Effect of a single administration of ketoconazole on total and physiologically free plasma testosterone and 17β-oestradiol levels in healthy male volunteers

Summary The effect of a single oral dose of 400 mg ketoconazole, given as an 80 mg/ml suspension, on total and physiologically free (i.e. non-sex hormonebound) testosterone and 17-oestradiol has been investigated in 6 healthy male volunteers. The two steroids fell to nadir levels of 18 and 60% of their respective initial concentrations 6 hours after drug intake, and then completely recovered. Although in vitro slight displacement of testosterone from the sex-hormone binding globulin, by high doses of ketoconazole was found, the physiologically free concentration of testosterone in vivo was closely correlated with that of the total hormone, suggesting that there is no direct interference with sex-hormone binding globulin in vivo. Plasma LH and FSH were not significantly modified by treatment. The effect of ketoconazole on plasma oestradiol levels was less pronounced and was not clearly related to a block of the aromatase system, as reported in vitro.This work was presented in part at the International Symposium on the Regulation of Androgen Actions, Montreal, 29th June–1st July 1984     

Effect of 17β-oestradiol and ginsenoside on osteoporosis in ovariectomised rats

To study the anti-osteoporosis effects and mechanism of action of oestradiol (E₂) and ginsenoside (tR), we measured the bone mineral densities (BMD) of lumbar vertebra and tibia and analysed the tibia histological morphological data, as well observed the activity and the number of osteoblasts and the activity of alkaline phosphatase (ALP) and the concentration of cAMP. Results showed that E₂ (400 μg kg^? 1 week^? 1) and tR (10, 20, 30 mg kg^? 1 day^? 1) were able to countervail the decreasing in BMDs of lumbar vertebra and tibia induced by OVX in rats (P < 0.05); E₂ (0.1 μmol l^? 1) and ginsenoside Rg₁ (1 μmol l^? 1 and 10 μmol l^? 1) were able to increase the number of osteoblasts, the activity of ALP and the concentration of intercellular cAMP in cultured osteoblast cells. The present findings suggest that E₂ and tR have an anti-osteoporosis effect in ovariectomised rats.     

Gender differences in response to chronic treatment with 17β-oestradiol and 17β-aminoestrogen pentolame on hemostasis in rats

Objectives:

This work evaluated chronic treatment with 17β-oestradiol (E₂) and 17β-aminoestrogen pentolame (AEP) on prothrombin time (PT), activated partial thromboplastin time (aPTT), thrombin time (TT), and fibrinogen concentration (FIB). Male (M) and ovariectomized (Ovx) Wistar rats were used to explore gender differences in the pharmacological response.

Materials and Methods:

Rats (n = 12-18) were treated every third day during three months with E₂ (1, 10, 100 μg/kg), AEP (1, 10, 100, 500 μg/kg) or vehicle (propylenglycol 1 ml/ kg). PT, aPTT, TT, and FIB were measured using standardized techniques.

Results:

Chronic treatment with E₂ in male rats increased PT (4-7%; P < 0.05), decreased aPTT (9%; 100 μg/kg; P < 0.05) and decreased TT (5% at 100 μg/Kg; P < 0.05). Chronic treatment with E₂ in ovariectomized female rats decreased PT (3-4%; P < 0.05), did not induce significant changes on aPTT and decreased TT in a dose dependent manner (12-27%; P < 0.05). Chronic treatment with AEP in male rats did not alter PT, increased aPTT in a dose dependent manner (5-16%; P < 0.05), and decreased TT (5%; 500 μg/Kg; P < 0.05) while in female ovariectomized rats it decreased PT (5-9%; P < 0.05), increased aPTT (8-13%; P < 0.05) and decreased TT (6-13%; P < 0.05). E₂ and AEP decreased FIB in M and Ovx animals. Decreases in FIB by E₂ were more pronounced in male (15-18% P < 0.05) than in ovariectomized rats (10-14% P < 0.05). E₂ showed more potency than AEP, lowering FIB at 1 and 10 μg/kg doses. Both estrogens decreased FIB in ovariectomized animals (E₂, 10-14%, P < 0.05; AEP, 9% P < 0.05) and were reverted by increasing dosage.

Conclusions:

Gender influenced response to chronic treatment with E₂ and AEP on hemostatic parameters. PT and aPTT were the most affected parameters, demonstrating non-equivalence in the pharmacological response of M and Ovx rats.KEY WORDS: 17β-aminoestrogens, gender, hemostasis, oestradiol, rat     

Allopregnanolone concentration and mood—a bimodal association in postmenopausal women treated with oral progesterone

Rationale Allopregnanolone effects on mood in postmenopausal women are unclear thus far.Objectives Allopregnanolone is a neuroactive steroid with contradictory effects. Anaesthetic, sedative, and anxiolytic as well as aggressive and anxiogenic properties have been reported. The aim of this study is to compare severity of negative mood between women receiving different serum allopregnanolone concentrations during progesterone treatment.Materials and methods A randomized, placebo-controlled, double-blind, crossover study of postmenopausal women (n=43) treated with 2 mg estradiol daily during four treatment cycles. Oral micronized progesterone at 30, 60, and 200 mg/day, and placebo were added sequentially to each cycle. Participants kept daily symptom ratings using a validated rating scale. Blood samples for progesterone and allopregnanolone analyses were collected during each treatment cycle.Results During progesterone treatment, women had significantly higher negative mood scores when allopregnanolone serum concentration was in the range of 1.5–2 nmol/l compared to lower and higher concentrations. In addition, women displayed a significant increase in negative mood during the progesterone treatment period, compared to the estradiol-only period when 30 mg progesterone daily was used. On the other hand, treatment with higher doses of progesterone had no influence on negative mood.Conclusions Mood effects during progesterone treatment seem to be related to allopregnanolone concentration, and a bimodal association between allopregnanolone and adverse mood is evident.     

Effect of 17β-oestradiol on cytokine-induced nitric oxide production in rat isolated aorta

1. Studies were performed on isolated aortic rings without endothelium to investigate the effect of 17β-oestradiol on cytokine-induced nitric oxide production by the inducible nitric oxide synthase (iNOS).
2. Treatment of the isolated aortic rings with interleukin-1β (IL-1β, 20 μ ml⁻¹) led to the expression of iNOS mRNA and protein, as well as significant nitrite accumulation in the incubation media and suppression of phenylephrine (1 nM–10 μM)-evoked contraction.
3. Cycloheximide (1 μM), a protein synthesis inhibitor, prevented iNOS protein expression, nitrite accumulation and the suppression of contractility by IL-1β on the isolated aortic rings. 17β-oestradiol (1 nM–10 μM) and the partial oestrogen receptor agonist 4-OH-tamoxifen (1 nM–10 μM) produced concentration-dependent inhibition of IL-1β-induced nitrite accumulation and restored vasoconstrictor responsiveness to phenylephrine, similar to the iNOS inhibitor aminoguanidine (100 μM).
4. Semiquantitative PCR demonstrated decreased iNOS mRNA in the IL-1β-induced and 17β-oestradiol-treated rings. Western blot analysis of rat aorta homogenates revealed that 17β-oestradiol treatment resulted in a reduction in IL-1ß-induced iNOS protein level.
5. Incubation with tumour necrosis factor α (TNFα, 1 ng ml⁻¹) resulted in significant nitrite accumulation in the incubation media and suppression of the smooth muscle contractile response to phenylephrine, similar to IL-1β. The effects of TNFα were also inhibited by co-incubation of the rings with 17β-oestradiol and 4-OH-tamoxifen (1 μM).
6. The anti-transforming growth factor-β1 (TGF-β1) antibody, which inhibited TGF-β1-induced suppression of nitrite production from IL-1β-treated vascular rings, did not affect the inhibitory action of 17β-oestradiol, suggesting that the effect of oestrogen on iNOS inhibition was not mediated by TGF-β1.
7. These results show that the ovarian sex steroid, 17β-oestradiol is a modulator of cytokine-induced iNOS activity in rat vascular smooth muscle and its mechanism of action involves decrease of iNOS mRNA and protein.

     

Exposure study to examine chemosensory effects of ɛ-caprolactam in healthy men and women

Context: ε-Caprolactam is an important industrial chemical with a relatively low human toxicity; of importance is the irritations that occur after exposure to ε-caprolactam as aerosols or vapors.

Objective: The aim of this study was to examine symptoms and objective effects, which occur on the mucous membranes of the eyes and the upper respiratory tract.

Methods: A total of 52 healthy volunteers (26 women and 26 men, aged between 19 and 50 years) were exposed by random to different ε-caprolactam concentrations (0.05, 0.5 and 5.0?mg/m³) and the control condition (0.0?mg/m³) for 6?h on four consecutive days. Eye blinking frequency, tear film break-up time, eye redness, nasal flows and resistance, olfactory function as well as total protein and interleukin-8 in nasal lavage fluid were determined daily before, during and after exposure. Questionnaires were used to record both subjective symptoms and personality factors.

Results: There were no significant specific effects on the subjective and objective endpoints examined. Statistical analysis yielded no evidence of concentration-response relationships. Evaluation of olfactory symptoms showed that the duration of the stay in the chamber and not the ε-caprolactam concentration was decisive for the perception of “impure air”. Personality factors had no significant influence on the reported symptoms.

Conclusions: Exposure to ε-caprolactam concentrations of 5.0?mg/m³ at maximum for 6?h did not cause chemosensory effects on the upper respiratory tract or eyes of healthy volunteers. Therefore, the concentration of 5.0?mg/m³ corresponds to the No Observed Adverse Effect Level (NOAEL).     

Dynamic oxidative stress and DNA damage induced by oestrogen deficiency and protective effects of puerarin and 17β-oestradiol in ovariectomized rats

Oxidative stress plays an essential role in the pathogenesis of cardiovascular diseases and osteoporosis resulting from oestrogen deficiency in the postmenopausal period. In this report, we observed a dynamic change of oxidative stress and DNA damage after ovariectomy in female rats. We then compared phytoestrogen puerarin and 17β‐oestradiol (E₂) in their effects on oestrogen deficiency‐induced oxidative stress and DNA damage. Serum total antioxidant capacity (TAC), malondialdehyde (MDA) and lymphocytes DNA damage (comet%) were measured. There was a gradual increase in oxidative stress in the ovariectomized (OVX) rats over time after ovariectomy, as compared to rats receiving sham operation. OVX rats that were on puerarin and E₂ showed increased TAC and decreased MDA in the serum, as well as decreased lymphocytes comet%. Puerarin appeared to have a more powerful protective effect on DNA oxidative damage than E₂. The study indicates that postmenopausal women may benefit from phytoestrogen puerarin.     

Effects of single doses of rabeprazole 20 mg and esomeprazole 40 mg on 24-h intragastric pH in healthy subjects

Objective To compare antisecretory effects of single doses of rabeprazole and esomeprazole.Methods Open, randomised, 2-way crossover, clinical pharmacology study. 24 healthy subjects (10 men; mean age 26.2 y) received a single dose of rabeprazole 20 mg or esomeprazole 40 mg, with a 14-day ‘washout’. Intragastric pH was recorded continuously from 24 h before to 24 h after dosing.Results Mean intragastric pH was higher after esomeprazole than rabeprazole during 0–5 h after dosing (P=0.0001); the reverse was true from 14–24 h (P=0.0002). Mean % time pH>3 and >4 was greater after esomeprazole than rabeprazole during 0–14 h (P=0.041 and 0.044), but the reverse was true during 14–24 h (P=0.0005 and 0.001). In the 0–24 h interval as a whole, there was no difference between treatments in mean pH or % time pH>3 or >4.Conclusion Single-dose rabeprazole 20 mg was as effective as esomeprazole 40 mg in increasing intragastric pH and maintaining pH>3 and >4, despite the 2-fold difference in dose.Supported by Eisai Ltd., London, UK     

The effect of progesterone and 17-β estradiol on membrane-bound HLA-G in adipose derived stem cells

Membrane-bound HLA-G (mHLA-G) discovery on adipose derived stem cells (ADSCs) as a tolerogenic and immunosuppressive molecule was very important. Many documents have shown that HLA-G expression can be controlled via some hormones such as progesterone (P4) and estradiol (E2). Therefore, this study was designed to evaluate progesterone and estradiol effects on mHLA-G in ADSCs at restricted and combination concentrations. Three independent cell lines were cultured in complete free phenol red DMEM and subcultured to achieve suffi cient cells. These cells were treated with P4, E2 and P4 plus E2 at physiologic and pregnancy concentrations for 3 days in cell culture conditions. The HLA-G positive ADSCs was measured via monoclonal anti HLA-G-FITC/MEMG-09 by means of flow cytometry in nine groups. Data were analyzed by one way ANOVA and Tukey''s post hoc tests. There were no signifi cant values of the mean percentage of HLA-G positive cells in E₂-treated and the combination of P4 plus E2-treated ADSCs compared to control cells (p value>0.05) but P4 had a signifi cant increase on mHLA-G in ADSCs (p value<0.05). High P4 concentration increased mHLA-G but E2 and the combination of P4 plus E2 could not change mHLA-G on ADSCs.     

The neuroprotective effects of progesterone on traumatic brain injury： current status and future prospects

Traumatic brain injury is the leading cause of morbidity and mortality in young adults. The secondary injury in traumatic brain injury consists of a complex cascade of processes that simultaneously react to the primary injury to the brain. This cascade has been the target of numerous therapeutic agents investigated over the last 30 years, but no neuroprotective treatment option is currently available that improve neurological outcome after traumatic brain injury. Progesterone has long been considered merely a female reproductive hormone. Numerous studies, however, show that progesterone has substantial pleiotropic properties as a neuroprotective agent in both animal models and humans. Here, we review the increasing evidence that progesterone can act as a neuroprotective agent to treat traumatic brain injury and the mechanisms underlying these effects. Additionally, we discuss the current progress of clinical studies on the application of progesterone in the treatment of traumatic brain injuries.     

Research on the pharmacokinetics of single dose and multipledose of gemifloxacin in healthy Chinese male subjects

AIM： To investigate the pharmacokinetics and safety of gemifloxacin in healthy Chinese subjects and provide some theoretic bases for its reasonable application in clinic. METHODS： （1） 12 healthy Chinese male subjects were enrolled in this study with an open label, 3-period crossover oral single dosing pharmacokinetic study. The subjects sequentially took 3 doses of gemifloxacin （ 160, 320 and 480 mg） according to the randomization schedule. （2）20 subjects were chosen to participate in a randomized, double blind, multiple dosing pharmacokinetic study. The subjects were orally given 320 mg gemifloxacin or matching placebo once daily for 7 consecutive days. Clinical observation and laboratory test were performed during the study for the assessment of adverse events. The concentrations of gemifloxacin in serum and urine were determined by high performance liquid chromatogram （HPLC）. The pharmacokinetic parameters were analysed by 31797 analysis software. RESULTS： （1）The pharmacokinetic courses following single crossover oral dose of 160, 320 and 480 mg were all in accordance with the two-compartment model.[第一段]     

Pharmacokinetics of guaifenesin,pseudoephedrine and hydrocodone in a combination oral liquid formulation,administered as single and multiple doses in healthy Chinese volunteers,and comparison with data for individual compounds formulated as Antuss®

1. A new oral liquid formulation combining guaifenesin, pseudoephedrine and hydrocodone is effective in improving the symptoms of common cold. The pharmacokinetic properties of the individual components were evaluated in a randomized, open-label, four-period study in 12 healthy Chinese volunteers following single and multiple doses. The data were compared with data for the individual ingredients in Antuss®.

2. In the single-dose period, exposure levels (AUC and C_max) for guaifenesin, pseudoephedrine and hydrocodone increased directly as the dose of the oral liquid formulation increased from 5 to 15?mL. Only minor amounts of guaifenesin and hydrocodone were excreted in urine (～0.10% and 4.66%, respectively). Pseudoephedrine was mainly excreted unchanged, with 44.95% of the dose excreted in urine within 24?h. After multiple dosing, there was no obvious accumulation of any drug, as assessed by AUC. When considering C_max, there was a trend toward accumulation of hydrocodone and pseudoephedrine. The pharmacokinetic profiles of guaifenesin and pseudoephedrine in the oral liquid formulation were similar to those in the branded preparation, Antuss®.

3. The newly developed oral liquid formulation combining guaifenesin, pseudoephedrine and hydrocodone was safe and well tolerated and might provide a reliable alternative to the branded formulation for patients with common colds.     

Metabolism of the plasticizer and phthalate substitute diisononyl-cyclohexane-1,2-dicarboxylate (DINCH®) in humans after single oral doses

Hexamoll^® DINCH^® (diisononyl-cyclohexane-1,2-dicarboxylate) is a new high-molecular-weight plasticizer and a phthalate substitute. In this study, the metabolism of DINCH^® was investigated by oral dosage of three male volunteers with approximately 50 mg Hexamoll^® DINCH^® (resulting in individual doses between 0.552 and 0.606 mg/kg body weight). Their urine samples were consecutively collected over 48 h. In analogy to di-iso-nonylphthalate (DINP) metabolism, we quantified the simple monoester mono-isononyl-cyclohexane-1,2-dicarboxylate (MINCH) and its secondary oxidized metabolites with HPLC–MS/MS via isotope dilution analysis. Additionally, we quantified the unspecific full breakdown product, cyclohexane-1,2-dicarboxylic acid (CHDA), via standard addition. All postulated metabolites were present in all samples analyzed. The unspecific CHDA was identified as the major urinary metabolite representing 23.7 % of the dose as the mean of the three volunteers (range 20.0–26.5 %). 14.8 % (11.3–16.7 %) of the dose was excreted as monoesters with oxidative modifications, in particular OH-MINCH 10.7 % (7.7–12.9 %), oxo-MINCH 2.0 % (1.5–2.6 %) and carboxy-MINCH 2.0 % (1.8–2.3 %). Less than 1 % was excreted as the simple monoester MINCH. In sum, 39.2 % (35.9–42.4 %) of the DINCH^® dose was excreted as these metabolites in urine within 48 h. Over 90 % of the metabolites investigated were excreted within 24 h after application. The secondary oxidized metabolites, with elimination half-times between 10 and 18 h, proved to be apt and specific biomarkers to determine DINCH^® exposure. With this study, we provide reliable urinary excretion factors to calculate DINCH^® intakes based on these metabolites in environmental and occupational studies.     

Non-genomic vasorelaxant effects of 17β-estradiol and progesterone in rat aorta are mediated by L-type Ca2+ current inhibition

Aim:

The sex hormones 17β-estradiol (βES) and progesterone (PRG) induce rapid non-genomic vasodilator effects which could be protective for the cardiovascular system. The purpose of this study was to analyze the mechanisms underlying their vasodilator effect in rat aortic smooth muscle preparations.

Methods:

Endothelium-denuded aorta artery rings were prepared from male Wistar rats and incubated in an organ bath. The contractions of the preparation were recorded through isometric transducers. The effects of the hormones on K⁺ current and L-type Ca²⁺ current (LTCC) were analyzed by using the whole cell voltage-clamp technique in A7r5 cells.

Results:

Both βES and PRG (1–100 μmol/L) concentration-dependently relaxed the endothelium-denuded aortic rings contracted by (–)-Bay K8644 (0.1 μmol/L) or by KCl (60 mmol/L). The IC₅₀ values of the two hormones were not statistically different. The K_V channel blocker 4-aminopyridine (2 mmol/L), BK_Ca channel blocker tetraethylammonium (1mmol/L) and K_ATP channel blocker glibenclamide (10 μmol/L) did not significantly modify the relaxant effect of the hormones. On the other hand, the blockage of the intracellular βES and PRG receptors with estradiol receptor antagonists ICI 182,780 (1 μmol/L) and PRG receptor antagonist mifepristone (30 μmol/L), respectively, did not significantly modify the relaxant action of the hormones. In A7r5 cells, both the hormones (1–100 μmol/L) rapidly and reversibly inhibited the basal and BAY-stimulated LTCC. However, these hormones had no effect on the basal K⁺ current.

Conclusion:

The vasorelaxant effects of βES and PRG are due to the inhibition of LTCC. The K⁺ channels are not involved in the effects.     

Combinatory effects of PBDEs and 17β-estradiol on MCF-7 cell proliferation and apoptosis

In the present work, we analyzed whether polybrominated diphenyl ethers (PBDEs) (47, 99, 100 and 209) interfere with the effect of 17β-estradiol on the proliferation and apoptosis of the MCF-7 cell line. MCF-7 cells were cultured in DMEM without phenol red; upplemented with 5% charcoal-treated fetal bovine serum for 3 days with 10 nM 17β-estradiol; with 0.1 μM, 0.5 μM or 1 μM of the tested PBDE congeners; or with both 17β-estradiol and a congener. Cell proliferation was determined by measuring BrdU incorporation, and cell apoptosis was measured by caspase-9 activity. No PBDE congener had an effect on basal cell proliferation, but they all significantly decreased basal caspase-9 activity. An additive anti-apoptotic activity and ability to induce cell proliferation was observed in the presence of 17β-estradiol.