bluuebunny/crossref_metadata_2025 huggingface dataset:
Molecular Scanning of β-3-Adrenergic Receptor Gene in Total Congenital Lipoatrophic Diabetes Mellitus*
Authors: Silver Kristi , Walston Jeremy , Plotnick Leslie , Taylor Simeon I. , Kahn C. Ronald , Shuldiner Alan R.
Published: January 2014

Total congenital lipoatrophic diabetes is characterized by absence of subcutaneous adipose tissue, hypertriglyceridemia, and insulin resistance. We hypothesized that mutations in the β-3-adrenergic receptor (β3AR) gene might result in the lipoatrophic phenotype by preventing triglyceride storage in adipocytes; thereby, resulting in secondary insulin resistance. We screened the β3AR gene in 7 subjects with total congenital lipoatropic diabetes. We found a heterozygous substitution of a guanine to cytosine at position −153 (G-153C) in the 5′-untranslated region of 3 African-American lipoatrophic siblings and 1 sibling without lipoatrophy but with insulin resistance. To determine whether the base change was related to the lipoatrophic phenotype, we genotyped 69 African-Americans without lipoatrophy and found the G-153C substitution in 2 control subjects (allele frequency = 0.01). No other single-stranded polymorphism variants were found in any of the 7 lipoatrophic subjects. Direct sequencing of both alleles of 1 lipoatrophic subject demonstrated a thymidine insertion at position− 300 in both alleles. All lipoatrophic subjects along with 20 African-American control subjects were homozygous for the base insertion, suggesting an error in the published sequence. In conclusion, mutations in the β3AR gene do not appear to be involved in the development of congenital total lipoatrophy.
Urinary Follicle-Stimulating Hormone for Normogonadotropic Clomiphene-Resistant Anovulatory Infertility: Prospective, Randomized Comparison between Low Dose Step-Up and Step-Down Dose Regimens1
Authors: van Santbrink Evert J. P. , Fauser Bart C. J. M.
Published: January 2014

A low dose step-up and step-down regimen for induction of ovulation using urinary FSH was compared in a prospective randomized fashion in 37 normogonadotropic clomiphene-resistant oligo- or amenorrheic infertile women. The objectives was to assess potential differences in duration of treatment, ovarian stimulation (serum FSH levels), and response [serum estradiol (E2) levels and number and size of follicles]. Monitoring (blood sampling and transvaginal sonography) took place on the day of initiation of treatment, the first day of ovarian response as assessed by ultrasound (i.e. the first day a follicle ≥10 mm could be recognized), the day of hCG administration to induce ovulation, and 3 days thereafter.

The median duration of treatment in the low dose step-up group was 18 (range, 7–41) days compared to 9 (range, 4–16) days in the step-down group (P = 0.003), and the total numbers of ampules administered were 20 (range, 7–69) and 14 (range, 7–33), respectively (P = NS). Serum FSH levels from the first day of sonographic ovarian response until the administration of hCG were constant (median increase, 2%/day) in patients receiving the low dose step-up protocol, but showed a decrease (median, 5%/day) in step-down cycles (P < 0.001). Monofollicular growth, defined as not more than one follicle 16 mm or larger on the day of hCG administration, was observed in 56% of low dose step-up and 88% of step-down cycles (P = 0.04). The percentage of patients with normal range periovulatory E2 serum levels (500–1500 pmol/L) was 33% in the low dose step-up group vs. 71% in the step-down group (P = 0.03).

We conclude that a step-down protocol for gonadotropin induction of ovulation exhibits a more physiological, late follicular phase FSH serum profile than a low dose step-up protocol. This results in a shorter duration of treatment, a greater number of monofollicular cycles, and more cycles with periovulatory E2 levels within the normal range in the step-down protocol.
Identification of a New Thyrotropin Receptor Germline Mutation (Leu629Phe) in a Family with Neonatal Onset of Autosomal Dominant Nonautoimmune Hyperthyroidism
Authors: Führer Dagmar , Wonerow Peter , Willgerodt Helmut , Paschke Ralf , Führer-Sakel Dagmar
Published: January 2014

Constitutively activating germline mutations in the TSH receptor (TSHR) gene have been identified as a cause of autosomal dominant nonautoimmune hyperthyroidism and sporadic congenital hyperthyroidism. We report a 10-yr-old boy and his 31-yr-old mother, both presenting with a history of recurring toxic thyroid hyperplasia and no evidence for autoimmune thyroid disease. In the boy, onset of hyperthyroidism and goiter was neonatal. In the mother, onset of thyroid disease dates back to early childhood. There was no history of thyroid disease in the rest of the family. Screening for germline mutations in exon 10 of the TSHR was performed by direct sequencing of genomic DNA extracted from peripheral blood leukocytes of both patients. In the boy and his mother, an identical heterozygous TSHR mutation was identified, exchanging leucine for phenylalanine at residue 629 of the TSHR (TTG→TTT). Transient expression of the mutated TSHR construct in COS-7 cells confirmed the constitutive activity of the new TSHR germline mutation. This is the second family displaying congenital manifestation of hyperthyroidism in familial nonautoimmune hyperthyroidism.
Somatostatin and Growth Hormone-Releasing Hormone in Normal and Tumoral Human Breast Tissue: Endogenous Content, in Vitro Pulsatile Release, and Regulation
Authors: Benlot Caroline , Lévy Laurence , Fontanaud Pierre , Roche Annick , Rouannet Philippe , Joubert Dominique
Published: January 2014

Endogenous production of SRIH and GHRH was analyzed in human breast tissue. SRIH precursor (pro-SRIH) was identified after Sephadex G-50 filtration of acetic acid extracts of normal and tumoral human breast samples. SRIH-(1–14) or -(1–28) could not be detected in breast tissue, whereas the immunoreactive SRIH released in vitro was characterized as SRIH-(1–28). Endogenous production of GHRH was assessed by identification of GHRH messenger ribonucleic acid by PCR followed by sequencing of the amplified complementary DNA and by high performance liquid chromatographic characterization of immunoreactive GHRH contained in the tissue and released in vitro. There were no differences in pro-SRIH or GHRH-(1–44) tissue contents between normal and tumoral samples. The release of both peptides was evidenced in perifusion and static incubation. Perifusion of normal breast tissue (n = 3) showed pulsatile release of SRIH and GHRH. Perifusion of tumors (n = 4) showed SRIH release in 50% of the cases. SRIH release was pulsatile in one case. GHRH release was observed in the four tumoral samples analyzed, but was pulsatile in only one case. In static incubation, tumors (n = 6) secreted 13 times more GHRH than did normal samples (n = 3; 383 ± 92 vs. 29.6 ± 4.6 fmol/mg protein; P < 0.05). Stimulation of GHRH release by exogenous SRIH was observed only with the normal tissue.

Together these data provide evidence for the existence of local production of SRIH and GHRH by human breast. Hypersecretion of GHRH by breast tumors indicates that this peptide could play a role in maintaining epithelial cell proliferation as is the case for other peptides produced locally.
The Pituitary-Adrenal Responses to Exogenous Human Corticotropin-Releasing Hormone in Preterm, Very Low Birth Weight Infants
Authors: Ng P. C. , Wong G. W. K. , Lam C. W. K. , Lee C. H. , Wong M. Y. , Fok T. F. , Wong W. , Chan D. C. F.
Published: January 2014

To evaluate the pituitary-adrenal reserve and to standardize the methodology of performing the human CRH (hCRH) stimulation test, we performed the hCRH test on 14 preterm (<32 gestational weeks), very low birth weight infants, who did not receive antenatal or postnatal corticosteroid treatment, on days 7 and 14 of life. Blood samples were obtained 0 (baseline), 15, 30, and 60 min after an iv dose of hCRH (1μ g/kg). The plasma ACTH concentration rose from a basal value of 5.7 ± 0.6 pmol/L (mean ± sem) to 11.9 ± 2.1 pmol/L (P < 0.005), 9.2 ± 1.2 pmol/L (P < 0.005), and 7.7 ± 0.8 pmol/L (P < 0.005) at 15, 30, and 60 min, respectively. The corresponding rises in serum cortisol from a basal concentration of 396 ± 67 nmol/L were 509 ± 71 nmol/L (P< 0.0001), 647 ± 62 nmol/L (P < 0.0001), and 578 ± 60 nmol/L (P < 0.0001). The plasma ACTH concentration consistently peaked early at 15 min, whereas the maximum cortisol response occurred 30 min post-hCRH stimulation. No significant differences were detected between the hCRH tests performed on days 7 and 14 (P > 0.15). Mechanical ventilation, infant gender, and mode of delivery did not significantly influence the hormonal responses (P > 0.25).

We have defined in this study the pattern, the magnitude of the pituitary-adrenal response, and the timing of the peak concentrations of plasma ACTH and serum cortisol in relation to a standard iv dose of hCRH. The hCRH test in very low birth weight infants appears to be safe and reproducible, and produces a pituitary-adrenal response comparable to that seen in older children and adults, indicating that pituitary-adrenal function is mature at these early stages of gestation.
Characterization of a New Case of Autoimmune Type I Hyperlipidemia: Long-Term Remission under Immunosuppressive Therapy1
Authors: Pruneta Valerie , Moulin Philippe , Labrousse Florence , Bondon Pierre-Jean , Ponsin Gabriel , Berthezene Francois
Published: January 2014

Only a few cases of type I hyperlipidemia occurring in patients with autoimmune disease have been reported. We describe the case of a 35-yr-old woman suffering from severe type I hyperchylomicronemia. A combination of various hypolipidemic treatments, including strict hypolipidemic dietary therapy and administration of fibrates or n-3 fatty acids, was inefficient. Because of a history of familial autoimmunity, we introduced an immunosuppressive therapy that resulted in consistent long term and stable remission. Two attempts to reduce the immunosuppressor dose resulted in major relapses. To explain the defect of chylomicron hydrolysis, we investigated the postheparin plasma lipase activities. Hepatic triglyceride lipase activity was normal, whereas that of lipoprotein lipase (LPL) was reduced to about 30% of normal. Immunosuppressive therapy resulted in a complete and durable normalization of LPL activity. Using Western blot analysis, we found in the plasma of the patient a circulating IgG specifically directed against LPL, which became undetectable during immunosuppressive therapy. Western blot analysis revealed that the whole circulating anti-LPL autoantibody was bound to chylomicrons. Proteins extracted from patient’s chylomicrons were able to induce a dose-related inhibition of LPL activity in vitro, whereas that of hepatic triglyceride lipase remained unchanged.

These data constitute the first description of autoimmune hyperchylomicronemia due to an exclusive defect of LPL activity, and they show that a complete remission has been obtained after immunosuppressive therapy. Finally, our finding that the anti-LPL autoantibody is bound to chylomicrons emphasizes their previously unrecognized ability to transport LPL, already described for other lipoprotein fractions.
Effect of Plasma Osmolality on Pituitary-Adrenal Responses to Corticotropin-Releasing Hormone and Atrial Natriuretic Peptide Changes in Central Diabetes Insipidus*
Authors: Elias Lucila L. K. , Antunes-Rodrigues José , Elias Paula C. L. , Moreira Ayrton C.
Published: January 2014

The objective of the present study was to examine the effect of changes in plasma osmolality (pOsm) on the responses of the pituitary-adrenal axis to CRH and atrial natriuretic peptide (ANP) release in patients with central diabetes insipidus (DI). Eight normal subjects and six DI patients were subjected to human CRH (hCRH) (1 μg/kg) stimulation alone or associated with isotonic volume loading (0.9% NaCl, 12 mL·kg·60 min) or an osmotic stimulus (5% NaCl, 0.06 mL·kg/min·120 min). The DI group showed significantly increased pOsm and undetectable or low plasma arginine vasopressin (AVP) during all tests. In the control group, pOsm and plasma AVP increased only during the osmotic stimulus. The DI group presented lower plasma ANP levels than controls during osmotic stimulus and isotonic volume loading. The lower ANP secretion in DI patients corroborates the importance of neurohypophyseal hormones in ANP regulation. Basal plasma ACTH and cortisol levels did not differ between controls and DI. The latter group presented a higher ACTH response than controls during stimulation with hCRH alone [area under the curve (AUC) 1138 ± 99 vs. 709 ± 62 pmol·L/min] and hCRH/5% NaCl (AUC 1602 ± 209 vs. 1158 ± 187 pmol·L·min). The DI cortisol AUC were higher than controls during stimulation with hCRH alone (65,471 ± 6,070 vs. 48,062 ± 3,476 nmol·L·min) and hCRH/5% NaCl (89,005 ± 10,043 vs. 62,105 ± 5,600 nmol·L·min). The highest ACTH and cortisol responses to hCRH in both groups were obtained with hCRH/5% NaCl. There was a significant correlation between mean pOsm and ACTH response to hCRH (r = 0.62). The increased responses to hCRH with increasing pOsm were present in control subjects and in patients with DI. However, at any given level of pOsm, there was no difference in ACTH response between controls and DI. These data indicate that the acute increases in pOsm augmented the ACTH and cortisol responses to hCRH that involve other factors besides magnocellular AVP.
A Novel Point Mutation in the Insulin Gene Giving Rise to Hyperproinsulinemia*
Authors: Warren-Perry Margaret G. , Manley Susan E. , Ostrega Diane , Polonsky Ken , Mussett Sandra , Brown Phil , Turner Robert C.
Published: January 2014

A 58-yr-old obese white Caucasian male type 2 diabetic, entered into the UK Prospective Diabetes Study, was found to have raised fasting total proinsulin levels 708 pmol/L−1 (normal range, 3–16 pmol/L−1) and normal specific plasma insulin level 29 pmol/L−1 (normal range, 21–75 pmol/L−1). Immunoreactive plasma insulin, measured by RIA, was 503 pmol/L−1. DNA was extracted, the insulin gene amplified by the PCR, and by direct sequencing, a novel point mutation, G1552C, was identified, which resulted in the substitution of proline (CCT) for arginine (CGT) at position 65. This prevented cleavage of the C-peptide A-chain dibasic cleavage site (lys-arg) by the processing protease in the pancreatic β-cells. The plasma proinsulin and insulin levels were in accord with expression of both the wild-type and the mutant alleles. The G1552C mutation was not linked with diabetes, because it was present in a 37-yr-old nondiabetic daughter and not in a 35-yr-old daughter who had had gestational diabetes.
Proteolysis of Insulin-Like Growth Factor-Binding Protein-3 by Human Skin Keratinocytes in Culture in Comparison to that in Skin Interstitial Fluid: The Role and Regulation of Components of the Plasmin System1
Authors: Xu Su , Savage Paul , Burton John L. , Sansom Jane , Holly Jeffrey M. P.
Published: January 2014

Proteolysis of insulin-like growth factor (IGF)-binding protein-3 (IGFBP-3) is an important determinant of IGF action on cells. We have investigated this in a human skin keratinocyte cell line HaCaT. Although these cells did not normally produce an active IGFBP-3 protease, addition of plasminogen resulted in a dose-dependent proteolysis of endogenous and exogenous IGFBP-3, producing fragments similar to those cleaved by skin interstitial fluid, but different from those generated by plasmin. Protease inhibitor profiles suggested the enzyme in the conditioned medium to be a calcium-dependent serine protease.

Exogenous IGFBP-3 either inhibited or slightly stimulated IGF-I-induced cell proliferation when it was coincubated or preincubated with the cells, respectively. Both effects were attenuated in the presence of plasminogen.

Preincubation of cells with IGF-I or long R3 IGF-I divergently changed plasminogen activator inhibitor-1 and -2 secretion, but only IGF-I blocked IGFBP-3 proteolysis. Such inhibition was also observed in a cell-free protease assay. IGF-I, however, had no effect on plasmin-induced IGFBP-3 degradation.

Together, these data indicate that an IGFBP-3 protease similar to that in skin interstitial fluid is generated in plasminogen-treated HaCaT cells, and it attenuates the effects of IGFBP-3 on IGF action. IGF-I, probably by coupling with IGFBP-3, can protect it from the action of this protease.
Long-Term Testosterone Administration Increases Visceral Fat in Female to Male Transsexuals1
Authors: Elbers Jolanda M. H. , Asscheman Henk , Seidell Jacob C. , Megens Jos A. J. , Gooren Louis J. G.
Published: January 2014

The amount of intraabdominal (visceral) fat is an important determinant of disturbances in lipid and glucose metabolism. Cross-sectional studies in women have found associations between high androgen levels and visceral fat accumulation. The causal relation between these phenomena is unknown. We, therefore, studied prospectively the effect of testosterone administration on body fat distribution in 10 young, nonobese, female to male transsexuals undergoing sex reassignment. Before, after 1 yr, and after 3 yr of testosterone administration, magnetic resonance images were obtained at the level of the abdomen, hip, and thigh to quantify both sc and visceral fat depots. After 1 yr of testosterone administration, sc fat depots at all levels showed significant reductions compared to baseline measurements. The mean visceral fat area did not change significantly, but subjects who gained weight in the first year after testosterone administration showed an increase in visceral fat. After 3 yr of testosterone administration, sc fat depots were no longer significantly lower compared to pretreatment measurements, but the mean visceral fat depot had increased significantly by 13 cm2 (95% confidence interval, 4–22 cm2), a relative increase of 47% (95% confidence interval, 8–91%) from baseline. The increase in visceral fat was most pronounced in those subjects who had gained weight.

We conclude that long term testosterone administration in young, nonobese, female subjects increases the amount of visceral fat. In addition, an increase in weight in this hyperandrogenic state leads to a preferential storage of fat in the visceral depot.
Dehydroepiandrosterone Treatment Is Not Beneficial to the Immune Response to Influenza in Elderly Subjects*
Authors: Danenberg H. D. , Ben-Yehuda A. , Zakay-Rones Z. , Gross D. J. , Friedman G.
Published: January 2014

Background. Dehydroepiandrosterone (DHEA) is a native steroid with an immunomodulating activity that was suggested to counterregulate some phenomena of immunosenescence. Recently, it was shown to reverse the age-associated decline of immune response against influenza vaccine in aged mice. The present study was designed to evaluate the effect of DHEA on the immunization of elderly volunteers against influenza.

Methods. Seventy-one elderly volunteers age 61–89 yr were enrolled in a prospective randomized, double-blind study to receive either DHEA (50 mg qd p.o. for four consecutive days starting two days before immunization) or placebo. Antibody response to the vaccine was measured before and 28 days after vaccination.

Results. DHEA treatment significantly increased serum DHEA-sulfate (DHEA-S). No enhancement in established immunity was observed. A significant decrease in attainment of protective antibody titer (1:40 or higher) against the A/Texas strain in subjects with nonprotective baseline antibody titer was recorded following DHEA treatment compared with placebo (52% vs. 84%, P < 0.05). Baseline DHEA-S serum levels were inversely related to attainment of immunization in DHEA-treated subjects. Influenza-like morbidity during the winter was low in the study group with no difference between the DHEA and placebo groups.

Conclusions. Although highly effective in aged rodents, a short course of DHEA treatment did not improve the age-related declined response to immunization against influenza in human subjects. Higher baseline DHEA-S levels are not predictive of better immunization against influenza in the elderly.
The Effect of Oral Glucose on Serum Free Insulin-Like Growth Factor-I and -II in Healthy Adults*
Authors: Frystyk Jan , Grøfte Thorbjørn , Skjærbæk Christian , Ørskov Hans
Published: January 2014

Insulin-like growth factor (IGF) binding protein-I (IGFBP-1) has been suggested to regulate the availability of free IGF and the glucose lowering activity of the IGF-system in relation to fuel supply. Our recent observations of significant inverse correlations between free IGF-I and IGFBP-1 in cross-sectionally collected fasting serum samples support a possible physiological association between the peptides. To further study the impact of IGFBP-1 on free IGF levels and the possible participation of the IGF-system in glucose homeostasis, we studied the time course of changes in IGFBP-1 and free IGFs in 13 healthy subjects undergoing an oral glucose tolerance test (OGTT). Serum was collected every 30 min for 330 min.

Glucose, insulin, and GH followed the expected patterns and had regained baseline levels at 270 min. Total IGF-I and free and total IGF-II remained unaltered. IGFBP-1 decreased significantly by 37–52% (P < 0.05) from 150 to 210 min, whereafter the concentration gradually increased by 75% to a level that tended to be above baseline (P = 0.052). Free IGF-I decreased by 29–38% (P < 0.05) at the end of the study (270–330 min). IGFBP-1 was inversely correlated to free IGF-I at baseline (r = −0.57; P < 0.05), as well as during the OGTT (r = 0.66; P < 0.0001). In contrast, free IGF-II was not correlated to IGFBP-1. Insulin, but not free IGF-I, correlated significantly with serum glucose (P < 0.05).

These results extend our previous findings of an inverse correlation between free IGF-I and IGFBP-1 in cross-sectional studies to include longitudinal observations, and thus further substantiates the hypothesis that IGFBP-1 is an important determinant of free IGF-I in vivo. Significant changes in free IGF-I were observed only in the late postprandial phase, when glucose and insulin were fully normalized, demonstrating that free IGFs probably do not participate in glucoregulation to any significant degree during an oral glucose load in healthy subjects.
A Spontaneous and Severe Hyperstimulation of the Ovaries Revealing a Gonadotroph Adenoma1
Authors: Christin-Maitre Sophie , Rongières-Bertrand Catherine , Kottler Marie-Laure , Lahlou Najiba , Frydman René , Touraine Philippe , Bouchard Philippe
Published: January 2014

We report an unusual case of a gonadotroph adenoma in a 34-yr-old woman, revealed by a dramatic rise in the plasma estradiol (E2) concentration (26,800 pmol/L; normal, <370), with nonsuppressed FSH and LH levels (4.9 and 2.4 mIU/mL, respectively). The PRL level was 503 ng/mL. The testosterone and progesterone levels were 7 and 17 nmol/L, respectively. The levels of inhibin α, inhibin A, and inhibin B were increased compared to normal values in both the follicular (fp) and luteal (lp) phases of the menstrual cycle[ inhibin α, 1986 IU/L (fp normal, <700; lp normal, <1650); inhibin A, 254 pg/mL (fp normal, <20; lp normal, <120); inhibin B, 246 pg/mL (fp normal, <150; lp normal, <30 lp)]. Pituitary magnetic resonance imaging revealed a huge pituitary adenoma. After transphenoidal surgery, the patient presented with pituitary insufficiency and diabetes insipidus. RT-PCR of the tumor tissue was positive for LHβ, FSHβ, α-subunit, and PRL. This case is of particular interest because 1) although the E2 level was extremely high, the patient did not present with ascitis, suggesting that chronic elevated E2 does not play a crucial role in the hyperstimulation symptoms; 2) the extreme rise in E2 was related to the cosecretion of FSH and LH, confirming the two-cell two-gonadotropin theory; and 3) the rise in inhibin B is associated with FSH secretion, whereas the rise in inhibin A is probably due to luteinization.
Immunohistochemical Localization of Somatostatin Receptor sst2A in Human Pancreatic Islets
Authors: Reubi J. C. , Kappeler A. , Waser B. , Schonbrunn A. , Laissue J.
Published: January 2014

Somatostatin and octreotide inhibit endocrine pancreatic functions in man, via specific somatostatin receptors. However, the cellular distribution of the different somatostatin receptor subtype proteins has not been determined in the human pancreas. Here, the immunohistochemical distribution of the sst2A receptor was investigated using the sst2A receptor specific anti-peptide antibody R2-88 in cryostat as well as in formalin-fixed paraffin-embedded sections of human pancreatic tissue, and compared with insulin, glucagon and somatostatin immunostaining of adjacent sections. All pancreatic islets were immunostained with R2-88. Most islet cells were labeled: the sst2A receptors were present in insulin as well as glucagon producing cells, but were not detected in intra-islet vessels nor in adjacent acinar tissue. Absorption of the sst2A antibody with 100 nM of the antigen peptide abolished specific staining in tissue sections. Immunohistochemical staining with 125I-Tyr3-octreotide. Therefore, the clinical efficacy of octreotide on glucagon and insulin release can be explained by the presence of sst2A receptors in human A and B pancreatic islet cells. Moreover, absence of sst2A receptors in human acinar tissue suggests that the action of somatostatin on pancreatic exocrine secretion is mediated either indirectly or through a different somatostatin receptor subtype on acinar cells.
Corticotropin-Releasing Hormone Receptor Subtype 1 Is Significantly Up-Regulated at the Time of Labor in the Human Myometrium1
Authors: Stevens M. Yvette , Challis John R. G. , Lye Stephen J.
Published: January 2014

Circulating concentrations of CRH rise late in human pregnancy, reaching a peak at labor. The presence of functional CRH receptors, CRH-R1 and CRH-R2, in the human myometrium suggests that CRH may modulate uterine activity. We hypothesized that the number of CRH receptors would be higher in myometrium than fetal membranes (FM) and would change during labor.

Myometrial samples were collected from the lower segment (LS) in nonpregnant, preterm (32 ± 2 weeks), and term (39 ± 1.6 weeks) pregnant patients before and at labor. Fundus and LS samples were also collected from nonpregnant, pregnant, laboring, and postpartum women. FM were collected at term and at labor. We identified CRH receptors in myometrium and FM by semiquantitative RT-PCR and immunohistochemistry.

CRH-R1 messenger ribonucleic acid (mRNA) in the LS was decreased in pregnancy and increased significantly in both preterm and term labor (P < 0.05), but remained unchanged in the fundus. CRH-R2 mRNA was present in 28% of LS myometrium with no change at labor. CRH-R1 and CRH-R2 protein was localized to myometrial smooth muscle in nonpregnant and laboring patients, with lower levels at term. CRH-R1 mRNA was present in chorion and decidua, but CRH-R2 was undetectable in these tissues. We conclude that CRH-R1 is expressed preferentially in myometrium and FM. Changes in CRH receptors during labor are consistent with CRH mediating effects on myometrial activity.
Both Inhibin A and B Respond to Exogenous Follicle-Stimulating Hormone in the Follicular Phase of the Human Menstrual Cycle1
Authors: Burger Henry G. , Groome Nigel P. , Robertson David M.
Published: January 2014

To ascertain whether changes in the concentrations of the dimeric inhibins A and/or B (INH-A and INH-B) contributed to the previously described dose-dependent increase in immunoreactive inhibin (INH) in response to FSH during the follicular phase of the human menstrual cycle, both dimers were measured by specific two-site assays in stored serum samples from regularly cycling normal volunteers who had received saline as a control (n = 5) or FSH [100 IU (n = 6) or 200 IU (n = 5)] between days 3–5 of the menstrual cycle. Both INH-A and INH-B showed a dose-dependent increase in response to administered FSH; INH-A rose from 13.5 to 35.9 ng/L (P < 0.01), and INH-B rose from 77.8 to 205 ng/L (P < 0.05) at 36 h after 200 IU FSH. Highly significant correlations were observed between INH and each of the specific inhibin dimers (A: r= 0.79, P < 0.001; B: r = 0.76, P < 0.001), and the responses of the two dimers were also highly correlated (r = 0.59, P < 0.001). The response of each inhibin was also highly correlated with the response of serum estradiol (A: r = 0.45, P < 0.001; B: r = 0.40, P< 0.001). When analyzed by ANOVA, the INH response of INH-B was significantly above the control value at 36 h after treatment with both 100 and 200 IU FSH, whereas the response of INH-A was significant only at 200 IU. It is concluded that the concentrations of both dimeric INH-A and INH-B are stimulated by increases in FSH within the physiological range in the follicular phase of the human menstrual cycle and that both contribute to the previously observed rise in INH.
Cortisol Secretion Is Related to Electroencephalographic Alertness in Human Subjects during Daytime Wakefulness1
Authors: Chapotot Florian , Gronfier Claude , Jouny Christophe , Muzet Alain , Brandenberger Gabrielle
Published: January 2014

To determine whether human hypothalamo-pituitary-adrenal axis activity is related to the alertness level during wakefulness, 10 healthy young men were studied under resting conditions in the daytime (0900–1800 h) after an 8-h nighttime sleep (2300–0700 h). A serial 70-sec gaze fixation task was required every 10 min throughout the daytime experimental session. The corresponding waking electroencephalographic (EEG) segments were submitted to quantitative spectral analysis, from which EEG β activity (absolute power density in the 13–35 Hz frequency band), an index of central alertness, was computed. Blood was collected continuously through an indwelling venous catheter and sampled at 10-min intervals. Plasma cortisol concentrations were measured by RIA, and the corresponding secretory rates were determined by a deconvolution procedure.

Analysis of individual profiles demonstrated a declining tendency for EEG β activity and cortisol secretory rate, with an overall temporal relationship indicated by positive and significant cross-correlation coefficients between the two variables in all subjects (average r= 0.565, P < 0.001). Changes in cortisol secretion lagged behind fluctuations in EEG β activity, with an average delay of 10 min for all the subjects. On the average, 4.6 ± 0.4 cortisol secretory pulses and 4.9 ± 0.5 peaks in EEG β activity were identified by a detection algorithm. A significant, although not systematic, association between the episodes in the two variables was found: 44% of the peaks in EEG β activity (relative amplitude, near 125%; P < 0.001) occurred during an ascending phase of cortisol secretion, cortisol secretory rates increasing by 40% (P < 0.01) 10-min after peaks in EEG β activity. However, no significant change in EEG β activity was observed during the period from 50 min before to 50 min after pulses in cortisol secretion.

In conclusion, the present study describes a temporal coupling between cortisol release and central alertness, as reflected in the waking EEGβ activity. These findings suggest the existence of connections between the mechanisms involved in the control of hypothalamo-pituitary-adrenal activity and the activation processes of the brain, which undergoes varying degrees of alertness throughout daytime wakefulness.
Activating Mutations of the Gsα Gene Are Associated with Low Levels of Gsα Protein in Growth Hormone-Secreting Tumors1
Authors: Ballaré Emilia , Mantovani Simona , Lania Andrea , Di Blasio Anna M. , Vallar Lucia , Spada Anna
Published: January 2014

Evidence suggests the existence of a direct relationship between cellular Gsα content and activation of the adenylyl cyclase system. Data on Gsα levels in endocrine tumors that depend on cAMP for growth, particularly pituitary adenomas, are still limited. The levels of Gsα protein were evaluated in 11 GH-secreting adenomas with Gsα mutations (gsp+) and 15 without (gsp). Complementary DNAs from gsp+ tumors contained very low amounts of wild-type Gsα sequences, indicating a preponderance of the mutant Gsα transcripts in these tumors. Immunoblotting of Gsα protein showed that the two isoforms were present at high levels in all gsp−, but were undetectable or barely detectable in gsp+. The low Gsα content in gsp+ tumors was not due to a reduction in ribonucleic acid synthesis or stability, as Gsα messenger ribonucleic acid levels were similar in wild-type and mutant tissues. Treatment of gsp− cells with cholera toxin caused a marked reduction of Gsα levels. As in other cell systems cholera toxin increases Gsα degradation, our data are consistent with an accelerated removal of mutant Gsα. This may represent an additional mechanism of feedback response to the constitutive activation of cAMP signaling in pituitary tumors with mutations in the Gsα gene.
Glucocorticoid Effects in the Human Placenta: Evidence That Dexamethasone-Mediated Inhibition of Fibronectin Expression in Cytotrophoblasts Involves a Protein Intermediate1
Authors: Yoon Diana Y. , Ma Yuehong , Krikun Graciela , Lockwood Charles J. , LaChapelle Linda , Guller Seth
Published: January 2014

Oncofetal fibronectin is an extracellular matrix protein that is suggested to play an important role in regulating adherence at uterine-placental interfaces. The purpose of the present study was to elucidate a mechanism through which glucocorticoids (GCs) inhibit the synthesis of FN in human placenta as part of their matrix-suppressive action near parturition. We observed that treatment of cytotrophoblasts isolated from human term placentas for 48 h with 10−7 mol/L dexamethasone (DEX) down-regulated levels of FN expression to 13–19% of control levels in immunoprecipitation, Northern blotting, and enzyme-linked immunosorbent assay experiments. Conversely, GC treatment increased FN expression in placental fibroblasts to 164–310% of control levels in Northern blotting and enzyme-linked immunosorbent assay procedures, suggesting that GC-mediated suppression of FN expression is specific to cytotrophoblasts. Results indicated that the DEX-mediated suppression of FN expression in cytotrophoblasts was not mediated through changes in the stability of FN messenger ribonucleic acid (mRNA). Run-on transcription assays using isolated nuclei suggested that GC treatment did not markedly affect transcription of the FN gene in cytotrophoblasts. To test whether the GC-mediated suppression of FN expression was mediated through a protein intermediate, levels of FN mRNA were examined by Northern blotting in cells treated for 48 h with and without 10−7 mol/L DEX and cycloheximide (CHX; 125 ng/mL). We observed that CHX treatment increased FN expression in DEX-treated cells to 91% of control values. We noted that whereas the presence of 100–300 ng/mL CHX reversed the DEX-mediated inhibition of FN mRNA expression in cytotrophoblasts, it did not alter the overall rates of protein synthesis in DEX-treated and control cells. These data suggest that suppression of FN mRNA expression by GC in cytotrophoblasts requires de novo protein synthesis and is mediated through a short lived intermediate, the synthesis of which is inhibited at low concentrations of CHX. Thus, GC-induced protein intermediates may influence uterine-placental adherence by modulating levels of oncofetal FN at sites of uterine-placental contact.
Cohort Size Rather Than Follicle-Stimulating Hormone Threshold Level Determines Ovarian Sensitivity in Polycystic Ovary Syndrome1
Authors: van der Meer Maartje , Hompes Peter G. A. , de Boer Jacoba A. M. , Schats Roel , Schoemaker Joop
Published: January 2014

The aim of the presented study was to compare FSH threshold levels and ovarian response to stimulation with one of two standard increments of exogenous FSH above the threshold in patients with polycystic ovary syndrome (PCOS) (n = 12) and eumenorrheic women (n = 11). The individual FSH threshold was determined by treatment according to a low-dose, step-up protocol with urinary FSH (Metrodin; Ares Serono, Geneva, Switzerland). In a subsequent treatment cycle, six PCOS patients and six eumenorrheic women were randomly assigned to double-blind treatment with the threshold dose plus 1/2 ampoule; the other six PCOS patients and five eumenorrheic women were treated with 1 ampoule above the threshold dose. Determination of threshold levels showed no significant differences in median and range between PCOS patients and eumenorrheic women. The number of follicles on the day of human chorionic gonadotropin administration showed no significant correlation with the increase in FSH level above the threshold level. Irrespective of the dose given, the number of follicles in the PCOS group was significantly higher than in eumenorrheic women. The higher sensitivity for gonadotropin stimulation in patients with PCOS compared with women with regular menstrual cycles therefore appears not to be dependent on differences in FSH threshold level, but rather on the larger size of the FSH sensitive cohort of small antral follicles.
Human Adrenocorticotropin-Secreting Pituitary Adenomas Show Frequent Loss of Heterozygosity at the Glucocorticoid Receptor Gene Locus1
Authors: Huizenga Nannette A. T. M. , de Lange Pieter , Koper Jan W. , Clayton Richard N. , Farrell William E. , van der Lely Aart Jan , Brinkmann Albert O. , de Jong Frank H. , Lamberts Steven W. J.
Published: January 2014

Corticotropinomas are characterized by a relative resistance to the negative feedback action of cortisol on ACTH secretion. In this respect there is a similarity with the clinical syndrome of cortisol resistance. As cortisol resistance can be caused by genetic abnormalities in the glucocorticoid receptor (GR) gene, we investigated whether the insensitivity of corticotropinomas to cortisol is also caused by de novo mutations in the GR gene.

We screened for the GR gene in leukocyte and tumor DNA from 22 patients with Cushing’s disease for mutations using PCR/single strand conformation polymorphism analysis. In a previous study, we identified 5 polymorphisms in the GR gene in a normal population. These polymorphisms were used as markers for the possible occurrence of loss of heterozygosity (LOH) at the GR gene locus.

Except for 1 silent point mutation, we did not identify novel mutations in the GR gene in leukocytes or corticotropinomas from these patients. Of the 22 patients, 18 were heterozygous for at least 1 of the polymorphisms. In 6 of these patients, LOH had occurred in the tumor DNA. Of 21 patients examined for LOH on chromosome 11q13, only 1, with a corticotroph carcinoma, showed allelic deletion. As controls we studied 28 pituitary tumors of other subtypes (11 clinically nonfunctioning, 8 prolactinomas, and 9 GH-producing adenomas) and found evidence for LOH in only 1 prolactinoma.

In six patients LOH was found at the GR gene locus (chromosome 5) in DNA derived from adenoma cells. Our observations indicate for the first time that LOH at the GR gene locus is a relatively frequent phenomenon in pituitary adenomas of patients with Cushing’s disease. This might explain the relative resistance of the adenoma cells to the inhibitory feedback action of cortisol on ACTH secretion. The specificity of the GR LOH to corticotropinomas supports this concept. Somatic mutations of the GR are not a frequent cause of relative cortisol resistance in these cells.
Absence of Germ-Line Mutations of the Multiple Endocrine Neoplasia Type 1 (MEN1) Gene in Familial Pituitary Adenoma in Contrast to MEN1 in Japanese1
Authors: Tanaka Chisato , Yoshimoto Katsuhiko , Yamada Shozo , Nishioka Hiroshi , Ii Setsuko , Moritani Maki , Yamaoka Takashi , Itakura Mitsuo
Published: January 2014

Germ-line mutations of the MEN1 gene were analyzed in five cases of familial and four cases of sporadic multiple endocrine neoplasia type 1 (MEN-1), six cases in three independent pedigrees of familial pituitary adenoma without MEN-1, and three cases of familial isolated primary hyperparathyroidism (FIHP) in Japanese. Eight different types of germ-line mutations in all nine cases of MEN-1 were distributed in exons 2, 3, 7, and 10 and intron 7 of the MEN1 gene. Loss of heterozygosity (LOH) on 11q13 was detected in all nine tumors of these cases with microsatellite analysis. No germ-line mutation of the MEN1 gene was detected in three pedigrees of familial pituitary adenoma and three cases of FIHP. LOH on 11q13 was detected in two cases in one pedigree of familial pituitary adenoma, and one of them showed a heterozygous somatic mutation of the MEN1 gene. No LOH on 11q13 was detected in three cases of FIHP. Based on these, we conclude that the loss of function of menin is etiological for familial or sporadic MEN-1, but not for FIHP or most familial pituitary adenoma without MEN-1.
Sex Differences in Endocrine and Psychological Responses to Psychosocial Stress in Healthy Elderly Subjects and the Impact of a 2-Week Dehydroepiandrosterone Treatment1
Authors: Kudielka Brigitte M. , Hellhammer Juliane , Hellhammer Dirk H. , Wolf Oliver T. , Pirke Karl-Martin , Varadi Enikö , Pilz Jürgen , Kirschbaum Clemens
Published: January 2014

Evidence from animal as well as human studies has suggested that significant sex differences exist in hypothalamus-pituitary-adrenal axis (HPA) activity. As gonadal steroids could be important modulators of HPA sex differences, stress responses were investigated in subjects of advanced age after dehydroepiandrosterone (DHEA) or placebo treatment.

After a 2-week treatment with 50 mg DHEA daily or placebo, 75 men and women (mean age, 67.6 yr) were exposed to the Trier Social Stress Test (TSST). The TSST is a brief psychosocial stress that consists of a free speech and mental arithmetic task in front of an audience. The results show that the TSST induced significant increases in ACTH, salivary free cortisol, total plasma cortisol, norepinephrine, and heart rates (all P < 0.0001) as well as decreased positive affect in the elderly (P = 0.0009). Men showed larger stress responses in ACTH (P = 0.004), salivary free cortisol (P = 0.044), and plasma total cortisol (P = 0.076) compared to women. No sex differences were observed in norepinephrine, epinephrine, or heart rate responses. In contrast to ACTH and cortisol response differences, women reported that they were significantly more stressed by the TSST than men (P = 0.0051).

Women treated with DHEA showed ACTH stress responses similar to those of men, but significantly enhanced compared to those of women taking placebos (P < 0.009). No other stress response differences emerged between DHEA and placebo groups. Finally, DHEA treatment did not result in an improvement of subjective well-being.

We conclude that elderly men show larger HPA responses than women to psychosocial stress, as studied in the TSST. Estrogen effects on hypothalamic CRF-producing neurons might be responsible for these sex differences.
Involvement of Ovarian Steroids in the Opioid-Mediated Reduction of Insulin Secretion in Hyperinsulinemic Patients with Polycystic Ovary Syndrome
Authors: Guido Maurizio , Pavone Virginia , Ciampelli Mario , Murgia Francesca , Fulghesu Anna Maria , Apa Rosanna , Caruso Alessandro , Mancuso Salvatore , Lanzone Antonio
Published: January 2014

To evaluate the possible involvement of ovarian steroids on the opioid-mediated disorders of insulin in patients affected by polycystic ovary syndrome (PCOS), we studied 40 PCOS women. All patients underwent an oral glucose tolerance test (OGTT; 75 g) and basal hormone assay; based on the insulin response to OGTT, 26 women were classified as hyperinsulinemic and continued the study protocol.

Patients were randomly divided into three groups characterized by different treatments: group A (nine patients) was treated with GnRH analog (one ampule every 28 days for 2 months), group B (eight patients) was treated with naltrexone (an oral opioid antagonist, 50 mg/day, orally) for 8 weeks, and group C (nine patients) was treated with GnRH analog plus naltrexone for 2 months. After continuation of treatment, all patients repeated the basal study in a second hospitalization.

Naltrexone treatment significantly reduced the insulin response to OGTT, whereas GnRH analogue administration did not significantly change the insulin secretion after the glucose load. The GnRH analog/naltrexone cotreatment was not able to influence the insulin secretory pattern; in fact, the insulin area under the curve was superimposable before and after therapy. These data could lead to the hypothesis that the opioidergic regulation of insulin secretion requires a normal steroidogenic pattern, thus suggesting that ovarian steroids modulate opioid activity also at peripheric districts.
Age-Related Perception of Stature, Acceptance of Therapy, and Psychosocial Functioning in Human Growth Hormone-Treated Girls with Turner’s Syndrome1
Authors: Lagrou Katrien , Xhrouet-Heinrichs Danielle , Heinrichs Claudine , Craen Margarita , Chanoine Jean-Pierre , Malvaux Paul , Bourguignon Jean-Pierre
Published: January 2014

This study evaluated the perception of stature, acceptance of therapy, and psychosocial functioning in relation to age at onset and time on treatment during 2 yr of GH therapy in 31 girls with Turner’s syndrome grouped by age (group A: 3.7–5.8 yr, n = 9; group B: 7.2–11.8 yr, n = 13; group C: 12.5–16.4 yr, n = 9). The growth response after 2 yr was significant in the 3 groups when calculated in terms of growth norms for untreated Turner girls (mean increase in height sd score: +1.2, +1.5, and +1.1, respectively). The effect was less marked in terms of growth norms for normal girls, particularly in group B (+0.5 sd score). Height was perceived as a problem by most patients, except in the youngest girls at the start of treatment (group A) and in the majority of the adolescents after 2 yr of GH therapy (group C), without evidence of relation to growth response during therapy. The GH injections were fairly well accepted by all patients, except those younger than 6 yr. In all patients, expected adult height was unrealistic and became more realistic with age, whereas no consistent changes were observed in relation to growth response to GH therapy. The Child Behavior Checklist revealed elevated mean scores at the behavioral subscales of attention problems (group A and B), social problems, withdrawal, and anxiety-depression (most obviously in group B). No significant changes were seen during GH therapy. In group C, an elevated mean social problem score at the Youth Self Report and a low mean social self-esteem score at the Self-Esteem Inventory were observed before therapy and showed a significant improvement during 2 yr of GH treatment. These results, however, might be biased due to an increase in social desirability during therapy. We conclude that the perception of height, acceptance of GH therapy, and psychosocial functioning in girls with Turner’s syndrome show important differences between age groups, with only slight changes observed during GH therapy.