bluuebunny/crossref_metadata_2025 huggingface dataset:
Budesonide Inhalation Powder: A Review of Its Pharmacologic Properties and Role in the Treatment of Asthma
Authors: Davis Kristin Coveny , Small Ralph E.
Published: October 2023

Budesonide inhalation powder, available as Pulmicort Turbuhaler, is a corticosteroid with a high ratio of local to systemic effects that is administered to treat persistent asthma. The Turbuhaler achieves lung deposition approximately twice that of a metered‐dose inhaler (MDI) with or without a spacer device. Budesonide inhalation powder has clinical efficacy equivalent to that of fluticasone and beclomethasone, but it has lower systemic bioavailability and fewer systemic side effects. As with other inhaled corticosteroids, dysphonia and oral candidiasis are the most frequent adverse effects, and systemic effects are infrequent. The initial starting dosage is 200 μg (1 puff) twice/day and may be increased to 800 μg twice/day in adults or 400 μg twice/day in children. Patients prefer the Turbuhaler to the MDI, Diskhaler, and Rotahaler because it is easier to use and more convenient to carry.
The Pharmacology and Therapeutic Utility of Bisphosphonates
Authors: Lourwood David L.
Published: October 2023

Bisphosphonates have clinical application in diseases associated with increased bone turnover that inhibit osteoclast‐mediated bone resorption by direct and indirect actions on osteoblasts and macrophages. Bisphosphonates are the treatment of choice for Paget's disease of bone, in which they return to normal the increased rate of bone turnover and slow radiographic disease progression. The agents reduce hypercalcemia associated with malignancy, and may reduce bone pain and prevent radiographic progression of metastatic bone disease. In patients with postmenopausal osteoporosis, they prevent further bone loss and reduce fracture rates. The drugs also ameliorate osteoporosis associated with long‐term corticosteroid treatment. Bisphosphonates are well tolerated; gastrointestinal disturbances are the most common adverse events. Potential bone mineralization defects that occur with first‐generation bisphosphonates are not of concern with therapeutic doses of newer ones.
Sedation in Children: Current Concepts
Authors: Bhatt‐Mehta Varsha , Rosen David A.
Published: October 2023

Sedation in children poses a great challenge, with the main concern one of safety. The importance of providing adequate sedation to children was realized only in the last decade and a half, and relevant data are severely lacking. Use of potent sedative agents is not without risk. Children are given sedative agents in a wide variety of settings by practitioners with different degrees of experience with the drugs and management of adverse effects. Controversial issues must be addressed in this area, and appropriate tools developed to measure sedation and individualize treatment based on the drugs' pharmacokinetic and pharmacodynamic properties.
Effect of Plasma TNF‐α on Filgrastim‐Stimulated Hematopoiesis in Mice and Humans
Authors: Petros William P. , Rabinowitz Josh , Gibbs John P. , Hall Iris H. , Stuart Ann R. , Peters William P.
Published: October 2023

Study Objective. To delineate possible explanations for a nonmonotone hematopoiesis, dose‐response curve with filgrastim therapy after high‐dose chemotherapy.

Design. Sequential two‐phase study.

Settings. University teaching hospital and basic pharmaceutical sciences laboratory.

Subjects. Thirty‐nine patients with breast cancer or melanoma and 15 normal CF‐1 male mice.

Interventions. Serial blood samples were obtained from patients after high‐dose chemotherapy to evaluate hematopoiesis and tumor necrosis factor‐α (TNF‐α) concentrations. Murine hematopoiesis was induced by filgrastim with or without coadministration of lipopolysaccharide.

Measurements and Main Results. Detection of plasma TNF‐α in patients corresponded to substantially slower recovery of granulocytes, erythrocytes, and platelets, and was directly proportional to the prescribed dosage of filgrastim. Lipopolysaccharide stimulated the secretion of TNF‐α in mice and totally aberrated filgrastim‐induced granulopoiesis.

Conclusions. This in vivo evidence suggests that regulatory pathways involving endogenous cytokines may override the effect of recombinant cytokines.
Angioedema and Photosensitive Rash Induced by Valsartan
Authors: Frye Carla B. , Pettigrew Timothy J.
Published: October 2023

A 71‐year‐old woman experienced an acute onset of angioedema and a photosensitive pruritic rash after 3 months of therapy with valsartan, an angiotensin II receptor antagonist. After discontinuing valsartan and treatment with subcutaneous epinephrine, intravenous methylprednisolone, diphenhydramine, and Eucerin cream, her symptoms dissipated and the rash resolved. Rash and angioedema have been reported with angiotensin‐converting enzyme inhibitors and, very rarely, with losartan, the other currently marketed angiotensin II receptor antagonist. To our knowledge, this is the first report of such a reaction with valsartan.
Pharmacotherapy of Primary Obsessive‐Compulsive Disorder: Review of the Literature
Authors: Ellingrod Vicki L.
Published: October 2023

Obsessive‐compulsive disorder (OCD) is a chronic illness that affects 2–3% of Americans during their lifetime. It is characterized by recurrent obtrusive thoughts (obsessions) that compel patients to perform repetitive behaviors that can be excessively time consuming and cause marked distress. More than 40 controlled trials have been published on the treatment of OCD. Of drugs available to treat the disorder, serotonin reuptake inhibitors (SRIs) are most studied. With SRIs, symptoms improve in 22–62% of patients, but complete remission is rare. An agent is often selected based on side effects and patient tolerance, since SRIs are all equally effective. If no response is seen with average dosages, dosages should be increased to the maximum within 4–8 weeks from starting treatment. In patients with partial response, the dosage should be increased to the maximum by 5–9 weeks. Before determining the effectiveness of therapy, a trial of 8–13 weeks is necessary.
Recombinant Follicle‐Stimulating Hormone: New Biotechnology for Infertility
Authors: Prevost Rebecca R.
Published: October 2023

The frequency of infertility in developed countries is approximately 8–10%. New drugs are available for assisted reproduction techniques. Two recombinant follicle‐stimulating hormone (FSH) products, follitropin‐β (Follistim in the United States, Puregon in Europe) and follitropin‐α (Gonal‐F), join compounds derived through transfecting nonhuman cell lines with genetic material capable of replicating identical amino acid sequences to human compounds. The cell line used for recombinant (r)‐FSH production is the Chinese hamster ovary (CHO). Previously, the only agents that showed benefit in controlled ovulatory stimulation were derived from the urine of menopausal women. Those compounds contain additional substances, such as urinary proteins and various amounts of luteininzing hormone. The amino acid sequence of r‐FSH is identical to that of human FSH, but the two recombinant products exist in many different isoforms and differ from each other and from human FSH due to varied carbohydrate side chains. Due to variation in the carbohydrate side chains, follitropin‐β in solution has a higher pH than urine‐derived FSH, which enhances receptor affinity and therefore is a greater inducer of folliculogenesis. Follitropin‐β does not cause endogenous production of anti‐CHO or anti‐FSH antibodies, and is well tolerated.
α1‐Acid Glycoprotein Concentrations and Cerebrospinal Fluid Drug Distribution after Subarachnoid Hemorrhage
Authors: Woodward Donald K. , Hatton Jimmi , Ensom Mary H. H. , Young Byron , Dempsey Robert , Clifton G. Dennis
Published: October 2023

Study Objective. To test the hypothesis that changes in α1‐acid glycoprotein (AAG) concentration alter central nervous system (CNS) drug distribution after subarachnoid hemorrhage.

Design. Two‐phase, prospective study.

Setting. University‐associated medical center.

Patients. Twenty‐one patients with subarachnoid hemorrhage.

Intervention. In phase I, serum AAG concentrations of patients with subarachnoid hemorrhage were measured serially and compared with those in 21 controls undergoing elective neurosurgical procedures. In phase II, nimodipine was the pharmacologic probe to determine the relationship between drug distribution into the CNS and changes in AAG concentration.

Measurements and Main Results. Serum and cerebrospinal fluid (CSF) samples were collected from patients with subarachnoid hemorrhage treated with nimodipine and used to measure total and unbound drug concentrations. Concentrations of AAG were 39% higher in patients than in controls preoperatively. They decreased significantly by 24 hours after surgery in patients and increased in controls. In both groups the concentrations were higher than reported normal values. During the period of reduced AAG concentration, calculated unbound nimodipine concentrations were 3‐fold higher (p<0.05) than at later periods, with a trend toward higher total concentrations. Overall, mean CSF nimodipine concentration was 6.4% of mean serum total concentration. The CSF concentrations decreased as AAG concentrations increased, independent of serum concentrations (r = −0.52, p<0.02).

Conclusion. Concentrations of AAG change after subarachnoid hemorrhage and are transiently influenced by surgery. Unbound drug concentration increases when AAG concentrations decrease, whereas CSF concentrations decrease when AAG concentrations increase. These preliminary findings suggest that changes in AAG concentrations can alter unbound serum nimodipine concentrations and may affect CSF drug distribution.
Evaluation of a Rat Model of Valproate‐Induced Obesity
Authors: Wolden‐Hanson Tami , Gidal Barry E. , Atkinson Richard L.
Published: October 2023

Long‐term treatment with the anticonvulsant valproate (VPA) leads to well‐documented weight gain and obesity in humans. In an attempt to develop an animal model of this condition, adult rats were given VPA 20 g/kg (high‐dose) or 2 g/kg (low‐dose) in their daily feeding or orally 120 mg/kg body weight/day in two divided doses, and food intake and body weight were assessed. Valproate resulted in lower body weights in all protocols. Food intake was lower (p<0.001) for rats receiving high‐dose VPA than for controls. Feed efficiency (change in weight divided by cumulative food intake for that period) was lower than that of controls for both high (p<0.0001) and low doses (NS). Metabolic rate and physical activity were not different between control and VPA animals, although decreased food intake would be expected to decrease metabolic rate. Valproate failed to produce obesity in rats in any treatment period. For reasons that are unclear, rats do not appear to be suitable as a model to study this adverse side effect of VPA in humans with epilepsy.
Frequency of Hospitalization after Exposure to Known Drug‐Drug Interactions in a Medicaid Population
Authors: Hamilton Robert A. , Briceland Laurie L. , Andritz Mary H.
Published: October 2023

A matched‐pair case‐control analysis of Medicaid claims was performed to determine the risks of hospitalization associated with drug‐drug interactions. Patients were hospitalized and controls were not. They were randomly matched based on contemporaneous eligibility for Medicaid benefits. Odds ratios for hospitalization in patients exposed to known drug‐drug interactions were compared with those in patients exposed to one of the interacting agents. When confidence intervals did not overlap, the odds ratio was considered to be significantly increased. Odds ratios were significantly increased for many interacting drug pairs, and were associated with commonly recognized interactions as well as less widely recognized ones. Cimetidine interactions achieved significance only with theophylline. In the Medicaid population, exposure to a number of drug‐drug interactions was associated with a significantly increased risk of hospitalization.
Introduction
Authors: Bauman Jerry L.
Published: October 2023

Since the beginning of this century cardiovascular disease has been the leading cause of death in the United States in every year except one. According to current estimates, almost 60 million Americans have one or more types of cardiovascular disease (Figure 1). Although death rates have declined, cardiovascular disease still claims more lives each year than the next seven leading causes of death combined (Figures 2A and 2B). In 1995, for example, they caused the death of 960,592 Americans, about 41% of all deaths in the United States. To put it another way, more than 2600 Americans die each day of the disease, an average of 1 every 33 seconds. The public health impact is reflected in recent data from the National Center for Health Statistics, which indicate that if all major forms of the disease were eliminated, life expectancy would rise by almost 10 years.
Optimal Management of Amiodarone Therapy: Efficacy and Side Effects
Authors: Hilleman Daniel , Miller Mary Ann , Parker Robert , Doering Paul , Pieper John A.
Published: October 2023

Objectives. To review management and dosing guidelines for amiodarone therapy, and discuss the drug's adverse event profile.

Methods. Review of relevant studies and reports.

Results. Amiodarone is a highly effective antiarrhythmic drug, but is associated with adverse effects involving several organs. Amiodarone‐induced arrhythmia is rare, with frequency of 0.3% in one study. Pulmonary toxicity is the most serious noncardiac side effect (2–17% of patients). Hypersensitivity pneumonitis can appear early in the course of therapy. Interstitial pneumonitis is a more common but insidious pulmonary reaction characterized by cough, low‐grade fever, and dyspnea that occurs after months or years of therapy. Clinically important hypothyroidism and hyperthyroidism occur in 2–10% of patients. Optic neuritis or neuropathy in which patients experience decreased or blurred vision may progress to permanent blindness. Abnormalities in liver function tests, especially elevated aminotransferase and alkaline phosphatase levels, are seen in 4–25% of patients. Neurologic side effects were reported in 20–40% of patients, at times associated with tremor, ataxia, peripheral neuropathy, malaise or fatigue, sleep disturbances, dizziness, and headaches. Several types of dermatologic reactions have been reported, including allergic rash, photosensitivity, and blue‐gray skin discoloration. The best strategy for early detection of pulmonary toxicity is vigilant clinical follow‐up with monitoring of cardiac status and liver and thyroid function, and prescription of the lowest effective dosage. After an initial loading dose, 200 mg/day in many patients maintains arrhythmia control and minimizes the frequency of side effects.

Conclusion. Amiodarone is a safe and efficacious antiarrhythmic agent when lower dosages are given to patients who are closely monitored and subject to careful follow‐up.
A Flow Cytometric Technique Using Peanut Agglutinin for Evaluating Acrosomal Loss From Human Spermatozoa
Authors: COOPER TREVOR G. , YEUNG CHING‐HEI
Published: October 2023

ABSTRACT: Flow cytometric methods for the quantification of acrosome‐reacted ejaculated human spermatozoa are described in which fluorescence‐labeled peanut agglutinin (Arachis hypogaea) binding to the outer acrosomal membrane is used after incubation with ethidium homodimer (EHD) as vital dye and membrane permeation using methanol. Fluorescein‐labeled fucoidan (F‐fucoidin) was shown to bind to sperm that were also stained by propidium iodide (PI) and EHD; therefore, the use of F‐fucoidin as vital dye was incorporated into the study. F‐fucoidin and EHD can withstand the acrosome‐staining procedure, unlike PI, which is leached out during cell processing. During incubation of spermatozoa, nonviable cells, as indicated by PI or F‐fucoidin staining, appear in parallel with acrosome‐reacted cells regardless of whether they are induced by the calcium ionophore A23187. The window of time in which viable, acrosome‐reacted sperm cells can be detected is narrow and could depend on the vital dye as well as the acrosomal marker used.
Two‐Day IUI Treatment Cycles Are More Successful Than One‐Day IUI Cycles When Using Frozen‐Thawed Donor Sperm
Authors: MATILSKY MOSHE , GESLEVICH YOEL , BEN‐AMI MOSHE , BEN‐SHLOMO IZHAR , WEINER‐MEGNAGI TZOFNAT , SHALEV ELIEZER
Published: October 2023

ABSTRACT: The difference in pregnancy rates following intrauterine insemination (IUI) for 1 vs. 2 days in the periovulatory period has been reported as either inconsequential or favoring the use of two consecutive inseminations, 24 hours apart. Our study compared the monthly fecundity and cumulative probability of pregnancy in a large group of women (n = 123) undergoing controlled ovarian hyperstimulation and 1‐ or 2‐day inseminations with donor sperm prepared from frozen‐thawed samples. All patients underwent controlled ovarian hyperstimulation employing either clomiphene citrate in 217 cycles or human menopausal gonadotropin in 185 cycles. The choice of single or double insemination was decided by the day of the week each patient received human chorionic gonadotropin for ovulation induction. Approximately 80% of all the patients underwent both single and double insemination treatments during the 2.5‐year study period. Ninety‐three patients received single inseminations in 180 cycles, whereas 103 patients received double inseminations in 222 cycles. Nine clinical pregnancies were achieved in the 1‐day group (5% per cycle, 9.7% per patient), while 39 pregnancies occurred in the 2‐day group (17.9% per cycle, 37.9% per patient). Two and five spontaneous abortions occurred in the 1‐ and 2‐day groups, yielding take‐home baby rates of 3.9% per cycle (7.5% per patient) and 15.3% per cycle (33.0% per patient), respectively. The cumulative probability of conception over 15 cycles of treatment was consistently twice as high or higher for the 2‐day group. The results of this study support the use of 2‐day IUI treatment cycles when using frozen‐thawed donor sperm.
Effects of Levodopa on Nocturnal Penile Tumescence: A Preliminary Study
Authors: HORTTA HIROKI , SATO YOSHIKAZU , ADACHI HIDEKI , SUZUKI NORIYOSHI , KATO RYUICHI , HISASUE SHINICHI , SUZUKI KAZUHIRO , TSUKAMOTO TAIJI
Published: October 2023

ABSTRACT: We studied the effects of levodopa (L‐dopa), which is reported to increase the dopamine level in the brain, on male erectile function. Of the 21 subjects studied, 12 subjects who were 50 years old or older showed significant increases of two nocturnal penile tumescence (NPT) parameters, NPT frequency and total tumescence time, with L‐dopa. On the other hand, in nine subjects who were younger than 50 years, maximum penile circumference increase showed a significant increment with L‐dopa. This significant increment of NPT with L‐dopa was not observed in the subjects who had low androgen levels. The results of this preliminary study show a positive relationship between administration of L‐dopa and erectile function. L‐Dopa administration may improve erectile function in subjects aged 50 years and older who have normal androgen levels.
The Use of Two Fluorescent Dyes to Identify Sperm in a Competitive Binding Assay to Oocytes
Authors: MILLER DAVID J. , DEMERS JILL M. , BRAUNDMEEER ANDREA G. , BEHRENS MELISSA L.
Published: October 2023

ABSTRACT: The relationship of most sperm laboratory assays to male fertility is inconsistent. Assays that measure traits required to fertilize oocytes are expected to have the most predictive value. A new assay that measures the competitive ability of two sperm samples to bind to oocytes was developed. Two populations of sperm were labeled using a pair of lipophilic dyes. A concentration of 75 μM of the two dyes, DiQ (4‐[4‐(dihexadecylamino)styryl]‐N‐methylquinolinium iodide; an orangered dye) and DiOC16 (3,3′‐dihexadecyloxacar‐bocyanine perchlorate; a yellow‐green dye), intensely stained 66 and 73% of sperm, respectively, without affecting sperm motility or oocyte‐binding ability. Because sperm could be stained with fluorescent dyes, sperm from two semen samples were mixed together in a droplet, and oocytes were added to allow sperm to bind oocytes competitively. Oocytebound sperm from each sample were counted. Binding was specific; nonspecific sperm binding was estimated by sperm bound to two‐cell mouse embryos and averaged one to three sperm per embryo. Staining with DiQ or DiOC16 did not affect oocyte‐binding ability since more than 80% of the sperm bound were stained with either dye. Furthermore, if different ratios of DiQ‐ or DiOC16‐stained sperm from the same ejaculate were prepared in droplets and oocytes were added, the percentage of sperm bound to the oocytes reflected the percentage of sperm in the droplet; there was no differential effect of either dye. This assay used fixed oocytes because sperm bound equally to fixed and fresh bovine oocytes. This competitive oocyte‐binding assay allows one to make a series of pairwise comparisons between a group of males or to include an internal control sample in spermoocyte binding assays. This assay may allow more accurate prediction of the oocyte‐binding ability of sperm.
Evidence for Cross‐Talk Between Sertoli and Germ Cells Using Selected Cathepsins as Markers
Authors: CHUNG SANNY S. W. , ZHU LI‐JI , MO MENG‐YUN , SILVESTRINI BRUNO , LEE WILL M. , CHENG C. YAN
Published: October 2023

ABSTRACT:To examine whether proteases are possibly involved in cellular migration and/or spermiation when developing germ cells translocate across the seminiferous epithelium during spermatogenesis,in situhybridization was used to localize messenger RNA (mRNA) transcripts of cathepsin L, D, and S in the epithelium at different stages of the spermatogenic cycle in the rat. Cathepsin L mRNA was found to localize almost exclusively near the basal lamina of the epithelium. At stages VI and VII of the cycle before spermiation, the signal of cathepsin L mRNA was so intense that it formed a complete dark precipitate near the basal lamina encircling the entire tubule. At stage VIII, the expression of cathepsin L was completely abolished, and no staining of cathepsin L mRNA was seen in the epithelium. The mRNA of cathepsin D and S was found near the basal lamina, a finding consistent with their localization in Sertoli cells and possibly primary spermatocytes in almost all stages, but peaked at stages VII‐IX and VII‐VIII of the cycle, respectively, at the time before and during spermiation. These results illustrate the possible involvement of these proteases in facilitating germ cell movement and spermiation. To examine whether germ cells express any of these cathepsin genes, spermatocytes with a purity of greater I than 95% were isolated from 15‐day‐old rat testes by Percoll gradient centrifugation for reverse transcriptase—polymerase chain reaction. It was found that primary spermatocytes expressed multiple cathepsin genes, including cathepsin B, C, D, H, L, and S. Furthermore, the expression of cathepsin L by germ cells isolated from 15‐day‐old rats (largely spermatocytes and spermatogonia) can be stimulated by Sertoli cell—enriched culture medium in a dose‐dependent manner, but not by germ cell—conditioned medium. These results reveal that germ cell function can be regulated by Sertoli cells. Moreover, these results suggest that germ cells may play an active role in the overall testicular protease expression. Also, we present evidence suggesting there is cross‐talk between Sertoli and germ cells, since the expression of cathepsin L in each cell type is regulated by one another via either soluble factors or cell‐cell contact.
Quantification of Apoptotic Testicular Germ Cells in Normal and Methoxyacetic Acid‐Treated Mice as Determined by Flow Cytometry
Authors: KRISHNAMURTHY HANUMANTHAPPA , WEINBAUER GERHARD E , ASLAM HUMAIRA , YEUNG CHING‐HEI , NIESCHLAG EBERHARD
Published: October 2023

ABSTRACT: Several studies have reported the occurrence and significance of programmed ceil death (apoptosis) of testicular germ cells in mammals. In those studies, apoptotic germ cells were identified by morphological criteria or by in situ end labeling (TUNEL) and were enumerated from histological sections by semi‐quantitative and time‐consuming techniques. In the present study, we have established a flow cytometric technique for quantification of TUNEL‐positive cells in the mouse testis. Groups of five adult mice each received 0, 650, or 1300 mg/kg (IP) of methoxyacetic acid (MAA), and testes were collected 24 hours later. MAA is known to induce germ cell apoptosis in rodent testes. MAA induced a significant (P < 0.01) dose‐dependent decline in the percentage of pachytene spermatocytes (4C cells). DNA strand breaks generated by the activation of endogenous endonuclease in the apoptotic germ cells were detected by the in situ labeling of the 3′‐OH termini with biotinylated dUTP in the presence of terminal deoxynucleotidyl transferase (TUNEL technique). Histologically, TUNEL‐positive germ cells were observed in control testes, and the number of these cells was visibly increased following MAA exposure. As determined by flow cytometry, four cell populations contained TUNEL‐positive cells: 1C cells (round spermatids), 2C cells (mainly spermatogonia), S‐ph cells (spermatogonia! cells and preleptotene spermatocytes synthesizing DNA [the S‐phase]), and 4C cells (primary spermatocytes). Analysis of the percentages of TUNEL‐positive cells within each population yielded values of 1.57 ± 0.23% for 1C cells, 1.65 ± 0.27% for 2C cells, 6.26 ± 1.03% for S‐ph cells, and 3.24 ± 0.39% for 4C cells. Hence, a substantial proportion of proliferating cells are undergoing apoptosis during normal spermatogenesis. The overall incidence of apoptotic cells among all testicular cells was around 2%. At 650 mg per kilogram of body weight, MAA induced a fourfold to eightfold increase (P < 0.001) in the percentage of TUNEL‐positive cells, compared with saline‐treated controls, and, overall, 17% of testicular cells were apoptotic. This effect of MAA was most pronounced for S‐ph and 4C cells, with 25–30% of cells being affected in each of those populations. At 1300 mg per kilogram of body weight, MAA had no further effect. These quantitative data demonstrate that 1) in the normal testis, it is mainly proliferating cells that undergo apoptosis, and 2) MAA induces primary spermatocyte loss by germ cell apoptosis.
The Primary Culture of Rat Prostate Basal Cells
Authors: ILIO KENNETH Y. , NEMETH JEFFREY A. , LANG SHARON , LEE CHUNG
Published: October 2023

ABSTRACT: The objective of this study was to identify the cells from rat prostate epithelium that attach and proliferate in primary culture. Minced ventral prostate was dissociated by DNAse/collagenase digestion, suspended in RPMI‐1640 containing 10% fetal bovine serum, and subjected to Percoll centrifugation to separate the epithelial cells from stromal cells. With the use of lectins, it became possible to identify and monitor the fate of the dissociated epithelial cells held in suspension at 37°C for several hours. In tissue sections of the rat prostate, Griffonia simplicifolia 1‐isolectin B4 (GSI‐B4) specifically bound to basal cells, while Glycine maximus (soybean agglutinin [SBA]) was specific for secretory cells. Double staining with lectins and propidium iodide of dissociated cells revealed a preponderance of GSI‐B4‐positive live cells. The cells were plated in WAJC‐404 medium supplemented with various factors, including insulin (5 ng/ml), transferrin (5 ng/ml), EGF (10 ng/ml), and bovine pituitary extract (30 tig/ml). Epithelial colonies that formed and proliferated from these cells also stained positively for GSI‐B4 marker and for cytokeratins specific for basal cells as assessed by immunocytochemical staining. Proliferation was greater in cells grown on a collagen Type I matrix. These findings suggest that the epithelial cells that survived in suspension and proliferated in culture originated from basal cells of the rat prostate epithelium.
Follicle‐Stimulating Hormone Is Required for the Initial Phase of Spermatogenic Restoration in Adult Rats Following Gonadotropin Suppression
Authors: MEACHEM SARAH J. , WREFORD NIGEL G. , STANTON PETER G. , ROBERTSON DAVID M. , McLACHLAN ROBERT I.
Published: October 2023

ABSTRACT: The role of follicle‐stimulating hormone (FSH) in adult rat spermatogenesis is unclear. Although exogenous testosterone (T) restores spermatogenesis following gonadotropin‐releasing hormone (GnRH) immunization or T plus estradiol (TE) treatments, an assessment of the independent action of T and FSH was not possible, as exogenous T treatment maintains serum FSH levels. We have used passive immunization against FSH to determine whether T alone is capable of reinitiating spermatogenesis after chronic and acute FSH withdrawal. Adult rats received T‐filled Silastic implants 6 cm (T6) or 8 cm (T24) in length for 7 days in combination with either a polyclonal sheep antisera raised against rat FSH (FSHAb, 2 mg/kg SC daily) or control sheep immunoglobulin (ConAb) after either GnRH immunization (12 weeks) or TE treatment (9 weeks). The neutralizing capacity of the FSHAb was determined using a FSH in vitro bioassay; this analysis demonstrated that administration of FSHAb in vivo reduced FSH levels by >90%. Testes were fixed and germ cell number per testis quantified using the optical dissector. GnRH immunization reduced spermatogonia, pachytene spermatocytes, and round spermatids to 50,13, and <1% of normal, respectively. T6 and T24 Silastic implants with the inclusion of the FSHAb did not increase the number of spermatogonia, pachytene spermatocytes, and round spermatids (50, 15, and 1% of normal, respectively). T6+ConAb treatment increased spermatogonial, pachytene spermatocyte, and round spermatid numbers to 74, 30, and 3% of normal, respectively (P < 0.05). No further increases were seen with T24 implants. TE treatment suppressed pachytene spermatocytes and round spermatids to 33 and 1% of normal, respectively (P < 0.05). T6+FSHAb treatment did not increase the number of pachytene spermatocytes and round spermatids (36 and 8%, respectively), whereas T6+ConAb treatment increased pachytene spermatocyte and round spermatid number to 50 and 28% of normal, respectively (P < 0.05). T24+FSHAb treatment increased the number of pachytene spermatocyte and round spermatids (56 and 22% of normal, respectively; P < 0.05), whereas T24+ConAb treatment increased these cells forms to 79 and 31% of normal, respectively. In conclusion, T alone is unable to restore spermatogenic cell populations in the setting of chronic FSH withdrawal. Although acute FSH withdrawal markedly impairs the restoration process, higher doses of T can partially compensate for the lack of FSH. These data suggest that FSH is important for the initial phase of spermatogenic restoration.
Protein Kinase CK2 Activities in Human Prostatic and Seminal‐Vesicle Secretions and Seminal Plasma
Authors: WILSON MICHAEL J. , DAVIS ALAN , ERCOLE CESAR , PRYOR JON L. , HENSLEIGH HUGH , KAYE KEITH W. , DAWKINS HUGH J. S. , WASSERMAN NEIL F. , REDDY PRATAP , AHMED KHALIL
Published: October 2023

ABSTRACT: Human prostatic secretion and seminal plasma contain certain protein kinase activities. Protein kinases play important roles in regulating a vast variety of cellular functions. The objective of this study was to determine whether one of these protein kinase activities in human prostatic secretion and seminal plasma is due to CK2, a messenger‐independent, serine/threonine protein kinase that has considerable potential as a regulatory enzyme. By employing an anti‐CK2 antibody and a CK2‐specific peptide substrate, we have established that CK2 is present in these secretions. Approximately 70% of the CK2 activity present in seminal plasma of normozoospermic men (n = 49) is correlated to the number of sperm originally present in the semen. Further, both the prostate gland and the seminal vesicles are sources of CK2 activity in the seminal plasma of vasectomized men (n = 38). Although there was considerable variation between individuals in CK2 activity, the variation in repeat semen samples of the same vasectomized men (n = 6) was within 21%. There was no correlation of CK2 activity in seminal plasma with age for vasectomized (27–48 years, n = 38), oligozoospermic (28–43 years, n = 24), or normozoospermic men (26–48 years, n = 49). These data suggest that the majority of CK2 activity in the seminal plasma of normozoospermic men originates from sperm but that the prostate and seminal vesicles are accessory sex‐gland sources of this enzyme.
Androgen Regulation of thePemHomeodomain Gene in Mice and Rat Sertoli and Epididymal Cells
Authors: SUTTON KEITH A. , MATH SOURINDRA , TRIBLEY WALTER A. , LINDSEY J. SUZANNE , MEISTRICH MARVIN L. , BUCANA CORAZON D. , SANBORN BARBARA M. , JOSEPH DAVID R. , GRISWOLD MICHAEL D. , CORNWALL GALL A. , WILKINSON MILES F.
Published: October 2023

ABSTRACT:Although the role of homeodomain transcription factors during embryogenesis is well known, their developmental function in postnatal animals is only beginning to be understood. We examined the regulation and expression pattern ofPem, a homeodomain protein that may regulate androgen‐dependent events in the testis and epididymis. Immunohistochemical analysis showed thatPemprotein is expressed selectively in the nuclei of Sertoli cells during the androgen‐dependent stage of the seminiferous epithelium cyclein vivo. RNase protection analysis revealed that a proximal promoter was responsible for androgen‐dependent mousePemexpression in testis and epididymisin vivo, whereas a distal promoter was used in placenta. The mousePemgene was expressed at ≅ 10‐fold higher levels in the testis than in the epididymis; conversely, the ratPemgene was expressed at > 10‐fold higher levels in the epididymis than in the testis. Because androgen‐binding protein has been proposed to transport androgens from the testis to the epididymis, we tested whether the ≥20‐fold higher levels of androgen‐binding protein expression in the rat, compared to that of mouse, are responsible for the differential expression ofPemin these two rodent species. Studies with androgen‐binding protein transgenic mice demonstrated that the species‐specific difference in androgen‐binding protein expression is unlikely to be responsible for the species‐specific difference inPemexpression. We found that androgen is necessary but not sufficient forPemexpression, since purified Sertoli cells rapidly down‐regulatedPemtranscripts in culture, regardless of the presence of testosterone. We conclude thatPemgene expression in Sertoli cells requires other cell types or cellular factors in addition to androgen.
Exposure of Thawed Frozen Bull Sperm to a Synthetic Peptide Before Artificial Insemination Increases Fertility
Authors: AMANN RUPERT P. , SEIDEL GEORGE E. , BRINK ZELLA A.
Published: October 2023

ABSTRACT: We evaluated the effect on fertility of in vitro exposure of thawed frozen bull sperm to synthetic FertPlus• peptide prior to artificial insemination (Al). The peptide represented a 60‐amino acid sequence within rat prosaposin. Commercial cryopreserved semen was from three Holstein bulls. Onset of estrus in groups of Holstein nulliparous heifers was synchronized via injection of prostaglandin F2‐α, and heifers were scheduled for Al 8–24 hours after estrus was detected. Semen was thawed, diluted to 2.4 × 105 sperm/ml with buffer, and split to provide control and exposed aliquots (0 or 30 μ.M peptide) that were incubated at 37°C 10 minutes and then were held at 32°C. The two aliquots of semen then were used on an alternate basis 2–65 minutes later to inseminate females. Each Al (one per female) involved the deposit of ∽250,000 sperm into each uterine horn. This procedure for Al was used to reduce the pregnancy rate with control semen to below the maximum value for a given bull and to facilitate detection of any beneficial effect of the peptide. For each bull, ∽32 heifers were inseminated with control semen, and ∽32 heifers were inseminated with peptide‐exposed semen. Pregnancy was evaluated ultrasonically ∽60 days after Al. After excluding one group of heifers with unusually low fertility, averaged across all animals, a 29% increase in pregnancy rate resulted from exposure of sperm to peptide (P < 0.04; one‐tailed chi‐square test; means were 48 vs. 62%). Pregnancy rates for the three bulls for control and peptide‐exposed semen, respectively, were 42 and 62%, 44 and 64%, and 56 and 61%: means in the first two pairs of values tended to differ (P ⋍ 0.10). These observations should be confirmed with sperm from other bulls used in a more conventional manner. However, with insemination of a limiting number of cryopreserved sperm, brief exposure of the thawed bull sperm to FertPlus® peptide appeared to improve fertility dramatically.
Suppression of Growth Hormone Does Not Affect Ongoing Spermatogenesis in Rats
Authors: AWONIYI CALEB A. , VEERAMACHANENI D. N. RAO , ROBERTS DAWN , TUCKER KATHLEEN E. , CHANDRASHEKAR VARADARAJ , SCHLAFF WILLIAM D.
Published: October 2023

ABSTRACT: Recent evidence suggests that growth hormone (GH) may enhance physiologic processes, such as spermatogenesis, in addition to causing classical anabolic effects. We have previously shown that testosterone restores spermatogenesis in rats that were made azoospermic by immunization against gonadotropin‐releasing hormone (GnRH). In this study, we investigated whether suppression of GH affects spermatogenesis and the ability of testosterone to restore spermatogenesis following immunization against GnRFI and/or growth hormone‐releasing hormone (GHRH). Twelve rats were actively immunized against GnRH (anti‐GnRH), twelve rats were actively immunized against GHRH (anti‐GHRH), six rats were immunized against both GnRH and GHRH (anti‐GnRH/GHRH), and six rats served as controls. Two weeks after the second booster, six rats each from the anti‐GnRH and anti‐GHRH groups as well as the six anti‐GnRH/GHRH rats received 24‐cm testosterone‐filled Silastic implants (1), and the remaining six rats from each of these groups received empty Silastic implants. All rats were euthanized 2 months later. Weights of testes and testicular sperm counts were determined Serum testosterone, luteinizing hormone (LH), follicle‐stimulating hormone (FSH), growth hormone (GH), and insulin‐like growth factor‐1 (IGF‐1) concentrations were determined by radioimmunoassays. Serum GH and IGF‐1 were suppressed in anti‐GHRH rats. IGF‐1 was partiallyrestored by testosterone in anti‐GHRH and in anti‐GnRH/GHRH rats, but GH was restored to control value in anti‐GnRH/GHRH rats. Serum LH and FSH were suppressed in anti‐GnRH and anti‐GnRH/GHRH rats, but only FSH was partially restored by testosterone. Suppression of GH did not affect maintenance of spermatogenesis. However, because 1 partially restored GH and IGF‐1 levels in anti‐GnRH/GHRH rats and because spermatogenesis was found to be restored in these rats, we conclude that GH does not play a role in the maintenance of spermatogenesis in adult rats, but it may be required for the replenishment of germ cells in experimentally induced regressed rat testes.
Immunocytochemical Localization of the Ya, Yb1, Yc, Yf, and Yo Subunits of Glutathione S‐Transferases in the Cauda Epididymidis and Yas Deferens of Adult Rats
Authors: ANDONIAN SERO , HERMO LOUIS
Published: October 2023

ABSTRACT: Glutathione S‐transferases (GSTs) are dimeric proteins grouped into five classes based on the degree of amino acid homology of their subunits. They are involved in cellular detoxification through the catalyzation of the conjugation of reduced glutathione with various electrophilic substances. In the present study, the distribution of Ya and Yc subunits from the alpha family, Yb1 and Yo subunits of the mu class, and the Yf subunit of the pi class were examined with light microscope immunocytochemistry in Bouin‐fixed, paraffin‐embedded tissue of different regions of the cauda epididymidis and vas deferens. In the cauda, principal cells showed high levels of expression of Ya, Yc, and Yo subunits, while in the vas deferens, staining decreased to moderate levels for the Ya and Yo subunits and to low levels for the Yc subunit. While Yf was maintained at low levels in principal cells of all cauda and vas deferens regions, Yb1 expression was more erratic, presenting a checkerboard‐like staining pattern in the proximal vas deferens and showing moderate cytoplasmic but intense nuclear reactivity in all other regions. Basal cells in the cauda were intensely reactive for Yf, while in the vas deferens, they became unreactive. Conversely, basal cells were unreactive for Ya in the cauda and proximal vas deferens, while in the middle and distal vas deferens, they became moderately reactive. In the case of Yb1 and Yo, some basal cells were reactive while others appeared unreactive in all cauda and vas deferens regions. Yc elicited the display of both reactive and unreactive basal cells in the cauda regions, and while the cells were moderately reactive in the proximal vas deferens, they became intensely reactive in the middle and distal vas deferens. In summary, both principal and basal cells show varying degrees of GST expression in the different regions of the cauda and vas deferens, suggesting that these cells are subjected to a complex, changing environment of substrates. Furthermore, while expression often differs from principal to basal cells, the absence of reactivity of a given GST in one cell type is usually compensated for by expression in the other cell type in any given region of the cauda or vas deferens. Taken together, the data suggest that ample protection from harmful circulating electrophiles can be provided for sperm during their storage in the cauda and vas deferens. In addition, since principal cells of the vas deferens are involved in steroid synthesis, the presence of GSTs in these cells may also serve to bind steroids, or this presence may be involved in steroid isomerization.
Principal Cells of the Vas Deferens Are Involved in Water Transport and Steroid Synthesis in the Adult Rat
Authors: ANDONIAN SERO , HERMO LOUIS
Published: October 2023

ABSTRACT: Principal cells show marked structural differences in the proximal, middle, and distal regions of the vas deferens, reflective of diverse functional activities. In the present study, we performed electron microscopy to examine the structural features of principal cells using glutaraldehyde‐fixed, Epon‐embedded material, while functional parameters were examined using light microscopic immunocytochemistry on Bouin‐fixed, paraffin‐embedded material. In the proximal region, the cuboidal principal cells resembled those of the cauda epididymidis, but few clear cells and occasional narrow cells were present. In the middle region, principal cells often contained blebs of their apical cytoplasm containing vesicular and tubular profiles. These blebs extended far from the cell surface and appeared to be liberated into the lumen, suggesting an apocrine type of secretion. In the distal region, dilated intercellular spaces containing numerous membranous profiles of different shapes and sizes were noted between adjacent principal cells and overlying basal cells. The use of an anti‐aquaporin‐1 antibody revealed an intense reaction over the endothelial cells of numerous vascular channels in the lamina propria. Taken together, these observations suggested water transport from the lumen of the vas deferens via the dilated spaces to underlying vascular channels, the function of which may be to concentrate sperm. The infranuclear cytoplasm of principal cells of this region showed whorls of smooth endoplasmic reticulum (sER). Large intracytoplasmic cavities were found within the sER aggregates, and these contained membranous profiles that appeared to peel off from the surrounding sER elements. Various images of such cavities closely juxtaposed to the lateral plasma membrane suggested that the membranous profiles of the intercellular spaces were derived from them. Use of anti‐3β‐hydroxysteroid dehydrogenase antibody revealed an intense reaction over principal cells of the vas deferens, as well as over the blebs in the lumen of the vas deferens, which is indicative of the steroid synthesis performed by these cells. The release of sER membranous profiles into the dilated spaces and the presence of blebs in the lumen may represent a means of transporting steroids that are destined for different sites out of the principal cells. Steroids in the blebs would be ultimately destined for utilization by luminal sperm, while those steroids in the dilated spaces are designed for utilization by muscle layers of the lamina propria. In summary, principal cells of the vas deferens appear to be involved in synthesis and secretion of steroids and in eliminating water from the lumen of the vas deferens.
Sodium Chlorate Induces Undersulfation of Cellular Proteoglycans and Increases in FSH‐Stimulated Estradiol Production in Immature Rat Sertoli Cells
Authors: PHAMANTU NHU‐THU , FAGNEN GAÉLLE , GODARD FLORENCE , BOCQUET JEAN , BONNAMY PIERRE‐JACQUES
Published: October 2023

ABSTRACT: The functional influence of cell proteoglycan (PG) undersulfation on estradiol synthesis by immature rat Sertoli cell cultures was investigated by using sodium chlorate, an inhibitor of the active sulfate donor for sulfotransferases. The addition of sodium chlorate to 20‐day‐old rat Sertoli cell cultures abolished [35S]‐sulfate incorporation into neosynthesized PG and consequently reduced the residence time of undersulfated PG in cell membrane. Simultaneously, follicle‐stimulating hormone (FSH)‐stimulated estradiol synthesis was increased by 45%. The effects of sodium chlorate upon Sertoli cell PG synthesis and steroidogenesis were not reproduced with the addition of sodium chloride. Addition of phosphodiesterase inhibitors (MIX or Ro20‐1724) decreased the magnitude of the chlorate effect on FSH‐stimulated steroidogenesis, suggesting that part of chlorate's effect on steroidogenesis resulted from a decrease in adenosine cyclic 3′,5′‐phosphate (cAMP)‐specific phosphodiesterase activity. Additionally, chlorate 1) increased Sertoli cell steroidogenesis at a step located beyond cAMP (restricted to Sertoli cell cultures exhibiting moderate steroidogenic response to (Bu)2cAMP) and 2) abolished the inhibition of steroidogenesis induced by transforming growth factor‐β. These results support our previous data, which showed that alteration in PG synthesis and the consequent decrease in cell membrane PG content induce an increase in FSH‐stimulated estradiol synthesis in Sertoli cell cultures. The identification of cAMP‐specific phosphodiesterase activity as a signal transduction step modified by PG undersulfation suggests the possible involvement of cell PG in the regulation of phosphodiesterase activity and, therefore, of FSH responsiveness during testicular development.
Stereological Evaluation of Mouse Prostate Development
Authors: SINGH JASKIRAT , ZHU QI , HANDELSMAN DAVBD J.
Published: October 2023

ABSTRACT: A stereological study of the development of the mouse prostate was undertaken between the second week of postnatal life and maturity. The aim was to quantify the progressive changes in the size and areal density of the ventral prostate gland components during development. Male mice were studied at weekly intervals from days of life 15 to 49 for organ and body weights, ductal branching, diameters of ventral prostate ducts and lumen and volume densities of epithelium, lumen, and stroma. Ductal branch‐tip numbers were maximal at 35 days of age, while prostate weights increased linearly with age and did not reach a plateau at 49 days. Prostatic glandular and luminal diameters both showed a continuous increase until day 49. At 5 weeks of age, there was a decrease in the volume density of prostatic epithelium accompanied by a simultaneous increase in the volume density of the lumen. This study indicates that prostate‐branching morphogenesis is complete by the fifth week in mice but that further growth of the prostate continues due to the increase in ductal dimensions. Qualitatively, the ventral prostate in mice is fully mature by 5 weeks, and this histological maturity coincides with the completion of branching morphogenesis.