialic acid, and 67% of the oligosaccharides are uncharged, indicating that immobilin is less acidic than most other mucins.
Androgens in Various Fluid Compartments of the Rat Testis and Epididymis after Hypophysectomy and Gonadotropin Supplementation
Authors: TURNER T. T. , EWING L. L. , JONES C. E. , HOWARDS S. S. , ZEGEYE B.
Published: December 2015

Hypophysectomized male rats were administered LH or LH + FSH for 14 days and subjected to in vivo micropuncture collection of reproductive tract fluids to determine if FSH alters the compartmentalization of testosterone in the rat testis or of 5α‐dihydrotestosterone (DHT) in the epididymis. Testosterone and DHT concentrations were determined in cardiac blood serum, testicular venous serum, testicular interstitial fluid, and seminiferous tubule fluid, and in intraluminal fluid and tissue extracts from the caput and cauda epididymidis. Testosterone is the predominant androgen in the testis, and compared with control values, concentrations in venous sera, interstitial fluid, and tubule fluid were returned to values indistinguishable from controls by supplementation with 24 μg LH/day. 24 μg LH + 24 μg FSH/day did not augment the intratubular partition of testosterone. Epididymal DHT values were returned to control levels by LH alone, but additional supplementation with FSH significantly increased DHT from the caput epididymidis even further. It is speculated that FSH does not alter the compartmentalization of androgens in the rat testis, but may play a role in retaining androgens in the epididymis.
In Vitro Production of a TRH‐Homologous Peptide and His‐Pro Diketopiperazine by Human Semen
Authors: PEKARY A. EUGENE , REEVE JOSEPH R. , SMITH VIERKA P. , FRIEDMAN STANLEY , HERSHMAN JEROME M.
Published: December 2015

Fresh human semen, diluted 1:1 v/v with 0.15 M NaCl‐0.05 M phosphate, pH 7.5, undergoes a 3‐fold increase in total thyrotropin‐releasing hormone (TRH) immunoreactivity on incubation at 4 C for 8 to 16 hours. To identify the mechanism for this increase, pooled human semen was incubated at 4, 37, and 60 C, and the change in composition of the immunoreactive TRH peptides was quantitated by high pressure liquid chromatography and radioimmunoassay of TRH. His‐Pro diketopiperazine, a biologically active metabolite of TRH consisting of a cyclic dipeptide of histidine and proline, also was measured by specific RIA. The concentration of TRH (pGlu‐His‐Pro‐NH2) dropped precipitously within the first hour after dilution and incubation at all temperatures studied. A hydrophobic TRH‐homologous peptide with the amino acid composition (Glu,X,Y,Pro), where X and Y are neutral, nonaromatic amino acids, increased 8‐fold during 16 hours of incubation at 4 C. This TRH‐homologous peptide is not derived from TRH because it lacks the histidine residue. A 3‐ to 23‐fold increase in His‐Pro diketopiperazine levels occurred after 4 hours at 37 C. This was not due primarily to enzymatic removal of the pyroglutamyl residue from TRH by pyroglutamate aminopeptidase, since about 1 hour after ejaculation the initial His‐Pro diketopiperazine levels were 9.7 ± 5.1 μg/ml, or approximately 1000‐fold greater than the corresponding levels of seminal TRH. Because cycloheximide, which blocks ribosomal protein biosynthesis, did not inhibit in vitro production of the TRH‐homologous peptide and the His‐Pro cyclic dipeptide, these peptides, like TRH, most likely arise from post‐translational cleavage and processing from pre‐existing macromolecular precursor proteins.
Factors Influencing the Success of Sperm‐Cervical Mucus Interaction in Patients Exhibiting Unexplained Infertility
Authors: AITKEN R. JOHN , WARNER PAMELA E. , REID CLARE
Published: December 2015

The factors regulating the success of sperm‐cervical mucus interaction in 46 patients exhibiting unexplained infertility were analyzed. Within this group of patients, considerable variation in the degree of mucus penetration, which appeared to be related to the properties exhibited by the spermatozoa rather than the quality of the mucus itself, was observed. The ability of the spermatozoa to fuse with zona‐free hamster oocytes reflected their capacity for mucus penetration, regardless of whether human or bovine cervical mucus was used as the target. Multiple regression analysis also indicated the importance of the movement characteristics exhibited by the spermatozoa, which alone could account for up to 85% of the variability in mucus penetration. The concentration of motile spermatozoa and their linear velocity of progression influenced the number of spermatozoa penetrating the mucus in unit time, presumably through an effect on the frequency of collisions at the cervical mucus interface. Whether these collisions resulted in mucus penetration depended upon the movement characteristics of the spermatozoa and was positively associated with a “rolling” mode of progression and the amplitude of lateral sperm head displacement.
Laser Light‐scattering Study of the Toxic Effects of Methylmercury on Sperm Motility
Authors: MOHAMED MOSTAFA K. , LEE WYLIE I. , MOTTET N. KARLE , BURBACHER THOMAS M.
Published: December 2015

An in vitro study was designed using the laser light‐scattering technique to obtain further information on the dose‐effect relationship of methylmercury on sperm motility. The technique provided a quantitative evaluation of sperm swimming speed. Semen samples were collected from normal male Macaca fascicularis monkeys by anal electroejaculation. Methylmercury was added to aliquots of sperm suspensions in BWW medium in doses of 10, 5, 2, and 1 ppm. After 3 hours, the relative speed was 35%, 59%, 69%, and 92% of the corresponding controls at doses of 10, 5, 2, and 1 ppm, respectively. The percentage of motile spermatozoa decreased significantly at 10 ppm. By microscopic observation abnormal motility was detected at 5 and 10 ppm, especially after 20 to 40 minutes. Head movement increased from side to side, and many spermatozoa developed coiled tails. The technique proved useful for defining the dose‐effect relationship of methylmercury and sperm swimming speed.
Germ Cell Degeneration in the Contralateral Testis of the Guinea Pig with Unilateral Torsion of the Spermatic Cord Quantitative and Ultrastructural Studies
Authors: JHUNJHUNWALA JAGADISH S. , HIKIM AMIYA P. SINHA , BUDD CHURTON A. , CHAKRABORTY JYOTSNA
Published: December 2015

This study evaluated the long term effects of unilateral torsion of the spermatic cord on the contralateral testis of guinea pigs, employing both fine structural and quantitative studies. Young, adult Hartley strain guinea pigs were divided into six experimental groups (12 animals per group). The first three groups consisted of 36 animals in which unilateral torsion was surgically induced. In group I (torsion maintained), unilateral torsion of the spermatic cord was maintained until the day of sacrifice; in group II (torsion and untwist), torsion of the spermatic cord was maintained for 8 to 12 hours, then the spermatic cord was untwisted and the testis was retained until the day of sacrifice. In group III (torsion and orchiectomy), testes were removed after 8 to 12 hours of spermatic cord torsion. The second three groups consisted of 36 animals: group IV (unilateral orchiectomy), group V (unilateral sham operation), and group VI (pentobarbital injection alone), which served as controls. One half of the animals from each group were killed after 4 months and the other half were killed after 8 months. The most frequently observed histologic changes in the contralateral testes of the experimental animals were focal disorganization and exfoliation of immature germ cells into the lumen. Severe damage, with almost complete absence of germ cells, was noted only in an occasional tubule. Quantitative evaluation of the germ cells of the contralateral testis revealed significant loss of germ cells in groups I, II, and III after 4 months, and in groups I and II after 8 months. Germ cell degeneration was progressive in groups I and II, as demonstrated by the lower germ cell count in the testes of animals of the 8‐month group in comparison with the 4‐month group. However, in group III animals, a higher germ cell count was recorded at 8 months, which was similar to those of control values. The present study confirms our earlier findings that unilateral torsion of the spermatic cord has adverse effects on the contralateral testis. However, since these effects are subtle and inconsistent, a systematic germ cell quantitation is needed for critical assessment of the deleterious effect of unilateral spermatic cord torsion on the contralateral testis.
Dose and Time Relationships in the Endocrine Response of the Irradiated Adult Rat Testis
Authors: DELIC JULIAN I. , HENDRY JOLYON H. , MORRIS IAN D. , SHALET STEPHEN M.
Published: December 2015

The dose‐ and time‐dependent responses for the interstitial and tubular compartments in irradiated adult rat testes are described. Leydig cell dysfunction, as indicated by increased serum LH (to a maximum of 385% of control after 5 Gy) and decreased serum T (to a minimum of 30% of control after 10 Gy), was observed at 8 weeks postirradiation. Subsequent recovery of Leydig cell function was then observed, so that after 9 months serum T was normal but LH was still marginally elevated. The dysfunction, with a threshold of about 4 to 5 Gy, was associated with a loss of Leydig cells from the testis. Spermatogenic damage was observed; after doses of 3 Gy and above a marked dose‐response was recorded as assessed by counts of tubule cross sections exhibiting spermatogenesis. Reduced serum levels of androgen binding protein indicated Sertoli cell dysfunction at 8 weeks after 3 Gy and above, with values of less than one half of those seen in the controls. Serum FSH also was elevated to between 150% and 200% of control, and after 9 months closely reflected androgen binding protein changes. Unlike the Leydig cell, no recovery with time was observed for this aspect of Sertoli cell function.
Spectrum of the Pulsatile Characteristics of LH Release in Normal Men
Authors: VELDHUIS JOHANNES D. , ROGOL ALAN D. , EVANS WILLIAM S. , IRANMANESH ALI , LIZARRALDE GERMAN , JOHNSON MICHAEL L.
Published: December 2015

To assess the spectrum of LH pulse characteristics in normal men, blood samples from 36 individuals were drawn at 20‐minute intervals for 8 hours. The subsequent immunoactive LH concentrations were analyzed by computer algorithms to delineate the frequency and amplitude characteristics of pulsatile LH secretion. The absolute range for LH pulse frequency estimated by a modified threshold method was 1–6 pulses/8 hr, with a mean (± SEM) of 3.36 ± 0.17 (median − 3) pulses/8 hr. The distribution differed significantly from a Gaussian pattern. The mean LH pulse amplitude expressed as a percent increase above nadir was 92.1 ± 6.1 (median‐91.5%). When LH pulse amplitude was defined as an increment (mIU/ml) above nadir, the mean value was 5.13 ± 0.4 (median − 4.8) mIU/ml. These two expressions of amplitude were positively correlated (P < 0.01), while the incremental (mIU/ml) pulse amplitude correlated inversely with pulse frequency (P < 0.01). To examine the influence of more intensified rates of venous sampling on the spectrum of LH pulse properties, blood was sampled at 4‐minute intervals for 8 hours in a subgroup of 13 men. Under these conditions, estimated LH pulse frequency was significantly higher, with a mean of 10.31 ± 1.87 (median − 9) pulses/8 hr compared with 20‐minute sampling in the same individuals (P < 0.001). Although the estimates of LH pulse frequency at 4‐minute and 20‐minute sampling intervals were significantly correlated (P < 0.01), the dispersion of the LH pulse frequency estimates was considerably larger at more rapid rates of sampling. There was an absolute range of 2–20 pulses/8 hr for the 4‐minute sampling, and 1–6 pulses/8 hr for the 20‐minute sampling in the same individuals. This increase in LH pulse frequency and the broader dispersion of the range of frequencies estimated at 4‐minute compared with 20‐minute sampling intervals were confirmed using either another pulse detection algorithm, or separate criteria designed to adjust false‐positive error rates in relation to sampling intensity. It was concluded that eugonadal men exhibit a broad spectrum of pulsatile LH characteristics, and the range of LH pulse attributes is even greater at more intensive rates of venous sampling. The results of this study in normal men demonstrate that a wider dispersion of physiologic LH pulse characteristics must be recognized in man. Such information is particularly important whenever possible departures from normal patterns of episodic gonadotropin secretion are sought in various clinical disorders. Moreover, these observations indicate that a wide range of LH pulse signal characteristics will maintain testosterone production effectively in the normal male.
Impaired Estrogen Production by Leydig Cells of the Naturally Retained Testis in Unilaterally Cryptorchid Boars and Stallions
Authors: RYAN PETER L. , FRIENDSHIP ROBERT M. , RAESIDE JAMES I.
Published: December 2015

Estrogen production in vitro was compared for Leydig cells from cryptorchid and scrotal testes in boars and stallions. Animals with natural and experimental cryptorchidism were used. Purified Leydig cells were prepared from testes of mature animals by collagenase treatment and Percoll density gradients. After incubation for 3 hours (1 × 106 cells), estrone sulfate and estrone in the media were measured by direct radioimmunoassay. Androstenedione and testosterone in media extracts also were determined. Cells from the abdominal testis of unilateral cryptorchid boars and stallions showed impaired estrogen production compared with that of the contralateral scrotal testis. Surgical translocation of the scrotal testis to the abdominal cavity in four unilaterally cryptorchid, prepubertal boars did not result in a reduced capacity for estrogen secretion by Leydig cells examined after puberty. Cells from the naturally retained testis in each of these four animals produced practically no estrogen. In a naturally bilateral cryptorchid stallion, there was a high rate of estrogen secretion by both testes. It was concluded that the scrotal testis of a unilaterally cryptorchid animal exerts a suppressive influence on estrogen formation by the abdominal testis.
Effects of Neonatal Estrogen Administration on Rat Testis Development with Particular Reference to Sertoli Cells
Authors: GAYTAN F. , PINILLA L. , AGUILAR R. , LUCENA M. C. , PANIAGUA R.
Published: December 2015

An ultrastructural and morphometric study of the testes in 15‐, 22‐, 45‐, and 90‐day‐old neonatally estrogenized rats was performed. At 45 days of age, the Sertoli cells appeared immature in estrogenized rats, whereas they were fully mature in the controls. This finding might be related to a deficiency in gonadotropins and androgens during the postnatal period. In 90‐day‐old estrogenized rats, however, Sertoli cell maturation had occurred, which might be attributed to a recovery of hormone levels. Cytoplasmic alterations, however, such as vacuolation, were present at this age. The morphometric study revealed decreased testicular and tubular volumes as well as decreased mean tubular diameters in the estrogenized animals. In contrast, the absolute tubular length increased more in these animals than in the controls during the period from 15 to 90 days of age. This lengthening process might be related to the large number of hypercurved tubules in the estrogenized rats.
Stability of Basic Semen Measures and Abnormal Morphology Within Individuals
Authors: POLAND MARILYN L. , MOGHISSI KAMRAN S. , GIBLIN PAUL T. , AGER JOEL W. , OLSON JANE M.
Published: December 2015

Semen from 15 healthy volunteers was assessed for basic semen measures every 2 weeks over a 6‐month period to determine the relative stability of these factors. The parameters were: sperm count, semen volume, sperm motility, and normal morphology, along with the type of abnormal morphologic forms. Basic semen measures were generally more stable than the morphologic forms. Using three samples, seven semen factors (sperm count, normal morphology, semen volume, sperm motility, microcephaly, tapering and round tail) were found to be stable and contributed to the identification of the fifteen individuals with a classification accuracy of 75.0%. While individuals do have characteristic patterns of spermatogenesis, the biologic basis for these findings is as yet undetermined.
The Effect of Selective Destruction and Regeneration of Rat Leydig Cells on the Intratesticular Distribution of Testosterone and Morphology of the Seminiferous Epithelium
Authors: BARTLETT JOHN M.S. , KERR JEFFREY B. , SHARPE RICHARD M.
Published: December 2015

This study was designed to explore the relationship between the intratesticular distribution of testosterone and spermatogenesis by completely destroying the Leydig cells of mature male rats with injection of a single i.p. dose of ethane dimethanesulphonate. After such treatment, testosterone levels in serum, testicular interstitial fluid, seminiferous tubules, and whole testis declined significantly 6 to 24 hours after injection and fell below assay detection limits between 3 and 7 days. At 3 and 7 days, serum LH and FSH levels rose significantly and remained elevated up to 4 and 6 weeks, respectively, in comparison with vehicle‐treated controls. Leydig cells disappeared from the interstitium by day 3, but between 2 and 4 weeks postinjection a new generation of fetal‐like Leydig cells repopulated the testicular interstitium and, during weeks 6 to 10, were transformed into, or replaced by, Leydig cells with an adult type of morphology. Histologic examination of the seminiferous tubules showed progressive disruption of spermatogenesis between 3 and 14 days post‐ethane dimethanesulphonate. The first histologic sign of spermatogenic damage was noted at day 3, with the occurrence of stage‐specific degenerating pachytene primary spermatocytes at stages VII to VIII of the spermatogenic cycle. On day 7, these cells and degenerating round, or step 19, spermatids often were observed during stages VII to XI, although qualitatively normal spermatogenesis also was seen in these and all other stages of the cycle. Maximum impairment of spermatogenesis occurred 2 weeks post‐ethane dimethane sulphonate, at which time the tubules commonly lacked one or more germ cell generations or, alternatively, showed accumulation of lipid inclusions, extracellular spaces, and variable numbers of degenerating germ cells. Following repopulation of the testis by Leydig cells during weeks 3 and 4, spermatogenesis recovered. By 10 weeks after treatment, qualitatively normal spermatogenesis was seen in the great majority of seminiferous tubules, although a few tubules still remained in which the germ cell complement was severely reduced, and contained only Sertoli cells and spermatogonia. The results suggest: 1) acute depression of intratesticular testosterone levels is followed by stage‐specific degeneration of germ cells; 2) following destruction of Leydig cells by ethane dimethanesulphonate, an interstitial cell population as yet unidentified is stimulated to replace the Leydig cell population; and 3) ethane dimethanesulphonate thus presents a novel opportunity to define in more detail the hormonal dependence of spermatogenesis and the intercommunication between the seminiferous tubules and the Leydig cells.
Effects of Diazacholesterol Dihydrochloride (SC‐12937), an Avian Antifertility Agent, on Rat Testis
Authors: HIKIM AMIYA P. SINHA , CHAKRABORTY J.
Published: December 2015

The present study was undertaken to evaluate the effectiveness of an avian chemosterilant, 20, 25‐diazacholesterol dihydrochloride (SC‐12937), on the rat testis. Adult male rats were injected intraperitoneally with 10 mg (Group 1) or 30 mg (Group 2) of SC‐12937/kg/d or with vehicle alone (Group 3) for 10 days, and were killed 24 hours after the last injection. A wide range of variation in the appearance of affected seminiferous tubules was observed in the testis of SC‐12937‐treated rats at both dose levels. This ranged from apparently normal‐looking seminiferous tubules to almost completely atrophied tubules with no cells. Affected tubules exhibited intraepithelial vacuoles of varying size, multinucleated giant cells, germ cell exfoliation, and tubular atrophy. The presence of severely damaged and entirely normal seminiferous tubules adjacent to one another in the same section was noteworthy. The changes appeared to be dose‐related. A greater number (34.6%) of affected tubules were observed in rats receiving 30 mg of SC‐12937 compared with the ones receiving 10 mg of this compound (19.6%). The Sertoli cells also were affected by this drug and exhibited cytoplasmic vacuolation, a marked increase in the accumulation of lipid droplets and myeloid bodies. Necrotic Sertoli cells also were observed in the severely affected tubules. The possible mechanism of antispermatogenic action of SC‐12937 in rats has been discussed briefly.
Altered Testosterone Feedback in Pubertal Male Rats Raised on Reduced Caloric Intake
Authors: PIACSEK BELA E. , BONIFER THOMAS M. , TAN ROSE C.
Published: December 2015

A study was conducted to explore the hypothesis that reduced caloric intake increases the negative feedback efficacy of testosterone on gonadotropin secretion. Daily subcutaneous injections of testosterone propionate in oil were administered to either age‐matched (37‐day‐old) or weight‐matched (mean = 178 g) ad libitum‐fed (control) or underfed rats that received 60% of normal daily food intake from the age of 20 days. Daily doses of testosterone propionate ranged from 25 to 400 μg/100 g body weight. Results indicate that reduced caloric intake increases the suppression of serum LH by testosterone propionate in both age‐matched and weight‐matched underfed rats. On the other hand, the response of serum FSH concentrations to testosterone propionate was affected by reduced diet only in the weight‐matched animals. Results suggest that the increased negative feedback of testosterone on LH secretion is a specific diet‐induced effect and not merely the result of reduced growth. The altered response in FSH secretion, however, may represent age‐associated changes in sensitivity to testosterone rather than diet‐induced effects.
Kinetics of Human Spermatozoa Long‐chain Fatty Acid CoASH Ligase
Authors: JONES ROBERT E. , PLYMATE STEPHEN R.
Published: December 2015

The kinetics of long‐chain saturated fatty acid activation were studied in the supernatant obtained from Triton‐treated human spermatozoa. Sperm long‐chain fatty acid: CoASH ligase (AMP) (E.C. 6.2.1.3) was able to activate myristic, palmitic, and stearic acids, but was incapable of utilizing lauric, arachidic, and behenic acids. Peak activity was obtained with palmitic acid. Although the Kms for each fatty acid were similar (4.3 to 5.0 μM), the Vmax was several‐fold higher for palmitate. In contrast, ligase from liver homogenates assayed under identical conditions activated lauric through stearic acids, with maximal rates being noted with myristic acid. When compared with nongerminal tissues, sperm ligase appears unique because of its narrower acyl substrate specificity.