bluuebunny/crossref_metadata_2025 huggingface dataset:
Diagnosis and Treatment of Skin and Soft Tissue Infections: Clinical Experience with Ticarcillin Disodium‐Clavulanate Potassium
Authors: Pankey George A.
Published: October 2023

The skin has highly effective mechanical, chemical, and immunologic defenses against microbial invasion. These defenses can be breached, however, when the surface of the skin is broken or when hematologic spread of infection reaches the skin or its underlying tissues. Host factors such as diabetes mellitus may also predispose individuals to skin and soft tissue infections, some of which may threaten limb or life. Management of serious infections of the skin and soft tissue often requires thorough drainage and surgical debridement, as well as aggressive antibiotic therapy. Empiric antibiotic therapy of life‐threatening skin structure infections should use an agent effective against a broad range of gram‐positive and gram‐negative, aerobic, and anaerobic organisms, including producers of ß‐lactamase enzymes. The combination of ticarcillin disodium and clavulanate potassium is such an agent, and is safe and effective in the treatment of serious skin and soft tissue infections.
Management of Intraabdominal Infections
Authors: Sirinek Kenneth R.
Published: October 2023

Intraabdominal bacterial infections (secondary peritonitis and intraabdominal abscesses) involve a large number of aerobic and anaerobic organisms and require a multifactorial therapeutic approach that consists of general supportive measures, surgical intervention, and antibiotic treatment. In spite of advances in antibiotic therapy, morbidity and mortality remain high. Well‐conducted studies using standardized classification of infections are necessary to determine the best antibiotic regimen, and to lower morbidity and mortality.
Age as a Factor Influencing the Power and Sensitivity of Experiments for Assessing Body Weight, Testis Size, and Spermatogenesis in Rats
Authors: BERNDTSON WILLIAM E. , THOMPSON TERRY L.
Published: October 2023

Data from 25 rats aged 60, 150, or 240 days (n = 75) were used to determine the number of observations per rat and or rats per treatment group needed to provide future experiments of known power and sensitivity. End points examined included body weight, testicular weight, number of resistant elongated spermatids, yields of germ cells from their younger progenitors, and germ cell: Sertoli cell ratios. Requirements differed in relation to the experimental power and sensitivity desired. However, the replication needed for detecting equivalent treatment effects on paired testes weight or the number of spermatids per gram of testis increased with age, and was twice as great for 240‐ vs 60‐day‐old rats. In contrast, approximately twice as many 60‐day‐old rats were required for detecting treatment effects on the number of spermatids per Sertoli cell than when 150‐ or 240‐day‐old rats were used.
Cellular Regulation of Follicle‐Stimulating Hormone (FSH) Binding in Rat Seminiferous Tubules
Authors: KANGASNIEMI MARKO , KAIPIA ANTTI , TOPPARI JORMA , PERHEENTUPA ANTTI , HUHTANIEMI ILPO , PARVINEN MARTTI
Published: October 2023

Stage‐specific binding of follicle‐stimulating hormone (FSH) was measured in rat seminiferous tubules. The binding in single‐point assays was over 3‐fold higher (P < 0.05) in stages XIII to I than in stages VI to VII of the epithelial cycle. No difference was found between the equilibrium association constants (Ka) of FSH binding in stages XIV to IV (10 ± 1.9 × 109 1/mol) and VII to VIII (9.2 ± 0.6 × 109 1/mol, mean ± SEM, n = 5). In another experiment, the testes were dosed locally with 3 Gy of 4 MV x‐irradiation to selectively lower the number of spermatogonia. After irradiation, FSH binding in staged seminiferous tubule segments was measured when the desired types of spermatogenic cells were reduced in number. Seven days after irradiation when differentiating spermatogonia and preleptotene spermatocytes were reduced in number, FSH binding was decreased in all stages of the cycle, but the cyclic variation remained. Seventeen days after irradiation when intermediate and type B spermatogonia and spermatocytes up to diplotene of stage XIII snowed low numbers, FSH binding was decreased in all stages of the cycle and the stage‐dependent variation disappeared. At 38 days when pachytene spermatocytes and early spermatids were reduced in number, similar results were found. But at 52 days postirradiation when all spermatids were low in number, FSH binding was slightly elevated compared with days 17 and 38. There were no significant differences in serum FSH or LH levels between irradiated and non‐irradiated animals. These findings suggest that all spermatogenic cell types may stimulate FSH binding in the Sertoli cells. Spermatocytes and spermatids seem to be responsible for the stage‐dependent variation.
Effects of Hypophysectomy and Alterations in Spermatogenic Function on Leydig Cell Volume, Number, and Proliferation in Adult Rats
Authors: KEENEY DIANE S. , EWING LARRY L.
Published: October 2023

The two major objectives of this study were to determine (i) whether the pituitary is required to maintain Leydig cell number per testis, and (ii) whether alterations in spermatogenic function in the absence of LH can affect Leydig cell volume, number, and 3H‐thymidine incorporation in vivo. Four experimental treatments tested the combinations of two factors: (i) the intact pituitary vs hypophysectomy (Hypox); and (ii) arrested vs active spermatogenesis. Subdermal Silastic capsules were used to deliver a low dosage of estradiol in addition to a low dosage of testosterone (TE), which arrested spermatogenesis, or a high dosage of testosterone (HTE) which maintained active spermatogenesis. All four treatments (TE, HTE, Hypox and Hypox‐HTE) inhibited LH secretion for 16 weeks. Control rats were sham hypophysectomized. Leydig cell volume per testis and the volume of an average Leydig cell decreased 70–85% (P < 0.01 vs controls) in all treated rats, whether deprived of LH (TE) or of all pituitary secretions (Hypox), and whether spermatogenesis was arrested (TE, Hypox) or maintained by exogenous testosterone (HTE, Hypox‐HTE). This result suggested that LH was the only factor required to maintain Leydig cell volume, since the absence of other pituitary factors or alterations in spermatogenic function could not override or modify the effect of LH deprivation. No significant differences were found in Leydig cell number per testis or the proportion of Leydig cells labeled with 3H‐thymidine among control and experimentally treated rats. In contrast to Leydig cell volume, which depended on LH, Leydig cell number and Leydig cell division were maintained at control values in the absence of pituitary factors and spermatogenic function for 16 weeks.
Spermatic Cord Torsion: Effects of Cyclosporine and Prednisone on Fertility and the Contralateral Testis in the Rat
Authors: PAKYZ RUTH E. , HEINDEL RACHEL M. , KALLISH MICHELE , COSENTINO M. JAMES
Published: October 2023

Unilateral spermatic cord torsion results in contralateral degeneration and reduced fertility in the prepubertal male rat. This study was conducted to investigate the use of immunosuppression with cyclosporine and prednisone to prevent these untoward effects. Thirty‐five‐day‐old male rats were subjected to 720° unilateral spermatic cord torsion of 9 hours duration. At the time of detorsion, animals were given a subcutaneous injection of i) cyclosporine, ii) prednisone, or iii) cyclosporine combined with prednisone. Control groups included: i) animals undergoing orchiectomy of the ipsilateral testis following the torsion period, ii) hemicastration in the absence of torsion and iii) sham surgery. Orchiectomy at the end of the torsion period prevented the torsion induced reduction of fertility, contralateral seminiferous tubule diameter and testis weight. Treatment with cyclosporine combined with prednisone significantly increased these parameters above detorsion alone. These data indicate that short term immunosuppression with cyclosporine alone or in combination with prednisone limits the adverse effects of unilateral spermatic cord torsion as does removal of the damaged organ at the end of the torsion period.
A Modified and Improved Assay for Sperm Amidase Activity
Authors: CROSS NICHOLAS L.
Published: October 2023

An assay for sperm amidase activity has been modified so that many samples can be measured rapidly, making it more suitable for use in a clinical setting. The modified assay gave the same results (P > 0.05) as its lengthier parent assay (Kennedy et al, 1989). The measured enzyme activity was proportional to the number of sperm assayed over the range tested (3.5 × 105 to 2.1 × 106 sperm). The within‐assay coefficient of variation was 7.8 ± 1.8% (average ± SE, n = 12), and the between‐assay coefficient of variation was 7.2 ± 1.2% (n = 3). The sensitivity was 7.6 μIU/well.
Differential Effects of FSH on the Activities of S‐Adenosyl‐L‐Methionine Decarboxylase and Ornithine Decarboxylase in Rat Sertoli Cells
Authors: SHUBHADA SANKARARAMAN , TSAI YU‐HUI
Published: October 2023

Follicle stimulating hormone (FSH) modulates testicular function via Sertoli cells. The effect of FSH on S‐adenosyl‐L‐methionine decarboxylase (AdoMetDC) activity was investigated in cultured Sertoli cells isolated from 18‐day‐old rats. In contrast to our previous finding that FSH inhibited Sertoli cell ornithine decarboxylase (ODC) activity, FSH stimulated AdoMetDC activity 160 to 300% above the level of the control cells during the initial 2 to 6 hours of treatment. The stimulated enzyme activity declined to 20 to 30% below the control values by 12 hours of exposure to FSH, and then rebounded to 120 to 125% of control values. To determine whether or not the initial opposite effects of FSH on AdoMetDC and on ODC activities are mediated by the same mechanism, various agents which increase intracellular cAMP level were used. All agents studied stimulated Sertoli cell AdoMetDC activity at 5 hours after their inclusion while they significantly inhibited ODC activity. In the presence of FSH, the stimulatory effect of these agents on AdoMetDC was either equal to or slightly greater than that caused by FSH or either agent alone. The combination of dbcGMP with FSH or with dbcAMP resulted in a synergistic or additive effect on AdoMetDC, as well as ODC, activity. The data suggest that the action of FSH on AdoMetDC activity in Sertoli cells is also mediated through cAMP, as concluded previously for ODC activity. Regardless of the effects of FSH on both enzymes, ODC always exhibited a relatively greater activity than did AdoMetDC in cultured rat Sertoli cells.
Fluid Movement in the Lumen of the Rat Epididymis: Effect of Vasectomy and Subsequent Vasovasostomy
Authors: TURNER T. T. , GLEAVY J. L. , HARRIS J. M.
Published: October 2023

Intraluminal fluid movement rate was measured in four regions of the rat epididymis. The fastest flow occurred in the proximal caput epididymis (18.5 ± 3.7 mm/hour) and the slowest in the distal cauda (2.5 ± 0.5 mm/hour). Vasectomy significantly reduced caput fluid flow rates unless a sperm granuloma was present at the vasectomy site. Thirty days after vasovasostomy, caput fluid movement remained reduced in animals unless a granuloma was present. Failures of this or other aspects of epididymal biology to return to normal after vasovasostomy could play a role in the frequent infertility which persists after the operation.
Relationship between Morphology and Motion Characteristics of Human Spermatozoa in Semen and in the Swim‐Up Sperm Fractions
Authors: OEHNINGER SERGIO , ACOSTA ROSITA , MORSHEDI MAHMOOD , PHILPUT CHRISTINE , SWANSON R. JAMES , ACOSTA ANIBAL A.
Published: October 2023

In this study, the authors evaluated the morphology pattern and motion characteristics of human spermatozoa before and after swim‐up separation. Samples were divided into two, morphologically different groups according to the percentage of normal sperm forms assessed by the strict criteria of the Norfolk laboratory: “good” (G) and “poor” (P) prognosis patterns. The percentages of normal forms, slightly abnormal forms, and severe head defects were significantly different in the two groups. Motile characteristics were analyzed by a computerized semen analyzer with constant parameter settings. Before swim‐up there were no significant differences in semen volume, percentage of neck and tail defects, concentration, or percentage of motility and linearity, but the mean velocity was higher in group G. After swim‐up the percentage of motility, total number of motile cells, and recovery rate were higher for group G, and the incidence of severe head defects correlated negatively with the percentage of cells with a velocity of >80 μm/sec. The results suggest that patients with a high incidence of sperm head defects have impaired original velocity, and swim‐up selects for velocity as well as normal forms and motility. Although motility and velocity improved substantially after swim‐up, the recovery rate and percentage of motility were significantly lower in the P group.
Participation of Human Epididymal Sperm Coating Antigens in Fertilization
Authors: SANJURJO CLAUDIA , DAWIDOWSKY ADRIANA R. , CAMEO MÓNICA S. , ECHEVERRIA FERNANDA GONZALEZ , BLAQUIER JORGE A.
Published: October 2023

A polyclonal antiserum directed against human sperm coating proteins of epididymal origin (anti‐KCl) was tested for its ability to alter sperm function. Spermatozoa from normal ejaculates were selected by swim‐up separation and capacitated by overnight incubation at room temperature. Exposure of these cells to anti‐KCl (0.39 mg protein/ml), prior to their use in the hamster ova penetration test, reduced the penetration of denuded oocytes by 65% (P < 0.005). Significant inhibitions of lesser magnitude were observed at lower serum concentrations (to 0.098 mg/ml). In an effort to understand the mechanism of this inhibition, other sperm function parameters thought to be related to oocyte penetration were studied. The inhibitory effect was exerted without noticeable changes in sperm motility (determined by the percentage of motile cells and their linear velocity), and in the absence of major sperm agglutination. Anti‐KCl did not inhibit the occurrence of spontaneous or induced (by human follicular fluid) acrosome reaction in capacitated spermatozoa. In contrast, exposure to anti‐KCl reduced the ability of capacitated spermatozoa to bind tightly to the hamster oolemma. None of these effects were elicited by a control preparation obtained from pre‐immune rabbit sera. Exposure of zona‐free oocytes to the antiserum did not alter their penetrability by normal sperm. These results suggest that the antigens recognized by anti‐KCl participate in some specific step of the sperm—ovum interaction.
Male Hypogonadism Due to Nontumorous Hyperestrogenism
Authors: ELIAS A.N. , VALENTA L.J. , DOMURAT E.S.
Published: October 2023

Male hypogonadism due to the nontumorous production of estrogen was studied in a patient with gynecomastia and bilateral small testicles. Both the gynecomastia and the decrease in testicular size developed in the 5‐year period before presentation. Peripheral serum concentrations of testosterone were in the low to low‐normal range, while those of 17β‐estradiol (E2) were significantly elevated, as were the urinary concentrations of total estrogen. Steroid hormone concentrations were measured in the left and right spermatic veins and the left and right adrenal veins in the basal state, and after stimulation with GnRH and ACTH. Spermatic vein concentrations of E2 were 3 to 20 times higher than concentrations previously reported in normal males. Spermatic vein concentrations of testosterone were normal. The spermatic vein concentrations of androstenedione were approximately three times higher than the mean concentration of androstenedione previously reported in the spermatic vein of normal males. The concentrations of E2 and androstenedione in the adrenal veins were also significantly elevated when compared to the concentrations previously reported in normal subjects. The authors postulate that the hyperestrogenism in this patient was due to increased aromatization of the precursor substrates, testosterone in the testes, and androstenedione in the adrenals to E2 and E1 in the testes and adrenals, respectively. Alternatively, an increased abundance or activity of the 17β‐hydroxysteroid dehydrogenase isoenzyme which converts estrone (E1) to E2 or a relative deficiency of the 17β‐hydroxysteroid dehydrogenase isoenzyme, which converts androstenedione to testosterone, could theoretically account for the reported abnormalities.
The Testicular and Epididymal Luminal Amino Acid Microenvironment in the Rat
Authors: HINTON BARRY T.
Published: October 2023

Concentrations of amino acids were measured in arterial and testicular venous blood, and in fluids from the seminiferous tubule, rete testis, and the caput, corpus, and cauda epididymidis. There were no significant differences in the concentrations of any amino acids between arterial and testicular venous blood, whereas there were significant differences between arterial/venous blood and testicular interstitial fluid. The predominant amino acids measured within seminiferous tubule fluid (STF) and rete testis fluid (RTF) were glycine, alanine, glutamate, and glutamine. RTF contained approximately equal concentrations of basic and total amino acids, but 17 times higher acidic amino acids and 1.2 and 1.3 times lower uncharged polar and nonpolar amino acids, respectively, compared to STF. The concentration of total amino acids within caput fluid reached over 50 mmol/L, but then declined to approximately 50% and 0.1% of caput for corpus and cauda, respectively. The predominant amino acids measured within epididymal luminal fluids were glutamate and taurine; glutamate contributed to ∼90% of the total amino acids measured in caput fluid. The presence of glutamate and taurine within the epididymal lumen is due primarily to a direct contribution from the epididymal epithelium, as measured using the split‐drop stopped‐flow microperfusion technique. Several other amino acids within the lumen also originate from the epididymal epithelium. Amino acids contribute approximately 20%, 9%, and 2% of the total osmolality of caput, corpus, and cauda fluid, respectively. Even when all ions and organic solutes are measured and compared to the total osmolality of luminal fluid within each epididymal region, there is still a difference of approximately 10, 70, and 35 mOsm/kg water for caput, corpus, and cauda, respectively.
Experimental Cryptorchidism in the Adult Mouse: I. Qualitative and Quantitative Light Microscopic Morphology
Authors: MENDIS‐HANDAGAMA S.M.L. CHAMINDRANI , KERR JEFFREY B. , de KRETSER DAVID M.
Published: October 2023

Morphologic changes in the testes of adult mice after experimentally induced cryptorchidism were studied by light microscopy and stereology. Increasing duration of cryptorchidism resulted in a gradual decrease in the volume of seminiferous tubules per testis, and this was associated with germ cell degeneration. The volumes of Sertoli cell lipid droplets increased, and dilations of the intercellular space between the Sertoli cell junctions was observed in the cryptorchid testis. The luminal volume of the seminiferous tubule was reduced by 50% after 28 days of cryptorchidism. However, the volumes of intertubular tissue and Leydig cells in control and cryptorchid testes were not significantly different. Leydig cell number per testis increased, and the average volume of a Leydig cell decreased gradually with the progression of the cryptorchid state. The volume of the connective tissue cells in the intertubular area increased, but no significant volume change was observed in the volume of intertubular macrophages. After 28 days, the cryptorchid testis contained a significantly increased volume of blood vessels and a reduced volume of lymphatic space per testis. These observations clearly demonstrate that, although the mouse is a species closely related to the rat, the morphologic changes that occur in the Leydig cell population after induction of experimental cryptorchidism in this species is different.
Experimental Cryptorchidism in the Adult Mouse: II. A Hormonal Study
Authors: MENDIS‐HANDAGAMA S. M. L. CHAMINDRANI , KERR JEFFREY B. , de KRETSER DAVID M.
Published: October 2023

Serum follicle‐stimulating hormone (FSH), luteinizing hormone (LH), and testosterone levels and secretory capacity of the in vitro stimulated testis were determined in control and bilateral cryptorchid mice after 7, 14, 21, and 28 days. In a separate study, serum FSH, LH, and testosterone levels were measured in unilateral and bilateral cryptorchid, hemicastrate, and bilaterally castrate adult mice after 28 days of treatment. Serum FSH levels were significantly increased in bilaterally cryptorchid mice compared to controls (0 days) at 7, 14, 21, and 28 days, but serum LH and testosterone levels did not change. At 28 days, the elevated serum FSH levels in the unilateral and bilateral cryptorchid mice were not different than those in hemicastrate mice. However, the FSH levels in bilaterally castrate mice after 28 days were significantly higher than all other groups. Serum LH and testosterone levels were significantly different only in the bilaterally castrate group, compared to control levels. Both the normal and cryptorchid testes of all ages studied were capable of producing equal levels of testosterone in vitro, both basally and with a human chorionic gonadotropin (hCG) dose of 700 mIU/ml (maximum stimulatory dose for both the normal and the 28 day cryptorchid testes). Changes that occur in mice with experimentally induced cryptorchidism are not identical to those seen in the rat. Serum FSH levels increase, but no changes occur in serum LH and testosterone levels. Additionally, a cryptorchid mouse testis is not hyperresponsive to hCG stimulation in vitro.
Gonadotroph and Leydig Cell Responsiveness in the Male Rat
Authors: TURNER T.T. , EVANS W.S. , LOPEZ T.J.
Published: October 2023

Previous experiments have found that experimental left‐sided varicocele (ELV) in rats is associated with significant bilateral reductions in intratesticular testosterone concentrations. The current experiments were performed to determine the source of this endocrinopathy. Sensitivity and responsivity of Leydig cells and gonadotrophs were determined in control male rats and in those with ELV. Initially, dose—response relationships were determined for luteinizing hormone (LH) stimulation of testosterone secretion by Leydig cells and for luteinizing hormone releasing hormone (LHRH) stimulation of LH secretion by gonadotrophs. Maximally (ED100) and half‐maximally (ED50) stimulating doses of LH and LHRH were selected from these studies and administered to sham‐operated and ELV rats 30 days after the operation to induce ELV. Leydig cell and gonadotroph sensitivity (response to ED50 and responsivity (response to ED100) to LH and LHRH, respectively, were determined. Responsivity of Leydig cells in control and ELV rats was similar. Responsivity of gonadotrophs to LHRH was significantly suppressed in ELV animals, but the physiologic relevance of this singular finding is unclear. It is possible that the previously determined ELV‐associated decrease in intratesticular testosterone concentrations is subsequent to a wash‐out phenomenon that follows the increased testicular blood flow that also is known to be associated with ELV.
Evidence for a Role of Cysteinyl Leukotrienes in Mouse and Human Sperm Function
Authors: BASURAY R. , de JONGE C. , ZANEVELD L.J.D.
Published: October 2023

The in vitro fertilizing capacity of mouse spermatozoa was inhibited by FPL‐55712, a receptor antagonist of the cysteinyl‐containing leukotrienes LTC4, LTD4, and LTE4. Inhibition occurred whether the compound was present in the medium during the capacitation period or during the fertilization period. Additionally, FPL‐55712 inhibited the penetration of human spermatozoa into zona‐free hamster oocytes. Inhibition of fertilization did not appear to be caused by an effect on sperm motility or on the oocyte. These observations suggest that mouse and human spermatozoa require cysteinyl leukotriene activity for both fertilization and oocyte penetration.
Onset of Spermatozoal Degeneration in Low‐Fertility Delaware Roosters and Test for Autoimmune Basis
Authors: FROMAN DAVID P. , KIRBY JOHN D. , LAWLER RITA M. , BERNIER PAUL E.
Published: October 2023

The objectives of this study were 1) to determine the onset of a heritable reproductive disorder in the rooster that is characterized by extensive spermatozoal degeneration within the ductus deferens, and 2) to determine if autoimmunity was associated with spermatozoal degeneration. Seventy‐five percent of the affected roosters did not ejaculate large percentages of degenerate spermatozoa at 20 wk of age, approximately the age of sexual maturity. Rather, seminal quality gradually declined over the next 6 wk, as both ejaculate volume and number of spermatozoa ejaculated increased. The evaluation of testicular and excurrent duct tissues via immunofluorescence failed to reveal either IgY or IgA associated with spermatozoa. While histological examination revealed greater lymphocyte numbers (P < .05) in the proximal ductus deferens, these cells were not associated with spermatozoa nor spermatozoal clumping. While spermatozoal degeneration tends to be latent at the onset of semen production, it does not appear to be due to spermatozoal autoimmunity.
Preparation and Short‐Term Culture of Enriched Populations of Guinea Pig Spermatocytes and Spermatids
Authors: JOSHI MADHU S. , ANAKWE ONYEAMA O. , GERTON GEORGE L.
Published: October 2023

In order to study acrosome biogenesis in vitro, the authors developed techniques for the isolation and culture of enriched populations of guinea pig spermatogenic cells using a modification of the technique of Romrell et al (1976). The modifications include changes in the medium, enzyme concentrations, cell loads for gradient separations, and sedimentation times. Three major cell populations were pooled: Pachytene spermatocytes (PS: 2.0 × 107, 80–85% pure), round spermatids, (RS: 1.2 × 108, 80–85% pure), and condensing spermatids (CS: 2.0 × 108, 50–60% pure, contaminated with residual bodies). The ultrastructural properties of the isolated cells appeared similar to those of cells in situ. PS were 3–4 times more active than RS and 10 times more active than CS in synthesizing proteins. These experiments demonstrate that highly enriched populations of guinea pig spermatogenic cells can be isolate, and that these cells can be cultured in the presence of radioactive precursors for studies of protein synthesis during spermatogenesis.
Initial Characterization of Factors From Testicular Fluid Which Alter In Vitro Androgen Secretion by Normal Rat Leydig Cells
Authors: JANSZ GWENDERLYN F. , COOK ROBERT A. , POMERANTZ DAVID K.
Published: October 2023

It is known that testicular interstitial fluid (TF) contains thermolabile factors that can alter in vitro production of androgens by the Leydig cells. The net stimulatory activity of this fluid increases in association with the disruption of spermatogenesis. The identity of the active agent(s) in TF is not known. Therefore, the authors used gel‐liquid chromatography to initially characterize TF from control and bilaterally cryptrorchid animals. The stimulatory activity of TF was retained on Concanavalin A Sepharose columns. Gel filtration on Ultrogel AcA 44 suggested a molecular size between 40 and 90 kD. The unfractionated fluid from control and bilaterally cryptorchid rats, as well as the chromatographic fractions containing stimulatory activity, were further resolved by SDS‐PAG electrophoresis. At least three bands representing glycoproteins with apparent molecular size between 57 and 75 kD were seen in all samples containing stimulatory activity. No difference in the pattern of protein bands was seen between TF from control and bilaterally cryptorchid testes. However, samples reduced with β‐mercaptoethanol showed protein bands with apparent molecular size of 78 and 118 kD which were present only in unpurified control TF. These data support the possibility that the stimulatory substance in TF from control and bilaterally cryptorchid testes is a glycoprotein with a molecular size between 57 and 75 kD. Differences in the bioactivity of the unfractionated TF may be due in part to the presence of additional larger protein molecules in the control TF.
Leydig Cell Cytoplasmic Content Is Related to Daily Sperm Production in Men
Authors: JOHNSON L. , GRUMBLES J.S. , CHASTAIN S. , GOSS H.F. , PETTY C.S.
Published: October 2023

The relationship between the abundance of specific Leydig cell organelles and daily sperm production (DSP) was determined. Testes from 10 men (26–53 years of age) were obtained at autopsy within 10 h of traumatic death or heart failure and fixed by vascular perfusion. Testicular tissue was processed for light and electron microscopy. DSP/testis and Leydig cell cytoplasmic volume/testis were determined by stereology of histologic sections. The Leydig cell organelle content was determined by point counting electron micrographs for smooth endoplasmic reticulum (SER), rough endoplasmic reticulum, mitochondria, lipofuscin pigment, lipid, Golgi bodies, and Reinke crystals. Men were divided equally into two groups based on DSP/testis. Men with low DSP/testis had less SER volume density (P < 0.01) and lower SER volume per testis (P < 0.05) than men with high DSP. Other organelles were unrelated to DSP. When all men were combined, the volume density of SER (r = 0.80; P < 0.01), the volume SER per testis (r = 0.69; P < 0.05), and the volume SER per Leydig cell (r = 0.84; P < 0.01) were significantly related to DSP. Hence, there appears to be a significant relationship between Leydig cell SER and the level of spermatogenesis in men.
Metabolism of 5α Reduced Androgens by Various Tissues of the Male Rat
Authors: PASUPULETI VIJAYA , HORTON RICHARD
Published: October 2023

Since an animal model for the study of peripheral androgen metabolism is needed, we studied the metabolism of 5α‐reduced androgens in various tissues of the rat. Labeled DHT and 3αandrostanediol (3αdiol) were added to tissue minces of male rat scrotal skin, muscle, prostate, or liver. Conversion ratios of the interconverting pair DHT→2 3 αdiol or the formation of the respective glucuronides (G) were determined over a 3 h period. Major differences in the activity and the oxidation/reduction relationship were observed between tissues. Scrotal skin was very active and balanced in the DHT→2 3αdiol interconversion (31 and 33%/100 mg/3 h, respectively, whereas liver was minimally active (3.1/2.7%). 3αreduction was prominent in muscle (37.0/2.7%), although 3αoxidation was more active in prostate (6.0/31.5%). Steroid glucuronidation also differed in the various tissues. Sexual skin formed about 2% 3αdiolG, but much smaller amounts of DHTG. Liver, muscle, and prostate formed minimal (< 0.2%) 3αdiolG, although liver synthesized 1.2% of DHTG. Addition of DHT or 3αdiol increased formation of the respective glucuronides by liver, whereas DHT blocked the synthesis of 3αdiolG, and 3αdiol markedly increased formation of 3αdiolG in skin. These studies indicate a similarity in DHT metabolism between rat and human sexual skin and a high rate of glucuronidation compared with other tissues. The pathway of 3αdiolG formation in skin is DHT ŕ 3αdiol ŕ 3αdiolG. Steroid 3αoxidase is more active than 3αreductase in muscle whereas 3αoxidase predominates in prostate. This may be a mechanism whereby DHT levels and action as a nuclear androgen is favored in prostate, whereas testosterone is the major androgen in muscle. Liver synthesized minimal 3αdiolG from DHT; this is not the result of selective glucuronidation to DHTG, but rather a relative deficiency in 3αoxidoreductase.
Lipid Transfer Activity in Human Follicular Fluid: Relation to Human Sperm Capacitation
Authors: RAVNIK STUART E. , ZARUTSKIE PAUL W. , MULLER CHARLES H.
Published: October 2023

A potentially important event during sperm capacitation is the loss of sperm membrane cholesterol. Although the exact mechanisms mediating this loss are not known, albumin and high density lipoprotein have been proposed as lipid acceptors. The authors propose that lipid transfer may be involved in capacitation as a specific mediator in the sequence of events leading to sperm membrane cholesterol loss. We present the first direct evidence of lipid transfer activity (LTA) in human follicular fluid (HFF). The redistribution of 14C‐cholesteryl ester among human plasma lipoproteins was used as a measure of LTA (% Transfer [%T]). The HFF was fractionated by S‐300 gel filtration chromatography and assayed for LTA. Three peaks of activity were consistently eluted from the column. Each peak of LTA also stimulated human sperm to penetrate zona‐free hamster oocytes after short capacitating incubations. The peak with highest LTA (12.75 ± 1.11 %T) with an Mr approximately 68,000, gave the greatest stimulation (penetration index, PI: 3.34 ± 0.96 fold increase above control, n = 4). The HFF also showed a significant dose response for both LTA and PI, whereas bovine serum albumin did not. These results demonstrate the existence of LTA in HFF and suggest that a specific lipid transfer protein may have a role in human sperm capacitation or acrosome reaction.
Seminal Alpha‐Glucosidase Activity as a Marker of Epididymal Pathology in Nonazoospermic Men Consulting for Infertility
Authors: GUERIN JEAN‐FRANÇOIS , ALI HABIB BEN , COTTINET DANIÈLE , ROLLET JACQUES
Published: October 2023

Glucosidase (αG) activity was measured in sperm free seminal plasma from 1200 patients consulting for primary infertility, in whom clinical examination of epididymides revealed some abnormalities and histories of genital infections. They constituted the group with epididymal pathology (P) that was compared with a reference group (R) of 246 men without any epididymal pathology. The distribution of αG was significantly different between the two groups, even if we considered only the subjects in group P with normal sperm count (PN: 353 men: p < 10−6). 15.9% of subjects in group PN exhibited αG values as low as vasectomized men, versus 1.2% in group R. A linear relationship was established between αG and sperm content in both groups, but αG activities were systematically lower in group P (y = 0.19 × + 64) than in group R (y = 0.30 × + 86). There was no correlation between αG and the percent of sperm motility. On the contrary, we found statistically more clinical epididymal abnormalities in cases of decreased αG activity than in cases of normal αG activity (p < .01).
Functional Aspects of Human Sperm Binding to the Zona Pellucida Using the Hemizona Assay
Authors: CODDINGTON CHARLES C. , FRANKEN DANIEL R. , BURKMAN LANI J. , OOSTHUIZEN W. T. , KRUGER THINUS , HODGEN GARY D.
Published: October 2023

The hemizona assay (HZA) has facilitated investigations of sperm function in relation to zona pellucida binding. In this study, the authors examined: 1) the association between hyperactivated sperm motility and HZA binding; 2) the binding kinetics and efficiency of sperm from subfertile men; and 3) the influence of sperm freezing and thawing on binding capacity. For each HZA, a nonviable human oocyte was cut into equal zona hemispheres. The mean number of bound sperm and the incidence of hyperactivation were significantly greater for samples of sperm from fertile men compared with sperm from subfertile men (P < 0.05). Subfertile sperm had a binding curve that paralleled the curve for fertile sperm, although the magnitude of binding was markedly reduced. Freezing and thawing of sperm from fertile samples impaired their capacity to bind to the zona pellucida. The HZA binding efficiency was reduced by 30%, although the binding curves for fresh versus frozen samples remained parallel.
Teratospermia in Domestic Cats Compromises Penetration of Zona‐Free Hamster Ova and Cat Zonae Pellucidae
Authors: HOWARD JOGAYLE , BUSH MITCHELL , WILDT DAVID E.
Published: October 2023

The ability of spermatozoa to bind and penetrate zona‐free hamster ova and the zonae pellucidae of domestic cat oocytes in vitro was compared between normospermic (>60% structurally normal spermatozoa per ejaculate) and teratospermic (<40% normal spermatozoa per ejaculate) domestic cats. The effects of culture media (Biggers, Whitten, Whittingham [BWW] versus modified Krebs Ringer bicarbonate [mKRB]) and simple dilution (DR), ejaculate centrifugation, and either resuspension (NS) or swim‐up processing (SU) on penetration also were examined. High percentages of structurally normal spermatozoa were bound to zona‐pellucida‐free hamster ova regardless of the morphological forms in the inseminant. Mean percent normal spermatozoa bound to ova in DR, NS, and SU sperm aliquots from teratospermic male cats were not different (P > 0.05) from similarly treated normospermic aliquots. However, the percent penetration of hamster ova by normospermic ejaculates (10.5%) was superior (P < 0.05) to that of teratospermic ejaculates (2.8%). Although swim‐up processing improved percent sperm motility, progressive motility, and normal morphology in teratospermic ejaculates (P < 0.05), no difference was observed in ovum penetration among the DR‐treated, NS‐treated, and SU‐treated spermatozoa (P > 0.05). Culture medium had no effect on sperm binding in the hamster assay, but ovum penetration rate by spermatozoa in the normospermic ejaculates was enhanced (P < 0.05) using mKRB (13.5%) when compared with BWW (7.6%) medium. Spermatozoa from teratospermic cats were capable of binding and penetrating cat zonae; however, sperm—zona interaction (defined as percent of oocytes with spermatozoa binding to or penetrating into the zona) was different (P < 0.05) between normospermic (65.3%) and teratospermic (24.2%) cats. The number of bound spermatozoa per ovum was five‐fold less in males producing high proportions of pleiomorphic spermatozoa (P < 0.05). Zona penetration was also different between normospermic (58.1%) and teratospermic (16.9%) males. Within cat populations, the DR, NS, and SU treatments had no influence on sperm binding or zona penetration. These results indicate that teratospermia in domestic cats has a detrimental impact on gamete interaction and penetration of zona‐free hamster ova and cat zonae pellucidae. The in vitro effect is not entirely dependent on absolute numbers of structurally normal spermatozoa but appears to be related to an unidentified characteristic of teratospermic ejaculates.
Response of Circulating Gonadotropin Levels to GnRH Agonist Treatment in Prostatic Cancer
Authors: HUHTANIEMI ILPO , VENHO PETRI , JACOBI GUNTHER , RANNIKKO SAKARI
Published: October 2023

Luteinizing hormone (LH), follicle‐stimulating hormone (FSH), and testosterone (T) responses to 6‐month treatments with a gonadotropin‐releasing hormone (GnRH) agonist (buserelin) and subsequent orchiectomy were studied in patients with advanced prostate cancer. For treatments, either an intranasal (600 μg, 3/day, n = 8) or subcutaneous depot preparation (6.6 mg every other month, n = 5) were used. A third group of patients received intranasal buserelin (400 μg, 3/day, n = 12) for 35 months. LH and FSH were measured using radioimmunoassay (RIA) and a sensitive (0.04 IU/L) immunofluorometric assay (IFMA). In addition, selected samples were analyzed for bioactive (bio) LH. The RIA‐LH levels decreased 70% with intranasal treatment. In contrast, when monitored by IFMA, the reduction was greater than 90%: 0.2 to 0.3 IU/L with intranasal and 0.044 to 0.052 IU/L with depot treatment (P < 0.01). Gonadotropin suppression was stable up to 35 months. Bio‐LH and IFMA‐LH levels decreased in parallel during treatment, with no apparent changes in the bio/immuno ratio. FSH levels were suppressed temporarily during the treatments. After castration and cessation of buserelin treatment, serum LH and FSH increased rapidly in the intranasal treatment group but only marginally during 3 months in the depot group. Serum T reached the castrate range when IFMA‐LH decreased below 0.5 IU/L. A further decrease in LH (<0.1 IU/L) still suppressed the intratesticular T concentration measured after orchiectomy. In conclusion, IFMA offers an improved method to monitor the antigonadotropic effect of GnRH agonist treatment. The results emphasize the necessity of profound LH suppression to achieve maximal inhibition of testicular androgen production.
Non‐Poisson Distribution of Sperm from Grandfathers in Zona‐free Hamster Ova
Authors: ZENZES MARIA TERESA , REED T. EDWARD , NIESCHLAG EBERHARD
Published: October 2023

Earlier reports indicated that sperm from 25% of patients from infertile couples, but not from normal or fertile donors, show deviations from the theoretical Poisson distribution of the number of sperm penetrating zona‐free hamster ova. Using semen samples from 15 grandfathers (aged 60 to 84 years) and 24 young fathers (aged 25 to 36 years), this study analyzed whether age also has an effect on the distribution. It was found that the overall fit to the Poisson distribution of the samples from grandfathers was very poor; in contrast, the samples from young fathers fit well. The observed deviations from the Poisson distribution among grandfathers may be a consequence of their long periods of sexual abstinence. Decrease in sexual activity produces age‐different populations of sperm that probably differ in penetrating ability. Samples from older fathers also show a worse fit to the Poisson distribution than do those from younger fathers. These results suggest that the duration of sperm storage in the genital tract after maturation has an effect on sperm function.
Correlation Between Changes in Rat Sperm Membrane Lipids, Protein, and the Membrane Physical State During Epididymal Maturation
Authors: HALL JOSEPH C. , HADLEY JILL , DOMAN THOMAS
Published: October 2023

An enriched plasma membrane fraction was isolated from caput, corpus, and cauda rat spermatozoa and analyzed for lipid and protein content, thermal phase transition temperature using electron paramagnetic resonance spectroscopy (EPR), and enzymatic assays of calcium‐dependent AT‐Pase activity. Based on sperm concentration, total membrane phospholipid, cholesterol, and protein content declined as sperm passed through the epididymis. A more refined analysis of the bulk plasma membrane phospholipid revealed that approximately 56% of the phospholipid consisted of choline (PC) and ethanolamine (PE) phosphoglycerides; the remainder consisted of sphingomyelin (SM), phosphatidylserine (PS), and diphosphatidylglycerol (DPG). The mole percent of PE increased in sperm proceeding from the caput to the corpus epididymis and then declined from the corpus to the cauda epididymis. The phospholipid‐bound fatty acids consisted primarily of palmitate (C16:0) and stearate (C18:0), with a significant increase in the mole percent of the docosapentenoyl acyl group (C22:5) in cauda sperm. Arrhenius' plots of the EPR peak height signals using the lipid soluble spin label, 5‐doxyldecane, and the calcium‐dependent ATPase activity as a function of temperature demonstrated a change in the apparent fluidity of the membrane and energy of activation of the calcium‐dependent ATPase associated with the three sperm membrane preparations. These data suggest that the apparent fluidity and biochemical composition of the sperm membrane change during epididymal maturation.
Rapid Development of Leydig Cell Tumors in a Wistar Rat Substrain
Authors: TEERDS KATJA J. , de ROOIJ DIRK G. , de JONG FRANK H. , ROMMERTS FOCKO F. G.
Published: October 2023

ABSTRACT: In 78% of the Wistar rats (substrain U) studied, spontaneous Leydig cell tumors developed between the ages of 12 and 30 months. The first signs of tumor development, in the form of nodules of Leydig cells, were already apparent in 1‐month‐old U‐rats. These nodules of Leydig cells were found in all animals studied. In no other strain of rats has this type of abnormality been observed at such an early age. The first Leydig cell tumors were noticed between the ages of 12 and 14 months. The tumor tissue appeared to have developed from a rapid, focal outgrowth of a nodule. The tumor Leydig cells were found to be sensitive to the cytotoxic action of the specific Leydig cell toxicant ethane dimethane sulphonate (EDS), although not all tumor cells were killed. Inhibin‐like immunoreactivity could be detected in both normal and tumor Leydig cells, and plasma levels varied considerably within the different groups of rats. Moreover, no significant changes in plasma levels of inhibin‐like immunoreactivity were found during the aging period when Leydig cell tumors were formed or after EDS administration when nearly all Leydig cells were killed. Therefore, the possible contribution of Leydig cells and tumor cells to the total content of inhibin‐like immunoreactivity in the testis and plasma may be of less importance than expected. Some significant fluctuations in plasma testosterone concentrations were found during aging; however, there appeared to be no correlation between plasma testosterone levels and the appearance of large Leydig cell tumors. This indicates that testosterone production by tumor cells is limited. Plasma luteinizing hormone (LH) levels did not vary significantly up to the age of 12 months and were undetectable in rats in which tumors had developed. This indicates that the tumor cells probably produce an LH‐suppressing product. This product is not likely to be estradiol because estradiol levels do not change significantly during tumor development. When isolated tumor cells were implanted intratesticularly, tumor development took place in U‐rats but not in another Wistar substrain. Since the abnormalities preceding tumor formation are already apparent at the age of 1 month in U‐rats, this Wistar substrain may provide a good model to study Leydig cell tumor development.
Diurnal Rhythm of Testosterone and Luteinizing Hormone in Hypogonadal Men
Authors: WINTERS STEPHEN J.
Published: October 2023

ABSTRACT: To determine the relationship between increasing luteinizing hormone (LH) production and the diurnal secretion of LH and testosterone (T) in adult men, studies were performed on five men with gonadotropin insufficiency associated with prolactinoma, five eugonadal men, and five men with primary testicular failure. Blood samples were drawn every 10 to 20 minutes for 24 hours beginning at 8:00 to 8:30 AM to evaluate diurnal periodicity. Mean (± SEM) LH levels in the three groups were 7.67 ± 1.46 mlU/ml, 13.9 ± 3.2 mlU/ml, and 62.3 ± 14.4 mlU/ml, respectively, and mean serum T levels were 8.05 ± 1.49 nmol/L, 13.9 ± 3.5 nmol/L, and 9.15 ± 1.3 nmol/L, respectively. Cosinor analysis revealed that each hyperprolactinemic man had a T rhythm with a significant 24‐hour periodicity; the mean acrophase was at 5:00 AM. Testosterone levels were 35.0 ± 10.6% less at 4:00 PM than at 8:00 AM. Eugonadal men also demonstrated a significant diurnal T rhythm with an acrophase at 6:00 AM, and T levels were 15.8 ± 5.3% less at 4:00 PM than at 8:00 AM. By contrast, there was no significant diurnal rhythm in T secretion among the men with testicular failure, although serum T levels were 11.5 ± 3.7% less at 4:00 PM than at 8:00 AM. For LH, hyperprolactinemic men demonstrated a significant 24‐hour rhythm with an acrophase at 1:30 AM, whereas no significant 24‐hour periodicity was identified among either eugonadal men or men with testicular failure. The diurnal LH rhythm characteristic of pubertal boys is a property of the gonadotropin‐receptor hormone (GnRH) pulse generator that is expressed in adult men with hyperprolactinemia and gonadotropin insufficiency. These data are consistent with the possibility that the expression of this rhythm reflects an enhanced gonadotropin‐suppressive action of T in these men. As gonadotropin secretion increases to and beyond normal levels, there is a progressive dampening of the diurnal patterns of T and LH secretion.