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1.
Mol Cell Endocrinol ; 362(1-2): 110-9, 2012 Oct 15.
Article in English | MEDLINE | ID: mdl-22687883

ABSTRACT

In the ovary, oocytes are surrounded by follicle cells and arrested in prophase of meiosis I. Although steroidogenic activity of follicle cells is involved in oogenesis regulation, clear qualitative and quantitative data about the steroid content of follicles are missing. We measured steroid levels of Xenopus oocytes and follicles by gas chromatography-mass spectrometry. We show that dehydroepiandrosterone sulfate is the main steroid present in oocytes. Lower levels of free steroids are also detected, e.g., androgens, whereas progesterone is almost undetectable. We propose that sulfatation is a protective mechanism against local variations of active steroids that could be deleterious for follicle-enclosed oocytes. Steroid levels were measured after LH stimulation, responsible for the release by follicle cells of a steroid signal triggering oocyte meiosis resumption. Oocyte levels of androgens rise slowly during meiosis re-entry whereas progesterone increases abruptly to micromolar concentration, therefore representing the main physiological mediator of meiosis resumption in Xenopus oocyte.


Subject(s)
Dehydroepiandrosterone Sulfate/metabolism , Meiosis , Oocytes/metabolism , Pregnenolone/metabolism , Xenopus laevis/physiology , Animals , Dehydroepiandrosterone Sulfate/isolation & purification , Female , Gonadal Hormones/isolation & purification , Gonadal Hormones/metabolism , Gonadal Hormones/physiology , Luteinizing Hormone/pharmacology , Luteinizing Hormone/physiology , Oocytes/drug effects , Oocytes/physiology , Ovary/cytology , Ovulation , Pregnenolone/isolation & purification , Pregnenolone/physiology , Steryl-Sulfatase/antagonists & inhibitors , Sulfonic Acids/pharmacology
2.
Age (Dordr) ; 33(4): 565-78, 2011 Dec.
Article in English | MEDLINE | ID: mdl-21222044

ABSTRACT

A major problem of ageing is progressive impairment of neuronal function and ultimately cell death. Since sex steroids are neuroprotective, their decrease with age may underlie age-related neuronal degeneration. To test this, we examined Purkinje cell numbers, plasma sex steroids and cerebellar neurosteroid concentrations during normal ageing (wild-type mice, WT), in our model of precocious ageing (Rora(+/sg), heterozygous staggerer mice in which expression of the neuroprotective factor RORα is disrupted) and after long-term hormone insufficiency (WT post-gonadectomy). During normal ageing (WT), circulating sex steroids declined prior to or in parallel with Purkinje cell loss, which began at 18 months of age. Although Purkinje cell death was advanced in WT long-term steroid deficiency, this premature neuronal loss did not begin until 9 months, indicating that vulnerability to sex steroid deficiency is a phenomenon of ageing Purkinje neurons. In precocious ageing (Rora(+/sg)), circulating sex steroids decreased prematurely, in conjunction with marked Purkinje cell death from 9 months. Although Rora(+/sg) Purkinje cells are vulnerable through their RORα haplo-insufficiency, it is only as they age (after 9 months) that sex steroid failure becomes critical. Finally, cerebellar neurosteroids did not decrease with age in either genotype or gender; but were profoundly reduced by 3 months in male Rora(+/sg) cerebella, which may contribute to the fragility of their Purkinje neurons. These data suggest that ageing Purkinje cells are maintained by circulating sex steroids, rather than local neurosteroids, and that in Rora(+/sg) their age-related death is advanced by premature sex steroid loss induced by RORα haplo-insufficiency.


Subject(s)
Aging/physiology , Cell Death/physiology , Cell Survival/physiology , Cerebellum/metabolism , Gonadal Steroid Hormones/physiology , Nuclear Receptor Subfamily 1, Group F, Member 1/physiology , Purkinje Cells/physiology , Animals , Castration , Cell Count , Cerebellum/cytology , Estradiol/blood , Female , Gonadal Steroid Hormones/metabolism , Hormone Replacement Therapy , Male , Mice , Mice, Mutant Strains , Mice, Neurologic Mutants , Progesterone/blood , Purkinje Cells/cytology , Testosterone/blood
3.
Am J Obstet Gynecol ; 203(5): 477.e1-9, 2010 Nov.
Article in English | MEDLINE | ID: mdl-20691412

ABSTRACT

OBJECTIVE: Experimental data have revealed the critical role played by 2-methoxy-estradiol, a metabolite of 17ß-estradiol, in the pathophysiology of preeclampsia. We used gas chromatography/mass spectrometry to measure a whole panel of hormonal steroids in the plasma from women during the third trimester of their pregnancy. STUDY DESIGN: The population study consists of 24 pregnant patients with different outcomes: normal, or complicated by isolated preeclampsia or by severe preeclampsia with Hemolysis Enzyme Liver Low Platelets (HELLP) syndrome. RESULTS: 17ß-estradiol was reduced by 50% in isolated preeclampsia, and by 70% in severe preeclampsia with HELLP syndrome (normal: 8.54 ± 0.9 ng/mL; isolated preeclampsia: 4.65 ± 1.0 ng/mL; severe preeclampsia with HELLP syndrome: 2.64 ± 0.4 ng/mL), as is estrone. Downstream, 2-methoxy-estradiol was decreased only in severe preeclampsia with HELLP syndrome. The concentrations of estrone and 17ß-estradiol precursors were comparable between groups, suggesting that placental aromatase is deficient in preeclampsia. CONCLUSION: The gradual decrease of estrogen levels with increasing severity of preeclampsia suggests an impairment of placental steroidogenesis.


Subject(s)
Aromatase/blood , Estradiol/blood , HELLP Syndrome/blood , Pre-Eclampsia/blood , Pregnancy Trimester, Third/blood , Adult , Analysis of Variance , Aromatase/deficiency , Estrone/blood , Female , Gas Chromatography-Mass Spectrometry , Humans , Pilot Projects , Pregnancy
4.
J Steroid Biochem Mol Biol ; 122(4): 149-58, 2010 Oct.
Article in English | MEDLINE | ID: mdl-20678573

ABSTRACT

The contribution of the two corticosteroid (mineralocorticoid and glucocorticoid) receptor (MR and GR) pathways to the function and regeneration of the sciatic nerve was investigated. We found that the corticosterone-inactivating enzyme 11ß-hydroxysteroid dehydrogenase type 2 (HSD2) was up-regulated 7 days after lesion in freeze-injured nerve. The maintenance of a low intracellular level of corticosterone by HSD2 activity in the regenerating nerve is concordant with the improvement of nervous function in injured animals (as measured by walking ability) after treatment by the GR antagonist mifepristone and with the reduction in GR participation in accumulation of the mRNA for numerous endogenous genes (from the renin-angiotensin system and other classical mineralocorticoid-responsive genes), in the same animals. Furthermore, using the MR antagonist spironolactone, we demonstrated that MR plays an active role in the function of the intact sciatic nerve: MR is required for walking ability and participates in the control of the accumulation of the mRNA for several endogenous genes. However, after injury, changes in gene expression cannot be fully explained by changes in MR/GR activity, due to an HSD2 effect, and other signalling pathway(s) induced by the lesion likely combine with the effect of the corticosteroid receptors.


Subject(s)
Nerve Regeneration , Receptors, Glucocorticoid/metabolism , Receptors, Mineralocorticoid/metabolism , Sciatic Nerve/physiology , 11-beta-Hydroxysteroid Dehydrogenase Type 2/genetics , 11-beta-Hydroxysteroid Dehydrogenase Type 2/metabolism , Animals , Cells, Cultured , Corticosterone/metabolism , Cyclic AMP/metabolism , Gene Expression , Mineralocorticoid Receptor Antagonists , RNA, Messenger/genetics , Rats , Rats, Sprague-Dawley , Receptors, Glucocorticoid/antagonists & inhibitors , Renin-Angiotensin System , Schwann Cells/cytology , Schwann Cells/metabolism , Sciatic Nerve/metabolism , Sodium Channels/genetics , Sodium Channels/metabolism , Sodium-Potassium-Exchanging ATPase/genetics , Sodium-Potassium-Exchanging ATPase/metabolism , Walking
5.
Arch Orthop Trauma Surg ; 130(1): 41-5, 2010 Jan.
Article in English | MEDLINE | ID: mdl-19333607

ABSTRACT

OBJECTIVE: Epoietin (EPO) effectively decreases perioperative blood transfusion requirements. We evaluated the feasibility of using EPO beta to increase haemoglobin levels before total hip replacement (THR) and reduce transfusion. MATERIALS AND METHODS: One hundred and seventy-one patients undergoing primary THR were included in this prospective observational study. Patients with initial haemoglobin level <130 g l−1 received EPO beta subcutaneously in a four-dose regimen. Haemoglobin levels were measured at baseline, before surgery, and at postoperative days 1 and 5. RESULTS: Twenty-eight patients received EPO beta preoperatively, 143 were not treated including 42 with initial haemoglobin level <130 g l−1. Haemoglobin increased by 29 ± 9 g l−1 in the low Hb + EPO group versus 2 ± 1 g l−1 for the other patients. In the low Hb + EPO group 3.6% were transfused, in the low HB group 45.2% (P = 0.001) and in the normal Hb group 11.9% (P = 0.2). CONCLUSIONS: Preoperative EPO beta increases haemoglobin level and reduces transfusion frequency in anaemic patients before total joint replacement. It could play a major role in the management of perioperative blood loss in orthopaedic surgery.


Subject(s)
Arthroplasty, Replacement, Hip , Erythropoietin/therapeutic use , Hematinics/therapeutic use , Adult , Aged , Aged, 80 and over , Blood Loss, Surgical/statistics & numerical data , Chi-Square Distribution , Female , Humans , Male , Middle Aged , Preoperative Care , Prospective Studies , Recombinant Proteins , Treatment Outcome
6.
J Lipid Res ; 50(12): 2430-44, 2009 Dec.
Article in English | MEDLINE | ID: mdl-19506304

ABSTRACT

Pregnenolone (PREG) and dehydroepiandrosterone (DHEA), and their respective sulfated forms PREGS and DHEAS, were among the first steroids to be identified in rodent brain. However, unreliable steroid isolation and solvolysis procedures resulted in errors, particularly in the case of brain steroid sulfates analyzed by radioimmunology or GC-MS of liberated free steroids. By using a solid-phase extraction recycling/elution procedure, allowing the strict separation of sulfated, free, and fatty acid esters of PREG and DHEA, PREGS and DHEAS, unlike free PREG, were not detected in rat and mouse brain and plasma. Conversely, considerable amounts of PREG and DHEA were released from unknown precursor(s) present in the lipoidal fraction, distinct from fatty acid ester conjugates. Chromatographic and mass spectrometric studies of the nature of the precursor(s) showed that autoxidation of brain cholesterol (CHOL) was responsible for the release of PREG and DHEA from the lipoidal fraction. When inappropriate protocols were used, CHOL was also the precursor of PREG and DHEA obtained from the fraction assumed to contain sulfated steroids. In contrast, free PREG was definitely confirmed as an endogenous steroid in rat brain. Our study shows that an early removal of CHOL from brain extracts coupled to well-validated extraction and fractionation procedures are prerequisites for reliable measurements of free and conjugated PREG and DHEA by GC-MS or other indirect methods.


Subject(s)
Brain/metabolism , Cholesterol/metabolism , Dehydroepiandrosterone/analysis , Pregnenolone/analysis , Animals , Brain Chemistry , Male , Mice , Oxidation-Reduction , Rats , Rats, Sprague-Dawley
7.
Anesth Analg ; 105(6): 1855-7, table of contents, 2007 Dec.
Article in English | MEDLINE | ID: mdl-18042894

ABSTRACT

BACKGROUND: Spinal anesthesia in elderly patients is frequently associated with significant technical difficulties. Thus, we compared the classical midline approach to the paramedian approach to perform continuous spinal anesthesia (CSA). METHODS: We prospectively studied 40 patients aged >75 yr who underwent open surgical repair of a hip fracture. These patients were randomly allocated to one of two groups: Group M: midline approach, and Group PM: paramedian approach. Patients were positioned in the lateral decubitus to receive CSA at L4-5 level. CSA was considered successful if cerebrospinal fluid was obtained through the needle. In case of initial failure in either approach, the same approach was repeated by the same operator. If two attempts were unsuccessful, the other anatomical approach was used by the same operator. If both approaches failed, a staff anesthesiologist performed a final attempt. In case of failure or insufficient block, the patient received general anesthesia. RESULTS: The success rate after the first attempt was 85% (17) for Group PM and 45% (9) for Group M (P = 0.02). All catheters were successfully introduced. No patient required general anesthesia. Vascular puncture after needle puncture was observed in six patients in Group M versus 0 in Group PM (P = 0.03), but none were of clinical consequence. No other clinically significant complications were observed. CONCLUSION: In summary, after the initial attempt, the paramedian approach is associated with an increased success rate, compared with the midline approach, during the performance of CSA in elderly patients.


Subject(s)
Anesthesia, Spinal/methods , Age Factors , Aged , Aged, 80 and over , Female , Hip Fractures/epidemiology , Hip Fractures/surgery , Humans , Male , Prospective Studies
8.
Reg Anesth Pain Med ; 32(2): 157-61, 2007.
Article in English | MEDLINE | ID: mdl-17350528

ABSTRACT

BACKGROUND: The authors describe a new lateral approach to the sciatic nerve (SN) block in the popliteal fossa by using magnetic resonance imaging (MRI) and assessed its clinical feasibility. METHODS: The authors reviewed the MRI of the SN of 40 patients to compare a new landmark with the classical one. For the modified technique, the landmarks were the upper edge of the patella and the tendon of the biceps femoris. A line was drawn vertically from the upper edge of the patella. The puncture site was located at the intersection of this line with the lower part of the tendon. For the MRI study from the puncture point, the authors determined simulated needle direction to access the neurovascular bundle and measured its depth. RESULTS: The mean +/- standard deviation distance from the skin puncture to the tibial nerve was 48 +/- 6 mm in the classic group versus 26 +/- 5 mm (P < .0001) and to the common peroneal nerve 42 +/- 6 mm in the classic group versus 19 +/- 5 mm (P < .0001). The success rate was 94% (95% confidence interval 89-99) in 100 patients. Ten patients required general anesthesia; 4 because of the saphenous nerve failure (not analyzed as a failure) and 6 because of SN block failure. CONCLUSIONS: Based on the MRI images, a needle inserted below the biceps femoris tendon provides an easy access point to the common peroneal and/or the tibial nerve. This modified lateral approach to the SN was easy to perform, had a high success rate, and was without complication in this small cohort.


Subject(s)
Knee/innervation , Magnetic Resonance Imaging , Nerve Block/methods , Sciatic Nerve/anatomy & histology , Adult , Feasibility Studies , Female , Humans , Male , Middle Aged
9.
Endocrinology ; 147(9): 4339-50, 2006 Sep.
Article in English | MEDLINE | ID: mdl-16763064

ABSTRACT

To investigate the role of steroid receptors in mediating the reported effects of steroids on Schwann cell (SC) myelination and growth, we determined mRNA contents and transcriptional activities of the corticosteroid (glucocorticosteroid and mineralocorticosteroid) receptors (GR and MR) and sex steroid (progesterone, androgen, and estrogen alpha and beta) receptors in rat SC cultured under proliferative (in the presence of insulin and forskolin, which induces a high intracellular cAMP content) and quiescent conditions. We found no or very low expression and activity of the sex steroid receptors, as shown by mRNA concentrations determined with real-time PCR and transcriptional activities using transient expression of reporter plasmids in SC. These data and binding studies in SC lines demonstrated that the levels of the sex steroid receptors were the limiting factors. GR was clearly expressed (approximately 8000 sequences/ng total RNA) and functional. No significant modification in GR mRNA levels was observed, but an increase in transcriptional efficiency was recorded in proliferating cells compared with quiescent cells. MR was also significantly expressed at the mRNA level (approximately 450 sequences/ng total RNA) under the two culture conditions. No MR transcriptional activity was observed in SC, but a low specific binding of aldosterone was detected in SC lines. 11 beta-Hydroxysteroid-dehydrogenase type 2 (HSD2), an enzyme that inactivates glucocorticoids, was strongly expressed and active in quiescent SC, although in proliferating cells, HSD2 exhibited a strong decrease in activity and mRNA concentration. These data support a physiological role for HSD2 regulation of glucocorticosteroid concentrations in nerve SC.


Subject(s)
11-beta-Hydroxysteroid Dehydrogenase Type 2/genetics , 11-beta-Hydroxysteroid Dehydrogenase Type 2/metabolism , Receptors, Steroid/genetics , Receptors, Steroid/metabolism , Schwann Cells/metabolism , Animals , Cell Division , Cells, Cultured , Colforsin/pharmacology , Estrogen Receptor alpha/genetics , Estrogen Receptor alpha/metabolism , Estrogen Receptor beta/genetics , Estrogen Receptor beta/metabolism , Gene Expression Regulation, Enzymologic/drug effects , Genes, Reporter/genetics , Glucocorticoids/pharmacology , Gonadal Steroid Hormones/metabolism , Insulin/pharmacology , Promoter Regions, Genetic/genetics , RNA, Messenger/analysis , Rats , Rats, Sprague-Dawley , Receptors, Androgen/genetics , Receptors, Androgen/metabolism , Receptors, Glucocorticoid/genetics , Receptors, Glucocorticoid/metabolism , Receptors, Mineralocorticoid/genetics , Receptors, Mineralocorticoid/metabolism , Receptors, Progesterone/genetics , Receptors, Progesterone/metabolism , Response Elements/genetics , Reverse Transcriptase Polymerase Chain Reaction , Schwann Cells/chemistry , Schwann Cells/cytology , Sciatic Nerve/cytology , Transcription, Genetic , Transfection
10.
Pharmacol Biochem Behav ; 82(4): 712-20, 2005 Dec.
Article in English | MEDLINE | ID: mdl-16388839

ABSTRACT

The peripheral benzodiazepine receptors (PBR) might be involved in certain pathophysiological events, such as anxiety, by stimulating the production of neuroactive steroids in the brain. A recent electrophysiological study has revealed an interaction between PK11195, a PBR ligand and the anxiolytic compound etifoxine at micromolar concentrations. The present work was aimed at further characterizing the etifoxine-PBR interaction. In membrane preparations from intact male rat forebrain, etifoxine uncompetitively inhibited the binding of [(3)H]PK11195 with an IC(50) = 18.3 +/- 1.2 microM, a value consistent with etifoxine plasma and brain concentrations measured after an anxiolytic-like dose (50 mg/kg). In vivo, that etifoxine dose was associated with increased concentrations of pregnenolone, progesterone, 5alpha-dihydroprogesterone and allopregnanolone in plasma and brain of sham-operated animals. In adrenalectomized and castrated rats, etifoxine enhanced the brain levels of these steroids, suggesting a stimulation of their local synthesis and/or a decrease of their disappearance rate, independently of peripheral sources. Finasteride, an inhibitor of 5alpha-reductase that converts progesterone into its 5alpha-reduced metabolites like allopregnanolone, attenuated the anti-conflict effect of etifoxine even though brain allopregnanolone contents were drastically reduced. These results indicate that following activation of the PBR in the brain, an increased cerebral production of allopregnanolone, a potent positive modulator of the GABA(A) receptor function, may partially contribute to the anxiolytic-like effects of etifoxine.


Subject(s)
Anti-Anxiety Agents/pharmacology , Brain/drug effects , Oxazines/pharmacology , Receptors, GABA-A/drug effects , Steroids/metabolism , Animals , Brain/metabolism , Isoquinolines/metabolism , Male , Pregnanolone/metabolism , Radioligand Assay , Rats , Rats, Wistar
11.
J Lipid Res ; 45(12): 2287-302, 2004 Dec.
Article in English | MEDLINE | ID: mdl-15342680

ABSTRACT

A new sample preparation method coupled to GC-MS analysis was developed and validated for quantification of sulfate esters of pregnenolone (PREG-S) and dehydroepiandrosterone (DHEA-S) in rat brain. Using a solid-phase extraction recycling protocol, the results show that little or no PREG-S and DHEA-S (<1 pmol/g) is present in rat and mouse brain. These data are in agreement with studies in which steroid sulfates were analyzed without deconjugation. We suggest that the discrepancies between analyses with and without deconjugation are caused by internal contamination of brain extract fractions, supposed to contain steroid sulfates, by lipoidal forms of PREG and DHEA (L-PREG and L-DHEA, respectively). These derivatives can be acylated very efficiently with heptafluorobutyric anhydride and triethylamine, and their levels in rodent brain (approximately 1 nmol/g) are much higher than those of their unconjugated counterparts. They are distinct from fatty acid esters, and preliminary data do not favor structures such as sulfolipids or sterol peroxides. Noncovalent interactions between steroids and proteolipidic elements, such as lipoproteins, could account for some experimental data. Given their abundance in rodent brain, the structural characterization and biological functions of L-PREG and L-DHEA in the central nervous system merit considerable attention.


Subject(s)
Brain/metabolism , Dehydroepiandrosterone Sulfate/metabolism , Dehydroepiandrosterone/metabolism , Pregnenolone/metabolism , Animals , Dehydroepiandrosterone/analogs & derivatives , Gas Chromatography-Mass Spectrometry , Male , Pregnenolone/analogs & derivatives , Rats , Rats, Sprague-Dawley
12.
Endocrinology ; 144(7): 2902-11, 2003 Jul.
Article in English | MEDLINE | ID: mdl-12810545

ABSTRACT

In the central nervous system, neurosteroids, in particular progesterone, have neurotrophic and neuroprotective effects. We thus decided to study the developmental expression of 3beta-hydroxysteroid-dehydrogenase/Delta(5)-Delta(4) isomerase (3betaHSD), an enzyme that converts pregnenolone to progesterone, in the male rat brain at 0, 7, 14, and 70 d after birth. 3betaHSD mRNA was widely distributed throughout the brain, as shown by in situ hybridization. At all ages, the same cerebral structures were labeled, but the intensity of the hybridization signal constantly decreased during postnatal development. As the hippocampus is of particular interest because of its neuronal plasticity, we chose to quantify the changes in 3betaHSD mRNA levels as well as progesterone and pregnenolone concentrations in this structure. Quantitative in situ hybridization confirmed a decrease in the expression of 3betaHSD mRNA with progressing age, as revealed by a significant reduction in the density of silver grains per cell in the CA1 layer. This decrease was confirmed by semiquantitative RT-PCR on hippocampal samples. Concentrations of hippocampal pregnenolone and progesterone measured by gas chromatography/mass spectrometry were highest on the day of birth and lower at the other ages. Plasma concentrations of these steroids were lower than those in the hippocampus, suggesting that they may have been mostly synthesized in situ since the day of birth. These results demonstrate variations in the expression of a gene coding for an enzyme critically involved in progesterone synthesis in the hippocampus throughout postnatal development.


Subject(s)
Hippocampus/enzymology , Hippocampus/growth & development , Multienzyme Complexes/genetics , Progesterone Reductase/genetics , Progesterone/genetics , Steroid Isomerases/genetics , Age Factors , Animals , Gene Expression Regulation, Developmental , Gene Expression Regulation, Enzymologic , Male , Pregnenolone/genetics , RNA, Messenger/analysis , Rats , Rats, Sprague-Dawley , Reverse Transcriptase Polymerase Chain Reaction
13.
J Clin Endocrinol Metab ; 87(11): 5138-43, 2002 Nov.
Article in English | MEDLINE | ID: mdl-12414884

ABSTRACT

Some neurosteroids have been shown to display beneficial effects on neuroprotection in rodents. To investigate the physiopathological significance of neurosteroids in Alzheimer's disease (AD), we compared the concentrations of pregnenolone, pregnenolone sulfate (PREGS), dehydroepiandrosterone, dehydroepiandrosterone sulfate (DHEAS), progesterone, and allopregnanolone, measured by gas chromatography-mass spectrometry, in individual brain regions of AD patients and aged nondemented controls, including hippocampus, amygdala, frontal cortex, striatum, hypothalamus, and cerebellum. A general trend toward decreased levels of all steroids was observed in all AD patients' brain regions compared with controls: PREGS and DHEAS were significantly lower in the striatum and cerebellum, and DHEAS was also significantly reduced in the hypothalamus. A significant negative correlation was found between the levels of cortical beta-amyloid peptides and those of PREGS in the striatum and cerebellum and between the levels of phosphorylated tau proteins and DHEAS in the hypothalamus. This study provides reference values for steroid concentrations determined by gas chromatography-mass spectrometry in various regions of the aged human brain. High levels of key proteins implicated in the formation of plaques and neurofibrillary tangles were correlated with decreased brain levels of PREGS and DHEAS, suggesting a possible neuroprotective role of these neurosteroids in AD.


Subject(s)
Alzheimer Disease/metabolism , Brain Chemistry , Steroids/analysis , Aged , Aged, 80 and over , Aging , Amygdala/chemistry , Amyloid beta-Peptides/analysis , Cerebellum/chemistry , Corpus Striatum/chemistry , Dehydroepiandrosterone Sulfate/analysis , Female , Frontal Lobe/chemistry , Gas Chromatography-Mass Spectrometry , Hippocampus/chemistry , Humans , Hypothalamus/chemistry , Male , Pregnanolone/analysis , Pregnenolone/analysis , Progesterone/analysis , Protein Structure, Secondary , tau Proteins/analysis , tau Proteins/chemistry
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