bluuebunny/crossref_metadata_2025 huggingface dataset:
Enteric Diseases of Homosexual Men
Authors: Baker Robert W. , Peppercorn Mark A.
Published: December 2015

Certain enteric ailments are particularly common among homosexual men. They are primarily infectious diseases and include not only such common veneral diseases as gonorrhea and syphilis but also infections not usually regarded as being sexually transmitted. Among the latter are shigellosis, salmonellosis, giardiasis, and amebiasis.

Patients' symptoms are non‐specific and seldom helpful in diagnosing particular diseases. The practitioner must be prepared to identify a number of infections with similar presentations that may occur singly or together in gay men.

Gonorrhea is probably the most common bacterial infection in gay men. Carriage rates as high as 50% have been reported, and extra‐genital carriage is common; this necessitates culturing the urethra, rectum, and pharynx. Procaine penicillin G is the treatment of choice for most patients; spectinomycin is probably the drug of choice in penicillin‐sensitive patients. In contrast to other venereal diseases, syphilis may have a characteristic protoscopic presentation. Benzathine penicillin G is the treatment of choice for most patients. Lymphogranuloma venereum causes penile lesions and inguinal lymphadenitis in heterosexual men, whereas homosexual men are more prone to proctitis. The disease may mimic Crohn's disease. Recommended treatment includes tetracycline or sulfamethoxazole‐trimethoprim. Shigellosis usually presents as an acute diarrheal illness. Patients generally require only supportive treatment with fluids.

Herpes simplex viral infection is difficult to diagnose and has several different presentations, including lumbosacral radiculomyelopathy. Symptomatic treatment with sitz baths, anesthetic ointment, and analgesics is recommended. Venereal warts are believed to be caused by the same virus that causes verrucous warts; they are usually found in the anal canal or around the anal orifice. They are commonly treated with 25% podophyllin solution.

Parasitic infections include giardiasis, amebiasis, and pinworm infections. Metronidazole may be used in the treatment of symptomatic giardiasis and amebiasis, but it is not approved for the former indication; quinacrine is approved for giardiasis. Pinworm infestation may be treated with pyrantel pamoate or mebendazole.

Cure of enteric diseases in homosexual men must be documented.
Pharmacology, Therapeutic Efficacy, and Adverse Effects of Bumetanide, A New “Loop” Diuretic
Authors: Flamenbaum Walter , Friedman Richard
Published: December 2015

Bumetanide is a recently developed natriuretic and diuretic agent, belonging to the “loop” class of diuretics. Since it is rapidly and almost completely absorbed after oral administration, oral and parenteral formulations have a similar pharmacokinetic profile. Peak plasma levels are achieved approximately 30 min after oral administration. The apparent half‐life is 1.2–1.5 hr, and the volume of distribution is about 25 liters. Plasma clearance is 228–255 ml/min. Bumetanide is promptly and almost completely eliminated by metabolism of the butyl side chain and urinary excretion of the parent drug and its metabolites. The principle renal site of action is the ascending limb of the loop of Henle, with a minor effect on the proximal tubule. The drug causes decreases in both free water clearance (during water diuresis) and solute free water reabsorption (during hydropenia), increased fractional delivery of sodium choloride to the distal tubule and a natriuresis approaching 20% of the filtered load of sodium, calciuria, phosphaturia, and minimal bicarbonaturia.

Extensive clinical studies have been conducted with both oral and parenteral bumetanide in patients with a variety of edematous conditions. The agent has been clearly demonstrated to be an effective diuretic in the treatment of edema due to cardiac disease (congestive heart failure) and edema, with or without ascites, due to hepatic disease. Bumetanide has also been shown effective in treating edema due to renal disease, even when modest to severe renal insufficiency is present, and it may be useful in the treatment of edema refractory to other loop diuretics.

As would be predicted for any potent diuretic, bumetanide administration has been associated with hypokalemia, hypochloremia, metabolic aklalosis, hyperuricemia, and prerenal azotemia. Alterations in glucose metabolism are an inconsistent finding. Transient thrombocytopenia and granulocytopenia have been noted, but no consistent or important alterations in biochemical parameters have been observed. In some patients, especially those with renal failure receiving high doses, myalgias and muscle tenderness have been described. To date only a very limited potential for ototoxicity has been observed. Bumetanide has been administered without difficulty to patients having side effects from other loop diuretics.
Long‐Term Comparison of Alprazolam, Lorazepam and Placebo in Patients with an Anxiety Disorder
Authors: Cohn Jay B. , Wilcox Charles S.
Published: December 2015

In a double‐blind, placebo‐controlled study of 200 patients with moderate to moderately severe anxiety we compared the anxiolytic efficacy and safety of alprazolam and lorazepam. Dosing was flexible and ranged from 1 to 4.5 mg/day of alprazolam and from 2 to 9 mg/day of lorazepam. The mean daily dose at the end of the 16 week study was alprazolam 3.3 mg and lorazepam 5.1 mg. Both active drugs were significantly more effective than placebo in relieving the symptoms of anxiety on the Hamilton Anxiety Rating Scale, with a trend toward more improvement in the alprazolam group in the later weeks of the study. Target Symptoms, Physician's and Patient's Global Impressions and the Self Rating Symptom Scale indicate that alprazolam and lorazepam were superior to placebo. The major side effects were sedation and drowsiness; the frequency was similar for alprazolam and lorazepam and twice as high for active drug as placebo. An overall rating of side effect severity was not significantly different among the three groups.
Amiodarone: Pharmacology and Antiarrhythmic and Adverse Effects
Authors: Naccarelli Gerald V. , Rinkenberger Robert L. , Dougherty Anne H. , Giebel Ruth A.
Published: December 2015

Amiodarone is a benzofuran derivative that has been effective for the treatment of both supraventricular and ventricular tachyarrhythmias. It has a large volume of distribution, moderate bioavailability and a long half‐life. Its pharmacokinetics are not well understood and its tissue distribution is not typical of a 2‐compartment model. Due to ocular, dermatologic, gastrointestinal, neurologic, cardiovascular, thyroid and pulmonary toxicity, amiodarone should be reserved for use in patients with refractory and/or life‐threatening arrhythmias.
Pharmacokinetics of Ciprofloxacin in Elderly Subjects
Authors: LeBel Marc , Barbeau Gilles , Bergeron Michel G. , Roy Denis , Vallée François
Published: December 2015

The pharmacokinetics of single‐dose oral ciprofloxacin 500 mg was ascertained in 12 elderly and 12 young subjects. Mean age of the elderly volunteers was 75.4 years and the mean measured creatinine clearance in this group was 40.7 ml/min. Serum and saliva were collected in serial order for 24 hours (elderly) and 10 hours (young), and assayed for ciprofloxacin by high‐performance liquid chromatography. The geriatric subjects had higher serum levels throughout the sampling period, with a peak level of 3.24 ± 0.79 versus 2.26 ± 0.75 μg/ml for the younger group (p < 0.005; one‐way analysis of variance). A twofold increase in the ciprofloxacin half‐life may be partly explained by a decrease in the glomerular filtration rate, as shown by slower ciprofloxacin renal clearance (152.4 ± 54.2 vs 395.6 ± 139.0 for elderly and young subjects respectively; p < 0.001). We concluded that in elderly patients, ciprofloxacin should be administered at an interval not less than every 12 hours to prevent accumulation and eventually toxicity.
Aztreonam
Authors: Childs Stacy J. , Bodey Gerald P.
Published: December 2015

Aztreonam is a new, totally synthetic beta‐lactamase agent — the first monobactam. It is highly resistant to hydrolytic inactivation caused by plasmid‐mediated (except PSE‐2 enzyme found in some Pseudomonas species) or chromosomally mediated beta‐lactamases (except for K1 produced by rare strains of Klebsiella oxytoca). Accordingly, aztreonam remains active against many pathogens that are resistant to other beta‐lactam antibiotics. The drug exhibits directed antibacterial activity against gram‐negative organisms and is effective as monotherapy against most Enterobacteriaceae and Hemophilus and Neisseria species, including beta‐lactamase‐producing strains; it is not active against anaerobes or gram‐positive organisms. Before culture results are known, it may be necessary to administer the agent empirically in combination with other antibiotics. Aztreonam is rapidly distributed to most body tissues and fluids when administered parenterally. Its serum half‐life is 1.7 hours, suggesting a dosing interval of 6–8 hours for severe or life‐threatening infections and 8–12 hours for moderately severe infections and urinary tract infections. It is primarily eliminated unchanged in the urine and in much lesser amounts as a microbiologically inactive metabolite; slight biliary excretion may occur. Aztreonam is well‐tolerated, lacking any serious adverse hematologic, otic, or renal system effects. Its lack of effect on anaerobes helps to maintain resistance against colonization. Particularly in light of its safety and unique properties, aztreonam promises to be a useful alternative to aminoglycoside therapy.
Neuroleptic‐induced Catatonia After Abrupt Withdrawal of Amantadine During Neuroleptic Therapy
Authors: Brown Candace S. , Wittkowsky Ann K. , Bryant Stephen G.
Published: December 2015

Catatonic reactions to high‐potency neuroleptic drugs have been described in the literature. Our patient developed neuroleptic‐induced catatonia (NIC) after abrupt withdrawal of amantadine, a dopaminergic agent that has been implicated in neuroleptic malignant syndrome (NMS) on its discontinuation. This case report adds support to the hypothesis that NIC and NMS are 2 syndromes on a continuum of dopamine blockade, each of which may be treated with or uncovered by amantadine therapy.
Pharmacologic and Pharmacokinetic Comparison of Antianginal Agents
Authors: Angaran David
Published: December 2015

Calcium channel blockers, nitrates, and beta blockers are the primary agents used for the treatment of angina. Calcium has a central role in excitation‐contraction, action potential generation, and ischemic cell death. The three currently available calcium antagonists are nifedipine, verapamil, and diltiazem. Second‐generation agents are in development, and a classification system of calcium channel blockers is used to place the currently available agents and those on the horizon in perspective. Nitrate pharmacology and pharmacodynamics are possibly related to nitrate tolerance; however, this is a matter of some controversy. The beta blockers are all equally effective in the treatment of angina; therefore, drug selection is based on ancillary properties
Clinical Management of Asymptomatic Myocardial Ischemia
Authors: Touchon Robert
Published: December 2015

Silent myocardial ischemia, an important contributor to myocardial infarction, sudden cardiac death, ischemic arrhythmias, and other aspects of cardiac morbidity and mortality, requires treatment. Two pathophysiologic mechanisms have been proposed to explain transient, acute episodes of myocardial ischemia: an increase in myocardial demand in coronary obstructive disease and a transient dynamic stenosis or increase in tone within the epicardial coronary arteries. Epidemiologic data suggest a high prevalence of asymptomatic myocardial ischemia, and that these episodes are associated with the morbid events of ischemic heart disease. Several choices exist for the treatment of the condition: conservative (risk factor modification), moderate (medical intervention, percutaneous transluminal angioplasty), and aggressive (coronary artery bypass graft). Further research will elucidate the mechanisms by which the dormant atherosclerotic plaque is transformed to a complicated and activated plaque, and by which endothelial dysfunctions contribute to coronary heart disease.
Duration of Effect of Gallopamil, a Calcium Channel Blocker, on Methacholine‐Induced Bronchoconstriction
Authors: Massey Kenneth L. , Harman Eloise , Hendeles Leslie
Published: December 2015

We previously demonstrated a modest but significant protective effect of inhaled gallopamil (D600), the methoxy derivative of verapamil, against methacholine‐induced bronchoconstriction; however, the duration of the protective effect of this and other calcium channel blockers is unknown. We therefore evaluated the duration of this protective effect in 15 asthmatic subjects in a prospective, placebo‐controlled trial. Methacholine challenges (Cockcroft method) were performed 2 hours before and 30 minutes after the administration of placebo, and 1 mg and 10 mg of inhaled gallopamil. Gallopamil did not alter resting airway caliber, but significantly increased the concentration of methacholine required to decrease the FEV 20% 30 minutes after the dose. Results with both the 10‐mg and 1‐mg doses were significantly different from placebo, but not from each other. The duration of this protective effect was transient in the group as a whole; the mean drug activity ratios were not significantly different with this sample 2.5 hours after the dose. Thus the short duration of effect limits the potential clinical usefulness of gallopamil in suppressing the signs and symptoms of chronic asthma.
The 4‐Quinolone Antibiotics: Past, Present, and Future
Authors: Stein Gary E.
Published: December 2015

During the past 5 years the 4‐quinolone antibiotics have progressed from relative obscurity to a highly visible and intensely studied class of compounds. The zeal for developing and marketing newer fluoroquinolones closely parallels that of the cephalosporins for the last 10 years. All of these newer agents appear to have similar mechanisms of action, but numerous derivatives of the basic 4‐quinolone structure have been synthesized in an effort to enhance the antimicrobial spectrum and pharmacologic properties of these antibiotics.
Further Studies on the Inhibitory Effect of [D‐A1a6, des‐Gly‐NH210]LHRH Ethylamide on Spermatogenesis and Steroidogenesis in the Rat: Reversibility and Effect of Androgen Administration
Authors: PELLETIER G. , CUSAN G. L. , BÉLANGER A. , SÉGUIN C. , KELLY P. A. , LABRIE F.
Published: December 2015

Following the recent observation that chronic treatment with [D‐Ala6, des‐Gly‐NH210]LHRH ethylamide, a potent LHRH agonist, leads to an almost complete degeneration of seminiferous tubules in the rat, the time‐course of recovery of spermatogenesis after cessation of treatment was investigated. Adult rats injected with the LHRH agonist (100 ng, every second day) for four weeks were studied four, eight, and 16 weeks later. Although progressive improvement was noted from four to 16 weeks after cessation of treatment, about 25% of tubules still appeared completely degenerated at the end of the recuperation period. One month following cessation of treatment, testicular LH and prolactin receptor concentrations, testicular testosterone (T) and dihydrotestosterone (DHT) levels, and ventral prostate and seminal vesicle weights had returned to normal values. Testis weight, however, which was decreased by 40% one month after treatment with the LHRH analog, increased more slowly toward normal and was still at 85% of control value four months after treatment. Plasma LH and FSH levels, which were increased approximately 100% after treatment with the LHRH analogue, returned to within normal values between one and two months following cessation of treatment, respectively. Administration of androgens (T or DHT) concomitantly with the LHRH agonist only partially prevented the deleterious effects of the peptide on spermatogenesis. On the other hand, administration of the LHRH agonist had no effect on testicular morphology in hypophysectomized rats. These data show a rapid return to normal of the hypophyseal testicular elements responsible for androgen formation after desensitization by LHRH agonist treatment in adult rats. These results suggest that the inhibitory effect of chronic LHRH agonist administration on spermatogenesis is at least partially reversible, and that a decrease in androgen production is probably not the only factor involved in the inhibition of spermatogenesis by LHRH agonist treatment.
Spermatogenesis in Men Treated with Subcutaneous Application of Levonorgestrel and Estrone Rods
Authors: FRICK J. , DANNER CH. , JOOS H. , KUNIT G. , LUUKKAINEN T.
Published: December 2015

The effects of a combination of a progestin, levonorgestrel (LNg), and estrone (E1), on exocrine and endocrine testicular function were examined in adult men. The mode of administration was subcutaneously implanted dimethyl polysiloxane rods impregnated with either of the two hormones. The technical data on the rods used are as follows: length 30 mm, outer diameter 2.4 mm, hormone content/rod approximately 40 mg of LNg or E1. The subjects enrolled in the study were divided into two regimen groups: group A received six levonorgestrel and three estrone rods; group B, ten levonorgestrel and six estrone rods. The rods were implanted subcutaneously in the submammillary region. Complete spermatogenic arrest of variable duration was achieved in eleven of 13 subjects. In the two nonresponders, spermatogenesis was completely unaffected by the treatment. All subjects complained of some decrease of libido, erections, and potency, particularly during the first three to four months after initiation of treatment when testosterone in serum was at Its lowest levels. Clinical evaluations and measurements of various urine and serum components revealed no significant changes during the treatment period. The combination of LNg and E1 administered in this fashion was not adequate to achieve spermatogenic arrest in all subjects, including the period when the hormone‐bearing rods were still in place. Consequently, this regimen is not judged to hold promise as a method of fertility regulation.
In Vitro Effect of Inhibin on Cyclic AMP‐Phosphodiesterase Activity in Rat Testes
Authors: SHETH ANIL R. , VIJAYALAKSHMI S. , SHETH PARUL R. , BANDIVDEKAR A. H. , MOODBIDRI SUDHIR B.
Published: December 2015

We have previously reported that incubation of testicular slices with inhibin in the presence of 6 × 10−8 M oF SH resulted in a dose‐related reduction in accumulation of cyclic AMP due to a decrease in adenyl cyclase activity. The present study demonstrates that inhibin enhances the cyclic AMP‐phosphodiesterase activity in rat testicular tissue. These results suggest an additional mechanism by which cyclic AMP levels in gonadal tissue could be regulated by inhibin.
Effect of Castration and Androgen Treatment on Sex‐Linked Reactivity of Rat Aorta to a Thromboxane‐Mimic
Authors: KARANIAN JOHN W. , RAMEY ESTELLE R. , RAMWELL PETER W.
Published: December 2015

In order to examine the possible role of androgens in the isotonic response of the isolated rat aorta, a highly active mimic of thromboxane was employed. The maximum response to this TXA2‐mimic U46619 was greater by 24 ± 3% (P <0.01) in the male, as compared with the female, In addition, the sensitivity of the male vessel was 35 ± 4% greater than that of the female vessel (P < 0.01). Orchiectomy decreased the maximum contractile response to the TXA2‐mimic; however, ovariectomy increased both the maximum response and sensitivity. Testosterone pretreatment (1 mg/kg intramuscularly, biweekly for two weeks) restored the response of the castrated male but did not further increase the response of the ovariectomized female. The maximum response and sensitivity of the aorta of the intact female pretreated with testosterone was increased (P <0.01). These results suggest that the response of rat aorta to a TXA2‐mimic may be regulated not only by androgens, but also by ovarian hormones.
Recovery of Normal Testicular Ultrastructure and Sperm Motility after Cessation of Gossypol Treatment in Rats
Authors: HOFFER ANITA P. , LISSER STEVEN P.
Published: December 2015

This study evaluates the reversibility of the effects of gossypol on testicular ultrastructure and the motility of epididymal spermatozoa. Adult male rats were treated 6 days weekly with the vehicle alone (Group A), or with 10 (Group B) or 20 (Group C) mg/kg of gossypol for 12 weeks, and then sacrificed six or 12 weeks after cessation of treatment. Although epididymal spermatozoa in Groups B and C were 100% immotile after gossypol treatment, little evidence of abnormality could be detected with the light microscope in the seminiferous tubules or interstitium. By contrast, at the ultrastructural level, there were demonstrable pathognomonic defects in the mitochondrial sheath and axonemes of step 18 and 19 spermatids which were identical to those reported earlier (Hoffer, 1983). In addition, an ultrastructural defect in the flagella of late testicular spermatozoa is described for the first time. This defect consists of an indentation, or constriction, of the mitochondrial sheath at outer dense fibers (ODFs) 1, 2, and 9, resulting in a separation of these 3 ODFs from the other fibers. This defect, though visible in an earlier ultrastructural study (Hoffer, 1983), was not described.

In Group B rats allowed to recover from gossypol treatment, ultrastructural defects in step 18 and 19 spermatids could not be detected at six or at 12 weeks after cessation of treatment, and sperm motility also did not differ significantly from controls by the end of either recovery period. In Group C rats, sperm motility returned to the normal range within six weeks after treatment ended, but a few morphological defects in the midpiece and axoneme of late spermatids could still be detected with the electron microscope. However, after 12 weeks of recovery, ultrastructural evidence of drug treatment could no longer be seen. The results of this study do not support the concern that use of gossypol leads to subtle irreversible damage in the testis which has previously escaped detection. These findings provide the first reported ultrastructural evidence that recovery from gossypol treatment is time and dose‐related, and that there is a complete reversal of its deleterious effects. The observations on the motility of epididymal spermatozoa at the end of the treatment and recovery periods support these conclusions.
Testicular Blood Flow in Young and Old Rats and Influence of hCG
Authors: PUNJABI U. , Van HOECKE J. , VERDONCK L. , VERMEULEN A.
Published: December 2015

To evaluate the influence of age, testicular capillary blood flow (TCBF) was measured, using the microsphere technique, in rats 3 to 24 months old, under basal conditions and after hCG stimulation (10 IU/d for two days). Despite a decline in plasma T levels, testicular capillary blood flow did not decrease with age, and hCG stimulation resulted in similar increases of approximately 50% (P < 0.01) in testicular capillary blood flow in all age groups. We concluded that the age‐associated decrease in testosterone secretion in rats is not the consequence of a decreased testicular blood flow.
Effects of AMSA, An Antineoplastic Agent, on Spermatogenesis in the Mouse
Authors: DA CUNHA MIGUEL F. , MEISTRICH MARVIN L. , FINCH‐NEIMEYER MARCIA V.
Published: December 2015

The stages of spermatogenic cells killed by the single and fractionated administration of AMSA, an acridine derivative used in cancer chemotherapy, have been identified in the mouse. A wide range of doses, up to a total of 30 mg/kg, which is the LD50 for AMSA given in three daily injections, was employed. Survival of differentiating (types Ai through Intermediate) and stem spermatogonia was measured by sperm counts performed 29 and 56 days after treatment, respectively. The sensitivity of germ cells to AMSA at other stages of differentiation was determined by semiquantitative histologic analysis at 11 days after treatment. Significant killing of differentiating spermatogonia, types A2 through B, but only minor killing of stem cells and no toxicity to post‐spermatogonial stages were observed with all treatment schedules. This pattern of differential sensitivity can explain the temporary azoospermia observed in man during AMSA treatment, which was followed by a return to normal sperm counts after cessation of therapy.
Activation of Palmitic Acid by Human Spermatozoa
Authors: JONES ROBERT E. , PLYMATE STEPHEN R. , FARISS BRUCE L.
Published: December 2015

Human spermatozoa were studied to determine if a long chain fatty acid, CoASH ligase (AMP) (E.C. 6.2.1.3), was present. Ligase activity was measured with a radioligand millipore filter technique and was readily detectable in spermatozoa or in the protein fraction extracted with Triton X‐100, but was not present in seminal plasma. The assay was optimized for pH, protein concentration, and incubation time. Activity was dependent upon palmitic acid, ATP, coenzyme A, and a divalent cation. Sperm ligase appeared similar to the ligase characterized from other tissues by sharing a common pH optimum (approximately 8.0–8.4), and a preference for magnesium over manganese in the incubation media.
Histopathology of Prepubertal Rat Testes Subjected to Various Durations of Spermatic Cord Torsion
Authors: COSENTINO M. JAMES , NISHIDA MASAO , RABINOWITZ RONALD , COCKETT ABRAHAM T.K.
Published: December 2015

Sixty prepubertal rats were subjected to unilateral spermatic cord torsion for a duration of 0,1,3,5,9 or 12 hours. At the end of this period, the ipsilateral testes either were removed for immediate processing or subjected to detorsion and orchiopexy, followed by a six‐week recovery period prior to histologic study. Twelve histologic parameters were each scored according to the degree of pathologic findings, thus allowing for a quantitative assessment of testicular damage. The sequence of specific histologic degeneration that occurred with spermatic cord torsion is described. These changes were found to be dependent on the duration of torsion, with the greatest damage occurring after three hours or more. In the animals undergoing detorsion followed by a six‐week recovery period, severe degeneration was noted for all durations of torsion studied. The extent of this degeneration was significantly correlated with a reduction in fertility.
Low Molecular Weight FSH Binding Inhibitor in Bovine Testis
Authors: KRISHNAN KRISHNASWAMY A. , SLUSS PATRICK M. , REICHERT LEO E.
Published: December 2015

Purification of low Mr(< 5000) inhibitors of FSH binding to receptors has been hampered by their low concentration in tissue extracts and physiologic fluids. The calf testis represents an accessible and economic source of large quanities of tissue, and was therefore studied as a source of FSH binding inhibitors. Supernatants (27,000 X g) of calf testis homogenates inhibited binding of [125I]‐hFSH as well as [125I]‐hCG to membrane‐bound receptor from the same source. FSH binding inhibitor was concentrated from large volumes of testis supernatant by precipitation of inert material with metaphosphoric acid, concentration/desalting of the resulting supernatant by ultrafiltration (Amicon UM‐05 membranes) and lyophilization. Separation of FSH binding inhibitor and LH(hCG) binding inhibitor activities could be achieved by molecular sieving on Sephadex G‐25. A partially purified fraction with inhibitors of FSH binding activity (ED50 = 44 μg protein) and free of LH(hCG) binding inhibitor activity (no activity at 800 μg protein) emerged with a Ve/Vo of 1.9, reflecting an apparent Mr of about 1500. Inhibitors of LH(hCG) binding activity emerged with the column outer volume. Rechromatography of the FSH binding inhibitor fraction on G‐25 indicated two closely associated peaks of activity. These could be further resolved by gel filtration through BioGel P2, to give a salt‐free fraction with an FSH binding inhibitor activity (ED50) of 24 μg protein. The inhibitor was heat‐labile, losing 80% of its activity after 2 hours at 100 C. The testicular low molecular weight FSH binding inhibitor is similar to bovine follicular fluid and serum FSH binding inhibitor by several parameters. Bovine testis should prove a useful source for binding inhibitor purification.
Intratesticular Factors and Testosterone Secretion. Effect of Treatments That Alter the Level of Testosterone Within the Testis
Authors: SHARPE RICHARD M. , KERR JEFFREY B. , FRASER HAMISH M. , BARTLETT JOHN M. S.
Published: December 2015

The authors recently have reported the presence of a nongonadotropic polypeptide factor in rat testicular interstitial fluid that can exert marked stimulatory effects on Leydig cell testosterone production. To assess the potential physiologic significance of this factor, its effective levels in rat interstitial fluid have been investigated in response to treatments that either markedly reduce interstitial fluid testosterone concentrations (anti‐LH treatment; transient or chronic experimental cryptorchidism; destruction of Leydig cells with ethane dimethanesulphonate) or that significantly elevate testosterone levels in interstitial fluid by injection of hCG. The possible relationship between this factor and changes in testicular weight, serum LH and FSH, and interstitial fluid volume also were monitored.

When testosterone levels in interstitial fluid were decreased by 75 to 99% either acutely (5–72 hours) or chronically (20–75 days), there was an accompanying increase (P < 0.001) in the levels of the interstitial fluid factor(s), as determined by the ability of charcoal‐stripped interstitial fluid from individual rats to enhance hCG‐stimulated testosterone production by Percoll‐purified Leydig cells in vitro. Anti‐LH treatment increased the levels of the interstitial fluid factor(s) over the ensuing 5 to 48 hours. In abdominal testes from rats made unilaterally or bilaterally cryptorchid for 20 or 55 days, a decrease in interstitial fluid testosterone levels was associated with increased levels of the interstitial fluid factor(s). The same inverse relationship was found 72 hours after treatment with ethane dimethanesulphonate in which Leydig cells had disappeared from the testis. When interstitial fluid testosterone levels were raised to supraphysiologic levels with hCG, however, levels of the factor(s) exhibited a stepwise decrease (P < 0.001) over 4 to 48 hours. No consistent correlation was found between changes in interstitial fluid volume and levels of the factor(s). In rats made bilaterally cryptorchid for 2 days, and in which unilateral or bilateral redescent of the testis occurred, redescended testes partially recovered in weight and interstitial fluid testosterone levels returned to normal, but levels of the factor(s) remained elevated. Raised serum FSH (but not LH) levels and/or presumed Sertoli cell dysfunction were always associated with increased levels of the interstitial fluid factor(s). Therefore, although the processes that control levels of the interstitial fluid factor(s) have not been identified unequivocally, our results suggest that modulation of its levels in interstitial fluid may be of physiologic significance.
Morphologic and Endocrine Changes in the Reproductive Organs in Rats Implanted with Gossypol Acetate Pellet in the Testis
Authors: AHLUWALIA BALWANT , HYPOLITE FRANK , ANDERSON WINSTON
Published: December 2015

Gossypol acetate pellets in concentrations ranging from 0.5, 1.0, 5.0, 10.0, to 20.0 mg were implanted in the testis of rats for a period up to 7 weeks. The implant was made in one testis only. In animals with a 10.0‐mg implant, the germinal epithelium showed degenerative changes following the 7‐week period, but spermatogonia and spermatocytes were unaffected. The effect of gossypol acetate appeared to be at the spermatid level. The electron microscopic study showed that the acrosomal membranes were disrupted. Blood testosterone and LH, but not FSH, decreased significantly (P < 0.01). The fertility studies showed 100% infertility but no loss in libido in the 10‐mg implant group. Up to the 7‐week period, implants of less than 10 mg produced no significant effects on any of the parameters mentioned above. Animals with 20‐mg implants had toxic symptoms. From the results in this study, it is concluded that 10.0 mg of gossypol acetate implanted in a single pellet in one testis causes antispermatogenic effects.
Steroidogenesis and Testosterone Metabolism in Cultured Principal Cells from the Ram Epididymis
Authors: AMANN RUPERT P. , MARENGO SUSAN R. , BROWN DAVID V.
Published: December 2015

To determine if ram principal cells can synthesize or metabolize testosterone, or metabolize other steroids present in rete testis fluid, principal cells from the initial segment, central caput, and proximal corpus epididymidis were isolated and cultured in a floating collagen matrix with medium containing 20% dialyzed rete testis fluid. In the first experiment, each matrix was washed twice in testosterone‐free medium on day 2.8, transferred into culture medium containing 100 nM of a tritiated steroid and incubated for 4 hours at 34 C. The tritiated steroids were pregnenolone, 5‐androstene‐3β,17β‐diol, progesterone, 4‐androstene‐3,17‐dione, testosterone, and dihydrotestosterone. Since testosterone was not formed from 5‐androstene‐3β,17β‐diol or 4‐androstene‐3,17‐dione, testosterone synthesis by ram principal cells is unlikely. Pregnenolone and 5‐androstene‐3β,17β‐diol were not metabolized and only slight metabolism of dihydrotestosterone occurred. Progesterone, 4‐androstene‐3,17‐dione, and testosterone were metabolized to 5α‐reduced products tentatively identified as 5α‐pregnane‐3,20‐dione and 5α‐pregnan‐3β‐ol‐20‐one and/or 5α‐pregnan‐20α‐ol‐3‐one; 5α‐androstane‐3,17‐dione and 5α‐androstan‐3α‐ol‐17‐one, and dihydrotestosterone, respectively. The second experiment evaluated testosterone metabolism by both cultured principal cells and minced epididymal tissue. On day 1 of culture, during 12 hours the accumulation of dihydrotestosterone in medium from cells of the central caput was 48 × and 1.1 × that in medium from cells of the initial segment and proximal corpus epididymidis. The difference (P > 0.01) persisted on days 3 and 6 of culture, although testosterone metabolism decreased. For 3‐hour incubations of minced tissue, testosterone metabolism by tissue from the central caput was 3.6 ×, 1.3 x, and 0.9 × that of tissues from the initial segment, distal caput, and proximal corpus, respectively. Thus, testosterone metabolism in the initial segment is low compared with more distal regions.
Diverticula of Human Seminiferous Tubules in the Normal and Pathologic Testis
Authors: NISTAL MANUEL , SANTAMARIA LUIS , REGADERA JAVIER , PANIAGUA RICARDO
Published: December 2015

The occurrence of diverticula in human seminiferous tubules was investigated in the adult human testis in normal as well as physiologic (aging) and pathologic (germ cell depletion, obstruction of male excretory ducts, varicocele and systemic arteriosclerosis) conditions. Diverticula, which are evaginations of the seminiferous epithelium surrounded by a thin tunica propria, were present in all groups studied. The number of diverticula per mm2 testis was higher in the testis with obstruction than in those without obstruction at each age considered. The number of diverticula increased with age in both the obstructed and nonobstructed testis. No changes in the number of diverticula per mm2 testis were found in relation to systemic arteriosclerosis or different degrees of germ cell depletion. Varicocele was only associated with increased numbers of diverticula when it was also associated with obstruction. The formation of diverticula in human seminiferous tubules seems to be an obstructive process related to increasing age.
Mechanism of LHRH‐Stimulated Steroidogenesis in Rat Leydig Cells: Lipoxygenase Products of Arachidonic Acid May Not Be Involved
Authors: DIDOLKAR ASHOK K. , SUNDARAM KALYAN
Published: December 2015

Luteinizing hormone releasing hormone agonist, [(imBzl)‐DHis6,Pro9,NEt]‐LHRH (LHRH‐A), caused a two to threefold increase in in vitro testosterone (T) secretion by rat Leydig cells. This LHRH‐A‐induced T secretion was completely blocked by quinacrine and chloroquine, inhibitors of phospholipase A2. Addition of phospholipase A2, however, was ineffective in stimulating basal or LHRH‐A‐induced T secretion. Phospholipase C, on the other hand, significantly stimulated both basal and LHRH‐A‐induced T secretion. Exogenously added arachidonic acid stimulated basal T secretion in a dose dependent manner, the maximum increase being about 100% over basal at a dose of 100 μM. Higher doses of arachidonic acid had no stimulatory effect. In the presence of LHRH‐A, the stimulatory effect of arachidonic acid was additive up to a concentration of 100 μM; but higher concentrations of arachidonic acid (200 μM) were inhibitory. LHRH‐A‐induced steroidogenesis was inhibited by 5, 8, 11, 14 Eicosatetraynoic acid (ETYA), an inhibitor of all the three known pathways of arachidonic acid metabolism, and by nordihydroguaiaretic acid, an inhibitor of the lipoxygenase pathway of arachidonic acid metabolism. LHRH‐A‐stimulated T secretion was not inhibited by indomethacin, an inhibitor of the cyclo‐oxygenase pathway of arachidonic acid metabolism. ETYA inhibited arachidonic acid‐induced T secretion. Nordihydroguaiaretic acid, on the other hand, augmented basal, arachidonic acid–, phospholipase C–, or phorbol 12, myristate 13 acetate‐induced testosterone secretion. These results suggest that arachidonic acid, whose release is influenced by phospholipase C, is involved in LHRH‐A‐induced T secretion by rat Leydig cells. Furthermore, LHRH‐A‐induced steroidogenic responses in Leydig cells seem to be mediated by arachidonic acid metabolites other than cyclo‐oxygenase or lipoxygenase products.
Cryopreservation in Different Concentrations of Glycerol Alters Boar Sperm and Their Membranes
Authors: BUHR MARY M. , FISER PAUL , BAILEY JANICE L. , CURTIS ELIZABETH F.
Published: December 2015

ABSTRACT: To test the hypothesis that glycerol would concomitantly affect sperm membrane structure and the function of the intact cells, boar semen (4 ejaculates from 4 boars) was cryopreserved in an egg yolk extender with 0%, 2%, 4%, or 8% glycerol in 0.5‐mL straws using previously derived optimal cooling and thawing rates. Increasing glycerol concentrations increased spermatozoal progressive motility immediately after thawing and after 2 hours at 43°C, but decreased the percentage of sperm with normal acrosomal morphology. The mathematical products of the motility and acrosomal integrity scores (MOT x NAR index) were low in 0% and 8% glycerol, and significantly higher in 2% and 4% glycerol. The fluidity of sperm‐head plasma membranes, a measure of molecular interaction, was assessed with the lipid probes trans‐parinaric acid and cis‐parinaric acid (tPNA, cPNA), during a 2.5‐hour incubation with or without 1 mM Ca2+. Membrane fluidity detected by each probe differed significantly, indicating the presence of at least 2 domains whose constituent molecules had unique dynamics. Behavior of each domain was radically altered by cryopreservation. Increasing glycerol concentration caused a variably faster loss of fluidity in the cPNA domain, and had highly variable effects on fluidity change over time in the tPNA domain. Normal acrosomal ridge (NAR) and the MOT x NAR index correlated significantly with the fluidity of the more mobile cPNA domain (±1mM Ca2+), supporting the hypothesis of an interrelationship of glycerol concentration during cryopreservation with sperm membrane structure and cell function. The MOT x NAR index may be a useful guide in choosing optimal cryoprotectant concentrations.
Y Chromosome Deletions in Azoospermic Men in India
Authors: Thangaraj Kumarasamy , Gupta Nalini J. , Pavani Kadupu , Reddy Alla G. , Subramainan Subbaya , Rani Deepa Selvi , Ghosh Bibaswan , Chakravarty Baidyanath , Singh Lalji
Published: December 2015

ABSTRACT: Genetic factors cause about 10% of male infertility. Azoospermia factors (AZFa, AZFb, AZFc) are considered to be the most important for spermatogenesis. We therefore made an attempt to evaluate the genetic cause of azoospermia, Y chromosome deletion in particular, in Indian men. We have analyzed a total of 570 men, including 340 azoospermic men and 230 normal control subjects. DNA samples were initially screened with 30 sequence‐tagged site (STS) markers representing AZF regions (AZFa, AZFb, AZFc). Samples, with deletion in the above regions were mapped by STS walking. Further, the deletions were confirmed by Southern hybridization using the probes from both euchromatic and heterochromatic regions. Of the total 340 azoospermic men analyzed, 29 individuals (8.5%) showed Y chromosome deletion, of which deletion in AZFc region was the most common (82.8%) followed by AZFb (55.2%) and AZFa (24.1%). Microdeletions were observed in AZFa, whereas macrodeletions were observed in AZFb and AZFc regions. Deletion of heterochromatic and azoospermic regions was detected in 20.7% of the azoospermic men. In 7 azoospermic men, deletion was found in more than 8.0 Mb spanning AZFb and AZFc regions. Sequence analysis at the break points on the Y chromosome revealed the presence of L1, ERV, and other retroviral repeat elements. We also identified a ∼240‐kb region consisting of 125 bp tandem repeats predominantly comprised of ERV elements in the AZFb region. Histological study of the testicular tissue of the azoospermic men, who showed Y chromosome deletion, revealed complete absence of germ cells and presence of only Sertoli cells.
Sex Chromosome Alignment at Meiosis of Azoospermic Men With Azoospermia Factor Microdeletion
Authors: Yogev Leah , Segal Shmuel , Zeharia Einav , Gamzu Ronni , Maymon Batia B. , Paz Gedalia , Botchan Amnon , Hauser Ron , Yavetz Haim , Kleiman Sandra E.
Published: December 2015

ABSTRACT: Deletions in the q arm of the Y chromosome result in spermatogenesis impairment. The aim of the present study was to observe the × and Y chromosome alignment in the spermatocytes of men with Y chromosome microdeletion of the azoospermia factor (AZF) region. This was performed by multicolor fluorescence in situ hybridization probes for the centromere and telomere regions. Testicular biopsies were performed in a testicular sperm extraction‐intracytoplasmic sperm injection set‐up in 11 azoospermic men: 8 (nonobstructive) with AZF deletions and 3 (obstructive) controls. Histological sections, cytology preparations of the testicular biopsies, and evaluation of the meiosis according to the percentage of XY and 18 bivalents formation were assessed. Spermatozoa were identified in at least one location in controls and specimens with AZFc‐deleted Y chromosomes. Complete spermatocyte arrest was found in those with a deletion that included the entire AZFb region. Bivalent formation rate of chromosome 18 was high in all samples (81%‐99%). In contrast, the rate of bivalent X‐Y as determined by centromeric probes was lower but in the range favorable with spermatozoa findings in controls and patients with the AZFc deletion (56%‐90%), but not in those with AZFb‐c deletions (28%‐29%). A dramatic impairment in the normal alignment of × and Y telomeres in the specimen with AZFb‐c deletion was shown (29%), compared to the specimens with AZFc deletion (70%‐94%). It is suggested that the absence of sperm cells in specimens with the entire AZFb and with AZFb‐c deletions is accompanied by meiosis impairment, perhaps as a result of the extent of the deletion or because of the absence of genes that are involved in the × and Y chromosome alignment.
Heat‐Induced Apoptosis of Mouse Meiotic Cells Is Suppressed by Ectopic Expression of Testis‐Specific Calpastatin
Authors: Somwaru Lily , Li Siming , Doglio Lynn , Goldberg Erwin , Zirkin Barry R.
Published: December 2015

ABSTRACT: Calpastatin is a naturally occurring inhibitor of calpain, a protease involved in apoptotic cell death. A testis‐specific isoform of calpastatin (tCAST) has been identified that is transcribed in haploid germ cells but not in spermatocytes. To investigate the possible function(s) of tCAST, we tested the hypothesis that the ectopic expression of calpastatin in spermatocytes would suppress the death of these cells in response to an apoptosis‐inducing stimulus in vivo. To this end, the 5′‐flanking region of the mouse ldhc gene was linked to tCAST, and transgenic mice were generated. Immunohistochemical analysis revealed that, in contrast to control sections in which the signal for tCAST was seen in round spermatids, intense staining was visualized in pachytene spermatocytes in the transgenic animals, indicating that the strategy we used to generate the transgenic animals resulted in the ectopic expression of tCAST in spermatocytes. We then tested the effect of a short period of heating on germ cell apoptosis in the testes of wild‐type and transgenic mice. Pachytene spermatocytes were the major germ cell type seen to undergo apoptosis after heat treatment. There were no differences in the number of apoptotic germ cells per seminiferous tubule between wild‐type and tCAST transgenic control mice; thus, there was no apparent effect of the transgene on normal apoptosis. Heating resulted in increased numbers of TUNEL‐positive germ cells in both wild‐type and tCAST transgenic mice, as well as increased testicular DNA fragmentation. Heating the tCAST transgenic mouse testes resulted in significantly fewer apoptotic cells per seminiferous tubule than in wild‐type mice at both 8 and 24 hours after treatment. Thus, as hypothesized, the ectopic expression of tCAST in pachytene spermatocytes suppressed germ cell apoptosis.
Cystatin C Is Highly Expressed in the Human Male Reproductive System
Authors: Jiborn Thomas , Abrahamson Magnus , Wallin Hanna , Malm Johan , Lundwall Åke , Gadaleanu Virgil , Abrahamsson Per‐Anders , Bjartell Anders
Published: December 2015

ABSTRACT: Cystatin C displays the strongest inhibitory activity of all cystatins toward lysosomal cysteine proteases in general and has a widespread distribution in human tissues and body fluids, including seminal plasma. The aim of this study was to investigate the distribution of cystatin C in the male reproductive system. Immunohistochemistry revealed a widespread distribution of cystatin C in normal tissues from the testis, epididymis, vas deferens, seminal vesicle, and prostate gland. Immunoreactive cystatin C was localized in basal and secretory epithelial cells, but also in neuroendocrine cells in the prostate, identified by immunostaining for chromogranin A. On adjacent tissue sections, we demonstrated local production of cystatin C utilizing nonradioactive in situ hybridization with a 201‐base‐long digoxigenin‐labeled antisense RNA probe specific for the cystatin C transcript. Staining patterns obtained by immunohistochemistry and in situ hybridization correlated well. Enzyme‐linked immunosorbent assay for quantitative analysis of cystatin C demonstrated that cystatin C was present at high concentrations in tissue homogenates from all locations investigated, compared to liver, muscle, spleen, and other general tissues. Western blotting of tissue homogenates revealed a predominant 15‐kd cystatin C immunoreactive component in accordance with previous findings in other organs. Quantitative real‐time polymerase chain reaction analysis to determine messenger RNA levels in whole tissue extracts showed that the cystatin C gene is highly expressed in the seminal vesicles and the prostate gland, indicating that the major amount of cystatin C in the male reproductive organs and seminal plasma is produced by cells in these 2 tissues. It is concluded that cystatin C is highly expressed and widely distributed throughout the male genital tract, suggesting that cystatin C is an important regulator for normal and pathological proteolysis in the male reproductive system.