Direct Measurement of the Return Current Instability in a Laser-Produced Plasma.

Measurements were made of the return current instability growth rate, demonstrating its concurrence with nonlocal transport. Thomson scattering was used to measure a maximum growth rate of 5.1×10^{9} Hz, which was 3 times less than classical Spitzer-Härm theory predicts. The measured plasma conditions indicate the heat flux was nonlocal, and Vlasov-Fokker-Planck simulations that account for nonlocality reproduce the measured growth rates. Furthermore, the threshold for the return current instability was measured (δ_{T}=0.017±0.002) to be in good agreement with previous theoretical models.

Aerobic capacity in speed-power athletes aged 20-90 years vs endurance runners and untrained participants.

We studied relationships between age and aerobic capacity in three groups of subjects adhering to different exercise modalities. A total of 203 men aged 20-90 years were examined: 52 speed-power track and field athletes (SP), 89 endurance runners (ER) and 62 untrained individuals (UT). Maximal exercise characteristics were obtained during a graded treadmill test until exhaustion: oxygen uptake (VO2max), heart rate (HRmax), oxygen pulse (O2 Pulsemax) and maximal distance (Distmax). Information about training history and weekly training amount was collected. A linear model of regression was adopted. VO2max in SP was lower than in ER, but significantly higher than in UT. The cross-sectional rates of decline in body mass-adjusted VO2max and Distmax were significantly smaller in SP than in ER and UT. About 80 years of age, the levels of VO2max and Distmax reached similar values in SP and ER. The decline in HRmax, but not in O2 Pulsemax was suggested as a cardiac adaptation accounting for between-group differences in VO2max loss. Weekly training volume was a significant positive predictor of age-related changes in aerobic capacity. In conclusion, not only endurance, but also speed-power exercise appears adequate to ensure an elevated aerobic capacity at old age.

In vitro metabolism of testosterone in the horse liver and involvement of equine CYPs 3A89, 3A94 and 3A95.

Testosterone (TES) 6-ß-hydroxylation is a significant metabolic step in the biotransformation of TES in human liver microsomes and reflects cytochrome P450 (CYP) 3A4/5 specific metabolic activity. Several CYP3A enzymes have been annotated in the horse genome, but functional characterization is missing. This descriptive study investigates TES metabolism in the horse liver in vitro and the qualitative contribution of three CYP3A isoforms of the horse. Metabolism of TES was investigated by using equine hepatocyte primary cultures and liver microsomes. Chemical inhibitors were used to determine the CYPs involved in TES biotransformation in equine microsomes. Single CYPs 3A89, 3A94, and 3A95, recombinantly expressed in V79 hamster lung fibroblasts, were incubated with TES and the fluorescent metabolite 7-benzyloxy-4-trifluoromethylcoumarin (BFC). The effect of ketoconazole and troleandomycin was evaluated on single CYPs. Testosterone metabolites were analyzed by HPLC and confirmed by GC/MS. In hepatocyte primary cultures, the most abundant metabolite was androstenedione (AS), whereas in liver microsomes, 6-ß-hydroxytestosterone showed the largest peak. Formation of 6-ß-hydroxytestosterone and 11-ß-hydroxytestosterone in liver microsomes was inhibited by ketoconazole, troleandomycin, and quercetin. Equine recombinant CYP3A95 catalyzed 11-ß-hydroxylation of testosterone (TES). Metabolism of BFC was significantly inhibited by ketoconazole in CYP3A95, whereas troleandomycin affected the activities of CYP3A94 and CYP3A95. Both inhibitors had no significant effect on CYP3A89. Metabolic reactions and effects of inhibitors differed between the equine CYP3A isoforms investigated. This has to be considered in future in vitro studies.

Bone density and neuromuscular function in older competitive athletes depend on running distance.

UNLABELLED: Individuals who are involved in explosive sport types, such as 100-m sprints and long jump, have greater bone density, leg muscle size, jumping height and grip strength than individuals involved in long-distance running. INTRODUCTION: The purpose of this study is to examine the relationship between different types of physical activity with bone, lean mass and neuromuscular performance in older individuals. METHODS: We examined short- (n = 50), middle- (n = 19) and long-distance (n = 109) athletes at the 15th European Masters Championships in Poznan, Poland. Dual X-ray absorptiometry was used to measure areal bone mineral density (aBMD) and lean tissue mass. Maximal countermovement jump, multiple one-leg hopping and maximal grip force tests were performed. RESULTS: Short-distance athletes showed significantly higher aBMD at the legs, hip, lumbar spine and trunk compared to long-distance athletes (p ≤ 0.0012). Countermovement jump performance, hop force, grip force, leg lean mass and arm lean mass were greater in short-distance athletes (p ≤ 0.027). A similar pattern was seen in middle-distance athletes who typically showed higher aBMD and better neuromuscular performance than long-distance athletes, but lower in magnitude than short-distance athletes. In all athletes, aBMD was the same or higher than the expected age-adjusted population mean at the lumbar spine, hip and whole body. This effect was greater in the short- and middle-distance athletes. CONCLUSIONS: The stepwise relation between short-, middle- and long-distance athletes on bone suggests that the higher-impact loading protocols in short-distance disciplines are more effective in promoting aBMD. The regional effect on bone, with the differences between the groups being most marked at load-bearing regions (legs, hip, spine and trunk) rather than non-load-bearing regions, is further evidence in support of the idea that bone adaptation to exercise is dependent upon the local loading environment, rather than as part of a systemic effect.

Equine cytochrome P450 2B6--genomic identification, expression and functional characterization with ketamine.

Ketamine is an anesthetic and analgesic regularly used in veterinary patients. As ketamine is almost always administered in combination with other drugs, interactions between ketamine and other drugs bear the risk of either adverse effects or diminished efficacy. Since cytochrome P450 enzymes (CYPs) play a pivotal role in the phase I metabolism of the majority of all marketed drugs, drug-drug interactions often occur at the active site of these enzymes. CYPs have been thoroughly examined in humans and laboratory animals, but little is known about equine CYPs. The characterization of equine CYPs is essential for a better understanding of drug metabolism in horses. We report annotation, cloning and heterologous expression of the equine CYP2B6 in V79 Chinese hamster fibroblasts. After computational annotation of all CYP2B genes, the coding sequence (CDS) of equine CYP2B6 was amplified by RT-PCR from horse liver total RNA and revealed an amino acid sequence identity of 77% and a similarity of 93.7% to its human ortholog. A non-synonymous variant c.226G>A in exon 2 of the equine CYP2B6 was detected in 97 horses. The mutant A-allele showed an allele frequency of 82%. Two further variants in exon 3 were detected in one and two horses of this group, respectively. Transfected V79 cells were incubated with racemic ketamine and norketamine as probe substrates to determine metabolic activity. The recombinant equine CYP2B6 N-demethylated ketamine to norketamine and produced metabolites of norketamine, such as hydroxylated norketamines and 5,6-dehydronorketamine. V(max) for S-/and R-norketamine formation was 0.49 and 0.45nmol/h/mg cellular protein and K(m) was 3.41 and 2.66µM, respectively. The N-demethylation of S-/R-ketamine was inhibited concentration-dependently with clopidogrel showing an IC(50) of 5.63 and 6.26µM, respectively. The functional importance of the recorded genetic variants remains to be explored. Equine CYP2B6 was determined to be a CYP enzyme involved in ketamine and norketamine metabolism, thus confirming results from inhibition studies with horse liver microsomes. Clopidogrel seems to be a feasible inhibitor for equine CYP2B6. The specificity still needs to be established with other single equine CYPs. Heterologous expression of single equine CYP enzymes opens new possibilities to substantially improve the understanding of drug metabolism and drug interactions in horses.

Hypoxanthine as a predictor of performance in highly trained athletes.

Purine metabolism reflects the exercise-induced muscle adaptations and training status. This study evaluated the utility of plasma hypoxanthine in the prediction of actual sport performance. We studied male athletes: 28 triathletes (21.4±2.9 years), 12 long-distance runners (23.2±1.9 years), 13 middle-distance runners (22.9±1.8 years) and 18 sprinters (22.0±2.7 years). Season-best race times were considered, achieved over standard triathlon, 5 000 m, 1 500 m and 100 m, respectively. Incremental treadmill test was administered to determine maximum and "threshold" oxygen uptake. Resting and post-exercise plasma concentrations of hypoxanthine, xanthine, uric acid and lactate were measured as well as resting erythrocyte hypoxanthine-guanine phosphoribosyltransferase activity. Simple and multiple regression analyses were used to identify significant contributors to the variance in performance. Hypoxanthine considered alone explained more variance in triathletes, long-distance runners, middle-distance runners and sprinters (r 2=0.81, 0.81, 0.88 and 0.78, respectively) than models based on aerobic capacity and lactate (R 2=0.51, 0.37, 0.59 and 0.31, respectively). Combining purine metabolites and cardiorespiratory variables resulted in the best prediction (R 2=0.86, 0.93, 0.93 and 0.91; r=0.93, 0.96, 0.96 and 0.95, respectively). In summary, hypoxanthine is a strong predictor of performance in highly trained athletes and its prediction ability is very high regardless of sport specialization, spanning the continuum from speed-power to endurance disciplines.

Development and evaluation of RADA-PDGF2 self-assembling peptide hydrogel for enhanced skin wound healing.

Background: Wound healing complications affect numerous patients each year, creating significant economic and medical challenges. Currently, available methods are not fully effective in the treatment of chronic or complicated wounds; thus, new methods are constantly sought. Our previous studies showed that a peptide designated as PDGF2 derived from PDGF-BB could be a promising drug candidate for wound treatment and that RADA16-I can serve as a release system for bioactive peptides in wound healing. Based on that, in this work, we designed a new self-assembling hydrogel RADA-PDGF2, connecting both peptides by a sequence specific for neutrophil elastase, and evaluated its activity in wound healing. Methods: The physicochemical properties of the designed scaffold were analyzed using transmission electron microscopy, atomic force microscopy, cryoSEM microscopies, and circular dichroism spectroscopy. The enzymatic cleavage was performed using human neutrophil elastase and monitored using high-performance liquid chromatography and MS spectroscopic techniques. The aforementioned techniques (HPLC and MS) were also used to assess the stability of the peptide in water and human plasma. The biological activity was analyzed on human skin cells using a colorimetric XTT test, collagen synthesis evaluation, and a migration assay. The biocompatibility was analyzed with LDH cytotoxicity assay and flow cytometric analysis of activation of immune cells. Finally, RADA-PDGF2 activity in wound healing was checked in a mouse dorsal skin injury model. Results: The analysis showed that RADA-PDGF2 can self-assemble, form a hydrogel, and release a bioactive sequence when incubated with human elastase. It shows pro-proliferative and pro-migratory properties and accelerates wound closure in the mouse model compared to RADA16-I. In addition, it is not cytotoxic to human cells and does not show immunogenicity. RADA-PDGF2 seems to be a promising drug candidate for wound management.

Recommendations for epidemiological studies on COPD.

The prevalence of chronic obstructive pulmonary disease (COPD) has been extensively studied, especially in Western Europe and North America. Few of these data are directly comparable because of differences between the surveys regarding composition of study populations, diagnostic criteria of the disease and definitions of the risk factors. Few community studies have examined phenotypes of COPD and included other ways of characterising the disease beyond that of spirometry. The objective of the present Task Force report is to present recommendations for the performance of general population studies in COPD in order to facilitate comparable and valid estimates on COPD prevalence by various risk factors. Diagnostic criteria in epidemiological settings, and standardised methods to examine the disease and its potential risk factors are discussed. The paper also offers practical advice for planning and performing an epidemiological study on COPD. The main message of the paper is that thorough planning is worth half the study. It is crucial to stick to standardised methods and good quality control during sampling. We recommend collecting biological markers, depending on the specific objectives of the study. Finally, studies of COPD in the population at large should assess various phenotypes of the disease.

The European Sleep Apnoea Database (ESADA): report from 22 European sleep laboratories.

The European Sleep Apnoea Database (ESADA) reflects a network of 22 sleep disorder centres in Europe enabled by a COST action B26 programme. This ongoing project aims to describe differences in standard clinical care of patients with obstructive sleep apnoea (OSA) and to establish a resource for genetic research in this disorder. Patients with suspected OSA are consecutively included and followed up according to local clinical standards. Anthropometrics, medical history, medication, daytime symptoms and sleep data (polysomnography or cardiorespiratory polygraphy) are recorded in a structured web-based report form. 5,103 patients (1,426 females, mean±sd age 51.8±12.6 yrs, 79.4% with apnoea/hypopnoea index (AHI) ≥5 events·h(-1)) were included from March 15, 2007 to August 1, 2009. Morbid obesity (body mass index ≥35 kg·m(-2)) was present in 21.1% of males and 28.6% of females. Cardiovascular, metabolic and pulmonary comorbidities were frequent (49.1%, 32.9% and 14.2%, respectively). Patients investigated with a polygraphic method had a lower AHI than those undergoing polysomnography (23.2±23.5 versus 29.1±26.3 events·h(-1), p<0.0001). The ESADA is a rapidly growing multicentre patient cohort that enables unique outcome research opportunities and genotyping. The first cross-sectional analysis reveals a high prevalence of cardiovascular and metabolic morbidity in patients investigated for OSA.

A pre-registered short-term forecasting study of COVID-19 in Germany and Poland during the second wave.

Disease modelling has had considerable policy impact during the ongoing COVID-19 pandemic, and it is increasingly acknowledged that combining multiple models can improve the reliability of outputs. Here we report insights from ten weeks of collaborative short-term forecasting of COVID-19 in Germany and Poland (12 October-19 December 2020). The study period covers the onset of the second wave in both countries, with tightening non-pharmaceutical interventions (NPIs) and subsequently a decay (Poland) or plateau and renewed increase (Germany) in reported cases. Thirteen independent teams provided probabilistic real-time forecasts of COVID-19 cases and deaths. These were reported for lead times of one to four weeks, with evaluation focused on one- and two-week horizons, which are less affected by changing NPIs. Heterogeneity between forecasts was considerable both in terms of point predictions and forecast spread. Ensemble forecasts showed good relative performance, in particular in terms of coverage, but did not clearly dominate single-model predictions. The study was preregistered and will be followed up in future phases of the pandemic.

Prevalence, severity and underdiagnosis of COPD in the primary care setting.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is a common disease with a steadily increasing prevalence and mortality. However, recent epidemiological estimates differ depending on the population studied and methods used. AIM: To investigate the prevalence, severity and burden of COPD in a primary care setting. METHODS: From 4730 patients registered in a single primary care practice, all 2250 patients aged 40 years or more were invited to participate. Participants completed a questionnaire on smoking, respiratory symptoms, education and social status. A physical examination was followed by pre- and post-bronchodilator (BD) spirometry. RESULTS: Of the eligible patients, 1960 (87%) participated. 92% of spirometric tests met the ATS criteria. Airflow limitation was demonstrated in 299 (15%) of the participants pre-BD and in 211 (11%) post-BD. COPD was diagnosed in 183 patients (9.3%). Of these patients, the degree of post-BD airflow limitation was mild in 30.6%, moderate in 51.4%, severe in 15.3% and very severe in 2.7%. Only 18.6% of these patients had previously been diagnosed with COPD; almost all of these had severe or very severe airflow limitation. As a result of the study, a diagnosis of asthma was made in 122 patients. CONCLUSIONS: The prevalence and underdiagnosis of COPD in adult patients in this primary care setting made case finding worthwhile. Large numbers of newly detected patients were symptomatic and needed treatment. Limiting investigations to smokers would have reduced the number of COPD diagnoses by 26%.

Medico-legal implications of sleep apnoea syndrome: driving license regulations in Europe.

BACKGROUND: Sleep apnoea syndrome (SAS), one of the main medical causes of excessive daytime sleepiness, has been shown to be a risk factor for traffic accidents. Treating SAS results in a normalized rate of traffic accidents. As part of the COST Action B-26, we looked at driving license regulations, and especially at its medical aspects in the European region. METHODS: We obtained data from Transport Authorities in 25 countries (Austria, AT; Belgium, BE; Czech Republic, CZ; Denmark, DK; Estonia, EE; Finland, FI; France, FR; Germany, DE; Greece, GR; Hungary, HU; Ireland, IE; Italy, IT; Lithuania, LT; Luxembourg, LU; Malta, MT; Netherlands, NL; Norway, EC; Poland, PL; Portugal, PT; Slovakia, SK; Slovenia, SI; Spain, ES; Sweden, SE; Switzerland, CH; United Kingdom, UK). RESULTS: Driving license regulations date from 1997 onwards. Excessive daytime sleepiness is mentioned in nine, whereas sleep apnoea syndrome is mentioned in 10 countries. A patient with untreated sleep apnoea is always considered unfit to drive. To recover the driving capacity, seven countries rely on a physician's medical certificate based on symptom control and compliance with therapy, whereas in two countries it is up to the patient to decide (on his doctor's advice) to drive again. Only FR requires a normalized electroencephalography (EEG)-based Maintenance of Wakefulness Test for professional drivers. Rare conditions (e.g., narcolepsy) are considered a driving safety risk more frequently than sleep apnoea syndrome. CONCLUSION: Despite the available scientific evidence, most countries in Europe do not include sleep apnoea syndrome or excessive daytime sleepiness among the specific medical conditions to be considered when judging whether or not a person is fit to drive. A unified European Directive seems desirable.

Case-control study of radon and lung cancer in New Jersey.

Radon is known to cause lung cancer in humans; however, there remain uncertainties about the effects associated with residential exposures. This case-control study of residential radon and lung cancer was conducted in five counties in New Jersey and involved 561 cases and 740 controls. A year long alpha-track detector measurement of radon was completed for approximately 93% of all residences lived in at the time of interview (a total of 2,063). While the odds ratios (ORs) for whole data were suggestive of an increased risk for exposures >75 Bq m(-3), these associations were not statistically significant. The adjusted excess OR (EOR) per 100 Bq m(-3) was -0.13 (95% CI: -0.30 to 0.44) for males, 0.29 (95% CI: -0.12 to 1.70) for females and 0.05 (95% CI: -0.14 to 0.56) for all subjects combined. An analysis of radon effects by histological type of lung cancer showed that the OR was strongest for small/oat cell carcinomas in both males and females. There was no statistical heterogeneity of radon effects by demographic factors (age at disease occurrence, education level and type of respondent). Analysis by categories of smoking status, frequency or duration did not modify the risk estimates of radon on lung cancer. The findings of this study are consistent with an earlier population-based study of radon and lung cancer among New Jersey women, and with the North American pooling of case control radon seven studies, including the previous New Jersey study. Several uncertainties regarding radon measurements and assumptions of exposure history may have resulted in underestimation of a true exposure-response relationship.

Testosterone metabolism of equine single CYPs of the 3A subfamily compared to the human CYP3A4.

Cytochrome P450 enzymes (CYPs) are responsible for the phase I metabolism of drugs, xenobiotics and endogenous substances. Knowledge of single CYPs and their substrates is important for drug metabolism, helps to predict adverse effects and may prevent reduced drug efficacy in polypharmacy. In this study, three equine isoenzymes of the 3A subfamily, the equine flavoprotein NADPH-P450 oxidoreductase (POR), and the cytochrome b5 (CYB5) were cloned, sequenced and heterologously expressed in a baculovirus expression system. Testosterone, the standard compound for characterization of the human CYP3A4, was used to characterize the newly expressed equine CYPs. The metabolite pattern was similar in equine and the human CYPs, but the amounts of metabolites were isoform-dependent. All equine CYPs produced 2-hydroxytestosterone (2-OH-TES), a metabolite never described in equines. The main metabolite of CYP3A4 6ß-hydroxytestosterone (6ß-OH-TES) was measured in CYPs 3A95 and 3A97 with levels close to the detection limit. Ketoconazole inhibited 2-OH-TES in the human CYP3A4 and the equine CYP3A94 and CYP3A97 completely, whereas a 70% inhibition was found in CYP3A95. Testosterone 6ß- and 2-hydroxylation was significantly different in the equine CYPs compared to CYP3A4. The expression of single equine CYPs allows characterizing drug metabolism and may allow prevention of drug-drug interactions.

The interrelationship between markers of inflammation and oxidative stress in chronic obstructive pulmonary disease: modulation by inhaled steroids and antioxidant.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is accompanied by both airway and systemic inflammation and by oxidative stress. This study aimed to characterise the relationship between oxidative stress and inflammatory components in induced sputum and blood. MATERIAL & METHODS: We studied blood and sputum samples from stable COPD patients (mean FEV1 60.5+/-7.5% predicted) at baseline (no treatment) and after 10 weeks treatment with either inhaled steroid, fluticasone propionate (FP) (1000 microg/d) or 10 weeks treatment with N-acetylcysteine (600mg/d) (NAC). We assessed the inflammatory markers (IL-8, ECP, sICAM-1, NE) in sputum and serum and we compared them with blood markers of oxidative stress (SOD, GPx, TEAC, albumin, vitamin E and A). RESULTS: At baseline blood sICAM-1 correlated with IL-8 levels (P<0.01, r = 0.62) and negatively with GPx (P<0.01, r = -0.63) and with TEAC (P<0.05, r = -0.53). TEAC correlated positively with GPx (P<0.01, r = 0.70). Correlation between sICAM and IL-8 disappeared after NAC treatment. The correlation between sICAM and GPx disappeared after FP treatment. The correlation between TEAC and GPx was maintained after both NAC and FP. CONCLUSIONS: The relationship between markers of inflammation, adhesion and antioxidant capacity is significantly modulated by treatment with N-acetylcysteine or inhaled corticosteroids.

16 Alpha-[125I]iodo-11 beta-methoxy-17 beta-estradiol: a radiochemical probe for estrogen-sensitive tissues.

We have synthesized an analog of 16 alpha-iodoestradiol, 11 beta-methoxy-16 alpha-iodo-estra-1,3,5-(10)triene-3,17 beta-diol (16 alpha-iodo-11 beta-methoxyestradiol), as a potential radiopharmaceutical for the in vivo imaging of estrogen-sensitive tissues. This steroid was synthesized labeled with 125I by halogen exchange of the stable intermediate 11 beta-methoxy-16 beta-bromo-17 beta-estradiol with Na125I. The halogen exchange reaction produces the radioiodinated steroid with a 65-80% yield in 3 h. This rapid synthesis and purification of the 125I-labeled estrogen permits a similar synthesis with 123I, a radioisotope with excellent properties for imaging. The 11-methoxy analog is a highly potent estrogen that binds to the estrogen receptor with an affinity equal to that of estradiol. In vivo, 11 beta-methoxy-16 alpha-[125I]iodoestradiol concentrates in an estrogen receptor-dependent manner in the uterus, producing remarkably sustained and much higher uterus to blood ratios than 16 alpha-[125I]iodoestradiol. Thus, this radiosteroid shows great promise, both as a research probe of the estrogen receptor and as a clinical tool for the imaging of estrogen-responsive tumors.

Characterization of 11 beta-methoxy-16 alpha-[125I]iodoestradiol binding: neuronal localization of estrogen-binding sites in the developing rat brain.

The binding properties of the gamma-emitting 125I-labeled 11 beta-methoxy analog of 16 alpha-iodoestradiol, 11 beta-methoxy-16 alpha-iodoestradiol (MIE2), were characterized for its use in vivo as a ligand for the measurement and localization of estrogen-binding sites. In binding displacement studies, MIE2 bound to rat, rabbit, and human estrogen receptors with high affinity. Association of MIE2 with uterine cytosol estrogen receptors reached maximum values within 30 min at 25 C. At 0-4 C, association was much slower, with maximum binding values not achieved until 16-24 h after the start of the incubation. Once formed, the MIE2-estrogen receptor complex was quite stable at 0-4 C (t1/2 much greater than 24 h). At 25 C, dissociation of MIE2-estrogen receptor complexes occurred nearly 3 times more slowly than that of E2-estrogen receptor complexes (t1/2, 3.3 vs. 1.2 h). The iodinated estrogen was highly specific for the estrogen receptor and did not bind appreciably to androgen, progestin, or glucocorticoid receptors or to either human sex hormone-binding globulin or rat alpha-fetoprotein. MIE2 is also not a ligand for human sex hormone-binding globulin. Dose-dependent uptake of [125I]MIE2 into pituitary and brain cell nuclei was observed after its in vivo administration to 25-day-old female rats. In 10-micron brain sections from immature female rats treated with [125I]MIE2 (7.5 microCi/g BW), regional localization of estrogen-sensitive brain areas could be obtained by autoradiography using LKB Ultrofilm with an exposure time of only 16 h. In comparison, after an identical dose of 16 alpha-[125I]iodoestradiol, an exposure time of 72 h was required to achieve an image of similar density. Combined autoradiographic and immunocytochemical studies in 5- to 11-day-old female rats demonstrated nuclear binding of [125I]MIE2 in cells immunoreactive for neurofilament protein but not glial fibrillary acidic protein, indicating that estrogen receptors in the developing postnatal brain are restricted to neurons and are not present in astroglial cells. The biological characteristics of [125I]MIE2 combined with its high specific activity make it an estrogenic probe with a wide range of possible uses for the study of estrogen action in the developing brain as well as other estrogen target tissues.

Regional sex differences in cell nuclear estrogen-binding capacity in the rat hypothalamus and preoptic area.

Estrogen binding was compared in cell nuclear KCl extracts from microdissected brain regions of gonadectomized-adrenalectomized male and female rats treated with a near-saturating dose of 17 beta-estradiol. Injection of 3.6 or 36.0 micrograms 17 beta-estradiol/kg BW, iv, 1 h before death resulted in a higher level of estrogen binding in the periventricular preoptic area (PVPOA), medial preoptic area, and ventromedial nucleus of the hypothalamus (VMN) of the female than in comparable tissue samples from the male. No significant sex differences in nuclear estrogen binding were observed in the arcuate-median eminence region, bed nucleus of the stria terminalis, or corticomedial amygdala. Scatchard analysis of saturation binding data revealed that the sex differences in cell nuclear estrogen binding in the PVPOA, medial preoptic area, and VMN reflect a difference in binding capacity rather than binding affinity. These in vitro biochemical findings were confirmed by autoradiographic studies. Gonadectomized-adrenalectomized animals were injected with 125I-labeled 11 beta-methoxy-16 alpha-iodoestradiol (2.0 micrograms/kg BW). Thin frozen sections (10 microns) through the preoptic area and hypothalamus were thaw-mounted onto microscope slides, then exposed against LKB Ultrofilm for 21 days. The autoradiographic images exhibited similar silver distributions and densities in males and females in the arcuate-median eminence region bed nucleus of the stria terminalis, and amygdala. However, 11 beta-[125I]methoxy-16 alpha-iodoestradiol uptake was lower in males than in females in the PVPOA and VMN. These results suggest that sex differences in responsiveness to estrogen stimulation in the rat may be due in part to sex differences in estrogen-binding capacity in specific regions of the hypothalamus that play important roles in the control of pituitary function and reproductive behaviors.