DES
Desmin
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Although chronic obstructive pulmonary disease (COPD) and idiopathic pulmonary fibrosis (IPF) seem to be opposite entities from a clinical perspective, common initial pathogenic steps have been suggested in both lung diseases. Emphysema is caused by an elastase/anti-elastase imbalance leading to accelerated elastin degradation. Elastinolysis is however, also accelerated in the IPF patients' lungs. The amino acids desmosine and isodesmosine (DES) are unique to elastin. During the degradation process, elastases liberate DES from elastin fibers. Blood DES levels consequently reflect the rate of systemic elastinolysis and are increased in COPD. This is the first report describing elevated DES levels in IPF patients. We also demonstrated that the age-related increment of DES concentrations is enhanced in IPF. Our current study suggests that elastinolysis is a shared pathogenic step in both COPD and IPF. Further investigation is required to establish the relevance of accelerated elastin degradation in IPF and to determine whether decelerating this process leads to slower progression of lung fibrosis and better survival for patients with IPF.
MeSH Terms
- Aged
- Aging
- Biomarkers
- Desmosine
- Elastin
- Female
- Humans
- Idiopathic Pulmonary Fibrosis
- Male
- Middle Aged
- Pulmonary Disease, Chronic Obstructive
To confirm genetic variation in low-dose effects of diethylstilbestrol (DES), two inbred strains of rats, which have been selectively bred for high- and low-avoidance learning (HAA and LAA, respectively), were used in this study. LAA rats characteristically show later sexual maturation, earlier reproductive senescence, and lower body weight as compared to HAA rats. Female neonates of each strain were daily administered DES by oral gavage at doses of 0 (vehicle only), 0.05 and 0.5μg/kg for the first 5days after birth. As a result, early onset of abnormal estrous cycles was observed during the same period in HAA and LAA rats treated with 0.5μg/kg. However, accelerated puberty and excessive body weight gains were observed only in LAA rats treated with 0.05 and 0.5μg/kg. These results suggest that the effects of neonatal DES exposure vary with the genetic background of the female rats used.
MeSH Terms
- Animals
- Animals, Newborn
- Avoidance Learning
- Body Weight
- Carcinogens
- Diethylstilbestrol
- Estrogens, Non-Steroidal
- Estrous Cycle
- Female
- Genetic Variation
- Pregnancy
- Rats
- Rats, Inbred Strains
- Sexual Maturation
Keywords
- Endocrine disruptor
- Genetic variation
- Puberty
- Reproductive senescence
- Sprague-Dawley rat
Aging and oxidative stress seem to play a key role in the onset and progression of ocular surface diseases. Dry Eye Syndrome (DES) is a multifactorial disease of the tears and ocular surface in which symptoms may interfere with the ability to work and carry out daily functions. This clinical trial was a pilot study to evaluate the effects of supplementation with mixture (Saccharomyces boulardii MUCL 53837 and Enterococcus faecium LMG S-28935) on the tear film. Following the run-in period subjects were randomized in two groups: group A (n.30 subjects) and group B (n.30 subjects). Group A (control) treated only with substitute tear and group B treated with substitute tear mixture (probiotic). The data obtained in the two study groups A and B were, respectively the following: Schirmer I: 9.2±0.2 vs. 12.8±0.4 (p< 0.001); Schirmer II: 3.6±0.1 vs. 4.6±0.2 (p<0.001); BUT 3.8±0.3 vs. 6.2±0.2 (p<0.001). Culture test showed initial bacterial growth in group "A" (placebo) 27 out of 60 samples tested, corresponding to 45.0% and "B" after treatment (probiotic) was found positive culture whit growth of bacteria in 18 tests equal to 30.0%. The total numbers of isolations of aerobic and anaerobic bacteria found group A and B after treatment. A reduction of 23 to 16 strains of aerobic and anaerobic isolates from 13 to 7 has been found. The administration of probiotics strains was effective in reducing DES. In light of these results, we have identified our probiotic (Saccharomyces boulardii MUCL 53837 and Enterococcus faecium LMG S-28935) activity integration with the action of tear substitutes, along with standardization of clinical parameters of the tear film and microbiological activity in restoring of the microbiota ocular surface subject with DES.
MeSH Terms
- Aged
- Aging
- Dry Eye Syndromes
- Enterococcus faecium
- Eye
- Female
- Humans
- Lubricant Eye Drops
- Male
- Microbiota
- Oxidative Stress
- Pilot Projects
- Probiotics
- Saccharomyces boulardii
- Treatment Outcome
Keywords
- Dry Eye Syndrome (DES)
- Enterococcus faecium LMGS-
28935
- MUCL 53837
- Saccharomyces boulardii
- ocular microbiota
- symbiotic
- tear film.
The influence of age upon the use of drug-eluting stents (DES) in patients aged ≥ 65 years is uncertain. The aim of this study is to investigate the impact of age increase in patients aged ≥ 65 years in the use of DES in patients undergoing percutaneous coronary intervention (PCI). The study cohort comprised 8,598 patients ≥ 65 years of age who underwent stent implantation from April 2003 to March 2014. We defined the first DES era as the period April 2003 to July 2008 and the second DES era as the period July 2008 to March 2014. Multivariable logistic regression was performed for both eras to assess the impact of age increase and analyze independent factors associated with DES implantation. In the first DES era cohort, the two groups of patients differed in their risk factor profile with lower rates of male sex, diabetes, smokers, and hypercholesterolemia in those aged ≥ 75 years. There were more Caucasian and less African-Americans in this age group. Furthermore, patients aged ≥ 75 years had lower left ventricular ejection fraction (LVEF) and baseline haematocrit concentration were more likely to present with an acute myocardial infarction (MI) than stable or unstable angina and had higher rates of a previous history for congestive heart failure (CHF), chronic renal insufficiency (CRI), and peripheral vascular disease (PVD). These differences were broadly similar for patients in the second DES era except for similarities in LVEF, presentation with unstable angina, and PVD, as well as a lower rate for previous PCI. DES use was reduced with increasing age in both the first (OR=0.78; 95% CI=0.69-0.89) and second DES era (OR=0.53; 95% CI=0.47-0.58). In both eras, DES use was less likely in current smokers, patients presenting with acute MI and cardiogenic shock, and those with a previous history of CHF. In patients aged ≥ 65 years, the use of DES decreased with increasing age. This observation was apparent in both the first and second DES era. © 2015 Wiley Periodicals, Inc.
MeSH Terms
- Age Factors
- Aged
- Aged, 80 and over
- Aging
- Coronary Artery Disease
- District of Columbia
- Drug-Eluting Stents
- Female
- Humans
- Logistic Models
- Male
- Multivariate Analysis
- Odds Ratio
- Percutaneous Coronary Intervention
- Prosthesis Design
- Retrospective Studies
- Risk Assessment
- Risk Factors
- Time Factors
- Treatment Outcome
Keywords
- coronary artery disease
- drug eluting
- elderly patients
- stent
Abdominal aortic aneurysm (AAA) repair aims to prevent premature death from AAA rupture. Elective repair is currently recommended when AAA diameter reaches 5.5 cm (men) and 5.0 cm (women). Applying population-based indications may not be appropriate for individual patient decisions, as the optimal indication is likely to differ between patients based on age and comorbidities. To develop an Aneurysm Repair Decision Aid (ARDA) to indicate when elective AAA repair optimises survival for individual patients and to assess the cost-effectiveness and associated uncertainty of elective repair at the aneurysm diameter recommended by the ARDA compared with current practice. The UK Vascular Governance North West and National Vascular Database provided individual patient data to develop predictive models for perioperative mortality and survival. Data from published literature were used to model AAA growth and risk of rupture. The cost-effectiveness analysis used data from published literature and from local and national databases. A combination of systematic review methods and clinical registries were used to provide data to populate models and inform the structure of the ARDA. Discrete event simulation (DES) was used to model the patient journey from diagnosis to death and synthesised data were used to estimate patient outcomes and costs for elective repair at alternative aneurysm diameters. Eight patient clinical scenarios (vignettes) were used as exemplars. The DES structure was validated by clinical and statistical experts. The economic evaluation estimated costs, quality-adjusted life-years (QALYs) and incremental cost-effectiveness ratios (ICERs) from the NHS, social care provider and patient perspective over a lifetime horizon. Cost-effectiveness acceptability analyses and probabilistic sensitivity analyses explored uncertainty in the data and the value for money of ARDA-based decisions. The ARDA outcome measures include perioperative mortality risk, annual risk of rupture, 1-, 5- and 10-year survival, postoperative long-term survival, median life expectancy and predicted time to current threshold for aneurysm repair. The primary economic measure was the ICER using the QALY as the measure of health benefit. The analysis demonstrated it is feasible to build and run a complex clinical decision aid using DES. The model results support current guidelines for most vignettes but suggest that earlier repair may be effective in younger, fitter patients and ongoing surveillance may be effective in elderly patients with comorbidities. The model adds information to support decisions for patients with aneurysms outside current indications. The economic evaluation suggests that using the ARDA compared with current guidelines could be cost-effective but there is a high level of uncertainty. Lack of high-quality long-term data to populate all sections of the model meant that there is high uncertainty about the long-term clinical and economic consequences of repair. Modelling assumptions were necessary and the developed survival models require external validation. The ARDA provides detailed information on the potential consequences of AAA repair or a decision not to repair that may be helpful to vascular surgeons and their patients in reaching informed decisions. Further research is required to reduce uncertainty about key data, including reintervention following AAA repair, and assess the acceptability and feasibility of the ARDA for use in routine clinical practice. The National Institute for Health Research Health Technology Assessment programme.
MeSH Terms
- Age Factors
- Aged
- Aortic Aneurysm, Abdominal
- Cost-Benefit Analysis
- Decision Support Techniques
- Elective Surgical Procedures
- Female
- Health Status
- Humans
- Kaplan-Meier Estimate
- Life Expectancy
- Male
- Middle Aged
- Models, Econometric
- Perioperative Period
- Quality-Adjusted Life Years
- Risk Factors
- Sex Factors
- State Medicine
- Vascular Surgical Procedures
Industrial growth has increased the exposition to endocrine disruptor compounds (EDC's), which are exogenous agents with agonist or antagonist action of endogenous steroid hormones that may affect the course of parasite infections. We wanted to determine if the exposure to diethylstilbestrol (DES), an estrogen agonist, to both male and female mice affected the immune response and their susceptibility to T. crassiceps cysticercosis. In all infected groups, females showed higher parasite loads than males, and neonatal DES administration did not modify this pattern. In the spleen, noninfected mice showed sex-related differences in the percentage of the CD8 subpopulation, but DES decreased the percentage of CD3 , CD19 , and CD8 subpopulations in infected mice. In the mesenteric lymphatic node (MNL), DES showed a dimorphic effect in the percentage of CD19 cells. Regarding estrogen receptor alpha (ER-α) expression, DES treatment induced a reduction in the expression of this receptor in both noninfected female and male mice in the spleen, which was decreased only in males in CD3 and CD8 lymphocytes in MNL cell subpopulations. Our study is the first one to demonstrate that DES neonatal treatment in male and female mice affects the immune cell percentage, without effect on the susceptibility to T. crassiceps cysticercosis.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- Cysticercosis
- Diethylstilbestrol
- Estrogen Receptor alpha
- Female
- Flow Cytometry
- Immunity
- Lymph Nodes
- Lymphocyte Count
- Male
- Mice, Inbred BALB C
- Parasite Load
- Spleen
To evaluate the relationship between androgen levels and subjective and objective measures of dry eye syndrome (DES). A total of 263 male patients from the Miami Veterans Affairs Medical Center eye clinic aged ≥50 were recruited for this prospective cross-sectional study. Patients completed Dry Eye Questionnaire 5, underwent tear film evaluation, and had serum androgen levels measured. The correlations between androgen levels, DES composite scores, DES symptoms, and global, lipid, and aqueous tear film parameters were evaluated. Two hundred sixty-three patients with a mean age of 69 (50-95) were examined. There was no linear association between composite DES scores (generated using latent class analysis) and androgen levels. However, eyes with high DES scores (0.95-1.0) had higher levels of sex hormone-binding globulin (P = 0.03) and lower levels of dehydroepiandrosterone sulfate (DHEAS) (P = 0.02), androstenedione (A) (P = 0.02), and androstane-3α,17β-diol glucuronide (P = 0.03) compared to eyes with intermediate (0.05-0.95) or low (0-0.05) scores. There were no strong correlations between tear film measures and androgen levels. Regarding global parameters, a weak inverse correlation was found between corneal staining and A (r = -0.17, P = 0.009). For lipid parameters, a weak correlation existed between tear breakup time (TBUT) and A (r = 0.15, P = 0.02). When considering aqueous and lipid deficiency independently, the association between TBUT and A existed only with aqueous tear deficiency (r = 0.66, P = 0.002). Regarding aqueous parameters, a weak correlation existed between Schirmer test and DHEAS (r = 0.13, P = 0.047) and A (r = 0.21, P = 0.001). There was a weak correlation between higher levels of androstenedione and healthier global, lipid, and aqueous tear film parameters.
MeSH Terms
- Aged
- Aged, 80 and over
- Aging
- Androgens
- Biomarkers
- Cross-Sectional Studies
- Dry Eye Syndromes
- Follow-Up Studies
- Humans
- Male
- Meibomian Glands
- Middle Aged
- Prospective Studies
- Tears
Keywords
- androgens
- dry eye syndrome
- meibomian gland
As early as the 1950s there were warnings about the toxicity of the synthetic oestrogen diethylstilbestrol (DES), but in the Netherlands this drug continued to be prescribed to pregnant women up to the late 1970s. Until recently the periodic check-ups of children born from these pregnancies - the so-called DES Daughters - was mainly focused on genital abnormalities and the sporadically occurring clear cell adenocarcinoma of the vagina. As they grow older, the risk of this and similar types of carcinoma decreases and the nature of the periodic check-ups changes. However, check-ups cannot be omitted as the 5-yearly population study for cervical cancer is not suitable for detecting clear cell adenocarcinoma. DES Daughters should continue to receive a 2-yearly cytological check-up from their general practitioner until at least the age of 60. Now more than ever, physicians must be particularly vigilant in checking for the non-oncological late effects of DES.
MeSH Terms
- Adenocarcinoma, Clear Cell
- Adult
- Aging
- Diethylstilbestrol
- Female
- Humans
- Netherlands
- Physical Examination
- Pregnancy
- Prenatal Exposure Delayed Effects
- Risk Factors
- Uterine Cervical Neoplasms
- Vaginal Neoplasms
BACKGROUND/STUDY CONTEXT: No emotion-induction procedure is clearly recommended to assess the emotional reactivity in the elderly. This study aimed to validate an emotional reactivity procedure in healthy old patients. Nineteen healthy elders (age range: 66-91 years old) were compared with 19 education- and sex-matched young adults (age range: 20-33 years old) using a cross-sectional design. The main outcome measure was the evaluation of emotional reactivity to commercial film excerpts used as stimuli (joy, anger, fear, sadness, disgust, or neutral state) according to Philippot's procedure and using a 5-point questionnaire assessing 10 emotion dimensions (Differential Emotions Scale, DES). In the elderly sample, targeted emotions of fear, disgust, anger, and sadness were significantly induced compared with the baseline status. The global emotional reactivity to each film showed that the elderly subjects rated the DES in the same manner as the young adults, but with significantly higher global intensity for the excerpts inducing fear, anger, disgust, and sadness. The Philippot procedure is accurate for studying emotional reactivity in healthy elderly. The simplicity and rapidity of this procedure makes it suitable for emotion studies in different elderly populations.
MeSH Terms
- Adult
- Aged
- Aged, 80 and over
- Aging
- Emotions
- Female
- Humans
- Male
- Motion Pictures
- Neuropsychological Tests
- Surveys and Questionnaires
- Young Adult
This study sought to investigate the long-term effectiveness of drug-eluting stents (DES) versus bare-metal stents (BMS). Improved recanalization techniques have increased interest in percutaneous coronary intervention (PCI) for chronic total coronary occlusion (CTO). The long-term effectiveness of DES and BMS is not known. We used data from 10,261 stable patients age ≥65 years at 889 U.S. hospitals who underwent CTO PCI from January 1, 2005, to December 31, 2008, in the NCDR (National Cardiovascular Data Registry) CathPCI Registry with linked Medicare inpatient claims for follow-up. Patient and procedural characteristics, and 30-month death, myocardial infarction, revascularization, and hospitalization for bleeding were evaluated by stent type. Outcomes following stenting were adjusted and compared using propensity score matching. DES were used for CTO PCI in 8,218 (80%) and BMS in 2,043 (20%). DES patients were younger (74.0 vs. 75.5 years, p < 0.001), had longer lesions (18.8 vs. 16.5 mm, p < 0.001), received more stents (≥2 stents in 45.7% vs. 37.9%, p < 0.001), and underwent multivessel PCI (18.9% vs. 15.1%, p < 0.001). DES implantation was associated with a lower hazard of mortality (hazard ratio [HR]: = 0.72, 95% confidence interval [CI]: 0.60 to 0.86, p < 0.001), a similar hazard for myocardial infarction (HR: 0.85, 95% CI: 0.61 to 1.19, p = 0.35), and subsequent revascularization (HR: 0.94, 95% CI: 0.79 to 1.12, p = 0.48), including PCI (HR: 0.98, 95% CI: 0.83 to 1.19, p = 0.87) and coronary artery bypass grafting (HR: 0.71, 95% CI: 0.46 to 1.10, p = 0.12). Hospitalization for bleeding was also similar for DES versus BMS (HR: 0.92; 95% CI: 0.61 to 1.39, p = 0.70). Compared with BMS, DES use in stable patients undergoing CTO PCI was associated with lower mortality, as well as similar myocardial infarction and repeat revascularization rates without an increase in subsequent bleeding requiring hospitalization.
MeSH Terms
- Age Factors
- Aged
- Aged, 80 and over
- Aging
- Angioplasty, Balloon, Coronary
- Comparative Effectiveness Research
- Confidence Intervals
- Coronary Occlusion
- Drug-Eluting Stents
- Female
- Humans
- Male
- Myocardial Reperfusion
- Propensity Score
- Registries
- Statistics as Topic
- Time Factors
- United States
Leaf senescence is the final phase of the leaf development, comprising several controlled complex physiological, biochemical and molecular events. From February to June it was possible to observe a rapid increase of Scirpus maritimus biomass accompanied by an increase in the overall pigment battery, photosynthetic efficiency and photoprotection capacity. With senescence progressing, the photosynthetic pigments decreased dramatically in a rather equal extent. With the exception of Zeaxanthin (90% decrease), all pigments suffered a 98-100% decreased during senescence. Overlooking the operational PSII quantum efficiency it was possible to observe that it suffered almost no changes during leaf maturation (with the exception of the senescent leaves), whilst the maximum quantum efficiency, showed more evident changes, decreasing with the leaf maturation. This observation coupled with the increased DES index may be an indication that the decrease in the PSII maximum yield may represent a mechanism to down-regulate the photosynthetic electron transport rate compensating the consequent decrease in CO(2) assimilation capacity. This fact allied with a decrease in the minimum light intensity for photosynthesis saturation in senescent leaves, suggest that the requirement for reducing power and photophosphorylation for the dark reaction is inevitably decreased and that photosynthesis in senescent leaves will be saturated.
MeSH Terms
- Aging
- Carbon Sequestration
- Chlorophyll
- Cyperaceae
- Photochemistry
- Photosynthesis
- Photosystem II Protein Complex
- Plant Leaves
The aim of this study was to estimate the incremental cost effectiveness of replacing bare metal stents (BMS) by drug-eluting stents (DES) when using trial data and registry data. We developed a Markov model (model of cost effectiveness of coronary artery disease) in which 60-year-old patients started by undergoing percutaneous coronary intervention for acute or subacute coronary artery disease. The patients are followed until death or 100 years of age. Data on the occurrence of events (revascularization, acute myocardial infarction, and death) were based on Scandinavian registry data. Separate analyses were conducted with data on effectiveness based on randomized controlled trials and patient registries. On using trial data, it was found that sirolimus-eluting stents (SES) yield 0.003 greater life expectancy and $3300 lower costs than do BMS (dominant strategy). Paclitaxel-eluting stents (PES) yield 0.148 more life years than do SES at additional lifetime costs of $2800 ($21,400 per life year gained). On using registry data, the cost per life year gained was found to be $4900 when replacing BMS with DES. Probabilistic sensitivity analyses, on the other hand, indicate that PES only has a 50%-75% probability of being cost effective, regardless of the type of effectiveness data. DESs are cost effective with current willingness to pay for life year gains. Whether PES or SES is the most effective DES remains uncertain.
MeSH Terms
- Coronary Artery Disease
- Cost-Benefit Analysis
- Drug-Eluting Stents
- Health Care Costs
- Humans
- Life Expectancy
- Male
- Markov Chains
- Middle Aged
- Paclitaxel
- Percutaneous Coronary Intervention
- Probability
- Randomized Controlled Trials as Topic
- Registries
- Sirolimus
- Stents
The effects of aging and an ovariectomy operation on the brain-disrupting actions caused by prenatal exposure to diethylstilbestrol (DES) were studied in mice. In the young DES-exposed female mice, the level of hippocampal phosphorylated CaM kinase II (pCaMKII) was not changed. However, at 8 months, the level of hippocampal pCaMKII in the DES-exposed female mice significantly increased compared to control. Moreover, the ovariectomy significantly increased the level of pCaMKII in the hippocampus but not the cortex of DES-exposed female mice. These findings suggest that the influence of prenatally-exposed DES on the hippocampal pCaMKII may be affected by the endogenous female sex hormones such as estrogen.
MeSH Terms
- Aging
- Animals
- Blotting, Western
- Brain Chemistry
- Calcium-Calmodulin-Dependent Protein Kinase Type 2
- Carcinogens
- Diethylstilbestrol
- Female
- Hippocampus
- Mice
- Ovariectomy
- Phosphorylation
- Prefrontal Cortex
- Pregnancy
- Prenatal Exposure Delayed Effects
The WNTs are secreted proteins that control essential developmental processes, such as embryonic patterning, cell growth, migration, and differentiation. In mice, three members of the Wnt gene family (Wnt4, Wnt5a, and Wnt7a) have been studied extensively in the female reproductive tract. The present study determined effects of postnatal day and exposure to diethylstilbestrol (DES) on Wnt and Fzd gene expression in the mouse uterus as well as the biological role of Wnt11 in postnatal mouse uterine development and function. Wnt4, Wnt5a, Wnt7a, Wnt7b, Wnt11, Wnt16, Fzd6, and Fzd10 were detected by in situ hybridization in the neonatal mouse uterus. In situ hybridization analyses revealed that Wnt4, Wnt5a, and Wnt16 were localized in the endometrial stroma, whereas Wnt7a, Wnt7b, Wnt11, Fzd6, and Fzd10 were in the uterine epithelia of neonatal mice. Exposure of mice to estrogen or estrogen receptor agonists during critical development periods inhibits endometrial adenogenesis. In the present study, DES-induced disruption of endometrial gland development was associated with reduction or suppression of Wnt4, Wnt5a, Wnt7a, Wnt11, Wnt16, and Fzd10. Ablation of Wnt11, an epithelial-expressed, DES-regulated gene, in the neonatal uterus did not affect endometrial adenogenesis or expression of other Wnt genes. Interestingly, Wnt11-deleted uteri had more endometrial glands on Postnatal Day 10. Although CTNNB1 expression was not affected by ablation of Wnt11, Vangl2 was inhibited in the uteri of Wnt11(d/d) mice. These results support the idea that a number of different Wnt genes are potential regulators for uterine morphogenesis; however, Wnt11 does not have a direct effect on uterine development.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- Diethylstilbestrol
- Endometrium
- Estrogens, Non-Steroidal
- Female
- Frizzled Receptors
- Gene Deletion
- Gene Expression
- In Situ Hybridization
- Mice
- Morphogenesis
- Nerve Tissue Proteins
- Receptors, G-Protein-Coupled
- Wnt Proteins
The aim of this study was to investigate the effect of fetal exposure to diethylstilbestrol (DES) on the induction of mammary tumors induced by 7,12-dimethylbenz[a]anthracene (DMBA) in female rats. Pregnant rats were fed only normal diet, diet mixed with 0.1 ppm DES throughout pregnancy period or diet mixed with 0.1, 1 or 10 ppm DES from day 13 of pregnancy till the end of pregnancy. Delivered pups were given 10 mg DMBA by gastric intubation at 50 days after birth and observed till 336 days after birth. Some rats exposed to DES throughout pregnancy and those from day 13 till the end of pregnancy showed endocrine disrupting effects such as absence of CL and active lactation in mammary glands at necropsy, while no abnormal estrus cycle such as persistent estrus was seen during the observation period until 88 days after birth. Fetal exposure to 0.1 ppm DES throughout pregnancy period, 0.1 and 1 ppm DES from day 13 of pregnancy increased the incidence and number of mammary carcinomas (MCs) at the earlier period while exposure to 0.1 ppm DES throughout pregnancy period enhanced the incidence and number of benign proliferative lesions (PLs) at the later period. MCs appeared earlier than benign PLs. These results suggest that exposure to DES throughout pregnancy and from day 13 of pregnancy could induce endocrine disrupting conditions and enhance the induction of MCs and that exposure to DES throughout pregnancy enhance PLs.
MeSH Terms
- Aging
- Animals
- Diethylstilbestrol
- Dose-Response Relationship, Drug
- Drug Administration Schedule
- Female
- Mammary Neoplasms, Animal
- Mammary Neoplasms, Experimental
- Pregnancy
- Prenatal Exposure Delayed Effects
- Rats
Natural and synthetic estrogens, Including diethylstilbestrol (DES), given during the critical period of newborn life Induce abnormalities in ovaries of mice. Induction of polyovular follicles (PFs) containing two or more oocytes in a follicle is one example. In this study, the involvement of estrogen receptor subtypes ERalpha and ERbeta in induction of PFs by neonatal treatment with DES was analyzed by using ERalpha knockout (alphaERKO) and ERbeta knockout (betaERKO) mice. Ovaries of mice injected with 3 microg DES for 5 days from the day of birth were examined histologically from 10 to 60 days of age, and the expression of genes involved in folliculogenesis was analyzed by real-time quantitative PCR. The PF Incidence (percent of PFs per 100 follicles greater than 50 microm in diameter) in the ovary of alphaERKO mice treated with DES was not different from that in the DES-treated wild-type mice. However, neonatal DES treatment did not increase the PF incidence in betaERKO mice, suggesting that PFs were induced by DES through ERbeta but not ERalpha. The expression of bone morphogenetic protein 15, growth differentiation factor 9, inhibin-alpha, Müllerian inhibiting substance, and other genes in the ovaries of DES-treated betaERKO mice was not different from that in the ovaries of DES-treated wild-type mice. These results indicate that ERbeta but not ERalpha is essential for DES to Induce PFs in mice.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- Diethylstilbestrol
- Estrogen Receptor alpha
- Estrogen Receptor beta
- Female
- Gene Expression Regulation
- Mice
- Mice, Knockout
- Ovarian Follicle
Our previous studies have demonstrated that prenatally administered diethylstilbestrol (DES) impairs testicular endocrine function in male offspring. The present study examined whether maternal DES treatment influences testicular steroidogenesis and spermatogenesis. DES was injected subcutaneously at 0.5 or 1.5 microg/kg/day (DES 0.5 and 1.5 groups, respectively) into pregnant SD rats on days 7-21 of gestation. Male offspring in the DES 0.5 and 1.5 groups were autopsied at 1, 3, 6 and 15 weeks after birth. At 1 week, DES treatment did not lead to a change in the volume of P450scc-positive cells (Leydig cells), suggesting that DES has no inhibitory effect on the development of Leydig cells. DES administration disrupted luteinizing hormone receptor (LHr) expression and exerted inhibitory effects on signal transduction from LHr to steroidogenic acute regulatory protein (StAR) in testicular steroidogenesis (P<0.05), although there were no changes in the mRNA expression levels of steroidogenic enzymes, such as P450scc, 3beta-hydroxysteroid dehydrogenase (3beta-HSD) and P450(17 alpha), which may have caused a decrease in the plasma testosterone level. DES treatment did not disrupt the cycle of spermatogenesis but did upregulate the expression levels of androgen receptor (AR) mRNA in both DES groups at 15 weeks (P<0.05). These results indicate that maternal DES treatment disrupts steroidogenesis but induces a high level of AR mRNA expression to counteract the low levels of testosterone during spermatogenesis.
MeSH Terms
- Aging
- Animals
- Body Weight
- Diethylstilbestrol
- Dose-Response Relationship, Drug
- Female
- Gene Expression Profiling
- Gene Expression Regulation, Developmental
- Male
- Organ Size
- Pregnancy
- Prenatal Exposure Delayed Effects
- RNA, Messenger
- Rats
- Rats, Sprague-Dawley
- Reverse Transcriptase Polymerase Chain Reaction
- Seminiferous Tubules
- Spermatogenesis
- Testicular Hormones
- Testis
We sought to evaluate the impact on long-term clinical outcomes of different types of drug-eluting stents (DES) in elderly patients. Elderly patients constitute a fast-growing portion of cardiovascular patients, however, they are not adequately represented in clinical trials. Moreover, few data comparing different type of DES in elderly patients are available. From a total of 2,330 consecutive patients treated at our institution with DES, we selected 207 elderly patients (> or = 75 years of age) who underwent, from May 2002 to December 2006, sirolimus-eluting stent (SES group, 116 patients [pts], 56%) or paclitaxel-eluting stent (PES group, 91 pts, 43.9%) implantation. We evaluated the 24-month incidence of major adverse cardiac events (MACE). Higher rates of hypertension (78.4% vs. 90.1%; p = 0.01), diabetes (37.9 vs. 45.1; p = 0.01) and previous coronary artery bypass grafts (10.3% vs. 19.4%; p = 0.04) in the PES group were observed, whereas in the SES group, there were more smokers (26.6% vs. 12.1%; p = 0.007) and a higher incidence of previous myocardial infarction (MI) (50% vs. 35.2%; p = 0.02). Procedural success and in-hospital MACE were similar in both groups. At follow up, there was a higher incidence of MACE (22.4% vs. 10.9%; p = 0.04) and target lesion revascularization (7.1% vs. 3.0%; p = 0.02) in the SES group compared to the PES group. The incidence of cardiac death and MI were comparable between the two groups, as well as the rate of stent thrombosis. After adjustment for clinical and angiographic characteristics, no significant differences in outcomes were observed between SES and PES. In this real-word experience, no significant differences were found in the safety and efficacy profiles between SES and PES use in elderly patients.
MeSH Terms
- Age Factors
- Aged
- Aged, 80 and over
- Aging
- Antineoplastic Agents, Phytogenic
- Coronary Angiography
- Coronary Restenosis
- Drug-Eluting Stents
- Female
- Humans
- Immunosuppressive Agents
- Male
- Multivariate Analysis
- Myocardial Infarction
- Paclitaxel
- Proportional Hazards Models
- Registries
- Retrospective Studies
- Sirolimus
- Time Factors
- Treatment Outcome
This study tested the hypothesis that the estrogen receptor (ESR) pathway, androgen receptor (AR) pathway, or both mediate estrogen-induced developmental penile disorders. Rat pups received diethylstilbestrol (DES), with or without the ESR antagonist ICI 182,780 (ICI) or the AR agonist dihydrotestosterone (DHT) or testosterone (T), from Postnatal Days 1 to 6. Testicular T concentration, penile morphology and morphometry, and/or fertility was determined at age 7, 28, or 150 days. DES treatment alone caused 90% reduction in the neonatal intratesticular T surge; this reduction was prevented by ICI coadministration, but not by DHT or T coadministration. Unlike the T surge, coadministration of ICI and coadministration of DHT or T mitigated penile deformities and loss of fertility. Generally, ICI, DHT, or T treatment alone did not alter penile morphology; however, fertility was 20% that of controls in ICI-treated rats vs. 70%-90% in DHT- or T-treated rats. The lower fertility in the rats treated with ICI alone could be due to altered sexual behavior, as these males did not deposit vaginal plugs. In conclusion, observations that both an ESR antagonist and AR agonists prevent penile deformities and infertility suggest that both pathways are involved in estrogen-induced penile disorders. Observations that coadministration of ICI, but not DHT or T, prevents the DES-induced reduction in the neonatal T surge suggest that, although ICI exerts its mitigating effect both at the level of Leydig cells and penile stromal cells, DHT and T do so only at the level of stromal cells.
MeSH Terms
- Aging
- Androgen Receptor Antagonists
- Androgens
- Animals
- Animals, Newborn
- Disorders of Sex Development
- Estrogens
- Female
- Hormone Antagonists
- Male
- Organ Size
- Penile Diseases
- Penis
- Rats
- Rats, Sprague-Dawley
- Receptors, Androgen
- Receptors, Estrogen
- Sexual Maturation
- Signal Transduction
- Testosterone
The internal anal sphincter (IAS) plays an important role in the pathophysiology of constipation and incontinence. We hypothesized that functional bowel obstruction in premature infants is because of a poorly developed IAS. We investigated the neuromuscular development of IAS in fetal, newborn, and adolescent pigs. Paraffin sections of IAS from 5 different age groups, E60, E90, 1 day, 4, and 12 weeks old, were stained with protein gene product 9.5 (PGP9.5), *-smooth muscle actin (*-SMA), caldesmon (CALD), calponin (CALP), and desmin (DES) antibodies. Quantification of results was performed by grading the density of immunostaining. The PGP9.5-positive ganglion cells were observed in the myenteric and submucosal region of the entire length of the IAS at E60. An increase in ganglion cell size and density was observed with increasing age. There were striking differences in the density of PGP9.5, alpha-SMA, DES, CALD, and CALP immunoreactive fibers between prenatal and postnatal period with gradient increase in the number of fibers from after birth to 12 weeks of age. This study shows for the first time that there are age-related differences in the distribution of neurons and smooth muscle cell components in the IAS. The decreased expression of contractile and cytoskeleton proteins in smooth muscle cells together with decreased expression of neurons in the IAS in the perinatal period may lead to motility dysfunction causing functional intestinal obstruction seen in premature infants.
MeSH Terms
- Aging
- Anal Canal
- Animals
- Animals, Newborn
- Constipation
- Fecal Incontinence
- Female
- Immunohistochemistry
- Models, Animal
- Muscle Contraction
- Muscle Relaxation
- Muscle, Smooth
- Nerve Fibers
- Nerve Tissue Proteins
- Neuromuscular Junction
- Pregnancy
- Proteins
- Sensitivity and Specificity
- Swine
- Tissue Culture Techniques
Limited data are available on contemporary percutaneous coronary intervention (PCI) practice patterns and outcomes in elderly patients. The objective of this study was to evaluate "real-world" PCI in elderly and nonelderly patients during the first year of availability of drug-eluting stents (DES) in the United States market (May 1, 2003-April 30, 2004). One thousand one hundred sixty-six consecutive patients (272 elderly [age > or =75 years] and 894 nonelderly [age <75 years]) having PCI for de novo coronary artery disease (CAD) at Dartmouth-Hitchcock Medical Center were included in this study. Primary outcome measures of this study were in-hospital major adverse cardiac events (MACE-death, new MI, urgent revascularization). Secondary end points included acute renal failure, respiratory failure, and vascular complications. Elderly patients had higher MACE (8.5% vs 1.5%, P < or = 0.001), unadjusted in-hospital mortality (7.4% vs 0.8%, P < or = 0.001), in-hospital cardiac arrest (1.5% vs 0.3%, P = 0.03), requirements for assisted blood pressure support (13.2% vs 7.0%, P = 0.0001), respiratory failure (2.2% vs 0.9%, P = 0.08), acute renal failure (2.9% vs 0.8%, P = 0.005), and vascular complications (10.3% vs 5.5%, P = 0.005) than their nonelderly counterparts. Higher MACE rates persisted in the elderly despite correction for baseline differences using multivariate regression modeling. Advanced age remains a predictor of adverse outcomes attending PCI even in the contemporary era in which DES are available. This study highlights the need for further progress and investigation to optimize outcomes of PCI in the elderly.
MeSH Terms
- Age Factors
- Aged
- Aging
- Analysis of Variance
- Angioplasty, Balloon, Coronary
- Blood Vessel Prosthesis Implantation
- Coated Materials, Biocompatible
- Combined Modality Therapy
- Coronary Artery Disease
- Female
- Hospital Mortality
- Humans
- Male
- Middle Aged
- Postoperative Complications
- Stents
- Treatment Outcome
- United States
The trajectory of posttraumatic stress disorder (PTSD) and PTSD-related symptoms in relation to aging is not well understood. We previously observed higher levels of dissociation as measured by the Dissociative Experiences Scale (DES) among older Holocaust survivors with, compared to those without, PTSD, though scores on the DES in Holocaust survivors were markedly lower than those that had been reported for younger cohorts. We undertook a longitudinal evaluation of dissociation in Holocaust survivors. Twenty-six Holocaust survivors with current PTSD, 30 Holocaust survivors without current PTSD, and 19 nonexposed were evaluated at the initial evaluation and subsequently 8.11 years later. Repeated measures analysis of variance (ANOVA) on the DES scores from these times demonstrated a significant main effect for time and a significant group by time interaction, reflecting a marked decline in Holocaust survivors, particularly those with PTSD. Controlling for age obliterated the effect of time, but not the group by time interaction. A similar pattern was shown with The Clinician Administered PTSD Scale (CAPS) scores. Different symptoms related to PTSD show different trajectories of change with age, with dissociation appearing to be less prominent with age.
MeSH Terms
- Aged
- Aging
- Dissociative Disorders
- Holocaust
- Humans
- Jews
- Longitudinal Studies
- Survivors
In this study, two selective estrogen receptor modulators (SERMs), tamoxifen (TAM) and toremifene (TOR) or two estrogens, ethinylestradiol (EE) and diethylstilbestrol (DES) were administered to newborn male and female Sprague-Dawley rats (days 1-5) to investigate the occurrence of developmental abnormalities in the adulthood. The compounds were dosed (s.c.) at an equimolar dose of 24.9 micromol/kg. During the follow-up period, mortality occurred mainly in DES-treated male rats (3/4), associated with obstructive urinary calculi and suppurative renal inflammation in 2/3 rats. Similar lesions were not evident in other groups. At the age of 15 months, the animals were necropsied and organs were collected for histopathology and histomorphometry. Treatment-related abnormalities were restricted to the reproductive organs. Chronic prostatitis and epithelial abnormalities in the vas deferens were observed in all treatment groups. The columnar epithelium of vas deferens showed hyperplasia and development of subepithelial glandular structures resembling epididymal cysts reported in humans exposed in utero to DES. Testicular atrophy was observed especially in estrogen-treated rats. Mainly in SERM-treated female rats, the uterus showed luminal dilation or obstruction, loss of endometrial glands and myometrium disorganization including foci of muscular disruption. TOR-treated female rats showed polyp-like nodules (incidence 4/15) and a high incidence (9/15) of a simple cuboidal epithelium in cervical regions normally occupied by multilayered epithelia. In conclusion, the vas deferens is a main target organ following neonatal administration of SERMs and estrogens. In addition, female rats were significantly more susceptible to SERM treatment than to treatment with estrogens.
MeSH Terms
- Animals
- Animals, Newborn
- Body Weight
- Diethylstilbestrol
- Estrogens
- Ethinyl Estradiol
- Female
- Female Urogenital Diseases
- Longevity
- Male
- Male Urogenital Diseases
- Nephritis, Interstitial
- Organ Size
- Rats
- Rats, Sprague-Dawley
- Selective Estrogen Receptor Modulators
- Survival Rate
- Tamoxifen
- Toremifene
- Urinary Calculi
- Urogenital System
- Vas Deferens
Chronic estrogen administration can lead to thymic atrophy in rodents. In this article we report that the Brown Norway (BN) rat is sensitive to thymic atrophy induced by the estrogen diethylstilbestrol (DES). By contrast, DES does not induce significant thymic atrophy in the August x Copenhagen-Irish (ACI) strain. The sensitivity of the BN rat to DES-induced thymic atrophy appears to segregate as an incompletely dominant trait in crosses between the BN and ACI strains. In a (BN x ACI)F(2) population, we find strong evidence for three major genetic determinants of sensitivity to DES-induced thymic atrophy on rat Chromosome (RNO) 10 and RNO2. Genotypes at these loci, termed Esta1, 2, and 3, do not have a significant impact on the ability of DES to induce pituitary tumorigenesis or inhibit growth of these F(2) rats. These data indicate that the genetic factors that control DES-induced thymic atrophy are distinct from those that control the effects of DES on pituitary mass and body mass. The Esta intervals on RNO10 and RNO2 overlap with loci that control sensitivity to radiation-induced thymocyte apoptosis, as well as susceptibility to a variety of allergic and autoimmune pathologies, including allergic encephalitis, arthritis, and glomerulonephritis in rodents. These observations suggest that common genetic determinants may control sensitivity to estrogen-induced thymic atrophy, maintenance of thymocyte homeostasis, and immune function.
MeSH Terms
- Aging
- Animals
- Atrophy
- Chromosome Mapping
- Crosses, Genetic
- Diethylstilbestrol
- Male
- Quantitative Trait Loci
- Rats
- Rats, Inbred ACI
- Rats, Inbred BN
- Thymus Gland
In this study, we compared the long-term effects of neonatal exposure to diethylstilbestrol (DES, 0.0125-50 microg), tamoxifen (TAM, 0.0125-50 microg), and toremifene (TOR, 53 microg) on mammary gland development and differentiation. Allometric growth of the mammary ducts was stimulated by neonatal DES exposure (12.5 microg) and impaired by exposure to TAM (25 microg). Neonatal treatment with high doses of DES resulted in mammary ducts that displayed extensive dilatation and precocious lactogenesis in postpubertal, nulliparous females. Initiation of this precocious differentiation coincided with the absence of corpora lutea, increased levels of serum prolactin (PRL), and the induction of Prl mRNA expression within the mammary glands. Neonatal exposure to 1.25 microg TAM increased alveolar development in postpubertal, nulliparous females similar to that recorded in females treated with low doses of DES. Lower doses of TAM did not affect alveolar development, whereas branching morphogenesis and alveolar development were impaired by higher doses. Increased alveolar development in females exposed to 1.25 microg TAM was associated with elevated serum progesterone (P) and increased alveolar development in response to exogenous P. Taken together, our findings demonstrate that neonatal exposure to both DES and TAM exerts long-lasting effects on the proliferation and differentiation of the mammary glands in female BALB/c, primarily as the result of endocrine disruption.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- Blotting, Western
- Diethylstilbestrol
- Dose-Response Relationship, Drug
- Estrogen Antagonists
- Estrogens, Non-Steroidal
- Female
- Genitalia, Female
- Hormones
- Immunohistochemistry
- In Situ Hybridization
- Lactation
- Mammary Glands, Animal
- Mice
- Mice, Inbred BALB C
- Organ Culture Techniques
- Ovariectomy
- RNA
- Tamoxifen
- Toremifene
Environmental contaminants with estrogenic properties have the potential to alter pubertal development. In addition to the reproductive system, other systems that mature under the influence of estrogen could be affected. This study examined the effect on immune, hematologic, and bone mass parameters of treatment with estrogenic agents (methoxychlor, MXC, 25 and 50 mg/kg/day; diethylstilbestrol, DES, 0.5 mg/kg/day) given in the peripubertal period to female rhesus monkeys. DES had striking effects on several parameters assessed measures CBC and clinical chemistry including hematocrit, hemoglobin, serum albumin, liver transaminases, and lipids. Circulating lymphocytes, particularly B cells, were depressed by DES, and a maturational shift in a memory T-cell population was altered. Bone mass and length, as measured after a 9-month recovery period, were significantly lower in the DES group and bone mass tended to be reduced in the femur of the MXC50 group relative to controls. In conclusion, the data indicate that DES had a clear effect on immunohematology and bone growth, while MXC influenced fewer parameters. Disruption in these systems during puberty could alter adolescent risk for anemia and infectious disease and subsequent adult risk for diseases such as osteoporosis, heart disease, and autoimmune disease.
MeSH Terms
- Absorptiometry, Photon
- Aging
- Animals
- Blood Cell Count
- Blood Chemical Analysis
- Bone and Bones
- Diethylstilbestrol
- Endocrine System Diseases
- Erythrocyte Count
- Estrogens, Non-Steroidal
- Female
- Flow Cytometry
- Immunity
- Insecticides
- Lymphocyte Count
- Macaca mulatta
- Methoxychlor
- Organ Size
Summary In utero or neonatal exposure to high levels of exogenous steroid hormones, such as the potent synthetic diethylstilbestrol (DES), incurs an increased risk of malfunctional male reproduction. In this study, we investigated whether neonatal exposure to DES induces the alteration of mRNA expression in adult mouse testis. Using a cDNA subtraction method, we isolated seven gene clones whose expression was changed in neonatally DES-treated mouse testis. Northern blot analysis revealed that five up-regulated genes (AF326230, AF356521, AK004975, AK006136 and BM237156) and two down-regulated genes (AK017044, AK017130) were predominantly expressed in testes of 8-week-old mice. Moreover, we confirmed that the expression of these seven genes was altered by neonatal DES-exposure using Northern blot analysis. Our results suggest that neonatal exposure to DES leads to the alteration of gene expression in the testis in the long term. These genes might be useful as biological markers of foetal or neonatal exposure to exogenous steroid hormones, such as DES.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- Blotting, Northern
- DNA, Complementary
- Diethylstilbestrol
- Down-Regulation
- Drug Administration Schedule
- Estrogens, Non-Steroidal
- Gene Expression
- Male
- Mice
- Mice, Inbred ICR
- Molecular Sequence Data
- RNA, Messenger
- Subtraction Technique
- Testis
- Up-Regulation
Objectives of the study were to determine developmental changes in morphology and expression of androgen receptor (AR) and estrogen receptor (ER)alpha in the body of the rat penis exposed neonatally to diethylstilbestrol (DES). Male pups received DES at a dose of 10 microg per rat on alternate days from Postnatal Day 2 to Postnatal Day 12. Controls received olive oil vehicle only. Tissue samples were collected on Days 18 (prepuberty), 41 (puberty), and 120 (adult) of age. DES-induced abnormalities were evident at 18 days of age and included smaller, lighter, and thinner penis, loss of cavernous spaces and associated smooth muscle cells, and increased deposition of fat cells in the corpora cavernosa penis. Fat cells virtually filled the entire area of the corpora cavernosa at puberty and adulthood. Plasma testosterone (T) was reduced to an undetectable level, while LH was unaltered in all treated groups. AR-positive cells were ubiquitous and their profile (incidence and staining intensity) did not differ between control and treated rats of the respective age groups. Conversely, ERalpha-positive cells were limited to the stroma of corpus spongiosus in all age groups of both control and treated rats, but the expression in treated rats at 18 days was up-regulated in stromal cells of corpora cavernosa, coincident with the presence of morphological abnormalities. Hence, this study reports for the first time DES-induced developmental, morphological abnormalities in the body of the penis and suggests that these abnormalities may have resulted from decreased T and/or overexpression of ERalpha.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- Diethylstilbestrol
- Estrogen Receptor alpha
- Estrogens, Non-Steroidal
- Female
- Immunohistochemistry
- Luteinizing Hormone
- Male
- Penis
- Rats
- Rats, Sprague-Dawley
- Receptors, Androgen
- Testosterone
- Tissue Distribution
The synthetic estrogen diethylstilbestrol (DES) is an established, estrogenic endocrine disruptor (ED). The Syrian golden hamster (Mesocricetus auratus) offers some unique advantages as an experimental system to investigate the perinatal ED action of DES and other estrogenic EDs. Previous analyses regarding the consequences of neonatal administration (100 microg) of DES versus estradiol-17beta (E2) showed that DES had a more potent disruptive effect on morphogenesis and gene expression in the uterus, oviduct, and ovary as well as in the testis and male accessory organs. The objectives of the present study were to describe the histopathological consequences of the two neonatal treatment regimens in the hamster cervix and to compare them with our previous observations in the hamster uterus. As previously found in the hamster uterus, DES was more potent than E2 as a neonatal disruptor of the hamster cervix in prepubertal animals and in ovarian-intact adult animals. However, the cervix-versus-uterus scenario diverged in animals that were ovariectomized prepubertally and then chronically stimulated with natural estrogen (E2). We confirmed previous observations that neonatal exposure to DES, but not to E2, permanently alters estrogen responsiveness in the adult hamster uterus, but neither neonatal treatment regimen affected estrogen responsiveness in the adult hamster cervix. These results suggest that an unidentified ovarian factor influences the extent of neonatal DES-induced disruption of the cervix, but not of the uterus, in hamsters.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- Cervix Uteri
- Cricetinae
- Diethylstilbestrol
- Estradiol
- Estrogens, Non-Steroidal
- Female
- Mesocricetus
- Ovariectomy
- Uterus
The immune system is a potential target for estrogenic endocrine disrupters. To date, there is limited information on whether estrogenic endocrine disruptors modulate the immune system of aged individuals. To address this issue, groups of 74-week-old mice were given nine oral doses of selected estrogenic endocrine disrupters: diethylstilbestrol (DES, 3 microg/100 g bw), alpha-zearalanol (0.5 mg/100 g bw), or genistein (0.15 mg/100 g bw) in corn oil, or corn oil alone, over 2.5 weeks. Both developmental (thymus) and mature (spleen) lymphoid organs were affected, although specific effects varied with the chemical. DES significantly decreased thymocyte numbers. However, relative percentages of thymocyte subsets were not altered. While splenic cellularity and percentages of T and B cells were unchanged, splenocytes from DES-exposed mice had significantly decreased ability to proliferate in response to Concanavalin-A (Con-A). Con-A-activated splenocytes from mice treated with genistein or alpha-zearalanol had decreased levels of interferon-gamma (IFNgamma) protein in their culture supernatants compared to similar cultures from oil-treated mice. RT-PCR analysis of Con-A-activated splenocytes revealed that the expression of IFNgamma gene is altered by DES or genistein treatment. Together, these results suggest that estrogenic endocrine disruptors modulate the immune system of aged mice.
MeSH Terms
- Administration, Oral
- Aging
- Animals
- Antigens, CD
- Cell Count
- Concanavalin A
- Diethylstilbestrol
- Endocrine System
- Estrogens, Non-Steroidal
- Female
- Genistein
- Immune System
- Interferon-gamma
- Lymphocytes
- Mice
- Mice, Inbred C57BL
- Organ Size
- RNA, Messenger
- Spleen
- Thymus Gland
- Uterus
- Zeranol
Concern has been raised that environmental contaminants with estrogenic properties can alter normal sexual maturation. Monkeys, like humans, undergo a long and complex period of development during adolescence, which makes them important models for understanding exogenous estrogen effects during this period. This study examined the consequences of treatment with estrogenic agents (methoxychlor, MXC, 25 and 50 mg/kg/day; diethylstilbestrol, DES, 0.5 mg/kg/day) given in the peripubertal period (6 months before and after the expected age at menarche) to female rhesus monkeys. These treatments increased estrogen activity of serum as determined with an in vitro estrogen receptor alpha (ERa) transcription assay. DES completely suppressed adolescent growth (weight and height) and menses in a reversible manner; smaller effects of MXC on the timing of growth and menarche were also detected. Both DES and MXC led to premature emergence of a secondary sex characteristic, reddening and swelling of skin, but retarded growth of the nipple. As evaluated by ultrasound after an 8-month recovery period, uterine size was not affected by exogenous estrogen, but there was some indication of increased incidence of ovarian cysts/masses in MXC- and DES-treated groups. Ovarian cyclicity, as reflected in urinary hormone metabolites, demonstrated shorter follicular stages in the MXC-treated monkeys. In conclusion, the data indicate that DES had a striking effect on adolescent maturation and that the estrogenic pesticide MXC also altered development during this period. The pattern of effects across agents and doses may be based on specifics of estrogenic action, such as relative ERalpha and ERbeta binding and activation. Long-term consequences of this disruption of pubertal development are being studied in this cohort of monkeys as adults.
MeSH Terms
- Aging
- Animals
- Body Weight
- Diethylstilbestrol
- Eating
- Estrogen Receptor alpha
- Estrogens
- Estrogens, Non-Steroidal
- Female
- Genitalia, Female
- Growth
- Insecticides
- Macaca mulatta
- Menstruation
- Methoxychlor
- Nipples
- Phenols
- Receptors, Estrogen
- Skin
- Ultrasonography
- Uterus
Vacuolar type H( )-ATPase is involved in lumenal acidification of the epididymis. This protein is highly expressed in narrow and clear cells where it is located in the apical pole, and it contributes to proton secretion into the lumen. We have previously shown that in rats, epididymal cells rich in H( )ATPase appear during postnatal development and reach maximal numbers at 3-4 wk of age. The factors that regulate the appearance of these cells have not been investigated, but androgens, estrogens, or both may be involved. This study examined whether neonatal administration of estrogens (diethylstilbestrol [[[DES]]] or ethinyl estradiol) or an antiandrogen (flutamide), or the suppression of androgen production via administration of a GnRH antagonist (GnRHa), was able to alter the appearance of cells rich in H( )-ATPase in the rat epididymis when assessed at age 25 days. Surprisingly, all of these treatments were able to significantly reduce the number of H( )-ATPase positive cells; this was determined by immunofluorescence and confirmed by Western blotting. In contrast, neonatal coadministration of DES and testosterone maintained the expression of H( )-ATPase in the epididymis at Day 25 despite the high level of concomitant estrogen exposure. These findings indicate that androgens, acting via the androgen receptor, are essential for the normal development of epididymal cells rich in H( )-ATPase, and that treatments that interfere directly or indirectly with androgen production (GnRHa, DES) or action (flutamide, DES) will result in reduced expression of H( )-ATPase. Our findings do not exclude the possibility that estrogens can directly suppress the postnatal development of cells in the epididymis that are rich in H( )-ATPase, but if this is the case, this suppression can be prevented by testosterone administration.
MeSH Terms
- Aging
- Androgen Antagonists
- Animals
- Animals, Newborn
- Blotting, Western
- Cell Count
- Diethylstilbestrol
- Epididymis
- Ethinyl Estradiol
- Fluorescent Antibody Technique
- Flutamide
- Gonadotropin-Releasing Hormone
- Hormones
- Male
- Organ Size
- Proton-Translocating ATPases
- Rats
- Rats, Wistar
- Testis
- Testosterone
Diethylstilbestrol (DES), the first orally active artificial estrogen ever developed, was prescribed to several million pregnant women during the 1940s through the 1960s in the mistaken belief that it reduced the risk of miscarriage. In 1971, the US Food and Drug Administration contraindicated its use in pregnancy when DES was associated with the development of vaginal clear cell adenocarcinoma (CCA) in daughters exposed in utero. In daughters whose mothers took DES during pregnancy, the drug has been associated with congenital malformations of the reproductive tract, fertility problems, a possible increased risk of cervical carcinoma in situ, and a presumed lifetime risk of vaginal and cervical CCA. DES mothers have an increased risk of breast cancer (RR = 1.3). DES sons have an increased prevalence of urogenital anomalies, and a possible increased risk of testicular cancer.
MeSH Terms
- Adenocarcinoma
- Aging
- Breast Neoplasms
- Carcinoma in Situ
- Diethylstilbestrol
- Estrogen Replacement Therapy
- Female
- Genitalia, Female
- Humans
- Mothers
- Nuclear Family
- Pregnancy
- Pregnancy Complications
- Pregnancy, High-Risk
- Prenatal Exposure Delayed Effects
- Risk Factors
- Uterine Cervical Neoplasms
- Vaginal Neoplasms
Newborn rats were treated with 10 microg of diethylstilbestrol (DES) on alternate days from the 2nd to the 12th postnatal day, and the testes were sequentially examined up to 105 days of age by light, electron, and confocal laser microscopy. In control rats, spermatozoa and step 19 spermatids were observed in stage VIII seminiferous tubules at 56 days of age. Spermatogenic cells in DES-treated rats differentiated normally from birth until 21 days of age, after which differentiation continued only to the pachytene-spermatocyte stage. From this age onward, spermatogenic cells older than pachytene spermatocytes were not found until 56 days of age. After this point, the cells resumed differentiation and finally became spermatozoa by 91 days of age; that is, 35 days later than control rats. Electron and confocal laser microscopy showed that in the normal rat, the formation of the ectoplasmic specialization between adjoining Sertoli cells was observed as early as 20 days of age. In contrast, the specialization was not formed until 56 days of age in DES-treated rats. Furthermore, the delay in functional maturation of this structure as the blood-testis barrier was confirmed by intercellular tracer experiments. It is clear that neonatal administration of DES delayed the establishment of the blood-testis barrier for 4 weeks. Consequently, during this period, pachytene spermatocytes were exfoliated from the seminiferous epithelium without completion of meiosis.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- Blood-Testis Barrier
- Cytochrome c Group
- Diethylstilbestrol
- Estrogens, Non-Steroidal
- Male
- Microscopy, Confocal
- Microscopy, Electron
- Rats
- Rats, Wistar
- Sertoli Cells
- Spermatogenesis
- Testis
This study in rats sought to 1) characterize immunoexpression of estrogen receptor alpha (ERalpha) and ERss in the efferent ducts, epididymis, and vas deferens during postnatal development; 2) establish whether ER expression changed after neonatal treatment with diethylstilbestrol (DES); and 3) determine whether ER changes coincided with abnormal epididymal/vas development. Rats were administered 10 microg DES or vehicle on days 2, 4, 6, 8, 10, and 12 and were sampled on days 10, 18, 25, 35, and 90 . At all ages, ERalpha was immunoexpressed intensely in the efferent ducts. On day 10, immunoexpression of ERalpha was absent from the epididymis and vas, but was detectable on day 18 in epithelial cells in the caput, corpus, and proximal cauda. Epithelial expression of ERalpha was absent from the distal cauda and in the proximal and distal vas was confined to a band of periductal stromal cells. Thus, on day 18, the site of ERalpha expression delineated the epididymis-vas boundary. On days 25-35, epithelial expression of ERalpha was absent, but stromal expression persisted in the vas and distal cauda. In adults, immunoexpression of ERalpha in the epididymis and vas was absent. In contrast, ERbeta was immunoexpressed in epithelial cells and some stromal cells in the efferent ducts, epididymis, and vas at all ages. In the vas, stromal expression of ERalpha and ERbeta was in different layers. DES treatment caused 1) underdevelopment of the epididymal duct and reduced epithelial height in epididymis and vas; 2) coiling of the extraepididymal vas; 3) thickening of the periductal actin-free stromal layer in the distal cauda and vas; and 4) reduced cell proliferation on day 18 in the epididymis and vas, based on incorporation of bromodeoxyuridine, especially in the epithelium. These changes coincided with abnormalities in cell- and region-specific immunoexpression of ERalpha, but not ERbeta. Thus, in DES-treated rats on day 18, epithelial expression of ERalpha occurred in all regions of the epididymis and vas instead of being confined to the caput, corpus, and proximal cauda as in controls. Similarly, stromal ERalpha expression in the vas of DES-treated rats was not confined to a periductal layer as in controls, but occurred diffusely in the muscle layer. It is suggested that 1) estrogens play a role in peripubertal development of the epididymis and vas; 2) the cellular site of expression of ERalpha either plays a role in or reflects demarcation of the epididymal/vas boundary; and 3) blurring of this boundary in DES-treated rats coincides with altered ERalpha immunoexpression.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- Cell Division
- Diethylstilbestrol
- Epididymis
- Estrogen Receptor alpha
- Estrogen Receptor beta
- Estrogens, Non-Steroidal
- Immunohistochemistry
- Male
- Rats
- Rats, Wistar
- Receptors, Estrogen
- Tissue Distribution
- Vas Deferens
This study investigated whether neonatal exposure of male rats to estrogenic compounds altered pubertal spermatogenesis (days 18 and 25) and whether the changes observed resulted in long-term changes in testis size, mating, or fertility (days 90-100). Rats were treated neonatally with a range of doses (0.01-10 microg) of diethylstilbestrol (DES; administered on alternate days from days 2-12), a high dose of octylphenol (OP; 2 mg administered daily from days 2-12) or bisphenol A (Bis-A; 0.5 mg administered daily from days 2-12), or vehicle, while maintained on a standard soy-containing diet. The effect on the same parameters of rearing control animals on a soy-free diet was also assessed as was the effect of administering such animals genistein (4 mg/kg/day daily from days 2-18). Testis weight, seminiferous tubule lumen formation, the germ cell apoptotic index (apoptotic/viable germ cell nuclear volume), and spermatocyte nuclear volume per unit Sertoli cell nuclear volume were used to characterize pubertal spermatogenesis. Compared with (soy-fed) controls, DES administration caused dose-dependent retardation of pubertal spermatogenesis on day 18, as evidenced by decreases in testis weight, lumen formation, and spermatocyte nuclear volume per unit Sertoli cell and elevation of the germ cell apoptotic index. However, the two lowest doses of DES (0.1 and 0.01 microg) significantly increased spermatocyte nuclear volume per unit Sertoli cell. Similarly, treatment with either OP or Bis-A significantly advanced this and some of the other aspects of pubertal spermatogenesis. Maintenance of control animals on a soy-free diet also significantly advanced lumen formation and spermatocyte nuclear volume per unit Sertoli cell compared with controls fed a soy-containing diet. Administration of genistein reversed the stimulatory effects of a soy-free diet and significantly retarded most measures of pubertal spermatogenesis. In general, plasma FSH levels in the treatment groups changed in parallel to the spermatogenic changes (reduced when pubertal spermatogenesis retarded, increased when pubertal spermatogenesis advanced). By day 25, although the changes in FSH levels largely persisted, all of the stimulatory effects on spermatogenesis seen on day 18 in the various treatment groups were no longer evident. In adulthood, testis weight was decreased dose dependently in rats treated neonatally with DES, but only the lowest dose group (0.01 microg) showed evidence of mating (3 of 6) and normal fertility (3 litters). Animals treated neonatally with OP or Bis-A had normal or increased (Bis-A) testis weights and exhibited reasonably normal mating/fertility. Animals fed a soy-free diet had significantly larger testes than controls fed a soy-containing diet, and this difference was confirmed in a much larger study of more than 24 litters, which also showed a significant decrease in plasma FSH levels and a significant increase in body weight in the males kept on a soy-free diet. Neonatal treatment with genistein did not alter adult testis weight, and although most males exhibited normal mating and fertility, a minority did not mate or were infertile. It is concluded that 1) neonatal exposure of rats to low levels of estrogens can advance the first wave of spermatogenesis at puberty, although it is unclear whether this is due to direct effects of the estrogen or to associated elevation of FSH levels; 2) the effect of high doses of OP and Bis-A on these processes is essentially benign; and 3) the presence or absence of soy or genistein in the diet has significant short-term (pubertal spermatogenesis) and long-term (body weight, testis size, FSH levels, and possibly mating) effects on males.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- Apoptosis
- Diet
- Environmental Exposure
- Estrogens
- Fertility
- Follicle Stimulating Hormone
- Inhibins
- Male
- Organ Size
- Rats
- Rats, Wistar
- Seminiferous Tubules
- Sertoli Cells
- Soybeans
- Spermatogenesis
- Testis
Oestrogen exposure of the male during fetal/neonatal life can fundamentally alter the structure and function of the reproductive system, though how is unknown. This study examined whether such treatment was able to induce a 'female' characteristic, namely immunoexpression of progesterone receptor (PR), in the reproductive system of the male. Rats were treated on postnatal days 2, 4, 6, 8, 10 and 12 with either 10, 1 or 0.1 microg diethystilbestrol (DES) or with the vehicle (20 microl corn oil). Groups of control and treated rats were killed on days 18, 25, 35 and 90 (= adults) and tissues fixed in Bouins for immunolocalisation studies using antisera to PR (recognises A and B forms) and oestrogen receptor-beta (ER beta). PR immunoexpression was absent from all tissues studied in control rats at all ages with the exception of the parasympathetic ganglia of the prostate. In rats treated with 10 microg DES, intense immunoexpression of PR was detected in the nuclei of stromal, but not epithelial, cells of the caput and cauda epididymis, the vas deferens, seminal vesicles and at the base of the dorsolateral prostatic complex (DLPC) at day 18, but was absent from the ventral prostate and from the testis. DES induction of PR immunoexpression was evident after a single injection (on day 3) and at 18-35 days the intensity of immunoexpression was DES dose-dependent; rats treated neonatally with 0.1 microg DES showed no detectable PR immunoexpression at any age. These findings were confirmed by Western analysis which indicated that most of the PR induced was probably the B form. Co-localisation studies, using confocal microscopy, demonstrated that PR and ER beta frequently co-localised to the same stromal cells in the DLPC, epididymis and seminal vesicles of DES-treated rats at day 18, whereas epithelial cells, which also expressed ER beta, did not express PR. In the tissues studied, only occasional stromal cells expressed ER alpha in comparison to the more widespread expression of ER beta, although epithelial cell expression of ER alpha was also detected in the epididymis on day 18 (but not on day 10). In DES-treated rats, immunoexpression of PR in the reproductive tract decreased progressively in intensity from days 18-35 and was non-detectable in adulthood. In conclusion, these findings are interpreted as evidence that neonatal oestrogen treatment exerts pervasive 'reprogramming' effects throughout the reproductive system of the developing male as indicated by the induction of PR immunoexpression. This induction was restricted to stromal tissue even though both stromal and epithelial cells at most sites expressed ER beta and/or ER alpha.
MeSH Terms
- Aging
- Animals
- Diethylstilbestrol
- Estrogens, Non-Steroidal
- Genitalia, Male
- Immunohistochemistry
- Male
- Rats
- Receptors, Progesterone
- Stromal Cells
The synthetic estrogen diethylstilbestrol (DES) is a potent neonatal endocrine disruptor in the hamster. To test the specificity of this phenomenon, newborn animals were treated with 100 microgram of either DES or the natural estrogen, estradiol-17beta (E2). Of the two, neonatal DES exposure caused greater morphological disruption throughout the female reproductive tract in prepubertal animals and in adults that either retained their ovaries or were ovariectomized and then given the same levels of chronic E2 stimulation. In the uterus, a characteristic histopathological profile, including enhancement of both hyperplastic and apoptotic activity, was initiated prepubertally and exclusively in the endometrial epithelial cell compartment from the neonatally DES-treated animals and then was promoted by E2 stimulation during adulthood. Interestingly, apoptotic activity was not detected in an area of endometrial epithelium that progressed to the neoplastic state in a DES-exposed animal. Lastly, chronic estrogen induction of lactoferrin was also restricted to the DES-exposed endometrium. We conclude that 1) DES is more active than E2 as a perinatal endocrine disruptor in the hamster and 2) this experimental system should be generally useful as a means to screen compounds for such activity and then probe their mechanism of action.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- Cricetinae
- Diethylstilbestrol
- Endometrium
- Estradiol
- Estrogens, Non-Steroidal
- Female
- Genitalia, Female
- Immunohistochemistry
- Mesocricetus
- Ovariectomy
- Uterus
The goal of this study was to determine the effects of aging and sex steroids on the frequency of T cells in hen ovary. Cryostat sections of ovarian tissues of immature and laying hens and those of immature hens treated with or without diethylstilbestrol (DES) or progesterone were immunostained for T cells using mouse anti-chicken CD3 (antigen of mature T cells), CD4 (antigen of helper T cells), and CD8 (antigen of cytotoxic T cells) monoclonal antibodies. Positive cells were observed under a light microscope and counted using a computer-assisted image analyzer. The frequency of CD3( ), CD4( ), and CD8( ) cells in the ovarian stroma and theca of primary follicles was significantly greater in young laying hens than in immature and old laying hens (P < 0.01). The CD4:CD8 ratio was significantly higher in the ovarian stroma of old laying hens than that of immature hens (P < 0.01), which was due to a greater decrease of CD8( ) cells than of CD4( ) cells. The frequency of CD3( ), CD4( ), and CD8( ) cells was significantly greater in the stroma and theca of primary follicles of DES-treated birds than in those of progesterone-treated and control birds (P < 0.01). Progesterone had no significant effect on the population of each subset of T cells. These results suggest that T cell frequency increases in association with sexual maturation and decreases thereafter during aging with an increase of CD4:CD8 ratio. Also, it is likely that estrogen is one of the factors which stimulates the influx of T cells in the hen ovary.
MeSH Terms
- Aging
- Animals
- CD3 Complex
- CD4 Antigens
- CD8 Antigens
- Chickens
- Diethylstilbestrol
- Female
- Gonadal Steroid Hormones
- Ovary
- Progesterone
- T-Lymphocyte Subsets
The role of macrophages in the function of the hen ovary has not yet been described, although these cells may be an important regulator of ovarian function in mammals. The aim of this study was to determine the changes in the frequency of macrophages during ageing and follicular atresia, and the effects of sex steroids on the macrophage population in the hen ovary. Cryostat sections of ovarian tissues of immature, young laying and old laying hens and those of immature hens treated with or without diethylstilboestrol (DES) or progesterone were immunostained for macrophage cells using mouse anti-chicken macrophage monoclonal antibody. Macrophages were observed under a light microscope and counted using a computer assisted image analyser. The frequency of macrophages in both the stroma and theca of primary follicles was significantly greater in young laying hens than in immature and old laying hens and these cells were more frequent in old laying hens than in immature hens (P < 0.01). Macrophages were more frequent in atretic follicles than in normal follicles (P < 0.01). The number of macrophages in both the stroma and theca of primary follicles of DES-treated birds was significantly greater than in those of progesterone-treated and control birds (P < 0.01). Progesterone had no significant effect on the population of macrophages. These results suggest that macrophages in the ovary increase in association with sexual maturation of birds and atresia of follicles and decrease during ageing. Oestrogen may be one of the factors that affect the population of macrophages in the hen ovary.
MeSH Terms
- Aging
- Animals
- Cell Count
- Chickens
- Diethylstilbestrol
- Estrogens, Non-Steroidal
- Female
- Macrophages
- Microscopy, Phase-Contrast
- Ovary
- Oviposition
The incidence of aneuploidy in male germ cells was evaluated by analyzing extra marker chromosome(s) signal(s) in round and/or hook spermatids of transgenic mice. Two types of transgenic mice were used as models. The inserted foreign DNA (lambda-gt10LacZ shuttle vector and/or pSVc-myc plasmid) was located at the middle of the long arms of chromosome 2 (lambda DNA) and/or chromosome 8 (c-myc). The number of marker chromosomes present could easily be detected after fluorescence in situ hybridization (FISH) in testicular cells. The frequency of spontaneous aneuploidy of chromosome 2 was similar in round spermatids of lambda and lambda-myc mice. Differential involvement of chromosomes 2 and 8 was observed in both round and hook spermatids. The frequency of spontaneous aneuploidy in round spermatids was higher than that in hook spermatids. The frequency of aneuploidy of marker chromosomes was significantly higher in older mice (2 years old) than in younger ones. Diethylstilbestrol (DES)-induced aneuploidy was dose dependent, and was not influenced by the stage at which germ cells were treated with DES. These results demonstrate the usefulness of a transgenic mouse model for the study of aneuploidy in germ cells.
MeSH Terms
- Aging
- Aneuploidy
- Animals
- Diethylstilbestrol
- Germ Cells
- In Situ Hybridization
- Male
- Mice
- Mice, Transgenic
- Spermatids
- Spleen
- Testis
Immunoglobulins in the chicken ovary are important for transfer of immunity to chicks through the egg and for protection of the ovary from infection. The aim of this study was to examine the effects of ageing and oestrogen on the population of Ig-containing cells in the chicken ovary. The ovarian tissue of immature, young laying and old laying hens and that of immature birds treated with diethylstilboestrol (DES), progesterone or sesame oil (vehicle) was processed for paraffin wax sections. The sections were stained for IgG, IgM and IgA by an indirect immunostaining method and the population of cells positive for each Ig was analysed under a light microscope. The number of cells positive for IgG, IgM and IgA was significantly greater in the ovarian stromal tissue of young laying hens than in immature or old laying hens (P < 0.01). The number of IgG- and IgM-positive cells in the thecal layer of primary follicles of young laying hens was significantly greater than that in immature and old laying hens (P < 0.01) and there were significantly more (P < 0.05) IgA-positive cells in young laying hens than in immature birds. The number of IgG-, IgM- and IgA-positive cells was significantly (P < 0.01) greater in both the stromal tissue and the thecal layer of DES-treated birds than in the vehicle-treated birds. Progesterone had no significant effect (P < 0.05) on the population of Ig-positive cells. These results indicate that the number of Ig-positive cells increases as chickens mature and decreases with ageing, and that oestrogen may be involved in this process.
MeSH Terms
- Aging
- Animals
- Chickens
- Diethylstilbestrol
- Female
- Immunoglobulin A
- Immunoglobulin G
- Immunoglobulin M
- Immunoglobulins
- Immunohistochemistry
- Ovary
- Progesterone
- Theca Cells
Light and electron microscopic techniques were used to study the morphometry and dynamic changes of macrophages in the postnatal and sex hormone-treated chicken oviduct, respectively. Abundant typical macrophages, containing clear vacuoles, well-developed mitochondria, Golgi complexes and lysosomal bodies in their cytoplasms, were observed in the lamina propria of all segments of the postnatal chicken oviduct, occurring more frequently in the vaginal part. When 7-day-old chickens were injected with diethylstilbestrol (DES), and DES plus progesterone, infiltration of a significant number of macrophages in both groups, but not in controls could be seen. The light and electron microscopic structures of the macrophages in both postnatal and sex hormone-treated chicken oviduct were similar. These results show that typical macrophages are present in the chicken oviduct; their frequency of occurrence varies with different oviductal segments, and they are influenced by sex hormones.
MeSH Terms
- Aging
- Animals
- Chickens
- Diethylstilbestrol
- Estrogens, Non-Steroidal
- Female
- Macrophages
- Organelles
- Oviducts
- Progesterone
Diethylstilbestrol (DES) was administered neonatally (Days 2-12; 10 microg on alternate days) to rats, and developmental changes in Sertoli cell function were evaluated at 18, 25, and 35 days of age and compared to those observed in rats administered a GnRH antagonist (GnRHa; Days 2 and 5; 10 mg/kg) or a vehicle (controls). DES and GnRHa treatments resulted in similar reductions in both Sertoli cell numbers (40% for DES, 48% for GnRHa) and suppression of testicular growth at 18 and 25 days, though by 35 days the suppression was more pronounced (p < 0.001) in DES-treated animals. Plasma FSH levels were suppressed markedly at 18 and 25 days, but not at 35 days, in GnRHa-treated rats, whereas in DES-treated rats the FSH levels were suppressed significantly only at 35 days. Both treatments suppressed plasma levels of inhibin B, though this was more pronounced (p < 0.05) in DES- than in GnRHa-treated rats. In controls, Sertoli cell immunoexpression of inhibin alpha, sulfated glycoprotein-1 (SGP-1), and androgen receptor (AR) increased in intensity and changed to an adult, stage-dependent pattern by 25 days. In GnRHa-treated rats these changes were reduced in intensity but were similar to those in controls at 35 days. In DES-treated rats, the increase in intensity and stage-dependent pattern of immunoexpression of inhibin alpha, SGP-1, and AR were virtually absent at 25 days but were present by 35 days. Germ cell volume per Sertoli cell was reduced in GnRHa- and DES-treated rats compared with controls at 18 and 25 days but was significantly greater (p < 0. 001) in DES- than in GnRHa-treated rats at 35 days. The proportion of apoptotic to viable germ cells was increased (p < 0.01) in GnRHa- and DES-treated rats compared with controls at 18 and 25 days; but at 35 days, values in GnRHa-treated rats had declined to control values whereas those for DES-treated rats remained 10-fold elevated (p < 0.001). In adulthood, testis weight and daily sperm production were reduced by 43% and 44%, respectively, in GnRHa-treated rats, but spermatogenesis was grossly normal. Comparable changes were observed in approximately 25% of DES-treated rats, but the majority exhibited > 60% reduction in testis weight with many Sertoli cell-only tubules and very low daily sperm production. Taken together, these data are interpreted as providing evidence for direct modulation of Sertoli cell (maturational) development by DES.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- Body Weight
- Cell Count
- Diethylstilbestrol
- Estrogens
- Follicle Stimulating Hormone
- Male
- Oligopeptides
- Organ Size
- Rats
- Rats, Wistar
- Sertoli Cells
- Spermatogenesis
- Testis
Recent data suggest that estrogens play a role in regulating fluid resorption from the efferent ducts, though the biochemical mechanisms involved are unknown. The present study has used immunocytochemistry to localize a water channel protein, Aquaporin-1 (AQP-1), to the efferent ducts of male rats and marmoset monkeys from perinatal life through to adulthood and has then investigated its potential hormonal regulation in neonatal/peripubertal life, via administration of a GnRH antagonist (GnRHa) or diethylstilbestrol (DES) to rats. AQP-1 was immunoexpressed intensely in the apical brush border of the epithelium lining the efferent ducts at all ages studied, from late fetal life through puberty to adulthood. In the marmoset, but not the rat, AQP-1 was also expressed in the epithelium of the rete testis. Once the cell types within the efferent duct epithelium had differentiated, it was clear that only nonciliated cells of the rat localized AQP-1. When gonadotropin secretion was suppressed in rats by neonatal administration of GnRHa, immunoexpression of AQP-1 at age 18 and 25 days was virtually unchanged in intensity, though the efferent ducts were reduced in size. In contrast, when DES was administered neonatally to rats (up to day 12), immunoexpression of AQP-1 was reduced at day 10, virtually abolished at day 18, reduced markedly at day 25 and to a small extent at day 35; these findings were confirmed by Western blot analysis at day 18. The DES-induced decrease in immunoexpression of AQP-1 was accompanied by pronounced distension of the efferent ducts and rete, consistent with reduced fluid resorption. The epithelial cells of the efferent ducts in DES-treated rats were cuboidal rather than columnar in shape as in controls and were reduced significantly in height compared with controls at all ages through to adulthood. These findings suggest that estrogens may play a role in regulating fluid resorption from the efferent ducts during fetal/neonatal development and/or a role in the gross and functional development of the efferent ducts and rete testis. The present data also suggest that AQP-1 is one of the elements involved in the regulation of fluid resorption in the efferent ducts. The importance of fluid flow in fetal/neonatal development of the excurrent duct system of the male is also suggested by these observations.
MeSH Terms
- Absorption
- Aging
- Animals
- Animals, Newborn
- Aquaporin 1
- Aquaporins
- Blotting, Western
- Body Fluids
- Callithrix
- Diethylstilbestrol
- Epididymis
- Estrogens
- Gonadotropin-Releasing Hormone
- Immunohistochemistry
- Ion Channels
- Male
- Rats
- Rats, Wistar
- Rete Testis
Exposure to both physiological and pharmacological doses of estrogenic compounds has been reported to alter immunologic responses in humans as well as in developmental and adult murine models. Despite the current therapeutic use of potent estrogens, including diethylstilbestrol (DES), in geriatric human disorders, elucidation of the effects of these agents on the aged immune system is limited. The present report describes highly significant alterations in the thymus and bone marrow of aged mice (21 /- 1 months) exposed subacutely to DES. Severe thymic hypocellularity developed in treated mice following five consecutive days of intraperitoneal injection with 1.5 or 6.0 mg kg(-1) DES. Cell maturation within the thymus was also affected, as indicated by a significant decrease in CD4 8 cells and a concomitant increase in CD4-8- cells. Cellularity of the bone marow was unchanged by DES. However, significant changes were observed in percentages of bone marrow cells expressing surface antigens CD45 (common leukocyte lineage), Mac-1 (macrophage lineage) and CD45R (B-lineage lymphocytes). Both percentages and total numbers of cells in the spleen expressing Thy 1.2 (T-lineage lymphocytes) were also reduced. These immune changes in geriatric mice exposed to DES were similar in direction but more severe than those reported in either young adult or perinatal models. These data may suggest a need for considering the geriatric immune system separately from other age groups when determining effects of immunosuppressive or immunotoxic compound exposure.
MeSH Terms
- Aging
- Animals
- Bone Marrow Cells
- CD4 Lymphocyte Count
- CD8-Positive T-Lymphocytes
- Diethylstilbestrol
- Estrogens, Non-Steroidal
- Female
- Immunity, Cellular
- Infusions, Parenteral
- Mice
- Thymus Gland
Contrary to the common description of estrogen-induced thymus atrophy we have observed a thymus enlargement after treatment of neonatal female mice with estrogen. We now describe an age-dependent difference in the estrogen response (enlargement, atrophy) as well as mechanisms relevant to the response. Groups of female NMRI mice were treated with estrogen (diethylstilbestrol, DES) at different 5-day periods in prepubertal (day 1-5, day 6-10, day 30-34) or postpubertal life (days 48-52). All the treatment groups showed a reduced thymus weight 4 days after the last treatment but later responses differed. Neonatal DES treatment resulted in an ovary-independent thymus enlargement 8 weeks after the treatment when the cortical part was relatively larger than in controls; treatment on days 30-34 was followed by a rebound type of regeneration; the acute weight reduction after treatment on days 48-52 was normalized 16 days later. Neonatal DES treatment transiently depressed the number of thymic S phase cells 4 days after the treatment while apoptosis was similar in controls and DES females. The estrogen receptor pattern was not affected by DES. The number of white blood cells was temporarily depressed while the bone marrow cellularity was still reduced in 8-week-old females. Neonatal treatment with an LH-releasing hormone antagonist reduced thymus weight at 8 weeks but had no effect on the DES-induced enlargement. The delayed-type hypersensitivity response developed differently in controls and DES females. The thymus enlargement after neonatal estrogen treatment could be the result of an increased immigration of precursor cells into the thymus and/or a defect maturation/emigration mechanism. Further studies on different cell subsets are necessary to explain the mechanism behind the thymus enlargement.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- Apoptosis
- Bone Marrow
- Bone Marrow Cells
- Cell Cycle
- Cell Division
- Diethylstilbestrol
- Drug Administration Schedule
- Estrogens, Non-Steroidal
- Female
- Gonadotropin-Releasing Hormone
- Hormone Antagonists
- Hypersensitivity, Delayed
- Hypertrophy
- Leukocytes
- Mice
- Organ Size
- Receptors, Estrogen
- Thymus Gland
Desmosine (DES) and isodesmosine (IDES) concentration in the urine can be used as a noninvasive method of assessing degradation of mature elastin in normal and pathologic states. The present study was undertaken to determine the distribution of elastin among organs and tissues of normal hamsters, and to determine the turnover rates of two elastin-containing organs (lung, thoracic aorta) as a reflection of their contributions to DES and IDES excretion in the urine. Hamsters were metabolically labeled at 5 days of age with 14C-lysine and studied at 1.5, 4.5, 8, and 12 months of age. The aorta DES IDES-associated radioactivity did not change significantly over the age span of 1.5-12 months. Lung DES IDES-associated radioactivity decreased with a half-life of 420 days. Measurement of DES IDES pools in other tissues, with relatively low concentrations of elastin, was carried out by the isotope dilution technique. At 12 months of age, the head and paws pool, skin, skeletal muscle, gastrointestinal tract, heart-liver-kidney-spleen pool, lungs, and thoracic aorta represented 37%, 28%, 13%, 11%, 6%, 4%, and 1%, respectively, of total body DES IDES. The organs with the highest DES IDES-specific radioactivity at 12 months were heart-liver-kidney-spleen, lung, and gastrointestinal tract, with 310, 217, and 217 dpm/nmol, respectively. Skin had the lowest specific radioactivity, with 90 dpm/nmol. The specific radioactivity of DES IDES in urine was 62 dpm/nmol at 12 months, down from 251 dpm/nmol at 1.5 months. These data clearly indicate that non-lung tissues contain a high proportion of the total body DES IDES and suggest that pathology in these other pools of DES IDES could result in significant elevation of urinary DES IDES. Nevertheless, the relatively high specific radioactivity of DES IDES in lung elastin as compared with urine makes monitoring labeled urinary DES IDES in this animal model a sensitive tool for assessing elastin degradation in experimental lung disease.
MeSH Terms
- Aging
- Animals
- Aorta
- Cricetinae
- Desmosine
- Elastin
- Isodesmosine
- Kinetics
- Lung
- Skin
It is well known that the elastic property of human aorta decreases gradually with age. Since the cross-linking structures are responsible for this elasticity, age-related changes of cross-linking amino acids in human aorta were studied using a high-performance liquid chromatography (HPLC). Non-atherosclerotic areas of thoracic aorta of 27 autopsy cases which had no particular aortic disease were obtained. After acid hydrolysis, SEP-PAK silica-gel column and Fe3 /activated charcoal column pretreatment were carried out for analysis of desmosine (DES), isodesmosine (ISDES), neodesmosine (NEO), oxodesmosine (OXO) and isooxodesomosine (ISOXO), and for analysis of aldosine (ALD), respectively. These prepared samples were applied to the reversed-phase HPLC column. We also analyzed pyridinoline (PYR), a major cross-linking amino acid of collagen as an index of fibrosis. All cross-linking amino acids of elastin rapidly increased in infancy and then gradually decreased with age. In the middle- and old-age, the amount of OXO showed marked variety. PYR was little detected at 0-year-old, and then gradually increased with age. The crosslinks of elastin were rapidly formed in childhood and then decreased with age. These findings suggest that the relative increase of NEO, OXO or ISOXO to DES and ISDES is associated with age-related weakening and/or damage of elastin, and that the gradual shift from elastin- to collagen-dominant state is a possible cause of the loss of elasticity and the gain of stiffness in the aging aorta.
MeSH Terms
- Adolescent
- Adult
- Aged
- Aged, 80 and over
- Aging
- Amino Acids
- Aorta, Thoracic
- Autopsy
- Child
- Child, Preschool
- Chromatography, High Pressure Liquid
- Cross-Linking Reagents
- Desmosine
- Elasticity
- Elastin
- Humans
- Infant
- Infant, Newborn
- Isodesmosine
- Middle Aged
- Muscle Development
- Muscle, Smooth, Vascular
- Piperidines
- Pyridines
It is hypothesized that smoking-related chronic obstructive pulmonary disease (COPD) results in part from excess lung elastin degradation. Taking advantage of spirometry performed over a 12-yr period at the Normative Aging Study, we conducted a nested case-control study of elastin and collagen degradation rates in current smokers with (n = 10) and without (n = 8) rapid decline of lung function, using a biochemical assay for urinary desmosine (DES), a specific marker for mature elastin degradation, and hydroxylysylpyridinoline (HP), a specific marker for mature fibrillar collagen degradation. Mean urinary excretion of DES in rapid decliners was 36% greater than in slow decliners (9.8 /- 0.7 [mean /- SE] versus 7.2 /- 0.4 microg/g creatinine, p < 0.01); after adjustment for age and lean body mass (LBM), DES excretion in rapid decliners was 30% greater than in slow decliners (9.6 /- 0.6 versus 7.4 /- 0.7 microg/g creatinine, p = 0.06). Among rapid decliners, there was no difference in DES excretion between those with and those without computed tomogaphic evidence of emphysema. There was no significant difference between rapid and slow decliners in mean urinary excretion of HP (24.7 /- 1.4 versus 21.6 /- 1.8 nmol/mmol creatinine, p = 0.18). Among all subjects, rate of decline of FEV1 was significantly correlated with DES excretion (r = 0.61, p < 0.01). In a linear regression model adjusting for age and LBM, an increase in DES excretion of 1 microg/g creatinine was associated with an excess decline of FEV1 of 10.6 ml/yr (p = 0.04). This study provides further evidence in support of the elastase-antielastase hypothesis of the pathogenesis of COPD, and it suggests a role for elastin degradation in both emphysema and small airways disease. Moreover, it suggests that urinary DES excretion may be a useful biochemical marker for the study of interventions designed to prevent the development or progression of COPD.
MeSH Terms
- Aged
- Aging
- Case-Control Studies
- Collagen
- Desmosine
- Elastin
- Humans
- Lung
- Male
- Middle Aged
- Respiratory Function Tests
- Smoking
Neonatal exposure to certain xenobiotics has been shown to alter hepatic metabolism in adult rats in a manner that indicates long-term changes in enzyme regulation. Previously, we have observed changes in adult testosterone metabolism and in cytochrome P-450 (P-450) mRNA levels in animals neonatally exposed to phenobarbital (PB) or diethylstilbestrol (DES). In order to test for other enzyme alterations, we used Western blot procedures for specific P-450s to analyze hepatic microsomes from adult rats (24 wk old) that had been exposed neonatally to DES, PB, 7,12-dimethylbenz[a]anthracene (DMBA), or pregnenolone 16 alpha-carbonitrile (PCN). The most striking effects were observed in the DES-treated males: P-4502C6 and an immunologically similar protein were increased 60 and 90%, respectively, relative to control values, but P-4503A2 was decreased by 44%. No changes were observed in the DES-treated males in levels of P-4502E1, P-4502B, or the male-specific P-4502C13. Adult males neonatally treated with PB had 150% increase in levels of anti-P4502B-reactive protein without significant changes in the other enzymes. The DES- and DMBA-treated females had increased levels of the female-specific P-4502C12 of 38 and 48%, respectively, but no other observed alterations. The results confirm that neonatal exposure to DES or PB can cause alterations in adult hepatic cytochrome P-450 levels but show that these chemicals act on different enzymes. Neonatal DMBA resulted in changes in adult females similar to those produced by the synthetic estrogen DES, but did so at about two-thirds lower dose.
MeSH Terms
- 9,10-Dimethyl-1,2-benzanthracene
- Aging
- Animals
- Blotting, Western
- Cytochrome P-450 Enzyme System
- Diethylstilbestrol
- Female
- Male
- Microsomes, Liver
- Phenobarbital
- Postpartum Period
- Pregnancy
- Prenatal Exposure Delayed Effects
- Rats
- Rats, Sprague-Dawley
Various studies have shown a direct effect of beta estradiol on cartilage and bone. Such effects point to the possibility that specific receptors to estradiol exist in the growth plate cartilage as well as in bone. 3H-estradiol specific binding (EB) was therefore investigated in the supernatant of cartilage homogenates from the epiphyses and ribs of young growing mice and rats. High levels of EB were observed in the cytosol fraction of cartilage homogenates in the late fetal stage and in young rats and mice. The EB levels decreased gradually from late fetal stage up to 14 days of age in both groups of animals independent of their sex. Nuclear binding of 3H estradiol was also demonstrated by autoradiography in the chondrocytes of proliferating and hypertrophic zones. Estradiol binding was inhibited by high doses of unlabelled beta-estradiol, but not by alpha estradiol. Binding was also inhibited by tamoxifen and DES but not by testosterone. High levels of estradiol binding (EBS) were also observed in serum from young animals, but not in animals 2 months of age or older. Study of estradiol binding in cartilage and in serum of rats of the same age showed a significant difference in estradiol binding between these two systems. The difference in estradiol binding between serum and cartilage was seen in the response to inhibitors, Scatchard analysis, and temperature dependence. The results of our study imply that there are specific receptors for 17 beta estradiol in growth plate cartilage; they originate from chondrocytes, and their amount decreases with age. The effects of estradiol on endochondral bone growth seems therefore to be receptor mediated.
MeSH Terms
- Aging
- Animals
- Autoradiography
- Diethylstilbestrol
- Estradiol
- Female
- Growth Plate
- Male
- Mice
- Protein Binding
- Rats
- Receptors, Estradiol
- Tamoxifen
- Time Factors
- Tritium
Treatment of neonatal female mice with DES markedly reduced the activity of NK cells in adult mice. This finding was most evident in C57BL/6 and BALB/c strains, but was also found in outbred NMRI mice. The mechanisms behind the reduced NK activity was further analyzed. No evidence of DES-induced cellular or humoral suppressors of natural killing could be detected. Pregnancy was found to be without effects on NK. Poly I:C augmented the NK activity in control females but even very high doses of Poly I:C failed to increase the level of NK activity in neonatally DES-treated animals. The lack of response to boosting with Poly I:C was not due to alterations in kinetics of NK induction.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- Bone Marrow
- Cytotoxicity, Immunologic
- Diethylstilbestrol
- Female
- Interferons
- Male
- Mice
- Mice, Inbred BALB C
- Mice, Inbred C57BL
- Mitogens
- Ovary
- Poly I-C
- Pregnancy
- Spleen
The progesterone receptor (PR) in the oviducts of aged, nonlaying hens was compared to that in laying hens. The PRs in the mature oviducts of chicks that were diethylstilbestrol (DES) primed and of chicks with DES withdrawn were similarly analyzed for comparative purposes. The oviducts from the DES-withdrawn chicks and from aged nonlaying hens showed marked atrophy. The PR concentrations in the oviducts of DES-withdrawn chicks and of aged nonlaying hens were reduced to one half and one fourth those of controls, respectively. The oviduct atrophy and reduced PR concentrations in the nonlaying hen were not caused by decreased plasma estrogen since estrogen concentrations were similar in aged nonlaying hen and in the laying hens. The stabilities of the PR from laying and nonlaying hen oviducts were identical. However, analysis in vivo and in vitro revealed that the PR from the oviducts of DES-withdrawn chicks and of aged, nonlaying hens had lost the capacity for nuclear translocation and binding. This was accompanied in the DES-withdrawn chick by the inability of progesterone to alter RNA synthesis in the oviduct in vivo. This loss in nuclear binding capacity of PR in both animal models was accompanied by a loss of one of the molecular species of the PR (the B species). The loss of the B species differs from the loss of the A species that occurs during the winter in the domestic fowl. Thus, two types of regulation of the receptor may exist. The results suggest that biologically inactive receptors may explain the cessation of reproductive function in these aged animals.
MeSH Terms
- Aging
- Animals
- Cell Nucleus
- Estrogens
- Female
- Isoelectric Focusing
- Models, Biological
- Organ Size
- Oviducts
- Progesterone
- Receptors, Progesterone
Intrauterine exposure to diethylstilbestrol (DES) has been associated with ths subsequent rare development of clear cell adenocarcinoma of the vagina and cervix in exposed female progeny. These tumors have been seen thus far in patients between the ages of 7 and 29 years. In addition, nonmalignant epithelial and structural genital alterations have been frequently observed. The epithelial changes of adenosis, ectropion, metaplasia, and potential neoplasia are considered. Thus far, no increased risk of squamous cell neoplasia has been demonstrated in DES-exposed subjects. However, increased pregnancy wastage, including premature birth, does appear to be more common among DES-exposed women; this increase may be related to the structural uterine and cervical changes that have occurred. DES-exposed men have been demonstrated to have anatomic genital changes, but the effects of these changes, if any, on male infertility are not known. Masculinization of the female external genitalia, including phallic enlargement and labioscrotal fusion, has been reported following intrauterine exposure to certain progestational agents and androgens.
MeSH Terms
- Abnormalities, Drug-Induced
- Adenocarcinoma
- Adolescent
- Adult
- Aging
- Animals
- Child
- Contraceptives, Oral
- Diethylstilbestrol
- Epithelium
- Female
- Gonadal Steroid Hormones
- Humans
- Male
- Maternal-Fetal Exchange
- Pregnancy
- Prenatal Exposure Delayed Effects
- Risk
- Spermatozoa
- Uterine Cervical Neoplasms
- Vaginal Neoplasms
Previous reports have indicated that 17 beta-estradiol is unable to stimulate DNA synthesis in the neonatal rat uterus. Possible causes for this unresponsiveness that have been suggested include the ontogenic acquisition of late responses to estrogen and the sequestering of injected estradiol by the higher concentration of serum alpha-fetoprotein. To test these hypotheses we have examined the mitogenic potency of diethylstilbestrol (DES), a nonsteroidal estrogen which has a lower affinity for alpha-fetoprotein. Our results reveal that 1.0 microgram DES administered in a single injection can stimulate DNA synthesis 24 h later in 5-day-old rats, the youngest tested. By comparison, a single injection of 1.0 microgram estradiol did not stimulate DNA synthesis until rats reached an age of 10-15 days. Higher doses of estradiol administered in a single injection stimulated DNA synthesis in neonates but to a lesser extent than DES. The potency of estradiol relative to DES increased with age up to 28 days, at which time they were equipotent. The magnitude of the stimulation of DNA synthesis increased with age even with DES because control levels of DNA synthesis are high in young rats and decreased as the rats matured. These results indicate that uterine cells of neonatal rats have no inherent inability to synthesize DNA in response to estrogen and are consistent with the hypothesis that serum alpha-fetoprotein is responsible for the decreased potency of estradiol.
MeSH Terms
- Aging
- Animals
- Cell Nucleus
- DNA Replication
- Diethylstilbestrol
- Estradiol
- Female
- Kinetics
- Rats
- Uterus
Three kinds of "white eggs" and red-blooded Bombyx mutants (W1, W2, W3 and rb) which lack enzymes in tryptophan metabolism showed almost no difference in their lifespans from that of normal stock. However, other Bombyx mutants, such as Nd, Nd-s and DES-Nd, unable to synthesize fibroin, had shorter lifespans than normal. Among these fibroin-deficient mutants, only the lifespan of DES-Nd was similar to the normal. However, further analysis of the amino acid composition in the cocoon of the mutant showed that its lifespan was also dependent on its ability to synthesize fibroin-like protein. It is very likely that, different from abnormalities in amino acid metabolism, a genetic abnormality in protein metabolism affects the maintenance of adult life in silkmoths and this results in the short lifespan. Generally speaking, an abnormality in amino acid metabolism seems to have no influence on silkmoth lifespan. However, an abnormality in protein metabolism results in shortening lifespan. The greater the abnormality in protein metabolism possessed by a silkmoth, the shorter the lifespan.
MeSH Terms
- Aging
- Animals
- Bombyx
- Female
- Fibroins
- Longevity
- Male
- Peptides, Cyclic
- Sericins
- Tryptophan
Female mice belonging to NMRI stock were inoculated neonatally with daily doses of 5 micrograms diethylstilbestrol (DES) or olive oil for the first 5 days after birth and in adult life with 20 or 100 micrograms 3-methylcholanthrene (MCA) or the vehicle tricaprylin only. The cytotoxic activity of spleen natural killer (NK) cells against YAC-1 target cells was studied in a 51Cr release assay at 2, 5, 10, or 15 days after the MCA injection. A dose of 20 micrograms MCA to neonatally olive oil-injected females did not influence the NK activity, whereas an injection of 100 micrograms MCA significantly depressed the NK activity to about half the value seen in controls. This suppression was transient, and the normal level was reached again 15 days after the injection. The depressed NK activity could not be related to humoral or cellular suppressor mechanisms. A study at the single-cell level revealed that the inhibition was due to interference with the lytic step of the NK cell without affecting target-binding capacity. In vitro exposure of spleen cells from MCA-treated animals to interferon fully restored the NK activity. Neonatal DES treatment resulted in a depressed NK activity in adult females to a level about half of that seen in olive oil-injected controls. The NK activity in DES-treated females was not influenced by either 20 or 100 micrograms MCA. The MCA-induced suppression of NK activity was discussed in relation to the earlier reported difference in incidence of MCA-induced sarcomas between DES-treated and control females after they were given 20 micrograms MCA in adult life, as well as in relation to the same incidence in the 2 groups treated with 100 micrograms MCA. The results are compatible with a significant role for NK cells during MCA carcinogenesis and indicate that a possible part of the tumorigenic effect of MCA is its early suppressive effect on NK cell activity.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- Cytotoxicity, Immunologic
- Diethylstilbestrol
- Killer Cells, Natural
- Leukemia, Experimental
- Methylcholanthrene
- Mice
- Mice, Inbred Strains
- Spleen
Pregnant strain CD-1 mice were treated with diethylstilbestrol (DES) or vehicle. Their female offspring were raised to old age and autopsied when terminally ill. Squamous metaplasia and adenomyosis were more common in uteri of these old mice exposed prenatally to DES than in control mice. Tumors of the uterine horns were seen in 17 of 143 DES-exposed mice and in 3 of 64 control mice. The controls had only leiomyomas, whereas 14 of the DES-exposed mice had adenocarcinomas. There were 5 cervical adenocarcinomas and 1 vaginal adenocarcinoma among treated mice but none in the control mice. Thus the effects of prenatal exposure to DES interacted with the effects of aging to produce a relatively high frequency of uterine adenocarcinoma.
MeSH Terms
- Adenocarcinoma
- Aging
- Animals
- Body Weight
- Diethylstilbestrol
- Endometriosis
- Female
- Fetus
- Gestational Age
- Litter Size
- Metaplasia
- Mice
- Organ Size
- Pregnancy
- Uterine Neoplasms
- Uterus
C3H/HeJ and C3H/HeN female mice were fed diets containing targeted concentrations of 320 or 640 ppb diethylstilboestrol (DES) starting at 7 or 11 wk of age and continuing throughout their remaining lifespan. Regardless of the DES concentration there was a faster rate of development and higher final incidence of mammary adenocarcinomas among the C3H/HeN mice than among the C3H/HeJ mice. In C3H/HeN mice started on DES when 11 wk old, mammary tumours developed more rapidly than when treatment was started at 7 wk of age. This was also true for C3H/HeJ mice given 320 ppb DES but not for those treated with 640 ppb DES. Both age at the start of treatment and strain of C3H mice are important factors to be considered in designing experiments to study the tumorigenic activity of oestrogens such as DES.
MeSH Terms
- Adenocarcinoma
- Administration, Oral
- Aging
- Animals
- Diethylstilbestrol
- Female
- Mammary Neoplasms, Experimental
- Mice
- Mice, Inbred C3H
- Species Specificity
Diethylstilbestrol (DES) pellets were implanted in female golden hamsters on day 22 after birth. Hamsters with or without the DES pellet were hypophysectomized on day 23. Starting from day 26, the hypophysectomized hamsters were injected daily with 2.3-40 micrograms NIH-LH-S19, 6 or 18 micrograms NIAMD-oFSH-13, 50 micrograms NIAMD-Rat-FSH-B-1, or saline for 3 days. Ovarian homogenates from these hamsters on day 29 were incubated with [14C]-4-androstene-3,17-dione and enzyme activity (nmol/g/h) was estimated. The 5 alpha- and 5 beta-reductase activities decreased significantly following hypophysectomy. In the hypophysectomized hamster ovary, a distinct response to LH but not to FSH or DES in the 5 alpha-reductase activity was found. On the other hand, the 17 beta-ol-dehydrogenase activity was stimulated by FSH but not by LH or DES. The 5 beta-reductase activity was stimulated by DES, FSH or 2.3 micrograms LH but not by 7-40 micrograms LH. In the DES-treated, hypophysectomized hamster ovary, LH and FSH stimulated the 5 alpha-reductase and 17 beta-ol-dehydrogenase activities, respectively, but FSH or LH treatment had no significant effect on the 5 beta-reductase activity. These results show that the 5 alpha-reductase activity is regulated by LH, while the 17 beta-ol-dehydrogenase activity is stimulated by FSH in immature golden hamster ovary. The 5 beta-reductase activity seems to be regulated predominantly by FSH but the effect of FSH is largely mediated by estrogen.
MeSH Terms
- 17-Hydroxysteroid Dehydrogenases
- 3-Oxo-5-alpha-Steroid 4-Dehydrogenase
- Aging
- Animals
- Cricetinae
- Diethylstilbestrol
- Female
- Follicle Stimulating Hormone
- Hypophysectomy
- Luteinizing Hormone
- Mesocricetus
- Organ Size
- Ovary
- Oxidoreductases
- Uterus
The plasma and tissue concentrations of 2-methoxyestrone (2-MeOE1) and 2-hydroxyestrone (2-OHE1) were measured in immature rats. The plasma levels of 2-MeOE1 were found to be high at birth and to decrease through puberty, when the low levels found in the adult rats were achieved. 2-OHE1 was undetectable in the plasma and brain, and barely detectable in the uterus and liver. 2-MeOE1 was undetectable in the brain and uterus, but high in the liver. The affinity of 2-MeOE1 and 2-OHE1 for rat alpha-fetoprotein was found to be low, while the affinity of estradiol, estrone, 4-hydroxyestrone, and 4-fluoroestradiol was high. This data suggests that 2-OHE1 and 2-MeOE1 would be available to estrogen target tissues in the fetal and neonatal rat. Although these metabolites lack uterotropic activity they are capable of acting in the liver. It is suggested that the plasma 2-MeOE1 of neonatal rats acts as a prohormone capable of stimulating the liver and other estrogen target tissues which possess demethylating enzymes. It is pointed out that unlike estradiol the non-steroidal estrogens such as diethylstibestrol (DES) lack the ability to form two sets of catechol and guaicol metabolites, i.e. "C-2" and "C-4" metabolites with their different biological characteristics are not formed by DES.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- Brain
- Estrogens
- Estrogens, Catechol
- Estrone
- Female
- Hydroxyestrones
- Liver
- Male
- Rats
- Receptors, Estrogen
- Structure-Activity Relationship
- Tissue Distribution
- Uterine Contraction
- Uterus
Sixteen-month-old ("aged") female rats were less susceptible than 6-month-old ("mature") females to parathyroid hormone (PTH)-mediated bone resorption induced by excess dietary phosphate. Ovariectomy enhanced 45Ca loss from the bones of mature rats but not of aged rats. In mature ovariectomized (OX) females, estradiol initially suppressed phosphate-induced bone resorption but its effectiveness decreased with continued administration. Diethylstilbestrol (DES) administered continuously or on a 3-weeks-on, 1-week-off schedule in the diet suppressed the increase in 45Ca loss in OX mature females fed a high (1.2%) P diet. In contrast, DES had no consistent effect on 45Ca loss by the aged animals. Urinary cyclic adenosine monophosphate (cAMP) was increased to a similar extent in both groups by feeding excess phosphate, indicating that the reduced effect of phosphate on 45Ca loss in the aged females was due to decreased bone response to PTH. Ovariectomies produced an increase in cAMP excretion which was suppressed by DES administration. DES also suppressed the increase in urinary cAMP induced by dietary phosphate, signifying that the decrease in bone resorption produced by estrogen is associated with an inhibition either of PTH synthesis or function.
MeSH Terms
- Aging
- Animals
- Bone Resorption
- Bone and Bones
- Calcium
- Castration
- Cyclic AMP
- Diethylstilbestrol
- Estradiol
- Female
- Parathyroid Hormone
- Phosphates
- Rats
C3H/HeN-MTV female mice were fed diets containing targeted concentrations of 320 or 640 ppb diethylstilboestrol (DES) starting at 3, 5, 7 or 11 wk of age and continuing throughout their remaining lifespan. Mice were housed in either a single-corridor conventional animal room or in a double-corridor barrier-type animal room. Mice housed in the conventional animal room and started on DES at 7 or 11 week of age developed palpable mammary tumours somewhat sooner than the corresponding groups of mice kept in the barrier animal room. In mice housed in the barrier animal room and exposed to a given DES concentration, there was very little difference between mice started on DES at 3, 5 or 7 wk of age in the exposure time required for the development of palpable mammary tumours. There was a striking difference, however, between mice started on DES at 7 wk and those started at 11 wk of age in the exposure time needed before mammary tumours appeared. Mice started at 11 wk of age developed tumours with, on average, about 4 wk less exposure than did those started at 7 wk. This suggests that treatment between 7 and 11 wk of age had little or no effect on mammary tumour development. In conclusion, both animal-room environment and age at the start of DES treatment influenced the mammary tumour response in female C3H/HeN-MTV mice.
MeSH Terms
- Adenocarcinoma
- Aging
- Animals
- Diethylstilbestrol
- Environment
- Female
- Housing, Animal
- Mammary Neoplasms, Experimental
- Mice
- Mice, Inbred C3H
Polyovular follicles (PF) occur in the ovary of 30-day-old offspring of ICR/JCL mice given 4 daily subcutaneous injections of 20-2,000 micrograms diethylstilbestrol (DES)/day from days 15 to 18 of gestation. PF containing 2-9 oocytes per follicle in the prenatally DES-exposed mice are increased 33- to 112-fold as compared to controls. In 5- to 25-day-old offspring of mothers given injections of 2,000 micrograms DES/day, PF are observed 17-65 times more frequent than in controls.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- Diethylstilbestrol
- Female
- Mice
- Mice, Inbred ICR
- Ovarian Follicle
- Pregnancy
- Prenatal Exposure Delayed Effects
A standardized procedure was developed for conducting the mouse bioassay for detecting estrogenic activity in rodent diets. Studies were conducted with CD-1 mice to determine the appropriate weaning age and length of bioassay period. Uterine growth curves were generated from mice weaned at 15 days of age and fed a negative control diet until 28 days of age. These mice showed slow regular increases in uterine weights from 15 22 days of age followed by rapid uterine growth in some mice from 24 to 28 days of age. Estrogenic bioassays using female mice weaned at 15 days of age and fed the positive control diets containing 4 or 6 ppb diethylstilbestrol (DES) demonstrated significant (P less than 0.05) increases in uterine weight and in uterus to body weight (U:BW) ratios over those of mice fed the negative control diet without DES for 5, 7 or 9 days after weaning. In contrast, mice weaned at 17 days of age showed significant (P less than 0.05) increases in uterine weight and in U:BW ratios only at 5 days after weaning. Six ppb DES was required in the positive control diet to produce a 1.5 fold increase in the U:BW ratio over those of mice fed the negative control diet. It was concluded that mice should be weaned at 15 days of age and that the bioassay period should be terminated at 7 days, when the mice are 22 days old, for best reproducible results. The criteria for a valid bioassay should include the demonstration of a significant statistical increase in the U:BW ratios of mice fed the DES positive diet over those of mice fed the negative control diet.
MeSH Terms
- Aging
- Animal Feed
- Animals
- Biological Assay
- Diethylstilbestrol
- Estrogens
- Female
- Mice
- Mice, Inbred Strains
- Organ Size
- Rodentia
- Uterus
Catecholamine histofluorescence was used to examine morphology of hypothalamic tuberoinfundibular dopaminergic (TIDA) median eminence-afferent neurons (areas A12 and A14) in male Fischer 344 rats bearing prolactin (PRL)-secreting tumors. These tumors were either spontaneous, discovered in rats aged 20-32 months, or were induced by chronic diethylstilbestrol (DES) treatment in younger animals. In the latter case, histofluorescence was examined in two groups of animals: (1) those which had been treated, beginning at 6 months of age, with 8 weeks of DES, and subsequently had had the treatment discontinued for 10 months, but which continued to exhibit hyperprolactinemia; (2) animals treated continuously with DES, for 50 or 70 days. Histofluorescence was induced using either the Falck-Hillarp, aluminum-formaldehyde ('ALFA') or the formaldehyde-glutaraldehyde ('FAGLU') technique and pituitaries were examined for PRL immunoreactivity, as permitted by the fixation method chosen for histofluorescence. For selected animals, pituitaries were enzymatically dissociated, maintained in primary culture, and were assessed for PRL cell quantification, intracellular PRL content, and in vitro release. Among animals treated chronically with DES, histofluorescence was assessed by the Falck-Hillarp technique, and counts of A12 and A14 perikarya were made in each hypothalamus. Median eminence zona externa fluorescence was diminished in aged rats with normal pituitaries, in aged rats bearing spontaneous tumors, and in rats treated continuously with DES, in comparison with fluorescence in younger rats with normal pituitaries or from which DES treatment had been with-drawn. Perikaryal fluorescence showed typical cell morphology and numbers in aged rats, aged rats bearing spontaneous tumors, and in rats from which DES had been withdrawn. In one animal, treated continuously for 70 days with DES, numbers of A12 and A14 perikarya decreased. In aged rats bearing spontaneous tumors, histofluorescence morphology indicated stimulatory effects on TIDA perikarya and terminals, including increased perikaryal and terminal fluorescence intensity. The morphological results support biochemical evidence of PRL stimulation of tuberoinfundibular dopaminergic neurons, and suggest that inhibitory effects of increased PRL secretion on these hypothalamic cells in DES-induced hyperprolactinemia reflect a direct and reversible effect of DES on these neurons.
MeSH Terms
- Aging
- Animals
- Cells, Cultured
- Chronic Disease
- Diethylstilbestrol
- Dopamine
- Histocytochemistry
- Hyperprolactinemia
- Immunoenzyme Techniques
- Male
- Median Eminence
- Pituitary Neoplasms
- Prolactin
- Rats
- Rats, Inbred F344
Exposure of rats to either estrogens or antiestrogens during early postnatal development reduces subsequent uterine growth as measured by uterine weight. However, individual uterine cell types respond differently to these agents and uterine weight alone cannot discern subtle or even large alterations in individual cell populations. Using a computerized planimetric technique, we estimated the prepubertal growth of the uterine luminal epithelium, endometrial stroma, glands, and circular and longitudinal muscle after exposure of neonatal rats (postnatal days 1-5) to the estrogens 17 beta-estradiol (E2), diethylstilbestrol (DES), or ethynylestradiol (EE), and the antiestrogens tamoxifen or clomiphene citrate. On postnatal day 26, the cross-sectional areas of the luminal epithelium, endometrial stroma, and circular muscle were reduced after estrogen exposure, compared to untreated controls, while longitudinal muscle cross-sectional area was not affected. Since cell densities (cell number/unit area) were increased, these estrogen-induced area reductions demonstrate a decrease in cell size. Total cell numbers, estimated as the product of cell type areas and their respective cell densities, were also reduced by neonatal estrogen exposure. The synthetic estrogens DES and EE were more potent than E2 with respect to reduction of uterine growth. Neonatal antiestrogen exposure caused large area reductions only in the uterine glands and luminal epithelium. Little change in cell density occurred in any cell population exposed to antiestrogen. These data demonstrate that the decreased uterine growth resulting from estrogen exposure during early postnatal development is a consequence of combined hypotrophy and hypoplasia in all cell types except longitudinal muscle while antiestrogen-induced morphological alterations were limited to hypoplasia having epithelial cell specificity.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- Clomiphene
- Diethylstilbestrol
- Endometrium
- Estradiol
- Ethinyl Estradiol
- Female
- Muscle Development
- Muscle, Smooth
- Rats
- Rats, Inbred Strains
- Reference Values
- Tamoxifen
- Uterus
The volume of the sexually dimorphic nucleus of the preoptic area (SDN-POA) of the male rat brain is larger than that of the female. In the female rat, treatment with diethylstilbestrol (DES), either perinatally (from day 16 of gestation to postnatal day 10), or postnatally (birth to day 10) was equally effective in increasing the volume of SDN-POA compared to controls. Prenatal treatment (day 16 of gestation to birth) with DES also increased the volume of the SDN-POA but this increase was significantly smaller than that achieved with the other treatments. These results confirm the effectiveness of DES in increasing the volume of the SDN-POA in the female rat brain, and prove that the differentiating SDN-POA is very receptive to hormone influences in the early postnatal period.
MeSH Terms
- Aging
- Animals
- Diethylstilbestrol
- Female
- Gestational Age
- Maternal-Fetal Exchange
- Pregnancy
- Preoptic Area
- Rats
- Reference Values
- Sex Differentiation
The effects of perinatal exposure to synthetic estrogens and progestins on mammary tumorigenesis were studied in female C3H/HeN/MTV mice. Mice were treated neonatally with 0.001 microgram/day diethylstilbestrol (DES), with 15 micrograms/day 17 alpha-hydroxyprogesterone caproate (HPC), or with oil on days 1-5 of life (birth = day 1). As adults, neonatally hormone-treated mice received long-term treatment with a synthetic estrogen and progestin combination or vehicle. Animals were palpated weekly for mammary gland tumors. The effect of treatment on the probability of tumor development was examined. Neonatal treatment with a low dose of DES increased the probability of mammary-gland tumor formation, whereas neonatal treatment with HPC had a slightly protective effect on tumorigenesis. Subsequent treatment of adult mice with synthetic steroids did not affect mammary gland tumorigenesis in neonatally DES-treated or oil-treated animals. There was a significant interaction between the effect of neonatal HPC treatment and subsequent steroid treatment on mammary tumorigenesis but examination of the data indicated that this interaction was due to the protective effect of HPC in the absence of subsequent exposure to synthetic steroids and the probability of tumor appearance in mice treated with both HPC and synthetic steroids as adults did not differ from that of neonatally oil-treated controls.
MeSH Terms
- 17 alpha-Hydroxyprogesterone Caproate
- Aging
- Animals
- Animals, Newborn
- Diethylstilbestrol
- Estrogen Antagonists
- Female
- Hydroxyprogesterones
- Mammary Neoplasms, Experimental
- Mice
- Mice, Inbred C3H
C3H/HeN female mice with low murine mammary tumor virus titer (MTV-) were fed diets containing a targeted concentration of 640 ppb diethylstilbestrol [(DES) CAS: 56-53-1; 4,4'-(1,2-diethyl-1,2-ethenediyl)bis-phenol]. Mice were started on DES at 3, 5, 7, or 9 weeks of age. Some continued on the diet throughout the rest of their life-spans, whereas others were killed as soon as they had been fed DES for 2, 4, 6, 8, 10, or 12 weeks. Controls were also examined throughout the study. Among mice killed early, the only observation significantly influenced by age at the start of DES treatment was the presence or absence of corpora lutea (CL). DES did not prevent CL from appearing in mice started on DES at 7 or 9 weeks of age, but it did prevent their appearance in about 25% of the mice started at 5 weeks and in up to 75% of the mice started at 3 weeks of age. In the life-span-exposure groups, CL either disappeared or were never formed in 88% or more of the mice, regardless of age at the start of treatment. Neoplastic or presumptive preneoplastic lesions apparently influenced by DES in the life-span-treatment groups included ovarian tubular adenomas; granulosa cell tumors and luteomas; pituitary cystoid degeneration, hyperplasia, and adenomas; uterine adenocarcinomas and cervical adenosis; mesotheliomas; and mammary hyperplastic alveolar nodules (HANs) and adenocarcinomas. Luteoma and granulosa cell tumor incidences were reduced by DES, regardless of age at the start of treatment. Influence of age at the start of treatment was minimal or not apparent for mesotheliomas, uterine adenocarcinomas, or pituitary adenomas; however, pituitary cystoid degeneration and hyperplasia and cervical adenosis occurred in higher frequency and/or with shorter duration of DES exposure the earlier that treatment was started. A delay in the start of DES treatment was associated with a remarkable delay in HAN and mammary adenocarcinoma development. This was especially apparent in young mice (3-7 wk old) in which a 2-week delay in treatment resulted in a 20-week delay in HAN or tumor onset. Age at the start of treatment was a major factor in susceptibility of C3H/HeN-MTV- female mice to DES-induced mammary tumorigenesis.
MeSH Terms
- Adenocarcinoma
- Aging
- Animals
- Corpus Luteum
- Diethylstilbestrol
- Female
- Granulosa Cell Tumor
- Mammary Neoplasms, Experimental
- Mammary Tumor Virus, Mouse
- Mice
- Mice, Inbred C3H
- Ovarian Neoplasms
- Pituitary Gland
- Pituitary Neoplasms
- Thecoma
- Uterine Neoplasms
Structural malformation of the oviduct has been reported in experimental animal models and women following prenatal exposure to diethylstilbestrol (DES). To study histological changes in the oviduct in the absence of gross structural malformation, neonatal CD-1 mice were treated with DES (2 micrograms/pup/day) on days 1-5 of age. Focal epithelial hyperplasia was present at 1 month of age in 16 out of 18 (89%) of the DES-treated mice. At 4 months of age, general epithelial hyperplasia with multiple gland-like structures into and through the muscle wall of the oviduct was observed in 90% of the treated mice; by 12 months of age, epithelial hyperplasia and pseudogland formation were seen in 100% of the DES-exposed animals. Epithelial hyperplasia and gland formation were not observed in control mice. The alteration induced by DES in the differentiation and proliferation of mouse oviductal epithelium suggests that the oviduct is a target for DES toxic effects. In addition, there was a progression of the epithelial changes with age. The histological changes described in this study may be partially responsible for the decreased fertility previously reported in this mouse model. Similar changes in the oviduct of DES-exposed women remain to be determined.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- Cell Division
- Diethylstilbestrol
- Epithelial Cells
- Fallopian Tubes
- Female
- Mice
Diethylstilbestrol-4',4"-quinone (DES Q) has previously been postulated to be a reactive intermediate in diethylstilbestrol (DES) metabolism. DES is oxidized to DES Q in vitro, but the occurrence of the quinone metabolite in vivo has not yet been demonstrated due to its instability and chemical reactivity. In this report, the characteristics of in vitro formation of DES Q and the isolation of 3H-labeled DES Q from tissue extracts of hamsters injected with radiolabeled DES is described. In vitro, the time-dependent formation of DES Q as a function of microsomal protein, cofactor or substrate concentrations was demonstrated. The microsome-mediated oxidation of DES to quinone was inhibited by various compounds that also effectively inhibit the peroxidatic activity of cytochrome P-450. In vivo, the formation of DES Q occurred in all tissues investigated, livers and kidneys of male and female adult hamsters, neonates and fetuses, and in uterus and placenta. Concentrations of quinone metabolite in liver and kidney of adult hamsters after injection of 75 mumol/kg DES were 76 and 20 pmol/g tissue respectively. In neonates and fetus, concentrations of DES Q after the same dose of DES were markedly less than those in adults (0.026 and 0.047% of adult levels in neonatal liver and kidney and 0.013 and 0.016% of adult levels in fetal liver and kidney respectively). Since DES Q was also formed by fetal liver homogenate in vitro, fetal oxidizing enzymes appear to be the source of the quinone metabolite in this tissue. DES Q concentrations were also examined after injection of DES into hamsters pretreated with vitamin C or alpha-naphthoflavone, substances known to inhibit DES-induced renal carcinogenesis. Quinone metabolite levels were cut in half in response to vitamin C in correlation with the approximately 50% decrease in DES-induced renal tumors reported previously. alpha-Naphthoflavone pretreatment decreased renal and hepatic DES Q concentrations by 70 and 17% respectively, also in correlation with the known prevention of kidney tumors by this flavone. These data support a role of DES Q in DES-induced carcinogenesis. Since there is no correlation between DES Q concentrations and target site specificity of DES induced tumors, the oxidation of DES to DES Q and the genotoxicity of this metabolite may be a necessary but not sufficient event in tumor development. Hormone-dependent growth of initiated cells may also be necessary for the occurrence of cancers.
MeSH Terms
- Aging
- Animals
- Biotransformation
- Cricetinae
- Diethylstilbestrol
- Female
- Kidney
- Liver
- Male
- Mesocricetus
- Microsomes, Liver
- Oxidation-Reduction
- Tritium
Neonatal female NMRI mice (n = 16) were treated with 5 micrograms diethylstilbestrol (DES) per day, for the first 5 days after birth and killed postpubertally. Control females (n = 52) were injected with vehicle only and killed in different stages of the estrous cycle. The plasma testosterone level was significantly lower in DES females than in control females in any of the estrous phases. Ovariectomy (n = 5), adrenalectomy (n = 5) or a combination of both ablations (n = 3) did not affect the plasma testosterone in DES treated females while it was significantly reduced in control females (ovariectomy n = 5; adrenalectomy n = 9); most effective was the combination ovariectomy-adrenalectomy (n = 7). Ovarian homogenates from DES treated females (n = 10) had a significantly lower testosterone content than homogenates from control females in any phase of the estrous cycle (6-10 females per phase), which held true on both a per ovary basis and when related to ovarian weight. After a 2 h incubation in vitro, the testosterone levels had increased significantly in DES homogenates (n = 6) and to a lower extent in homogenates from control females in estrus (n = 9). No similar effect was found in homogenates from diestrus (n = 10) or proestrus (n = 9) females. The results are discussed in relation to the special ovarian morphology of adult but neonatally DES treated females and also with respect to endocrine control mechanisms.
MeSH Terms
- Adrenalectomy
- Aging
- Animals
- Animals, Newborn
- Diestrus
- Diethylstilbestrol
- Estrus
- Female
- Mice
- Ovariectomy
- Ovary
- Proestrus
- Testosterone
Animals of several species exposed perinatally to diethylstilboestrol (DES) have been evaluated for anomalies and tumours. In male offspring, anomalies of the testis and epididymis have been reported, but evidence for tumours has been very limited. Many anomalies and tumours have been recorded in female offspring, and some of these duplicate the anomalies and tumours reported in DES-exposed women, whereas others either have not yet been discovered or else do not occur in the human species. A variety of abnormal physiological responses has been identified in animals exposed perinatally to DES. There were altered levels of hormones and receptors; responses to postnatal injection of hormones were often modified; and an increased susceptibility to other carcinogens has been established. Several mechanisms have been postulated to explain tumour production later in life after perinatal exposure to DES. Deficiencies in immune function indicate a mechanism of impaired immune surveillance. The presence of DES and its metabolites in the fetus and neonate raise the issue of somatic mutation. Evidence for sister chromatid exchange, cell transformation in tissue culture and other toxic effects on chromosomes support the somatic mutation hypothesis. A third hypothesis is involvement of abnormal differentiation of the hypothalamus. Structural, hormonal and behavioural changes support this idea. Possible additional problems in humans after exposure to DES, on the basis of animal model studies, are increased tumour frequency with ageing and transmission of cancer risk to the third generation. The multigeneration effect of DES provides a model to test the mechanism of transmission of cancer risk from one generation to the next. The outcome of such experiments could have considerable impact on the understanding of the association between DES and cancer specifically and transplacental cancer generally.
MeSH Terms
- Aging
- Animals
- Carcinogens
- Diethylstilbestrol
- Disease Models, Animal
- Female
- Genital Neoplasms, Female
- Genital Neoplasms, Male
- Genitalia, Female
- Genitalia, Male
- Male
- Pregnancy
- Prenatal Exposure Delayed Effects
Estrogens are teratogens and developmental carcinogens in several species. We have used uterine growth to quantitate the potency of three estrogens [estradiol (E2), diethylstilbestrol (DES), ethynylestradiol (EE2)] during four postnatal periods (days 1-5, 10-14, 20-24, and 60-64) in the rat. Alphafetoprotein (AFP), present at high levels in neonatal serum, is thought to regulate estrogen bioavailability. Association constants for DES and EE2 were 2.7% and 4.9% of that for E2 binding to AFP, determined in a batch Sephadex equilibrium binding assay. On days 1-5, DES and EE2 were about 80-fold more potent than E2 in increasing uterine weight. As AFP levels fell, potency differences between E2 and the synthetic estrogens decreased. In the adult, which essentially lacks AFP, the three estrogens were nearly equipotent. These data are consistent with AFP regulation of estrogen potency. On days 10-14, uterine growth was less sensitive than at other ages to all three estrogens, perhaps related to uterine differentiation and/or the high endogenous serum E2 levels reported at this age. However, when we examined another uterine estrogen response, ornithine decarboxylase (ODC) induction at 6 h following estrogen injection, all three hormones were about equipotent in both neonatal and adult animals. This apparently AFP-independent event shows dissociation of ODC induction and uterine growth, which could be due to separate mechanisms for hormone entry to target tissue or subsequent intracellular events.
MeSH Terms
- Aging
- Animals
- Diethylstilbestrol
- Estradiol
- Estrogens
- Ethinyl Estradiol
- Female
- Kinetics
- Organ Size
- Ornithine Decarboxylase
- Pregnancy
- Protein Binding
- Rats
- Uterus
- alpha-Fetoproteins
The stimulative effects of both oestradiol-17 beta (E2) and diethylstilbestrol (DES) injections on the proliferation of uterine cells of mice were investigated by using 5-[125I]iodo-2'-deoxyuridine [( 125I]IdUrd) incorporation as an index. Female mice of (WB X C57BL/6)F1 were neonatally castrated, and the [125I]IdUrd uptake by the whole uterus was determined on days 1, 5, 10, 20, 30 and 40 after birth. The relative minimal dose (a minimal dose expressed per body weight, microgram/g b.wt) of E2 necessary for the maximal [125I]IdUrd uptake was much higher than the relative minimal dose of DES on days 1, 5, and 10, when the serum concentration of alpha-fetoprotein was relatively high. However, the relative minimal dose of E2 was comparable to that of DES on days 30 and 40, when the concentration of alpha-fetoprotein was negligible. The difference between the effect of injected E2 and DES during neonatal and suckling periods seems to be attributable to the presence of alpha-fetoprotein in serum, which binds to E2 but not to DES with high affinity. When the relative minimal dose of DES necessary for the maximal [125I]IdUrd uptake was examined in detail, the value was lower on days 10 and 20 of age than on days 30 and 40 of age. This might represent the increased sensitivity of uterine cells to oestrogen on days 10 and 20.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- Cell Division
- Diethylstilbestrol
- Estradiol
- Estrogens
- Female
- Idoxuridine
- In Vitro Techniques
- Mice
- Ovariectomy
- Uterus
- alpha-Fetoproteins
The presence of sex differences in hippocampal morphology and function suggests that this brain region may be sensitive to the organizational actions of gonadal steroids. We therefore examined the postnatal development of estrogen receptor (ER) in the rat hippocampal formation. ER was measured by the in vitro binding of [3H]estradiol to a cytosolic preparation. Radioinert R2858 (moxestrol) was used to determine nonspecific binding. Hippocampal ER concentrations increased from birth through postnatal day (PND) 4 when levels peaked (10.05 /- 1.2 fmol/mg protein); these were maintained through PND-7 (9.45 /- 1.4) and declined thereafter to low levels characteristic of the adult (2.05 /- 0.35). This ontogenic profile is similar to that found in several neocortical regions, as well as in the cingulate cortex, but is distinct from that observed in the hypothalamus, where ER levels remain high in the adult. Saturation analysis of PND-7 hippocampal cytosols demonstrated a single, high affinity binding site (Kd: 5.51 /- 1.7 X 10(-10) M). [3H]Estradiol binding was specific in that it was displaced by radioinert R2858, diethylstilbestrol (DES), and 17 beta-estradiol but not by nonestrogenic steroids. Significantly greater ER levels were found in hippocampal nuclear extracts from DES-treated PND-7 animals compared to controls (9.74 /- 2.27 vs. 0.49 /- 0.24 fmol/mg DNA, P less than 0.01). The presence of functional ER was also shown by the ability of receptors to be retained on DNA cellulose. DNA cellulose column chromatography elution profiles for PND-7 hippocampal and medial basal hypothalamic (MBH) cytosols following incubation with [3H]estradiol were similar. The presence of elevated hippocampal ER levels during the perinatal critical period and evidence of functional transformation to the DNA binding state following DES treatment in vivo or estrogen incubation in vitro suggests that the hippocampus is a potential substrate for estrogen-mediated organizational events.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- Cell Nucleus
- Chromatography, Affinity
- Cytosol
- Diethylstilbestrol
- Estradiol
- Estrogen Antagonists
- Ethinyl Estradiol
- Female
- Hippocampus
- Kinetics
- Male
- Pregnancy
- Rats
- Rats, Inbred Strains
- Receptors, Estrogen
- Sex Factors
Prolonged exposure to hyperoxia markedly inhibits normal lung development (alveolarization and respiratory surface area expansion) in immature animals. Since (a) hyperoxia results in excess hydroxyl radical (OH.) formation, (b) (OH.) is implicated in O2-induced lipid peroxidation and DNA alterations, and (c) both OH. formation and its interaction with DNA are Fe dependent; chelation of Fe should act to protect against pulmonary O2 toxicity and hyperoxic inhibition of lung development. We therefore treated litters of newborn rats with the iron chelator Deferoxamine mesylate (DES) (150 mg/kg/day) during a 10-day exposure to greater than 95% O2. Morphometric analysis demonstrated that compared to the mean airspace size in air control rat pups (Lm = 44.5 microns), hyperoxic exposure resulted in a 34% larger mean air space diameter in O2-saline rat lungs (59.5 microns) versus only an 11% enlargement in O2-DES lungs (51.1 microns*). Lung internal surface area (cm2) per 100-g body weight were air control = 4480, O2-saline = 3570 (decreases 20.3%), and O2-DES = 4125* (decreases 7.9%) (*p less than 0.05 versus O2-saline group). DES-treated animals also had significantly decreased lung conjugated diene levels during hyperoxic exposure and increased lung elastin content (reflective of preserved lung alveolar formation) compared to O2-saline rats. These results indicate that DES treatment substantially ameliorated the inhibitory effects of neonatal hyperoxic exposure on normal lung development.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- Deferoxamine
- Lung
- Oxygen
- Rats
- Rats, Inbred Strains
Previous results have shown that when compared to male Syrian hamsters, female Syrian hamsters have a distinct predisposition to acquire amyloidosis either normally with aging or experimentally with sodium caseinate or diethylstilbestrol (DES) treatments. In the present study, we tested the influence of testosterone on expression of amyloid to determine if this hormone was solely responsible for the sex-limited amyloidosis of the Syrian hamster. Males deprived of testosterone by castration acquired amyloid at an unusually young age, an age of onset similar to that in female hamsters. Also, the amyloidogenic effect of DES in male Syrian hamsters was inhibited by concomitant injections of testosterone, indicating that estrogens induce amyloid in male hamsters by inhibiting testosterone synthesis. When administered to female hamsters, testosterone inhibited expression of amyloid in aging female Syrian hamsters and extended the life span of this gender. Of the two components of amyloid, the major component Amyloid A-derived fibril or the minor constituent, Amyloid P component, only the P component is under sex hormone control in the Syrian hamster; testosterone inhibits the hepatic synthesis of the P component homologue (called female protein), which is normally expressed 100-200-fold greater in female vs. male Syrian hamster. In general, the serum level of female protein under various experimental conditions correlated with the presence of amyloid and indicated that in the Syrian hamster the P component homologue is of primary importance in the deposition of amyloid.
MeSH Terms
- Aging
- Alpha-Globulins
- Amyloid
- Amyloidosis
- Animals
- C-Reactive Protein
- Cricetinae
- Diethylstilbestrol
- Female
- Kidney
- Liver
- Male
- Mesocricetus
- Orchiectomy
- Reference Values
- Sex Factors
- Spleen
- Testosterone
Diethylstilbestrol (DES) treatment of female rats on postnatal days (PND) 1-5 reduces uterine growth, estrogen receptor (ER) level and gland number by PND 25, while daily DES treatment on PND 1-25 increases uterine growth 4-fold, further reduces ER level and completely suppresses gland formation. We now report the persistence of these effects in adults. By PND 60, uterine weight was 70% of controls in rats injected with DES on PND 1-5 but only 10% of controls in rats injected PND 1-10 or longer. In fact, uterine weights were the same on PND 10 and 60. Uterine gland numbers were reduced to 30% of controls in all DES-treated rats regardless of exposure length; however, luminal and glandular epithelial cell heights were reduced to less than 50 and 70%, respectively, of controls when DES was given on PND 1-25 but not when given on PND 1-5. Ovariectomy 7 days prior to sacrifice on PND 60 reduced uterine weight in controls by 67% and in rats injected with DES on PND 1-5 by 53%, but had no effect in rats injected with DES on PND 1-10. DES exposure at either PND 1-5 or 1-10 lowered ER levels by 35-50% at both 60 and 90 days. Treatment with a high dose of estradiol (E2) 1 week before sacrifice significantly down-regulated ER to the same concentration in all treatment groups at PND 60 and 90. Following E2 treatment, all groups also showed increased uterine weight at PND 60 and 90. These data show there is a short period of development (PND 5-10) in which further DES exposure indirectly inhibits uterine growth.
MeSH Terms
- Aging
- Animals
- Diethylstilbestrol
- Down-Regulation
- Female
- Organ Size
- Ovariectomy
- Rats
- Rats, Inbred Strains
- Receptors, Estrogen
- Uterus
Several Federal agencies share responsibility for enforcing laws designed to control human exposures to carcinogens. This enforcement activity often has immense effects on other factors of our environment. Absolute safety is not possible, but increased protection is obtainable. Improvements are urgently needed in our ability not only to detect but also to quantify relative risk. Resources should be directed for maximum overall effect. Relative risk must be quantified if we are to reasonably compare risk and benefit. Examples of the inexactness of current toxicological, epidemiological, and mathematical models for estimating risk due to exposures to DDT, aflatoxinb1, DES, and benzidine are presented. The impact of different laws and regulations applicable to the control of these agents is compared. Reference is made to major programs in toxicological methods for risk estimation.
MeSH Terms
- Aflatoxins
- Animals
- Benzidines
- Carcinogens, Environmental
- DDT
- Diethylstilbestrol
- Disease Models, Animal
- Dose-Response Relationship, Drug
- Environmental Exposure
- Evaluation Studies as Topic
- Goals
- Humans
- Legislation, Drug
- Life Expectancy
- Mice
- Neoplasms
- Research Design
- Risk
- Safety
- Time Factors
- United States
Hepatic histidase was used as an enzyme marker for the study of neonatal programming in intact rats. Diethylstilbestrol (DES) or 17 beta-estradiol (E2) treatment for days, 2, 4, and 6 post partum resulted in decreased histidase activities in the adult female, but no effect was seen in prepubertal male and female rats or in adult males. In contrast, similar neonatal doses of testosterone propionate (TP) had no effect on histidase. Dose-response experiments demonstrate a 3-fold greater neonatal sensitivity to DES than to E2. The action of neonatal estrogen treatment is demonstrated to be permanent and irreversible. Neonatal treatment with E2, DES, or TP resulted in decreased uterine wet weights in adult females (E2 less than DES less than TP less than controls). Circulating sera estrogen levels were lower in adult E2- and DES-treated females than in TP-treated and control females. Our results suggest that these alterations may be due to direct toxic effects on the postnatal development of the female reproductive tract and endocrine system and/or to organizational effects on nerve endings in the hypothalamus that result in programming for altered sexual differentiation of hepatic metabolism.
MeSH Terms
- Aging
- Ammonia-Lyases
- Animals
- Animals, Newborn
- Castration
- Diethylstilbestrol
- Estradiol
- Female
- Histidine Ammonia-Lyase
- Liver
- Male
- Rats
- Sex Factors
- Sexual Maturation
The changes of DNA synthesis as well as the alterations of DNA polymerase alpha and beta have been determined in oviducts from immature (35 days old), mature (about 300 days old) and senescent (about 3 years old) quails in response to diethylstilbestrol (DES). The DNA synthesis in oviducts from immature quails is strongly stimulated by DES; after 12 days, values equivalent to those determined in mature and senescent animals are reached. The DNA synthesis in immature oviducts can be reversibly blocked by progesterone administration; no influence is observed in the case of DNA synthesis in mature as well as senescent animals. During DNA synthesis in immature oviducts, stimulated with DES, DNA polymerase alpha is strongly induced. Co-administration of progesterone blocks this enzyme induction in a reversible and strong way. The activity of DNA polymerase alpha is identical in mature and in immature animals (after DES treatment for 15 days); the activity of the same enzyme in senescent animals is about 40% lower than the values found in the younger quails. The activity of DNA polymerase beta is not altered if the animals are treated with DES or with DES and progersterone; however, the basic level of the enzyme in senescent animals in 50% lower than in immature or in mature animals.
MeSH Terms
- Aging
- Animals
- Coturnix
- DNA
- DNA Polymerase I
- DNA Polymerase II
- DNA-Directed DNA Polymerase
- Diethylstilbestrol
- Female
- Genes
- Oviducts
- Progesterone
- Quail
High diethylstilboestrol (DES) binding has been demonstrated in fetal and adult sera from man, rat and mouse by equilibrium dialysis and electrophoretic techniques. In the adults of the three species and in the human fetus only albumin shows an elevated binding capacity for DES. By contrast, in the case of rat and mouse embryos there are two proteins, namely albumin and alpha-fetoprotein, which afford major and quantatively similar contributions to the binding. Human alpha-fetoprotein does not bind DES. These phenomena are analysed in relation to the estrogen binding characteristics of the alpha-fetoproteins of the three species.
MeSH Terms
- Aging
- Animals
- Binding, Competitive
- Dialysis
- Diethylstilbestrol
- Electrophoresis, Polyacrylamide Gel
- Estradiol
- Female
- Fetus
- Humans
- Mice
- Pregnancy
- Protein Binding
- Rats
- alpha-Fetoproteins