bluuebunny/crossref_metadata_2025 huggingface dataset:
Impact of Managed Care in Pharmacy
Authors: Scott Bruce
Published: October 2023

Implementation of managed care results in decreased reimbursement, decreased use of specialty services, and consolidation of providers. With these changes, pharmacy services have been affected in all settings. However, pharmacists do have opportunities to implement change in this new health care environment. Employee compensation usually represents over half of hospital expenses and is therefore often the target of efforts to decrease costs. For pharmacy, this translates to increased automation and expanded technician responsibilities. In response to these changes, pharmacists have broadened their scope of responsibility to improve drug use, with the goal of decreasing drug‐related hospitalizations and providing quality care at a reasonable cost.
Lack of Effect of Olanzapine on the Pharmacokinetics of a Single Aminophylline Dose in Healthy Men
Authors: Macias William L. , Bergstrom Richard E , Cerimele Benito J. , Kassahun Kelem , Tatum David E. , Callaghan John T.
Published: October 2023

Study Objective. To test whether olanzapine, an atypical antipsychotic, is an inhibitor of cytochrome P450 (CYP) 1A2 activity, we conducted a drug interaction study with theophylline, a known CYP1A2 substrate.

Design. Two‐way, randomized, crossover study.

Setting. Clinical research laboratory.

Subjects. Nineteen healthy males (16 smokers, 3 nonsmokers).

Interventions. Because the a priori expectation was no effect of olanzapine on theophylline pharmacokinetics, a parallel study using cimetidine was included as a positive control. In group 1, 12 healthy subjects received a 30‐minute intravenous infusion of aminophylline 350 mg after 9 consecutive days of either olanzapine or placebo. In group 2, seven healthy subjects received a similar aminophylline infusion after 9 consecutive days of either cimetidine or placebo.

Measurements and Main Results. Concentrations of theophylline and its metabolites in serum and urine were measured for 24 and 72 hours, respectively. Plasma concentrations of olanzapine and its metabolites were measured for 24 hours after the next to last dose and 168 hours after the last olanzapine dose. Olanzapine did not affect theophylline pharmacokinetics. However, cimetidine significantly decreased theophylline clearance and the corresponding formation of its metabolites. Urinary excretion of theophylline and its metabolites was unaffected by olanzapine but was reduced significantly by cimetidine. Steady‐state concentrations of olanzapine (15.3 ng/ml), 10‐N‐glucuronide (4.9 ng/ml), and 4′‐N‐desmethyl olanzapine (2.5 ng/ml) were observed after olanzapine 10 mg once/day and were unaffected by coadministration of theophylline.

Conclusion. As predicted by in vitro studies, steady‐state concentrations of olanzapine and its metabolites did not affect theophylline pharmacokinetics and should not affect the pharmacokinetics of other agents metabolized by the CYP1A2 isozyme.
Retrospective and Prospective Analyses of the Treatment of Overanticoagulated Patients
Authors: Duong Thu‐Mai , Plowman Brian K. , Morreale Anthony P. , Janetzky Katherine
Published: October 2023

Study Objective. To compare withholding warfarin therapy with low‐dose (2.5 mg) oral vitamin K therapy in excessively anticoagulated patients without bleeding complications.

Design. Prospective and retrospective studies.

Setting. Anticoagulation clinic at a Veterans Affairs institution.

Patients. Twenty‐eight men were matched according to initial international normalized ratio (INR) and INR goal ranges.

Interventions. The retrospective arm of the study consisted of chart reviews of overanticoagulated patients whose warfarin doses were held until therapeutic INR values were reached. The prospective arm included overanticoagulated patients who were administered a single 2.5‐mg dose of oral vitamin K.

Measurements and Main Results. Mean days to therapeutic INR values were 2.3 ± 0.6 and 1.4 ± 0.6 (p=0.001), and mean reduction in INR 1 day after treatment intervention was 1.32 ± 0.79 and 3.46 ± 1.31 (p<001) U for the withholding and vitamin K groups, respectively.

Conclusion. Compared with withholding the warfarin dose, administration of 2.5 mg of oral vitamin K to excessively anticoagulated patients receiving warfarin significantly reduced the time required to reach a therapeutic INR as well as final INR.
Characteristics Associated with Ability to Prevent Adverse Drug Reactions in Hospitalized Patients
Authors: Seeger John D. , Kong Sheldon Xiaodong , Schumock Glen T.
Published: October 2023

We conducted a retrospective analysis to identify characteristics of preventable adverse drug reactions (ADRs). We reviewed reports on 612 ADRs occurring in hospitalized patients over 4 years, identified by the hospital's spontaneous ADR reporting program, and classified the events as potentially preventable or not preventable. Characteristics related to ADR preventability in the univariate analysis were the patient's clinical service, organ system involved in the ADR, class of drug causing the ADR, relationship to dosage, type of ADR, and probability that the reaction was due to the drug. Among these, relationship to dosage (p<0.001) and type of ADR (p<0.001) appeared to be most strongly related to preventability. In a multivariate analysis, preventable ADRs were associated with dosing (OR 3.82, 95% CI 2.42‐6.03) and previous allergy to the drug (OR 3.46, 95%CI 1.01–11.88). An ADR that was classified as an allergic (OR 0.50, 95% CI 0.27‐0.94) or idiosyncratic reaction (OR 0.44, 95% CI 0.28‐0.71) was unlikely to be considered preventable. Preventable ADRs in hospitalized patients are likely to be dosage related or to occur among patients allergic to the specific agent.
Peripheral Blood Progenitor Cell Mobilization: A Clinical Review
Authors: Kennedy M. John
Published: October 2023

High‐dose chemotherapy appears to confer a survival advantage on certain patients with lymphohematopoietic malignancies and may have a role in some solid tumors, notably germ cell and, possibly, breast cancer. Autologous bone marrow grafts adequately support high‐dose therapy in many patients, but more‐rapid engraftment is observed in those who are given mobilized peripheral blood progenitor cell grafts. The optimal method for mobilizing progenitor cells remains to be defined, but adequate numbers of cells can be obtained by apheresis after treatment with cytokines, chemotherapy, or a combination of both. The minimum number of progenitor cells that will result in rapid and durable engraftment from mobilized peripheral blood cell collections is probably between 2.5 times 106 and 5.0 times 106 CD34+ cells/kg, as determined by flow cytometry, a method not yet fully standardized among laboratories. Tumor cell contamination of peripheral blood progenitor cell grafts is less common, and there are fewer tumor cells than in matched bone marrow collections. Strategies for producing large numbers of stem cells by ex vivo expansion of small collections may be feasible in the near future. Such approaches may also reduce tumor cell contamination of the product.
The Association between Cryptococcal Variety and Immunocompetent and Immunocompromised Hosts
Authors: Peachey Pamala R. , Gubbins Paul O. , Martin Rebecca E.
Published: October 2023

The varieties of Cryptococcus neoformans serotypes are neoformans and gattii. Data suggest an association between cryptococcal variety and host immune status. In addition, the two varieties differ in their epidemiology and pathogenicity. Furthermore, symptoms, outcome, and response of cryptococcosis to antifungal therapy may vary. The two varieties also differ in immune‐modulating effects. Sparse clinical data suggest var. gattii is more virulent and may be more recalcitrant to antifungal therapy. Also, its infections produce more sequelae. A better understanding of how cryptococcal variety influences the clinical course and response to the treatment of cryptococcosis is needed. Clinicians should be aware of the association, especially in patients with refractory disease. It may be useful to type the isolate to the variety level and administer prolonged antifungal therapy.
Cyclosporine‐Induced β‐Adrenergic Receptor Down‐Regulation in Bovine Pulmonary Artery Smooth Muscle Cells: A Pilot Study
Authors: Dickens George R. , Morris Peter E.
Published: October 2023

We attempted to determine the effects of cyclosporine on β‐adrenergic receptors in bovine pulmonary artery smooth muscle cells. Bovine pulmonary artery smooth muscle cells were exposed to cyclosporine at a concentration of 100 ng/ml in culture media for 5 days, and control bovine pulmonary artery smooth muscle cells were exposed to only culture media for the same 5‐day period. β‐Adrenergic receptors were measured as total binding capacity (Bmax) by nonlinear least squares fit of the specific binding curve. In a separate experiment β1‐versus β2‐adrenergic receptor subtypes were identified by computer modeling (LIGAND) of 17–19 point CGP20712A‐125ICYP competition curves. Cyclosporine significantly (p = 0.02) decreased bovine pulmonary artery smooth muscle β‐adrenergic receptor density by 54% ± 7%. The Bmax for control versus treated cells was 38.9 ± 18 versus 17.7 ± 12 fmol/mg protein, respectively. Subtype determination of β‐receptors revealed 70% or more β2‐ and 30% or less β1‐adrenergic receptors. Cyclosporine caused a 54% reduction in overall β‐adrenergic receptor density in bovine pulmonary artery smooth muscle cells. The reduction in Bmax is suspected not to be a result of selective down‐regulation of β1‐adrenergic receptors alone. We believe that cyclosporine may also contribute to a decrease in β2‐adrenergic receptors.
Circadian Variations in the Pharmacokinetics of a New Microemulsion Formulation of Cyclosporine in Cardiac Transplant Recipients
Authors: Châteauvert Nathalie , Côté Hélène
Published: October 2023

We attempted to characterize circadian variations in pharmacokinetic parameters of a new formulation of cyclosporine (CsA) in nine cardiac allograft recipients. A secondary objective was to determine the sampling time that correlated best with exposure of patients to the drug. This was a two‐period study with each period lasting 12 hours. All patients received two equal doses of a new microemulsion of CsA 12 hours apart. Blood samples to measure drug levels were obtained at administration and 1, 2, 3, 4, 6, 9, and 12 hours after each dose. We found no statistically significant difference in pharmacokinetic parameters (areas under the curve, minimum blood concentration, oral clearance) measured during the day and during the night. However, maximum blood concentrations during the day were 30% higher than those at night (p<0.05). We found a good correlation between minimum concentrations in the morning and overall exposure of patients to CsA (r = 0.89). This new microemulsion appears to have few circadian variations of blood concentrations in cardiac transplant recipients. The clinical significance of higher maximum blood concentration during daytime remains to be elucidated. Our results support the most widely accepted method for monitoring CsA, which is based on minimum concentrations in the morning.
Nefazodone: Single versus Twice Daily Dose
Authors: Voris John C. , Shaurette Glen N. , Sebastian Praxedes S. , Will‐Wallace Louise A.
Published: October 2023

This pilot study involved six men with major depression treated with nefazodone dosed either twice/day or once/day at bedtime. Depression was rated before nefazodone therapy and at 4 weeks by the self‐report version of the Hamilton Depression Rating Scale. A simple 10‐cm visual analog side effect scale for daytime drowsiness was completed at the latter time. Dosages of nefazodone were at least 400 mg/day. The results suggest that nefazodone given once/day at bedtime may be as effective as the currently accepted twice/day regimen, with less daytime drowsiness.
The Stroke Pharmacopeia: Current Medical Therapies
Authors: Wittkowsky Ann K.
Published: October 2023

Drug therapies that inhibit or reverse thrombus formation are important components of the management of acute ischemic stroke. The role of antiplatelet and anticoagulant therapies in stroke prevention has been defined, but further research is needed to confirm the possible benefits of aspirin, heparin, and low‐molecular‐weight heparin products in acute ischemic stroke. Recently, double‐blind, placebo‐controlled studies have evaluated the role of the thrombolytic agents streptokinase and tissue plasminogen activator (t‐PA) in patients with acute ischemic stroke. Intravenous t‐PA, administered within 3 hours of symptom onset at a dose of 0.9 mg/kg, is safe and effective in carefully selected patients.
The Stroke Pharmacopeia: Promising Experimental Therapies
Authors: Gengo Francis M.
Published: October 2023

The development of promising acute stroke treatments is the fruit of the growing appreciation for the complex biochemical processes within neuronal tissue that commence immediately after the onset of ischemia. These biochemical cascades can be modified either by direct pharmacologic mitigation or by rapid restoration of perfusion and oxygenation. With both interventions, the ischemic tissue can remain viable and regain neurologic function rather than progress to infarction. Today the two major pharmacologic approaches to stroke therapy are neuroprotectants and thrombolytics. Neuroprotectants enable neuronal tissues to tolerate periods of minimal perfusion better, whereas thrombolytics facilitate reperfusion by disrupting the fresh thrombus. Important classes of neuroprotectants include calcium channel antagonists, N‐methyl‐d‐aspartate (NMDA) receptor antagonists, benzothiazoles, and free radical scavengers. Pro‐urokinase is a potentially important investigational thrombolytic. Ancrod, a defibrinogenating agent, is also currently being evaluated in acute stroke.
Thrombolytic Therapy for Acute Myocardial Infarction
Authors: Meister Fred L. , Stringer Kathleen A. , Spinler Sarah A. , Gonzalez Edgar , Talbert Robert L. , Weiner Bruce , Guerrero Rawley M. , Smith Richard
Published: October 2023

Numerous factors must be considered when determining the formulary status of thrombolytic agents for the treatment of acute myocardial infarction. Defined treatment options, predicted outcomes, and the economic consequences of this disorder continue to evolve from clinical trials. Pharmacists have a major role in delivering patient care, with responsibility for evaluating, procuring, and monitoring thrombolytic agents and drug therapy in general. By participating in the development and implementation of treatment guidelines, evaluating economic and therapeutic outcomes, providing timely optimal drug therapy, and educating health care providers and the public, they contribute significantly to the health care team.
Cross‐Reactivity in HIV‐infected Patients Switched from Trimethoprim‐Sulfamethoxazole to Dapsone
Authors: Holtzer Christopher D. , Flaherty John F. , Coleman Rebecca L.
Published: October 2023

Study Objective. To evaluate the cross‐reactivity of dapsone after a documented hypersensitivity reaction to trimethoprim‐sulfamethoxazole (TMP‐SMX) during prophylaxis for Pneumocystis carinii pneumonia.

Design. Retrospective, chart review, cohort study.

Setting. Two university‐affiliated teaching hospitals.

Patients. Sixty patients infected with the human immunodeficiency virus.

Measurements and Main Results. Thirteen patients (21.7%) had cross‐reactivity to dapsone after the reaction to TMP‐SMX. No significant risk factors for this response were identified. Most reactions were of mild or moderate severity and rated as possibly or probably caused by one of the agents. Of the 13 patients, 4 (30.8%) continued therapy.

Conclusions. Although cross‐reactivity can occur, dapsone may be considered in patients with mild hypersensitivity reactions to TMP‐SMX.
Hyperkalemia Associated with Rapid Infusion of Conventional and Lipid Complex Formulations of Amphotericin B
Authors: Barcia John P.
Published: October 2023

A child developed hyperkalemia and cardiac arrest with infusion of an amphotericin B lipid complex 5.0 mg/kg over 1 hour. Another child, with chronic renal failure, developed hyperkalemia after infusion of conventional amphotericin B deoxycholate 1.0 mg/kg over 2 hours. Rapid infusion of the agent causes hyperkalemia in dogs and humans that is exacerbated in the setting of renal failure. A lipid formulation of amphotericin B is commercially available, and no reports of hyperkalemia are associated with its administration.
Levofloxacin, a Second‐Generation Fluoroquinolone
Authors: North Donald S. , Fish Douglas N. , Redington J. Jay
Published: October 2023

Levofloxacin, levo‐isomer of the D,L‐racemate ofloxacin, is a new fluoroquinolone antibiotic approved for use in the United States in December 1996. It has an extended spectrum of activity compared with older‐generation fluoroquinolones (ciprofloxacin, ofloxacin), with improved activity against gram‐positive bacteria and excellent activity against gram‐negative bacteria and atypical organisms. Although its activity against anaerobic organisms is improved over that of earlier fluoroquinolones, levofloxacin should not be considered a first‐line anaerobic agent. It is available in an injectable form, as well as an oral formulation with virtually 100% oral bioavailability. The plasma elimination half‐life ranges from 6–8 hours in individuals with normal renal function. Approximately 80% of drug is eliminated unchanged in urine through glomerular filtration and tubular secretion. The pharmacokinetics are not appreciably affected by age, gender, or race when differences in renal function and body mass and composition are taken into account. Levofloxacin had impressive efficacy in clinical studies of community‐acquired pneumonia, acute bacterial exacerbations of chronic bronchitis, acute sinusitis, skin and skin structure infections, and complicated urinary tract infections and pyelonephritis. It is well tolerated; its adverse event profile is similar to that of other fluoroquinolones, with gastrointestinal and central nervous system effects reported most commonly. Drug interactions are uncommon with levofloxacin; however, coadministration with antacids or with other agents containing divalent or trivalent cations reduces levofloxacin absorption. The agent should prove to be more effective than older fluoroquinolones, especially for infections caused by pneumococci highly resistant to penicillin.
Monoclonal Antibodies in the Treatment of Steroid‐Resistant Acute Graft‐versus‐Host Disease
Authors: Tse John C. , Moore Theodore B.
Published: October 2023

Acute graft‐versus‐host disease (GVHD) remains the major obstacle for successful allogeneic bone marrow transplantation (BMT). The frequency of grade II or higher acute GVHD ranges from 30–50% in human leukocyte antigen (HLA)‐matched sibling transplants and 50–80% in HLA‐matched unrelated transplants. The mortality and morbidity associated with this complication are substantial. Corticosteroid and polyclonal antibodies such as antithymocyte globulin (ATG) have had little success in treating the disease; however, advances have been made in hybridoma technology and understanding its immunopathophysiology. Based on these new insights, monoclonal antibodies, either murine or “humanized,” were tested as rescue treatment for acute GVHD in human trials. Complete response rates ranged from 20–40%, with relapse occurring often. Side effects consisted of constitutional symptoms such as fever, chills, hypotension, thrombocytopenia, and leukopenia. Limitations of monoclonal antibody treatment included low response rate and patient survival, high relapse rate, risk of infectious complication, and leukemic relapse. Future study should focus not only on improved side effects and efficacy of monclonal antibodies but also on better patient survival.
Does Aspirin Interfere with the Therapeutic Efficacy of Angiotensin‐converting Enzyme Inhibitors in Hypertension or Congestive Heart Failure?
Authors: Nawarskas James J. , Spinler Sarah A.
Published: October 2023

We conducted a MEDLINE search of published literature from 1966 to January 1998 regarding the impact of aspirin (ASA) on the therapeutic effect of angiotensin‐converting enzyme (ACE) inhibitors in hypertension and congestive heart failure. Selected references from these articles and results of recent clinical trials were also included. By inhibiting cyclooxygenase, ASA may interfere with the prostaglandin‐mediated hemodynamic effects of ACE inhibitors. Although other nonsteroidal antiinflammatory drugs may increase blood pressure in hypertensive patients taking an ACE inhibitor, low‐dosage (≤ 100 mg/day) ASA does not. However, higher dosages of ASA may attenuate the benefits of ACE inhibitors in patients with hypertension and/or congestive heart failure (CHF). Low‐dosage ASA appears to interact little with ACE inhibitors, whereas higher dosages may produce a more significant interaction. Patients with CHF may also be more susceptible to this interaction because of underlying disease.
Clinical Efficacy of Amiodarone
Authors: Nolan Paul E. , Nappi Jean , Pollak P. Timothy
Published: October 2023

Objectives. To review the clinical efficacy and role of amiodarone in the management of supraventricular and ventricular arrhythmias and its effects on mortality.

Methods. Review of relevant studies and reports.

Results. Amiodarone exerts significant effects on atrial tissue. In most studies it was completely or partly effective in preventing recurrences of atrial fibrillation or flutter in up to 80% of patients. Amiodarone may be superior to class Ia agents for maintaining normal sinus rhythm. Large randomized trials indicate that it is a potent suppressor of ventricular arrhythmia and reduces arrhythmic death after myocardial infarction. In patients with cardiomyopathy, it suppresses asymptomatic arrhythmias and increases left ventricular ejection fraction. Meta‐analysis of relevant studies indicated that amiodarone reduces the risk of arrhythmic and sudden death by 29% in high‐risk patients with recent myocardial infarction or congestive heart failure. This translates into an overall 13% reduction in total mortality.

Conclusion. Because of its effectiveness against a broad range of arrhythmias, amiodarone is a valuable addition to the antiarrhythmic pharmacopeia.
The Acid‐Fast Stain Is a Superior Stain for Use in the Mean Mature Spermatid Count for Testicular Biopsies
Authors: MAGID MARGRET S. , MA ZHI‐WEI , GIRARDI SARAH K. , GOLDSTEIN MARC
Published: October 2023

ABSTRACT: The mean mature spermatid count (MMSC) provides a useful, simplified quantitative evaluation of human spermatogenesis that is based on the number of mature spermatids in histological sections of testicular biopsies. Here, the activity of the acid‐fast (AF) stain was compared to that of the usual hematoxylin and eosin (H&E) stain in performing the MMSC. Thirty bilateral testicular biopsies showing normal spermatogenesis were chosen retrospectively from 15 subfertile patients with obstructive azoospermia or severe oligospermia. The MMSC was determined on each biopsy by utilizing both H&E and AF stains. The AF stain proved to be specific for the mature spermatids normally counted for the MMSC. It simplified recognition of mature spermatids, thereby shortening the overall time required for the procedure. The mean AF MMSC was lower than the mean H&E MMSC, and the mean interobserver differences were decreased. The AF stain is a superior stain for the MMSC when used in conjunction with the H&E stain for descriptive histology.
Hormonal Regulation of Spermatogenesis in the Hypophysectomized Rat: FSH Maintenance of Cellular Viability During Pubertal Spermatogenesis
Authors: RUSSELL LONNIE D. , KERSHAW MIKE , BORG KURT E. , SHENNAWY AHMED EL , RULLI SUSANA S. , GATES ROBERT J. , CALANDRA RICARDO S.
Published: October 2023

ABSTRACT: The potential for follicle‐stimulating hormone (FSH) to promote germ‐cell survival and the cellular sites of FSH action were studied using a gonadally maturing (pubertal), hypophysectomized (Hx) rat model in which residual testosterone (T) activity was blocked by injections of an androgen‐receptor antagonist, flutamide. Recombinant human FSH was given to androgen‐deprived and androgen‐blocked male rats at 27 days of age to determine maintenance of individual germ‐cell types at 35 days of age. Follicle‐stimulating hormone significantly increased testis weights and tubular diameters as compared with Hx and Hx‐flutamide controls, although testis weights in FSH‐treated animals were significantly lower than in pituitary‐intact animals. Morphometry assays to determine ratios of germ cells to Sertoli cells and to determine the number of germ cells pre

sent per hour of development showed that the population of type A spermatogonia in the early stages of the cycle was not responsive to FSH. Follicle‐stimulating hormone had a marked ability to maintain cell viability in the rapid, successive divisions that begin in the latter part of the cycle and that continue through the next cycle (i.e., from type A1 to A4 and from intermediate spermatogonia to type B spermatogonia to preleptotene spermatocytes to leptotene/zygotene spermatocytes to young pachytene spermatocytes). The data also suggest T responsiveness of these cell types since the Hx‐FSH‐flutamide group showed lower cell viability at the aforementioned steps when compared with the Hx‐FSH group. Too few cell types were present at subsequent phases of spermatogenesis to allow a sensitive determination of FSH activity in the maintenance of cell viability. The data show the potential of FSH in the absence or relative absence of T activity to maintain cell viability. These data support the concept of overlapping and synergistic (or additive) effects of T and FSH in the immature rat and identify the cellular sites of FSH action.
Expression of 140‐kDa Neural Cell Adhesion Molecule in Developing TestesIn Vivoand in Long‐Term Sertoli Cell—Gonocyte Cocultures
Authors: LI LING‐HONG , JESTER WILLIAM F. , ORTH JOANNE M.
Published: October 2023

ABSTRACT:The basis for cell—cell adhesion in the seminiferous epithelium of the developing testis is doubtless critical in supporting events that are essential for the onset and maintenance of normal spermatogenesis. In this study, we applied immunoblotting and immunolocalization approaches for the following reasons: 1) to ask whether neural cell adhesion molecule (NCAM) underlies cell—cell interactionsin vivo, as we previously showed for cellsin vitro, 2) to characterize the isoform or isoforms of NCAM expressed during testicular development, and 3) to study NCAM expression in long‐term Sertoli cell—gonocyte cocultures and to compare and contrast this pattern of expression with thatin vivo. Our findings indicate that NCAM is found ubiquitously at cell—cell interfaces within the semi‐niferous cord from birth through day 10 and thereafter is restricted to interstitial cells. Moreover, only polysialic acid—negative 140‐kDa NCAM is expressed in the testis or in coculture, an isoform whose properties are compatible with the concept of NCAM as both a direct modifier of cell function and an indirect influence on cell responses mediated by other external factors. In addition, we found that germ cells, potentially gonocytes or Type A spermatogonia, persist in long‐term cocultures maintained for 15 days after isolation from 5‐day‐old rat pups and that NCAM continues to be expressed at high levels in these cultures. This observation is in marked contrast to our observation that NCAM gradually decreases and eventually disappearsin vivoby postnatal day 15. Thus, our findings indicate that 140‐kDa NCAM is prominent in neonatal testes but is down‐regulated by as yet unidentified mechanisms thereafter. Our findings also indicate that down‐regulation of NCAM fails to occur in hormone‐ and serum‐free Sertoli cell—germ cell cocultures.
Androgen Concentrations and Their Receptors in the Periurethral Region Are Higher Than Those of the Subcapsular Zone in Benign Prostatic Hyperplasia (BPH)
Authors: MONTI SALVATORE , DI SILVERIO FRANCO , TOSCANO VINCENZO , MARTINI CHIARA , LANZARA STEFANIA , VARASANO PAOLA A. , SCIARRA FRANCESCO
Published: October 2023

ABSTRACT: Benign prostatic hyperplasia (BPH) is an androgen‐dependent disease that initially develops in the inner prostate, where the highest concentrations of testosterone (T) and dihydrotestosterone (DHT) are found. In this study, we have evaluated the cytosolic androgen receptors (ARc), the nuclear androgen receptors (ARn), and the concentrations of T, DHT, and 3α‐androstanediol (3αDiol) in BPH tissue to verify the existence of a possible correlation between androgens and their receptor concentrations. Prostatic samples, removed by suprapubic prostatectomy in 15 untreated patients, were sectioned in periurethral, intermediate, and subcapsular zones. Testosterone, DHT, and 3αDiol were evaluated by radioimmunoassay after extraction and purification on celite microcolumns, and ARc and ARn were evaluated by means of dextran‐coated charcoal method. In total tissue, mean levels of DHT, T, and 3αDiol were 2,531 ± 308, 260 ± 36, and 403 ± 35 pg/mg of DNA (mean ± SE), respectively. Cytosolic androgen receptors, detectable in all cases, were 16 ± 2.8 fmol/mg of protein (mean ± SE), and ARn, detectable in 12 cases, were 108 ± 15 fmol/mg of DNA (mean ± SE). A linear correlation between DHT and 3αDiol, T and DHT, and 3aDiol and ARn was found. If the different regions are considered, the periurethral zone, site of the primitive BPH nodule, presents the highest levels of androgens and ARn with respect to the other regions. This relative hyperandrogenism may be responsible for the growth‐promoting processes of this area, leading to urinary obstruction.
The Influence of Castration on Pharmacologically Induced Penile Erection in the Cat
Authors: BIVALACQUA TRINITY J. , RAJASEKARAN MAHADEVAN , CHAMPION HUNTER C. , WANG RUN , SIKKA SURESH C. , KADOWTTZ PHILIP J. , HELLSTROM WAYNE J. G.
Published: October 2023

ABSTRACT: The purpose of this study was to investigate the in vivo effects of intracavemosal injections of adrenomedullin (ADM), calcitonin gene‐related peptide (CGRP), nociceptin, vasoactive intestinal polypeptide (VIP), sodium nitroprusside (SNP), and prostaglandin E1 (PGE,) on penile erection in castrated and intact (control) anesthetized cats. Erectile responses to ADM, CGRP, nociceptin, VIP, SNP, and PGEi were compared with responses to a standard triple‐drug combination (1.65 mg of papaverine, 25 μg of phentolamine, and 0.5 μg of PGE1) in both castrated and control cats. In control animals, ADM, CGRP, nociceptin, VIP, SNP, and PGE, induced penile erections similar to those elicited by the triple‐drug combination. However, in castrated animals, there was a significant decrease in erectile response; the response to intracavemosal injection of the standard triple‐drug combination in castrated cats was 28% of that of the control group of animals. Serum testosterone levels demonstrated a significant (P < 0.0001) positive correlation (r= 0.52) with intracavemosal pressure in response to the standard combination. A marked reduction in serum testosterone levels was observed in castrated cats when measured by radioimmunoassay (0.34 ±0.1 ng/dl in castrated cats, compared with 31.15 ± 6 ng/dl in control cats). These data suggest that the presence of testosterone is a necessary prerequisite to sustain a pharmacologically induced penile erection in the cat.
Involvement of Reactive Oxygen Species in Human Sperm Arcosome Reaction Induced by A23187, Lysophosphatidylcholine, and Biological Fluid Ultrafiltrates
Authors: de LAMIRANDE EVE , TSAI CONYEE , HARAKAT AZIZ , GAGNON CLAUDE
Published: October 2023

ABSTRACT: Although recent evidence indicated that the production of reactive oxygen species (ROS) by human spermatozoa may be involved in the regulation of capacitation, very little is known about the role of ROS in the acrosome reaction. To address this issue, Percoll‐washed spermatozoa were incubated in Ham's F‐10 medium in the absence (no capacitation) or presence (capacitation) of fetal cord serum ultrafiltrate (FCSu) or progesterone. The effects of the ROS scavengers, superoxide dismutase (SOD), and catalase were then tested on the acrosome reaction induced by lysophosphatidylcholine (LPC), A23187, and ultrafiltrates from follicular fluid (FFu) and FCSu, as well as on the protein tyrosine phosphorylation associated with this process. 2‐Methyl‐6‐(p‐meth‐oxyphenyl)‐3,7‐dihydroimidazo [1,2‐a] pyrazin‐3‐one (MCLA)‐amplified chemiluminescence was used to determine the extracellular superoxide (02*‐) production from spermatozoa. The observations that both SOD and catalase reduced (in the case of LPC) or totally prevented (in the other cases) the acrosome reaction of capacitated spermatozoa and that hydrogen peroxide (H202) or ROS generated by the combination of xanthine and xanthine oxidase (02*‐, which dismutates to H202) triggered the acrosome reaction indicated the involvement of ROS in this process. In fact, capacitated spermatozoa in which the acrosome reaction was induced by LPC, A23187, and FFu produced more 02‐than noncapacitated spermatozoa treated with the same agents. A23187 and LPC had minor effects on protein tyrosine phosphorylation of noncapacitated spermatozoa. However, these inducers caused a decrease in tyrosine phosphorylation of Triton‐soluble proteins (mainly those of 37,42, and 47 kDa) from capacitated spermatozoa, a decrease more pronounced in the presence of SOD. On the other hand, there was a marked increase in tyrosine phosphorylation of few proteins (70 to 105 kDa) from the Triton‐insoluble fraction, which was partly reversed by SOD (in the case of LPC and A23187) or catalase (in the case of A23187), or abolished in the presence of the two antioxidants (in the case of A23187). These data indicate that the acrosome reaction is associated with an extracellular O2*‐generation by spermatozoa and that both O2‐ and H2O2 may be involved in the regulation of this process. The mechanism by which these ROS act is unknown but may involve tyrosine phosphorylation of sperm proteins.
Regulation of Sperm Function by Protein Tyrosine Phosphorylation in Diverse Wild Felid Species
Authors: PUKAZHENTHl BUDHAN S. , LONG JULIE A. , WILDT DAVID E. , OTTINGER MARY ANN , ARMSTRONG DOUGLAS L. , HOWARD JOGAYLE
Published: October 2023

ABSTRACT: Protein tyrosine phosphorylation is associated with sperm capacitation and the acrosome reaction in several mammalian species. Changes in phosphorylation of a 95‐kDa protein in human, mouse, and domestic cat spermatozoa are known to be influenced by capacitation and exposure to zona pellucida (ZP) proteins. We previously reported diminished phosphorylation of 95‐ and 160‐kDa proteins in spermatozoa from teratospermic cats, compared with normospermic domestic cats. To determine if these proteins and mechanisms are present in other species in the phenotypically diverse Felidae family, we examined the relationship between tyrosine‐phosphorylated sperm proteins and sperm morphology in the leopard cat (∼65% normal sperm/ejaculate), tiger (∼65%), clouded leopard (∼15%), and cheetah (∼30%). Furthermore, we investigated the involvement of cyclic adenosine monophosphate (cAMP) in the regulation of sperm protein tyrosine phosphorylation. Specifically, we assessed the following: 1) presence of tyrosine‐phosphorylated proteins in sperm extracts; 2) changes in protein tyrosine phosphorylation after sperm capacitation and swim‐up separation; 3) impact of tyrosine kinase inhibition on leopard cat sperm protein phosphorylation and ZP penetration; and 4) involvement of a cAMP‐dependent pathway in the regulation of protein tyrosine phosphorylation. Immunoblotting analysis with anti‐phosphotyrosine antibody (PY20) indicated that a 95‐kDa protein was present in all four species. Additional phosphorylated proteins were detected in the leopard cat (145‐ and 175‐kDa proteins), tiger (185‐kDa protein), clouded leopard (160‐ and 190‐kDa proteins), and cheetah (115‐ and 155‐kDa proteins). Sperm capacitation in vitro increased phosphorylation of one or more proteins in the leopard cat, tiger and clouded leopard, but not in the cheetah. Although swim‐up separation increased the proportion of morphologically normal spermatozoa in the clouded leopard and cheetah, no changes were observed in phosphorylation of the 95‐kDa sperm protein. Thus, phosphorylation of the 95‐kDa protein appeared to be related to the condition of teratospermia. Exposing leopard cat spermatozoa to the tyrosine kinase inhibitor, tyrphostin, reduced (P < 0.05) phosphorylation of the 95‐and 145‐kDa proteins, as well as ZP penetration, without affecting sperm motility. Similarly, when spermatozoa were incubated in the presence of cAMP analogs or active and inactive stereoisomers of cAMP, phosphorylation of sperm proteins was either stimulated or inhibited. Together, these data suggest that protein tyrosine kinase mechanisms appear conserved within the family Felidae and are regulated by a cAMP/protein kinase A pathway.
Analysis of Promoter and Androgen Regulatory Sequences Required for Optimal Transcription of the Rat Androgen‐Binding Protein Gene
Authors: FENSTERMACHER DAVID A. , JOSEPH DAVID R.
Published: October 2023

ABSTRACT: The androgen‐binding protein (ABP) gene P1 promoter directs cell‐specific gene regulation of ABP secreted by Sertoli cells. A recent study using the mouse Sertoli cell line (MSC‐1) with a luciferase reporter system demonstrated Sertoli cell—specific gene expression with 619 bp of P1 DNA. Furthermore, based on studies of the rat and human genes, several controversies developed over the promoter characteristics, including the promoter type, the transcription start site, and whether the gene is regulated directly by androgens. In this study, the answers to several of these controversies were deciphered using the MSC‐1 cell model. The results of mutagenesis experiments were consistent with the presence of the major transcription start site at 36 bp upstream of the initiating Met residue. Modification of the initiator sequence at the start site reduced activity in MSC‐1 and NIH3T3 fibroblast cells. Mutation of a putative modified TATA sequence or conversion to the consensus TATA sequence had no effect on activity. Modification of a consensus RNA splice sequence at the start site also had no effect on activity. Furthermore, a minor start site was localized 179 bp up‐stream of the major site using reverse transcriptase—polymerase chain reaction with various P1 primers (primer walking), primer extension, and cDNA cloning. RNA transcripts from the minor site contain an untranslated 5′ exon but apparently encode the same protein as the major transcript. The effect of androgens on P1 expression was also investigated. Cotransfection experiments with pCMVAR, which encodes the androgen receptor, demonstrated that dihydro‐testosterone had no effect on the activity in MSC‐1 cells. Taken together, these experiments and previous studies indicate that the rat ABP promoter P1 is regulated at the major start site by an initiator element without a TATA sequence, and the gene appears not to be directly regulated by follicle‐stimulating hormone or androgens.
Exposure of Human, Boar, or Bull Sperm to a Synthetic Peptide Increases Binding to an Egg‐Membrane Substrate
Authors: AMANN RUPERT P. , HAMMERSTEDT ROY H. , SHABANOWITZ ROBERT B.
Published: October 2023

ABSTRACT: We evaluated the effects of in vitro exposure of sperm to synthetic FertPlus® peptide, which represents a 60‐amino acid sequence within rat prosaposin, using a microwell sperm‐binding assay (SBA), in which an extract of hen's egg served as the binding substrate. Sperm suspensions were incubated with FertPlus® peptide (six to eight concentrations: 0 and 20–1,280 pM) at 37°C for 10 minutes, diluted ≥20 times, and placed onto SBA plates. After 60 minutes at 37°C for 10,bound sperm were washed away and the DNA of bound sperm was quantified. Percentage of sperm bound was independent of the percentage of motile sperm, but immotile sperm did not bind. For fresh human sperm (25 ejaculates), the percentage of sperm bound was increased by exposure to 640 pM peptide (P < 0.01). For 11 of 25 samples, the percentage of sperm bound for the aliquot exposed to 640 pM peptide was ≥ 1.4 times the value for a 0 pM control aliquot. With frozen‐thawed human sperm, for six of seven samples, binding was ≥ 1.4 times greater after exposure to 640 pM peptide. For boar sperm held for ∽24 hours at ∽18°Cbefore use (28 ejaculates), there was a higher percentage of sperm bound for aliquots previously exposed to 1,280 pM peptide than there was for control aliquots (P < 0.01). For 16 of 28 samples, exposure to peptide increased the percentage of sperm bound by ≥1.4 times. For frozen‐thawed bull sperm, percentage of sperm bound was ≥1.4 times greater for 4 of 10 samples that were briefly exposed to 160 pM peptide. Clearly, human, boar, and bull sperm were beneficially modified by brief in vitro exposure to FertPlus® peptide, so that for many samples a greater percentage of sperm was bound in vitro. As presented in an accompanying paper, fertility of bull sperm was increased by brief exposure to FertPlus® peptide.
Hormonal Regulation of Inhibin B Secretion by Immature Rat Sertoli Cells In Vitro: Possible Use as a Bioassay for Estrogen Detection
Authors: DEPUYDT CHRISTOPHE E. , MAHMOUD AHMED M. , DHOOGE WILLEM S. , SCHOONJANS FRANK A. , COMHAIRE FRANK H.
Published: October 2023

ABSTRACT: The influences of follicle‐stimulating hormone (FSH), gonadal steroids, and culture time were studied in relation to inhibin B production by Sertoli cells of immature rats cultured in vitro. Sertoli cell‐enriched cultures were established from 18‐day‐old rats and were maintained in medium supplemented with insulin, transferrin, and epidermal growth factor at 34°C.A recently developed ELISA for the measurement of inhibin B was used to assess the effects of recombinant human FSH (rh FSH), testosterone (T), and estradiol (E2) on inhibin B production and accumulation in the culture media of Sertoli cell‐enriched cultures and to optimize the cell culture system to serve as a bioassay for the detection and quantification of estrogens and estrogenlike substances. Prolonging the incubation time (24, 48, or 72 hours) of Sertoli cells with control medium without rh FSH, T, or E2 resulted in a time‐dependent increase of inhibin B production. Incubation with rh FSH (1, 2.5, 5, or 10 U/L) caused a dose‐ and time‐dependent increase of inhibin B production by Sertoli cells (but not by cultured Leydig cells), reaching a plateau at 5 U/L rh FSH. Addition of T in concentrations of 2.88, 5, or 50 ng/ml to medium without rh FSH and E2 significantly lowered the daily production rate of inhibin B (P < 0.05). In contrast, addition of E2 (0.01 and 0.1 ng/ml) caused a dose‐responsive increase in inhibin B production after 24 and 48 hours. The relative increment of inhibin B production induced by E2 was maximal after 24 hours in the presence of 2.5 U/L rh FSH (acting synergistically) and in the absence of T. When these conditions are implemented, the Sertoli cell culture system may serve as a bioassay for estrogenic substances, and it may reflect the possibly harmful effect they may have on spermatogenesis.
Fertility and Its Relationship to Motility Characteristics of Spermatozoa in Ewes After Cervical, Transcervical, and Intrauterine Insemination With Frozen‐Thawed Ram Semen
Authors: SÁNCHEZ‐PARTIDA L. GABRIEL , WINDSOR DAVID P. , EPPLESTON JEFF , SETCHELL BRIAN P. , MAXWELL W. M. CHISHOLM
Published: October 2023

ABSTRACT: The fertility of ewes after artificial insemination and the relationship between fertility and motility characteristics assessed by a computerized motility analysis system were examined with ram semen frozen in diluents reported to improve postthaw motility. The percentages of motile and progressive spermatozoa were better when frozen in proline—or glycine betaine‐containing or HEPES‐based, rather than Trisbased, diluents (P < 0.01). The fertility of spermatozoa frozen in diluents containing proline or glycine betaine was slightly reduced, whereas when both compatible solutes were present, the reduction was more pronounced, in comparison with semen frozen in Tris‐ or HEPES‐based diluents (9.5 versus 71.1 and 66.6%; P < 0.01). Fertility of frozen—thawed spermatozoa was higher after laparoscopic insemination than after cervical or transcervical insemination (P < 0.01). Similarly, higher fertility was obtained after cervical insemination with fresh than with frozen‐thawed semen (32.4 versus 11.3%; P < 0.01). Furthermore, loss of embryos was lower after laparoscopic insemination of ewes with semen frozen in a Tris diluent than with semen frozen in proline diluents, in glycine betaine diluents, or in proline‐plus‐glycine betaine diluents (0.0 versus 26.0, 38.5, and 60.0%; P < 0.001). A wide variation in the postthaw percentage of motile (31.6–59.7%) and progressive (22.6–43.1%) spermatozoa and in the fertility of spermatozoa from individual rams was also observed after laparoscopic (29.2–59.7%) or cervical insemination (8.7–30.5%). Postthaw motility results from immediately after thawing and fertility results from experiments where intrauterine insemination was performed with semen frozen in proline‐ or glycine betaine—containing or HEPES‐ or Tris‐based diluents were pooled and subjected to a pairwise correlation procedure. The correlation analysis showed relationships between some of the motility characteristics (P < 0.01), but there were no relationships between the motility characteristics and fertility.
Sex‐ and Age Group‐Specific Fracture Incidence Rates Trends for Type 1 and 2 Diabetes Mellitus
Authors: Nasser Mohamad I , Kvist Annika Vestergaard , Vestergaard Peter , Eastell Richard , Burden Andrea M , Frost Morten
Published: October 2023

The incidence of major osteoporotic fractures has declined in men and women in Western countries over the last two decades. Although fracture risk is higher in persons with diabetes mellitus, trends of fractures remain unknown in men and women with diabetes. We investigated the trends in fracture incidence rates (IRs) in men and women with type 1 diabetes mellitus (T1D) and type 2 diabetes mellitus (T2D) in Denmark between 1997 and 2017. We identified men and women aged 18+ years who sustained a fracture (excluding skull and facial fractures) between 1997 and 2017 using the Danish National Patient Registry. We calculated sex‐specific IRs of fractures per 10,000 person‐years separately in persons with T1D, T2D, or without diabetes. Furthermore, we compared median IRs of the first 5 years (1997–2002) to the median IRs of the last 5 years (2012–2017). We identified 1,235,628 persons with fractures including 4863 (43.6% women) with T1D, 65,366 (57.5% women) with T2D, and 1,165,399 (54.1% women) without diabetes. The median IRs of fractures declined 20.2%, 19.9%, and 7.8% in men with T1D, T2D, and without diabetes, respectively (p‐trend <0.05). The median IRs decreased 6.4% in women with T1D (p‐trend = 0.35) and 25.6% in women with T2D (p‐trend <0.05) but increased 2.3% in women without diabetes (p‐trend = 0.08). Fracture IRs decreased in men with both diabetes types and only in women with T2D, highlighting the need for further attention behind the stable trend observed in women with T1D. © 2023 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.