Non-targeted profiling of circulating microRNAs in women with polycystic ovary syndrome (PCOS): effects of obesity and sex hormones.

PURPOSE: Circulating micro-ribonucleic acids (miRNAs) are small noncoding RNA molecules that influence gene transcription. We conducted the present profiling study to characterize the expression of circulating miRNAs in lean and obese patients with polycystic ovary syndrome (PCOS), the most common endocrine and metabolic disorder in premenopausal women. BASIC PROCEDURES: We selected 11 control women, 12 patients with PCOS and 12 men so that they were similar in terms of body mass index. Five control women, 6 men and 6 patients with PCOS had normal weight whereas 6 subjects per group were obese. We used miRCURY LNA™ Universal RT microRNA PCR for miRNA profiling. MAIN FINDINGS: The expression of 38 miRNAs and was different between subjects with PCOS and male and female controls. The differences in 15 miRNAs followed a pattern suggestive of androgenization characterized by expression levels that were similar in patients with PCOS and men but were different compared with those of control women. The expression of 13 miRNAs in women with PCOS was similar to that of control women and different compared with the expression observed in men, suggesting sexual dimorphism and, lastly, we observed 5 miRNAs that were expressed differently in women with PCOS compared with both men and control women, suggesting a specific abnormality in expression associated with the syndrome. Obesity interacted with the differences in several of these miRNAs, and the expression levels of many of them correlated with the hirsutism score, sex hormones and/or indexes of obesity, adiposity and metabolic dysfunction. PRINCIPAL CONCLUSIONS: The present results suggest that several serum miRNAs are influenced by PCOS, sex hormones and obesity. Our findings may guide the targeted search of miRNAs as clinically relevant markers for PCOS and its association with obesity and metabolic dysfunction in future studies.

Serum Bioavailable Vitamin D Concentrations and Bone Mineral Density in Women After Obesity Surgery.

INTRODUCTION: Low bone mass after obesity surgery may arise as a consequence of chronic malabsorption of calcium and vitamin D. However, we have not found any role of serum 25-hydroxyvitamin D or of polymorphisms in the vitamin D receptor gene in previous studies. PURPOSE: To investigate the circulating bioavailable 25-hydroxyvitamin D in women after bariatric procedures and its association with bone mass. PATIENTS AND METHODS: The study consisted of 91 women on follow-up for 7 ± 2 years after bariatric surgery. We measured bone mineral density (BMD), serum parathormone (PTH), 25-hydroxyvitamin D, and vitamin D binding protein (VDBP). All patients were genotyped for two variants in the coding region of VDBP (rs4588 and rs7041). Bioavailable 25-hydroxyvitamin D was calculated in double homozygotes. RESULTS: We found a negative correlation between bioavailable 25-hydroxyvitamin D and PTH (r = -0.373, P = 0.018), but not with BMD at lumbar spine (r = -0.065, P = 0.682) or hip (r = -0.029, P = 0.857). When adjusting by age, similar results were found for PTH (r = -0.441, P = 0.005), BMD at lumbar spine (r = -0.026, P = 0.874) and hip (r = -0.096, P = 0.561). After multivariate linear regression, forcing bioavailable 25-hydroxyvitamin D into the model resulted in a weak significant association with BMD at the lumbar spine (ß = - 0.247, P = 0.025). CONCLUSIONS: Serum bioavailable 25-hydroxyvitamin D concentrations are not associated with bone mass loss after bariatric surgery in women. The negative association with serum PTH levels suggests that vitamin D supplementation partly improves secondary hyperparathyroidism, yet other mechanisms may contribute to low bone mass after bariatric surgery.

Effects of polycystic ovary syndrome (PCOS), sex hormones, and obesity on circulating miRNA-21, miRNA-27b, miRNA-103, and miRNA-155 expression.

CONTEXT: MicroRNAs (miRNAs) are small, noncoding RNA sequences that negatively regulate gene expression at the post-transcriptional level. miRNA-21, miRNA-27b, miRNA-103, and miRNA-155 have been associated with metabolic disorders such as obesity and diabetes, which are also associated with polycystic ovary syndrome (PCOS). OBJECTIVE: We aimed to evaluate the effects of sex, sex hormones, and PCOS and their interactions with obesity on the expression in the circulation of these miRNAs. DESIGN: This was a case-control study. SETTINGS: The setting was an academic hospital. PARTICIPANTS: We included 12 control women, 12 patients with PCOS, and 12 men selected as to have similar body mass index (BMI) and age. Six subjects per group had normal weight (BMI < 25 kg/m(2)), and six subjects per group were obese (BMI ≥ 30 kg/m(2)). INTERVENTIONS: Blood samples were collected early in the morning after a 12-hour fast. MAIN OUTCOME MEASURES: We measured whole blood expression of miRNA-21, miRNA-27b, miRNA-103, and miRNA-155. RESULTS: Obesity significantly reduced the expression of miRNA-21, miRNA-27b, and miRNA-103. However, there was a significant interaction between obesity and the group of subjects in the expression of miRNA-21, miRNA-27b, miRNA-103, and miRNA-155 consisting of obesity reducing the expression of these miRNAs in control woman and men, but tending to increase their expression in women with PCOS. These differences paralleled those observed in serum T concentrations. CONCLUSIONS: The present results suggest that miRNAs that play an important role in metabolic and immune system processes are influenced by obesity and circulating androgen concentrations.

Detección de la deleción F508 del gen CFTR por la técnica de mutagénesis dirigida mediante PCR en pacientes con enfermedad fibroquística / Detection of F508 Deletion in the CFTR Gene by PCR Directed Mutagenesis in Patients with fibrocystic Ddsease

La Fibrosis quística es la enfermedad autosómica recesiva más común en la población blanca y se caracteriza por la obstrucción de conductos, principalmente en pulmón, páncreas y tracto genital. Se presenta en uno de cada 2000 a 2500 nacidos vivos y tiene una frecuencia de portadores de uno cada 20 a 25 nacidos vivos. La enfermedad es causada por diferentes mutaciones en el gen regulador de la conductancia transmembrana de la fibrosis quística (CFTR). La mutación más frecuente en el gen CFTR es la deleción de tres pares de bases (CTT) denominada F508. Este trabajo se realizó con el fin de estandarizar la técnica de mutagénesis dirigida mediante PCR (PSM) para la detección de.la mutación F508 en pacientes con fibrosis quística. El método utilizado fue validado mediante secuenciación del DNA del exón 10 en todos los individuos. Mediante este análisis genético se detectaron seis individuos con las mutaciones F508 e I507. El método implementado en nuestro laboratorio podría servir para realizar un sondeo poblacional de portadores de mutaciones para la FQ.

Impact of the storage temperature on human plasma proteomic analysis: implications for the use of human plasma collections in research.

PURPOSE: To analyze the impact of storage temperature on the 2D-DIGE profile of human plasma. EXPERIMENTAL DESIGN: 2D-DIGE and MS were used to identify the differences in the proteomic profiles of aliquots of eight human plasma samples stored at either - 30 or - 80°C for 18 months. RESULTS: After the depletion of albumin and IgG, 2D-DIGE identified four spots significantly and reproducibly increased, and five spots that were decreased, in samples stored at - 30°C compared with those stored at - 80°C. These nine spots were manually excised, digested ingel and analyzed by MALDI-TOF/TOF. MASCOT database search using the PMF spectra allowed the identification of the proteins present in eight of the nine spots. All the spots corresponded to the complement C3 precursor protein. CONCLUSIONS AND CLINICAL RELEVANCE: Our present results indicate that plasma collections stored at -30°C, and not only those stored at - 80°C, may be used for 2D-DIGE analyses of albumin-and IgG-depleted human plasma without loosing the essential information about highly abundant proteins. This finding expands the applicability of 2D-DIGE to the study of human disease by permitting the analysis of human plasma samples stored at temperatures between - 30 and - 80°C.

Proteomic analysis of plasma in the polycystic ovary syndrome identifies novel markers involved in iron metabolism, acute-phase response, and inflammation.

CONTEXT: Hypothesis-free approaches such as proteomic analysis may identify novel biomarkers for disease. OBJECTIVE: The objective of the study was to compare the plasma proteome of patients presenting with the polycystic ovary syndrome (PCOS) with that of women without hyperandrogenism. DESIGN: This was a case-control study. SETTINGS: The study was conducted at an academic hospital. PATIENTS: Patients included 12 PCOS patients and 12 women without hyperandrogenism. INTERVENTIONS: Interventions included basal blood sampling. MAIN OUTCOME MEASURES: Two-dimensional difference in-gel electrophoresis, matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry, Western blot, and ELISA analyses were measured. RESULTS: Two-dimensional difference in-gel electrophoresis and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry analyses identified haptoglobin beta-chain and alpha2-macroglobulin as proteins underexpressed in PCOS samples, whereas transferrin and kappa-free light chain were overexpressed. We were able to confirm only the underexpression of haptoglobin beta-chain in subsequent Western blot and ELISA analyses. CONCLUSIONS: Proteomic analysis of plasma from PCOS patients revealed changes in protein expression in several acute-phase response proteins including isoforms of plasma haptoglobin, alpha2-macroglobulin, and transferrin and in kappa-free light chain. In addition to their role as inflammatory markers, some of these molecules play major roles in iron metabolism, further suggesting that iron metabolism and low-grade chronic inflammation may be involved in the pathogenesis of insulin-resistant disorders such as PCOS.

Vitamin D receptor polymorphisms in secondary hyperparathyroidism after Scopinaro's biliopancreatic diversion.

BACKGROUND: Secondary hyperparathyroidism is a frequent metabolic complication of bariatric surgery. Individual differences in calcium absorption determine chronic secondary hyperparathyroidism after biliopancreatic diversion in half of the patients who have normal levels of 25-hydroxyvitamin D. We aimed to evaluate if certain vitamin D receptor polymorphisms may be responsible for the latter. Cases and controls study including 57 patients after biliopancreatic diversion with a mean serum 25-hydroxyvitamin D above 20 ng/mL, separated into those with secondary hyperparathyroidism (n = 26, cases) and those without it (n = 31, controls). METHODS: Genotyping for restriction-length-fragment polymorphisms of the vitamin D receptor gene was carried out for FOK1, BSM1, APA1, and TAQ1, and haplotype structure was also constructed. RESULTS: There were no differences in the allelic or genotypes distribution of the four studied polymorphisms between patients and controls (P = 0.352 and P = 0.301 for FOK1, P = 0.733 and P = 0.924 for BSM1, P = 0.974 and P = 0.992 for APA1, and P = 0.995 and P = 0.928 for TAQ1, respectively). Haplotype analysis showed no differences between patients and controls (P = 0.495 for BAT, P = 1.000 for BAt, P = 0.508 for Bat and P = 0.924 for bAT haplotypes, respectively). Furthermore, haplotypes were not associated with serum PTH levels or with the ratio between serum PTH and 25-hydroxyvitamin D levels. CONCLUSION: Chronic secondary hyperparathyroidism after biliopancreatic diversion in patients with normal levels of 25-hydroxyvitamin D is not dependent on vitamin D receptor gene polymorphisms.

The role of genetic variation in peroxisome proliferator-activated receptors in the polycystic ovary syndrome (PCOS): an original case-control study followed by systematic review and meta-analysis of existing evidence.

OBJECTIVE: To study the association of polymorphisms in the genes encoding peroxisome proliferator-activated receptors (PPARs) with the polycystic ovary syndrome (PCOS). DESIGN: Case-control study and meta-analysis of published evidence. PATIENTS: One hundred and sixty-one polycystic ovary syndrome patients and 113 non-hyperandrogenic women. MEASUREMENTS: Genotyping for PPAR-gamma coactivator-1 gene (PPARGC1A) Gly482Ser, PPAR-alpha Leu162Val, PPAR-delta rs2267668A/G, PPAR-delta-87T/C, PPAR-gamma2 Pro12Ala and PPAR-gamma2 -681C/G variants and systematic review of the literature using the Entrez-PubMed search engine, followed by meta-analysis whenever possible. RESULTS: Polycystic ovary syndrome patients carried the Gly482Ser variant in PPARGC1A more frequently than controls (72%vs. 58%, chi(2 )=( )5.54 P = 0.019), whereas carriers of the PPAR-alpha Leu162Val, PPAR-delta rs2267668A/G, PPAR-delta-87T/C, PPAR-gamma2 Pro12Ala and PPAR-gamma2 -681C/G variants were distributed similarly among both groups. The interaction between the PPARGC1A Gly482Ser and PPAR-delta-87T/C variants was also associated with PCOS (OR = 1.24, 95% CI 1.05-1.50, P = 0.008). The systematic review identified 31 studies addressing associations between PPARs variants and PCOS; meta-analysis was possible for nine studies focusing on the PPAR-gamma2 Pro12Ala variant. Although the individual studies did not reveal any statistically significant association, meta-analysis uncovered that carrying the PPAR-gamma2 Pro12Ala variant was associated with a reduced probability of having PCOS (OR = 0.77, 95% CI 0.61-0.96, P = 0.025), and that this association may be mediated by an effect on insulin sensitivity. CONCLUSIONS: Common polymorphisms in the PPARGC1A, PPAR-delta and PPAR-gamma2 loci are associated with PCOS.

Role of haptoglobin in polycystic ovary syndrome (PCOS), obesity and disorders of glucose tolerance in premenopausal women.

BACKGROUND: Hp(2) alleles of the haptoglobin alpha-chain polymorphism reduce the anti-oxidant properties and increase the pro-inflammatory actions of this acute-phase protein in a gene-dosage fashion. We hypothesized that the haptoglobin polymorphism might contribute to the increased oxidative stress and low-grade chronic inflammation frequently associated with polycystic ovary syndrome, obesity, and abnormalities of glucose tolerance. METHODOLOGY/PRINCIPAL FINDINGS: Serum haptoglobin and the haptoglobin alpha-chain polymorphism were determined in 141 patients with polycystic ovary syndrome and 102 non-hyperandrogenic women. Of the whole group of 243 premenopausal women, 117 were obese and 51 showed abnormal glucose tolerance. Although serum haptoglobin concentrations were similar in PCOS patients and controls, the former presented with an increased frequency of Hp(2) alleles (62% vs. 52%, P = 0.023). Circulating haptoglobin levels increased with obesity (P<0.001), yet no association was found between obesity and haptoglobin genotypes. No differences were observed in haptoglobin levels or genotype frequencies depending on glucose tolerance. Fifty percent of the variation in serum haptoglobin concentrations was explained by the variability in serum C-reactive protein concentrations, BMI, insulin sensitivity and haptoglobin genotypes. CONCLUSIONS/SIGNIFICANCE: Serum haptoglobin concentrations in premenopausal women are largely dependent on the haptoglobin polymorphism and on the presence of obesity, with insulin resistance and chronic inflammation possibly modulating this relationship. The association of polycystic ovary syndrome with Hp(2) alleles suggests that the anti-oxidant and anti-inflammatory properties of haptoglobin may be reduced in these patients.

Body iron stores and glucose intolerance in premenopausal women: role of hyperandrogenism, insulin resistance, and genomic variants related to inflammation, oxidative stress, and iron metabolism.

OBJECTIVE Increased serum ferritin levels and iron stores may be involved in the development of abnormal glucose tolerance in women presenting with obesity and/or polycystic ovary syndrome (PCOS). We aimed to study the determinants of serum ferritin levels in premenopausal women among indexes of insulin resistance, adiposity, hyperandrogenism, and genotypes pertaining to inflammation, oxidative stress, and iron metabolism. RESEARCH DESIGN AND METHODS A total of 257 premenopausal women, classified depending on the presence or absence of PCOS, obesity, and/or abnormal glucose tolerance, underwent a complete metabolic evaluation, serum ferritin, haptoglobin, and C-reactive protein (CRP) measurements, and genotyping for proinflammatory and prooxidant variants and mutations in the HFE gene. RESULTS Serum ferritin concentrations were increased in women presenting with PCOS and/or abnormal glucose tolerance, independent of obesity. A stepwise multivariate linear regression analysis (R(2) = 0.18, P < 0.0001) retained menstrual dysfunction (beta = 0.14, P = 0.035), free testosterone (beta = 0.14, P = 0.052), insulin sensitivity index (beta = -0.12, P = 0.012), the His63Asp variant in HFE (beta = 0.16, P = 0.008), and abnormal glucose tolerance (beta = 0.15, P = 0.015) as significant predictors of the logarithm of ferritin levels, whereas CRP, haptoglobin, waist-to-hip ratio, or variants in the TNFalpha, TNFRSF1B, IL6, IL6ST, IL6Ralpha, PON1, and HFE Cys282Tyr mutation exerted no influence. CONCLUSIONS Androgen excess (partly because of hyperandrogenemia and partly because of menstrual dysfunction), insulin resistance, abnormal glucose tolerance, and the HFE His63Asp variant correlate with ferritin levels in premenopausal women.

Unified criteria for ultrasonographic diagnosis of ADPKD.

Individuals who are at risk for autosomal dominant polycystic kidney disease are often screened by ultrasound using diagnostic criteria derived from individuals with mutations in PKD1. Families with mutations in PKD2 typically have less severe disease, suggesting a potential need for different diagnostic criteria. In this study, 577 and 371 at-risk individuals from 58 PKD1 and 39 PKD2 families, respectively, were assessed by renal ultrasound and molecular genotyping. Using sensitivity data derived from genetically affected individuals and specificity data derived from genetically unaffected individuals, various diagnostic criteria were compared. In addition, data sets were created to simulate the PKD1 and PKD2 case mix expected in practice to evaluate the performance of diagnostic criteria for families of unknown genotype. The diagnostic criteria currently in use performed suboptimally for individuals with mutations in PKD2 as a result of reduced test sensitivity. In families of unknown genotype, the presence of three or more (unilateral or bilateral) renal cysts is sufficient for establishing the diagnosis in individuals aged 15 to 39 y, two or more cysts in each kidney is sufficient for individuals aged 40 to 59 y, and four or more cysts in each kidney is required for individuals > or = 60 yr. Conversely, fewer than two renal cysts in at-risk individuals aged > or = 40 yr is sufficient to exclude the disease. These unified diagnostic criteria will be useful for testing individuals who are at risk for autosomal dominant polycystic kidney disease in the usual clinical setting in which molecular genotyping is seldom performed.

Serum osteoprotegerin concentrations are decreased in women with the polycystic ovary syndrome.

OBJECTIVE: Osteoprotegerin (OPG), an inhibitor of osteoclastic bone resorption, has a variety of functions including anti-inflammatory effects and a possible cardiovascular protective role. Both low-grade chronic inflammation and cardiovascular risk are increased in women with the polycystic ovary syndrome (PCOS). We aimed to study serum OPG concentrations in PCOS patients. DESIGN: Case-control study including 40 PCOS patients matched with 40 non-hyperandrogenic women for age and body mass index. METHODS: Basal serum sampling and standard oral glucose tolerance test, and measurement of serum OPG concentrations by commercial ELISA. RESULTS: Serum OPG concentrations were lower in women with PCOS compared with those of controls (304+/-120 vs 363+/-105 pg/ml respectively; F=7.641, P=0.007) independently of obesity. No differences were observed in serum receptor activator of nuclear factor-kappaB ligand (RANKL) levels and in the RANKL/OPG molar ratio. A multivariate linear regression model (R(2)=0.208, F=6.579, P=0.001) showed that PCOS (beta=-0.281, P=0.008), obesity (beta=-0.245, P=0.022) and age (beta=0.296, P=0.006) were predictive of serum OPG concentrations. CONCLUSIONS: Serum OPG concentrations are reduced in PCOS patients independently of obesity. Considering the anti-inflammatory effects of OPG, its reduced serum concentrations might contribute to the proinflammatory state and cardiovascular risk of PCOS patients.

The increase in serum visfatin after bariatric surgery in morbidly obese women is modulated by weight loss, waist circumference, and presence or absence of diabetes before surgery.

BACKGROUND: Previous studies addressing the changes in serum visfatin levels after bariatric surgery yielded conflicting results. METHODS: We measured serum visfatin levels in 41 morbidly obese women before bariatric surgery and after losing at least 15% of the initial weight, and analyzed the results taking into account the type of surgery, reproductive and diabetic status, among others. Body mass index, waist circumference, lipid profile, and insulin resistance determined by homeostasis model assessment (HOMA-IR) were also measured. RESULTS: Patients lost 30.3 +/- 6.1% of the initial body weight, and serum visfatin levels increased from 22.2 +/- 20.9 to 32.2 +/- 27.6 ng/ml (P = 0.031). A multiple regression model (R (2) = 0.314, F = 3.555, P = 0.017) including the percentage of weight loss, changes in waist circumference, HOMA-IR, high-density lipoprotein-cholesterol, and triglycerides (also expressed as percentage from baseline), the surgical procedure, time elapsed since surgery, and previous diabetic status as independent variables showed that weight loss (beta = -0.670, P = 0.010), previous diabetic status (beta = -0.330, P = 0.036), and change in waist circumference (beta = 0.556, P = 0.031) were the main determinants of the percentual increase in serum visfatin levels observed after bariatric surgery. CONCLUSION: Serum visfatin increased after bariatric surgery in relation to the amount of weight lost and to the changes in waist circumference, and this increase was higher in diabetic patients.

Proteomic analysis of human omental adipose tissue in the polycystic ovary syndrome using two-dimensional difference gel electrophoresis and mass spectrometry.

BACKGROUND: Our aim was to study the protein expression profiles of omental adipose tissue biopsies obtained from morbidly obese women with or without polycystic ovary syndrome (PCOS) at the time of bariatric surgery to evaluate the possible involvement of visceral adiposity in the development of PCOS. METHODS: Ten PCOS patients and nine control samples were included. We used two-dimensional difference gel electrophoresis (2D-DIGE) followed by in-gel digestion, and mass spectrometry (MS) of selected protein spots. RESULTS: The 2D-DIGE technology allowed the analysis of approximately 1840 protein spots in the comparative study of control and patient proteomes, revealing 15 statistically significant spot changes (>2-fold, P < 0.05). Unambiguous protein identification was achieved for 9 of these 15 spots by MS. This preliminary study revealed differences in expression of proteins that may be involved in lipid and glucose metabolism, oxidative stress processes and adipocyte differentiation; they include proapolipoprotein Apo-A1, annexin V, glutathione S-transferase M3 (GSTM3), triosephosphate isomerase, peroxiredoxin 2 isoform a, actin and adipocyte plasma membrane-associated protein. The most relevant finding was an increase of GSTM3 in the omental fat of PCOS patients confirming previous studies conducted by our group. CONCLUSIONS: Proteomic analysis of omental fat reveals differential expression of several proteins in PCOS patients and non-hyperandrogenic women presenting with morbid obesity. The application of this novel methodology adds further evidence to support the role of visceral adiposity in the pathogenesis of PCOS.

Proteomics and genomics: A hypothesis-free approach to the study of the role of visceral adiposity in the pathogenesis of the polycystic ovary syndrome.

The polycystic ovary syndrome (PCOS) is possibly the most common endocrine disorder in premenopausal women. The primary defect in PCOS consists of an exaggerated androgen secretion by the ovaries and the adrenal glands of affected women, which is amplified by several mechanisms including abdominal adiposity and insulin resistance. Abdominal adiposity contributes to hyperandrogenism by favoring insulin resistance and compensatory hyperinsulinism, because insulin facilitates ovarian and adrenal androgen synthesis, among other mechanisms. Furthermore, mounting evidence suggest that androgen excess may also contribute to a predominantly abdominal disposition of body fat in women, suggesting that women with PCOS suffer from a vicious circle whereby androgen excess favoring the abdominal deposition of fat further facilitates androgen secretion by the ovaries and adrenals. Familial aggregation of PCOS cases suggests an inherited component in PCOS, yet the genetic mechanisms underlying this inheritance remain elusive. The present manuscript reviews the hypothesis-free approaches - such as genomics and proteomics - that have been used recently to study PCOS, focusing on studies from our group addressing the gene expression profiles and the proteome of visceral adipose tissue of morbidly obese women presenting with or without PCOS.

A prospective study of the prevalence of nonclassical congenital adrenal hyperplasia among women presenting with hyperandrogenic symptoms and signs.

CONTEXT: The diagnosis of the polycystic ovary syndrome requires the exclusion of nonclassical congenital adrenal hyperplasia (NCAH). OBJECTIVE: Our objective was to evaluate the actual prevalences of 21-hydroxylase and 11beta-hydroxylase deficiencies among women presenting with hyperandrogenic complaints. SETTINGS: This study was performed at an academic hospital. PATIENTS: A total of 270 consecutive unselected women presenting with hyperandrogenic symptoms were prospectively recruited. INTERVENTIONS: Basal and ACTH-stimulated 11-deoxycortisol and 17-hydroxyprogesterone concentrations were measured. MAIN OUTCOME MEASURES: The prevalences of 21-hydroxylase and 11beta-hydroxylase deficiencies were calculated, and the diagnostic performance of basal serum 17-hydroxyprogesterone levels for the screening of NCAH was evaluated by receiver operating characteristic curve analysis. RESULTS: Six of the 270 patients had 21-hydroxylase-deficient NCAH that was confirmed by CYP21 genotyping, whereas no patient was diagnosed with 11beta-hydroxylase deficiency, for an overall NCAH prevalence of 2.2% (95% confidence limits 0.5-3.9%). According to receiver operating characteristic analysis, a single basal serum 17-hydroxyprogesterone determination has a 0.97 (95% confidence interval: 0.934-1.008) chance of detecting NCAH in hyperandrogenic women. In our experience, the most appropriate cutoff value for the detection of NCAH is a 17-hydroxyprogesterone above 1.7 ng/ml, showing a 100% sensitivity and a 88.6% specificity. Five of the six 21-hydroxylase-deficient NCAH patients carried a severe CYP21 allele requiring genetic counseling and highlighting the importance of excluding this disorder among hyperandrogenic patients. CONCLUSIONS: The prevalence of NCAH among hyperandrogenic patients from Spain is 2.2%. Basal serum 17-hydroxyprogesterone measurements have an excellent diagnostic performance, yet the cutoff value should be established in each laboratory to avoid false-negative results.

The decrease in serum IL-18 levels after bariatric surgery in morbidly obese women is a time-dependent event.

BACKGROUND: We have evaluated the impact of the reproductive status of morbidly obese women, and of the time elapsed since surgery, on the response of the proinflammatory serum cardiovascular risk marker interleukin-18 (IL-18) to the sustained and marked weight loss achieved after bariatric surgery. METHODS: Serum IL-18 levels were measured in 33 morbidly obese women before bariatric surgery and after losing at least 15% of the initial weight, irrespective of the time needed to achieve this goal (5 to 33 months). RESULTS: Patients lost 30.7 +/- 7.8% of the initial weight, with a concomitant reduction of serum IL-18 concentrations (P<0.001). A stepwise multiple regression analysis showed that the percentual decrease in serum IL-18 levels was determined by the interaction between the time elapsed since surgery and the percentual reduction of waist circumference (R2 = 0.333, F = 15.500, beta = 0.577, P<0.001), but not by the individual effects of the time elapsed since surgery, percentual body weight loss, percentual reduction of waist circumference, menopausal status or type of surgical procedure, or by the interaction between the time elapsed since surgery with the percentual body weight loss or with menopausal status. CONCLUSION: Serum IL-18 levels decrease after bariatric surgery in a time-dependent manner, in relation to the reduction in waist circumference. The fact that the amelioration of the obesity-associated inflammatory process requires time and not only weight loss, might contribute to explain early non-surgical cardiovascular complications of bariatric surgery.

Abdominal adiposity and the polycystic ovary syndrome.

Abdominal adiposity, overweightness and obesity are frequently present in patients with polycystic ovary syndrome (PCOS). A large body of evidence suggests that abdominal adiposity and the resulting insulin resistance contribute to ovarian and, possibly, adrenal hyperandrogenism. However, androgen excess itself might also contribute to abdominal fat deposition in hyperandrogenic women. Recent genomic and proteomic analyses of visceral fat from PCOS patients have detected differences in gene expression and protein content compared with those of non-hyperandrogenic women. Here we review the existing evidence for a vicious circle whereby androgen excess favoring the abdominal deposition of fat further facilitates androgen secretion by the ovaries and adrenals in PCOS patients.

Increased body iron stores of obese women with polycystic ovary syndrome are a consequence of insulin resistance and hyperinsulinism and are not a result of reduced menstrual losses.

OBJECTIVE: Increased serum ferritin levels, indicating increased body iron stores, have been found in overweight and obese women with polycystic ovary syndrome (PCOS). This finding might result from reduced menstrual losses secondary to oligo- or amenorrhea or from hyperinsulinism secondary to insulin resistance, because insulin favors the intestinal absorption and the tissue deposition of iron. To explore which of these mechanisms is responsible for the increase in body iron stores in women with PCOS, we have monitored the changes in serum ferritin levels during treatment with an antiandrogenic oral contraceptive or an insulin sensitizer. RESEARCH DESIGN AND METHODS: Thirty-four consecutive PCOS patients were randomized to an oral contraceptive containing 35 microg ethinyl-estradiol plus 2 mg cyproterone acetate (Diane(35) Diario) or metformin (850 mg twice daily), and their serum ferritin levels were evaluated at baseline and after 12 and 24 weeks of treatment. RESULTS: Despite the fact that treatment with Diane(35) Diario restored regular menstrual cycles in all the patients, whereas metformin only did so in 50% of them, serum ferritin levels decreased at 12 and 24 weeks of treatment only with metformin, in association with a marked increase in insulin sensitivity. On the contrary, no changes in ferritin and insulin sensitivity were observed with Diane(35) Diario. CONCLUSIONS: Our present results suggest that insulin resistance and hyperinsulinism, and not the reduced menstrual losses secondary to from oligo- or amenorrhea, are responsible of the increased ferritin levels and body iron stores found in overweight and obese women with PCOS.