bluuebunny/crossref_metadata_2025 huggingface dataset:
Anthropogenic climate change and the nature of Earth System science
Authors: Oldfield Frank , Steffen Will
Published: January 2014

One of the criticisms made by those sceptical of the majority scientific consensus on climate change and its likely future consequences is that the Earth System science upon which it is based is fundamentally flawed. This contention is challenged here by an outline of the nature of the science needed to make future projections possible. The classic Popperian approach to science, in which potentially refutable hypotheses are defined and tested is not well suited to the challenges posed by an Earth System that is characterised by high degrees of complexity, non-linearity and a lack of definable cause–consequence relationships. A science based on model–data comparisons and interactions is the only effective approach both to increasing our understanding of the Earth System and developing a well substantiated basis for future projections.
Resuscitation of a Neonate With Medium Chain Acyl-Coenzyme A Dehydrogenase Deficiency Using Extracorporeal Life Support
Authors: Kumar Gaurav , Mattke Adrian C. , Bowling Frank , McWhinney Avis , Alphonso Nelson , Karl Tom R.
Published: January 2014

We report a neonate with medium chain acyl-coenzyme A dehydrogenase deficiency (MCAD) who had a cardiac arrest due to ventricular tachycardia and fibrillation. Extracorporeal life support (ECLS) was deployed, from which the baby was subsequently separated and discharged from hospital. This case was a rare neonatal presentation of MCAD and an uncommon indication for ECLS. We discuss the presentations of patients with MCAD and the use of ECLS for patients with possible inborn errors of metabolism and other unknown primary diagnoses.
High Incidence of Abdominal Venovenous Collaterals After Kawashima Operation
Authors: Rahmath Muhammed Riyas K , Masud Faraz , Sallehuddin Ahmad , Kamal Reema Yousef , Bhat Akhlaque Nabi , Dilawar Muhammad
Published: January 2014

Introduction:

Systemic venous collaterals have been found at different stages of single-ventricle palliation, specifically after Kawashima operation. In this study, we present the incidence, clinical features, associated risk factors, and management of such venovenous collaterals (VVCs) after Kawashima procedure.

Materials and Methods:

A retrospective review of all the patients who underwent Kawashima procedure prior to December 2011 at Hamad General Hospital, Qatar, was performed. Data were collected and reviewed from medical records.

Results:

The study group involved a total of six patients with Kawashima procedure. The median age of patients was 112.7 months (range 46-336 months), and median age at the time of Kawashima operation was 31.7 months (range 15-187 months). Mean systemic arterial oxygen saturation (Spo2) after Kawashima operation was 94.5% ± 1.5%. In five patients, during a median follow-up of 53.4 months (range 16.9-147.9 months), the Spo2 declined to a mean of 78.8% ± 8.2%. A total of 16 venous collaterals were observed in the study group, and majority (70%) of these collaterals were subdiaphragmatic.

Conclusion:

In our study, abdominal VVCs were observed in 100% of the patients who were followed after Kawashima operation. We therefore recommend that thorough evaluation for such VVCs should be part of the ongoing evaluation of patients after Kawashima operation, especially in those with low Spo2. We also recommend early surgical rerouting of hepatic veins to pulmonary arteries in all post-Kawashima patients.
Left Main Coronary Artery Stenting in a Neonate After Arterial Switch Operation for Transposition of the Great Arteries
Authors: Sinelnikov Yuri S. , Kornilov Igor A. , Redkin Dmitry A. , Strelnikova Marina S. , Ivantsov Sergey M. , Kalinin Roman A. , Gorbatyh Yuri N.
Published: January 2014

Anatomical correction of transposition of the great arteries (TGA) with an intramural coronary artery is associated with high risk of coronary complications such as vessel injury and stenosis. Here, we report on a case of left main coronary artery stenting in a neonate with single coronary artery after repair for TGA.
Relationship Between Risk-Adjustment Tools and the Pediatric Logistic Organ Dysfunction Score
Authors: Russell Rebecca A. , Ghanayem Nancy S. , Kuhn Evelyn M. , Jeffries Howard E. , Scanlon Matthew C. , Rice Tom B.
Published: January 2014

Background:

The Risk-Adjusted Classification for Congenital Heart Surgery (RACHS-1) method and Aristotle Basic Complexity (ABC) scores correlate with mortality. However, low mortality rates in congenital heart disease (CHD) make use of mortality as the primary outcome measure insufficient. Demonstrating correlation between risk-adjustment tools and the Pediatric Logistic Organ Dysfunction (PELOD) score might allow for risk-adjusted comparison of an outcome measure other than mortality.

Methods:

Data were obtained from the Virtual PICU Systems database. Patients with postoperative CHD between 2009 and 2010 were included. Correlation between RACHS-1 category and PELOD score and between ABC level and PELOD score was examined using Spearman rank correlation. Consistency of PELOD scores across institutions for given levels of case complexity was examined using Kruskal-Wallis nonparametric analysis of variance.

Results:

A total of 1,981 patient visits among 12 institutions met inclusion criteria. Positive correlations between PELOD score and RACHS-1 category ( rs = .353, P < .0001) as well as between PELOD score and ABC level ( rs = .328, P < .0001) were demonstrated. Variability in PELOD scores across individual centers for given levels of case complexity was observed ( P < .04).

Conclusions:

Risk-Adjusted Classification for Congenital Heart Surgery categories and ABC levels correlate with postoperative organ dysfunction as measured by PELOD. However, the correlation was weak, potentially due to limitations of the PELOD score itself. Identification of a more accurate metric of morbidity for the congenital heart disease population is needed.
Medial Epicondyle Morphology in Elite Overhead Athletes
Authors: Makhni Eric C. , Khanna Krishn , Simpson Michael T. , Redler Lauren H. , Anakwenze Oke A. , Li Richard , Ahmad Christopher S.
Published: January 2014

Background:

Prior studies have attempted to determine morphological characteristics of the medial epicondyle in overhead athletes, but no study has reported on precise quantitative differences between elite overhead athletes and control patients.

Hypothesis:

The medial epicondyle in overhead athletes is larger in volume than those of control patients.

Study Design:

Cross-sectional study; Level of evidence, 3.

Methods:

Computer simulation modeling from advanced (computed tomography/magnetic resonance imaging) imaging of the elbow of 37 patients (22 elite overhead athletes, 15 control patients) was performed to provide detailed assessment of the morphological characteristics of the medial epicondyle. Several quantitative metrics regarding the medial epicondyle were measured and compared across both cohorts, including that of epicondyle width (medial-lateral), height (superior-inferior), thickness (anterior-posterior), volume, percentage cortical volume, and morphology of the inferior slope of the epicondyle.

Results:

The medial epicondyle in overhead athletes was significantly larger than that found in nonathlete controls (4976 vs 3682 mm3; P = .001). There was no significance between the 2 cohorts in medial-lateral width (16.8 vs 16.6 mm; P = .68), but there was a difference in anterior-posterior thickness (16.96 vs 14.40 mm; P = .001) and superior-inferior height (39.55 vs 35.86 mm; P = .09) in athletes versus controls. The epicondyle volume was 97.9% cortical bone in athletes compared with 82.3% in control patients ( P < .001). There were no differences in the morphology of the inferior epicondyle slope between the 2 groups.

Conclusion:

The medial epicondyle in overhead athletes is larger in volume and anterior-posterior thickness than those of control patients. Additionally, the medial epicondyle is comprised nearly entirely of cortical bone in overhead athletes.

Clinical Relevance:

These quantitative findings support the theory of adaptive remodeling in skeletally immature overhead athletes.
Characterization of Rhizobium grahamii extrachromosomal replicons and their transfer among rhizobia
Authors: Althabegoiti María Julia , Ormeño-Orrillo Ernesto , Lozano Luis , Torres Tejerizo Gonzalo , Rogel Marco Antonio , Mora Jaime , Martínez-Romero Esperanza
Published: January 2014

Background

Rhizobium grahamii belongs to a new phylogenetic group of rhizobia together with Rhizobium mesoamericanum and other species. R. grahamii has a broad-host-range that includes Leucaena leucocephala and Phaseolus vulgaris, although it is a poor competitor for P. vulgaris nodulation in the presence of Rhizobium etli or Rhizobium phaseoli strains. This work analyzed the genome sequence and transfer properties of R. grahamii plasmids.

Results

Genome sequence was obtained from R. grahamii CCGE502 type strain isolated from Dalea leporina in Mexico. The CCGE502 genome comprises one chromosome and two extrachromosomal replicons (ERs), pRgrCCGE502a and pRgrCCGE502b. Additionally, a plasmid integrated in the CCGE502 chromosome was found. The genomic comparison of ERs from this group showed that gene content is more variable than average nucleotide identity (ANI). Well conserved nod and nif genes were found in R. grahamii and R. mesoamericanum with some differences. R. phaseoli Ch24-10 genes expressed in bacterial cells in roots were found to be conserved in pRgrCCGE502b. Regarding conjugative transfer we were unable to transfer the R. grahamii CCGE502 symbiotic plasmid and its megaplasmid to other rhizobial hosts but we could transfer the symbiotic plasmid to Agrobacterium tumefaciens with transfer dependent on homoserine lactones.

Conclusion

Variable degrees of nucleotide identity and gene content conservation were found among the different R. grahamii CCGE502 replicons in comparison to R. mesoamericanum genomes. The extrachromosomal replicons from R. grahamii were more similar to those found in phylogenetically related Rhizobium species. However, limited similarities of R. grahamii CCGE502 symbiotic plasmid and megaplasmid were observed in other more distant Rhizobium species. The set of conserved genes in R. grahamii comprises some of those that are highly expressed in R. phaseoli on plant roots, suggesting that they play an important role in root colonization.
Factors associated with chronic musculoskeletal pain in patients with chronic kidney disease
Authors: Hsu Heng-Jung , Yen Chiung-Hui , Hsu Kuang-Hung , Wu I-Wen , Lee Chin-Chan , Hung Ming-Jui , Sun Chiao-Yin , Chou Chia-Chi , Chen Yung-Chih , Hsieh Ming-Fang , Chen Chun-Yu , Hsu Chiao-Ying , Tsai Chi-Jen , Wu Mai-Szu
Published: January 2014

Background

Chronic musculoskeletal (MS) pain is common in patients with chronic kidney disease (CKD) undergoing haemodialysis. However, epidemiological data for chronic MS pain and factors associated with chronic MS pain in patients with early- or late-stage CKD who are not undergoing dialysis are limited.

Method

A cross-sectional study to evaluate the prevalence of chronic MS pain and factors associated with chronic MS pain in patients with early- and late-stage CKD who were not undergoing dialysis, was conducted. In addition, the distribution of pain severity among patients with different stages of CKD was evaluated.

Results

Of the 456 CKD patients studied, 53.3% (n = 243/456) had chronic MS pain. Chronic MS pain was independently and significantly associated with hyperuricemia as co-morbidity, as well as with the calcium × phosphate product levels. In CKD patients with hyperuricemia, chronic MS pain showed a negative, independent significant association with diabetes mellitus as a co-morbidity (odds ratio: 0.413, p = 0.020). However, in the CKD patients without hyperuricemia as a co-morbidity, chronic MS pain showed an independent significant association with the calcium × phosphate product levels (odds ratio: 1.093, p = 0.027). Furthermore, stage-5 CKD patients seemed to experience more severe chronic MS pain than patients with other stages of CKD.

Conclusion

Chronic MS pain is common in CKD patients. Chronic MS pain was independently and significantly associated with hyperuricemia as co-morbidity, and with the calcium × phosphate product levels in early- and late-stage CKD patients who were not on dialysis.
Genetic diversity of Plasmodium vivax population in Anhui province of China
Authors: Huang Bo , Huang Shiguang , Su Xin-zhuan , Guo Hong , Xu Yucheng , Xu Fei , Hu Xuchu , Yang Yaming , Wang Shanqing , Lu Fangli
Published: January 2014

Background

Although the numbers of malaria cases in China have been declining in recent years, outbreaks of Plasmodium vivax malaria were still being reported in rural areas south of the Yellow River. To better understand the transmission dynamics of P. vivax parasites in China, the extent of genetic diversity of P. vivax populations circulating in Bozhou of Anhui province of China were investigated using three polymorphic genetic markers: merozoite surface proteins 1 and 3α (pvmsp-1 and pvmsp-3α) and circumsporozoite protein (pvcsp).

Methods

Forty-five P. vivax clinical isolates from Bouzhou of Anhui province were collected from 2009 to 2010 and were analysed using PCR/RFLP or DNA sequencing.

Results

Seven and six distinct allelic variants were identified using PCR/RFLP analysis of pvmsp-3α with Hha I and Alu I, respectively. DNA sequence analysis of pvmsp-1 (variable block 5) revealed that there were Sal-I and recombinant types but not Belem type, and seven distinct allelic variants in pvmsp-1 were detected, with recombinant subtype 2 (R2) being predominant (66.7%). All the isolates carried pvcsp with VK210 type but not VK247 or P. vivax-like types in the samples. Sequence analysis of pvcsp gene revealed 12 distinct allelic variants, with VK210-1 being predominant (41.5%).

Conclusions

The present data indicate that there is some degree of genetic diversity among P. vivax populations in Anhui province of China. The genetic data obtained may assist in the surveillance of P. vivax infection in endemic areas or in tracking potential future disease outbreak.
Microwave-assisted synthesis of layered basic zinc acetate nanosheets and their thermal decomposition into nanocrystalline ZnO
Authors: Tarat Afshin , Nettle Chris J , Bryant Daniel TJ , Jones Daniel R , Penny Mark W , Brown Richard A , Majitha Ravish , Meissner Kenith E , Maffeis Thierry GG
Published: January 2014

We have developed a low-cost technique using a conventional microwave oven to grow layered basic zinc acetate (LBZA) nanosheets (NSs) from a zinc acetate, zinc nitrate and HMTA solution in only 2 min. The as-grown crystals and their pyrolytic decomposition into ZnO nanocrystalline NSs are characterized by scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), atomic force microscopy (AFM), X-ray diffraction (XRD) and photoluminescence (PL). SEM and AFM measurements show that the LBZA NSs have typical lateral dimensions of 1 to 5 μm and thickness of 20 to 100 nm. Annealing in air from 200°C to 1,000°C results in the formation of ZnO nanocrystalline NSs, with a nanocrystallite size ranging from 16 nm at 200°C to 104 nm at 1,000°C, as determined by SEM. SEM shows evidence of sintering at 600°C. PL shows that the shape of the visible band is greatly affected by the annealing temperature and that the exciton band to defect band intensity ratio is maximum at 400°C and decreases by a factor of 15 after annealing at 600°C. The shape and thickness of the ZnO nanocrystalline NSs are the same as LBZA NSs. This structure provides a high surface-to-volume ratio of interconnected nanoparticles that is favorable for applications requiring high specific area and low resistivity such as gas sensing and dye-sensitized solar cells (DSCs). We show that resistive gas sensors fabricated with the ZnO NSs showed a response of 1.12 and 1.65 to 12.5 ppm and 200 ppm of CO at 350°C in dry air, respectively, and that DSCs also fabricated from the material had an overall efficiency of 1.3%.

PACS

81.07.-b; 62.23.Kn; 61.82.Fk
Neurobiological Mechanisms of the Onset of Puberty in Primates*
Authors: Terasawa Ei , Fernandez David L.
Published: January 2014

An increase in pulsatile release of LHRH is essential for the onset of puberty. However, the mechanism controlling the pubertal increase in LHRH release is still unclear. In primates the LHRH neurosecretory system is already active during the neonatal period but subsequently enters a dormant state in the juvenile/prepubertal period. Neither gonadal steroid hormones nor the absence of facilitatory neuronal inputs to LHRH neurons is responsible for the low levels of LHRH release before the onset of puberty in primates. Recent studies suggest that during the prepubertal period an inhibitory neuronal system suppresses LHRH release and that during the subsequent maturation of the hypothalamus this prepubertal inhibition is removed, allowing the adult pattern of pulsatile LHRH release. In fact,γ -aminobutyric acid (GABA) appears to be an inhibitory neurotransmitter responsible for restricting LHRH release before the onset of puberty in female rhesus monkeys. In addition, it appears that the reduction in tonic GABA inhibition allows an increase in the release of glutamate as well as other neurotransmitters, which contributes to the increase in pubertal LHRH release. In this review, developmental changes in several neurotransmitter systems controlling pulsatile LHRH release are extensively reviewed.
Estrogen and Spermatogenesis*
Authors: O’Donnell Liza , Robertson Kirsten M. , Jones Margaret E. , Simpson Evan R.
Published: January 2014

Although it has been known for many years that estrogen administration has deleterious effects on male fertility, data from transgenic mice deficient in estrogen receptors or aromatase point to an essential physiological role for estrogen in male fertility. This review summarizes the current knowledge on the localization of estrogen receptors and aromatase in the testis in an effort to understand the likely sites of estrogen action. The review also discusses the many studies that have used models employing the administration of estrogenic substances to show that male fertility is responsive to estrogen, thus providing a mechanism by which inappropriate exposure to estrogenic substances may cause adverse effects on spermatogenesis and male fertility. The reproductive phenotypes of mice deficient in estrogen receptors α and/or β and aromatase are also compared to evaluate the physiological role of estrogen in male fertility. The review focuses on the effects of estrogen administration or deprivation, primarily in rodents, on the hypothalamo-pituitary-testis axis, testicular function (including Leydig cell, Sertoli cell, and germ cell development and function), and in the development and function of the efferent ductules and epididymis. The requirement for estrogen in normal male sexual behavior is also reviewed, along with the somewhat limited data on the fertility of men who lack either the capacity to produce or respond to estrogen. This review highlights the ability of exogenous estrogen exposure to perturb spermatogenesis and male fertility, as well as the emerging physiological role of estrogens in male fertility, suggesting that, in this local context, estrogenic substances should also be considered “male hormones.”
Free Fatty Acids Inhibit Growth Hormone/Signal Transducer and Activator of Transcription-5 Signaling in Human Muscle: A Potential Feedback Mechanism
Authors: Møller Niels , Gormsen Lars C. , Schmitz Ole , Lund Sten , Jørgensen Jens Otto L. , Jessen Niels
Published: January 2014

Context:

Stimulation of lipolysis, leading to increased blood concentrations of free fatty acids (FFAs), is a primary effect of GH and phosphorylation of intracellular signal transducer and activator of transcription (STAT)-5 is a primary mediator of the effects of GH.

Objective:

Based on preliminary results, we intended to test whether FFAs exert a negative feedback inhibition of STAT5 phosphorylation in skeletal muscle.

Design and Participants:

Eight healthy young men were investigated for 8 h on four occasions at four different FFA levels in a single blind, randomized manner. Acipimox was used to suppress FFA levels and Intralipid was infused to obtain appropriate FFA concentrations. Somatostatin was infused to control GH levels and GH, insulin, and glucagon were replaced. Muscle biopsies were taken after 8 h and compared with a fifth biopsy taken under normal basal conditions.

Setting:

The study was conducted at a university clinical research unit.

Results:

GH concentrations remained steady and comparable in all studies and FFA concentrations varied between 0.01 and 1.71 mmol/liter on the four occasions (P < 0.05). We observed a dose-dependent 40% decrease of STAT5 phosphorylation in skeletal muscle with increasing concentrations of FFAs.

Conclusions:

Our results strongly suggest the existence of a negative feedback loop, whereby effects of GH may be dampened by FFA inhibition of GH-dependent STAT5 phosphorylation. The mechanisms behind and biological consequences of this finding awaits additional studies.
Impact of Resistance to Thyroid Hormone on the Cardiovascular System in Adults
Authors: Pulcrano Melania , Palmieri Emiliano , Mannavola Deborah , Campi Irene , Covelli Danila , Lombardi Gaetano , Biondi Bernadette , Beck-Peccoz Paolo
Published: January 2014

Background

The clinical manifestations of resistance to thyroid hormone (RTH) are highly variable, and the impact of RTH on the cardiovascular system has been poorly investigated.

Aim

The objective of the study was to evaluate the cardiovascular characteristics of 16 untreated and asymptomatic patients with RTH compared with 16 euthyroid healthy controls to define the cardiovascular involvement in RTH syndrome.

Patients and Methods

Sixteen untreated and asymptomatic RTH patients (eight males; aged 33 ± 12 yr, range 21–45 yr) and 16 controls (nine males; aged 33 ± 5 yr, range 24–42 yr) were enrolled. Clinical data, thyroid status, and echocardiographic results were recorded.

Results

Heart rate was comparable with that of controls, whereas arterial pressure was higher than controls. Mean interventricular septum diastolic thickness and mean left ventricular (LV) posterior wall diastolic thickness were significantly lower in RTH patients than controls with a consequent significant decrease of the mean LV mass and LV mass indexed by body surface area. Patients also had abnormalities of myocardial relaxation as indicated by a significant increase of peak A and consequent reduction of the early to late ratio. Finally, systemic vascular resistance was significantly higher in RTH patients than controls.

Conclusions

Our results suggest the presence of cardiovascular alterations in asymptomatic and untreated RTH patients similar to those reported in hypothyroid patients. Our strict selection likely created a bias in the inclusion of a particular type of RTH patients, who could represent a minority of patients with RTH. However, no correlation was found between the type of mutation and cardiovascular characteristics of RTH patients.
Increased 5α-Reductase Activity and Adrenocortical Drive in Women with Polycystic Ovary Syndrome
Authors: Vassiliadi Dimitra A. , Barber Thomas M. , Hughes Beverly A. , McCarthy Mark I. , Wass John A. H. , Franks Stephen , Nightingale Peter , Tomlinson Jeremy W. , Arlt Wiebke , Stewart Paul M.
Published: January 2014

Context

Polycystic Ovary Syndrome (PCOS) is characterized by hyperandrogenism, anovulation and susceptibility to the metabolic syndrome. Altered peripheral cortisol metabolism has been reported in PCOS but also in simple obesity.

Objective

To describe cortisol metabolism and metabolic characteristics of a large PCOS cohort and to delineate the effect of obesity by comparison to BMI-matched controls.

Design

Observational, cross-sectional study.

Setting

Outpatient clinics of two secondary/tertiary care centres

Patients or Other Participants

178 PCOS patients fulfilling Rotterdam criteria and 100 BMI-matched controls.

Intervention

24-h urine collection for steroid metabolite excretion, fasting blood samples followed by an OGTT.

Main Outcome Measures

Urinary steroid metabolites including glucocorticoids and androgens and the ratios reflecting enzymatic activities involved in peripheral cortisol and androgen metabolism, 5α-reductase and 11β-hydroxysteroid dehydrogenase type 1 and 2. Circulating levels of glucose, insulin, DHEA, DHEAS and testosterone, calculation of HOMA.

Results

Total androgen metabolites were higher in PCOS compared to BMI-matched controls (4105 ± 2047 vs. 2532 ± 1610 μg/24h for the non-obese, 5547 ± 2911 vs. 2468 ± 1794 μg/24hr for the obese, both p < 0.001). Total glucocorticoid metabolites were higher in obese PCOS vs. controls (10786 ± 3852 vs. 8834 ± 4487 μg/24hr, p = 0.001). 5α-reductase activity correlated with BMI, insulin levels and HOMA. Both obese and non-obese PCOS patients had higher 5α-reductase activity than controls (all p < 0.05). 11β-hydroxysteroid dehydrogenase activities did not differ between PCOS and controls.

Conclusions

PCOS is associated with enhanced androgen and cortisol metabolite excretion and increased 5α-reductase activity that cannot be explained by obesity alone. Increased adrenal corticosteroid production represents an important pathogenic pathway in PCOS.
TAC3/TACR3 Mutations Reveal Preferential Activation of GnRH Release by Neurokinin B in Neonatal Life Followed by Reversal in Adulthood
Authors: Gianetti Elena , Tusset Cintia , Noel Sekoni D. , Au Margaret G. , Dwyer Andrew A. , Hughes Virginia A. , Abreu Ana Paula , Carroll Jessica , Trarbach Ericka , Silveira Leticia FG , Costa Elaine MF , de Mendonça Berenice Bilharinho , de Castro Margaret , Lofrano Adriana , Hall Janet E. , Bolu Erol , Ozata Metin , Quinton Richard , Amory John K. , Stewart Susan E. , Arlt Wiebke , Cole Trevor R. , Crowley William F. , Kaiser Ursula B. , Latronico Ana Claudia , Seminara Stephanie B.
Published: January 2014

Context

Mutations in TAC3 and TACR3 (encoding neurokinin B and its receptor) have been identified in Turkish patients with hypogonadotropic hypogonadism (IHH), but broader populations have not yet been tested and genotype-phenotype correlations have not been established.

Objective

A broad cohort of normosmic IHH probands was screened for mutations in TAC3/TACR3 to evaluate the prevalence of such mutations and define the genotype/phenotype relationships.

Design

Sequencing of TAC3/TACR3; in vitro functional assays, neuroendocrine phenotyping.

Setting

Tertiary care centers world-wide.

Patients or other participants

345 probands, 18 family members, 292 controls.

Intervention

Examination of reproductive phenotypes throughout reproductive life and pre/post therapy.

Main Outcome Measure

Rare sequence variants in TAC3/TACR3.

Results

In TACR3, 19 probands harbored 13 distinct coding sequence rare nucleotide variants (3 nonsense mutations, 6 non-synonymous, 4 synonymous [one predicted to affect splicing]). In TAC3, one homozygous single base pair deletion was identified, resulting in complete loss of the neurokinin B decapeptide. Phenotypic information was available on 16 males and 7 females with coding sequence variants in TACR3/TAC3. Of the 16 males, 15 had microphallus; none of the females had spontaneous thelarche. Seven of the 16 males and 5/7 females were assessed after discontinuation of therapy and 6/7 males and 4/5 females demonstrated evidence for reversibility of their hypogonadotropism.

Conclusions

Mutations in the neurokinin B pathway are relatively common as causes of hypogonadism. While the neurokinin B pathway appears essential during early sexual development, its importance in sustaining the integrity of the hypothalamic-pituitary-gonadal axis appears attenuated over time.
XOMA 052, an Anti-IL-1β Monoclonal Antibody, Improves Glucose Control and β-Cell Function in the Diet-Induced Obesity Mouse Model
Authors: Owyang Alexander M. , Maedler Kathrin , Gross Lisa , Yin Johnny , Esposito Lin , Shu Luan , Jadhav Jaaee , Domsgen Erna , Bergemann Jennifer , Lee Steve , Kantak Seema
Published: January 2014

Recent evidence suggests that IL-1β-mediated glucotoxicity plays a critical role in type 2 diabetes mellitus. Although previous work has shown that inhibiting IL-1β can lead to improvements in glucose control and β-cell function, we hypothesized that more efficient targeting of IL-1β with a novel monoclonal antibody, XOMA 052, would reveal an effect on additional parameters affecting metabolic disease. In the diet-induced obesity model, XOMA 052 was administered to mice fed either normal or high-fat diet (HFD) for up to 19 wk. XOMA 052 was administered as a prophylactic treatment or as a therapy. Mice were analyzed for glucose tolerance, insulin tolerance, insulin secretion, and lipid profile. In addition, the pancreata were analyzed for β-cell apoptosis, proliferation, and β-cell mass. Mice on HFD exhibited elevated glucose and glycated hemoglobin levels, impaired glucose tolerance and insulin secretion, and elevated lipid profile, which were prevented by XOMA 052. XOMA 052 also reduced β-cell apoptosis and increased β-cell proliferation. XOMA 052 maintained the HFDinduced compensatory increase in β-cell mass, while also preventing the loss in β-cell mass seen with extended HFD feeding. Analysis of fasting insulin and glucose levels suggests that XOMA 052 prevented HFD-induced insulin resistance. These studies provide new evidence that targeting IL-1β in vivo could improve insulin sensitivity and lead to β-cell sparing. This is in addition to previously reported benefits on glycemic control. Taken together, the data presented suggest that XOMA 052 could be effective for treating many aspects of type 2 diabetes mellitus.
Missense Mutations in the Melanocortin 2 Receptor Accessory Protein That Lead to Late Onset Familial Glucocorticoid Deficiency Type 2
Authors: Hughes C. R. , Chung T. T. , Habeb A. M. , Kelestimur F. , Clark A. J. L. , Metherell L. A.
Published: January 2014

Background

Familial glucocorticoid deficiency (FGD) is an autosomal recessive disorder characterized by isolated glucocorticoid deficiency. Mutations in the ACTH receptor [melanocortin 2 receptor (MC2R)] or the MC2R accessory protein (MRAP) cause FGD types 1 and 2, respectively. Typically, type 2 patients present early (median age, 0.1 yr), and no patient reported to date has presented after 1.6 yr.

Aim

The aim of this study was to investigate the cause of disease in two families with late-onset FGD.

Patients

The proband in family 1 was diagnosed at age 4 yr. Family review revealed two older siblings with undiagnosed FGD. One sibling was well, whereas the second had cerebral palsy secondary to hypoglycemic seizures. The proband in family 2 was diagnosed at age 18 yr with symptoms of fatigue, weight loss, and depression.

Methods

The coding exons of MC2R and MRAP were sequenced. ACTH dose-response curves were generated for MC2R when transfected with wild-type or mutant MRAP constructs using HEK293 cells. MC2R trafficking with both mutant MRAPs was investigated using immunocytochemistry.

Results

MRAP gene analysis identified two novel homozygous missense mutations, c.175T>G (pY59D) in family 1 and c.76T>C (p.V26A) in family 2.

In vitro analysis showed that the Y59D mutant had significant impairment of cAMP generation, and both mutants caused a shift in the dose-response curve to the right when compared to wild type. Immunocytochemistry showed normal trafficking of MC2R when transfected with both mutant MRAPs, indicating a probable signaling defect.

Conclusion

These results indicate that missense MRAP mutations present with a variable phenotype of ACTH resistance and can present late in life.
Targeted Disruption of Mapk14 (p38MAPKα) in Granulosa Cells and Cumulus Cells Causes Cell-Specific Changes in Gene Expression Profiles that Rescue Cell-Oocyte Complex Expansion and Maintain Fertility
Authors: Liu Zhilin , Fan Heng-Yu , Wang Yibin , Richards JoAnne S.
Published: January 2014

MAPK14 (p38MAPKα) is critical for FSH and prostaglandin E (PGE)2 signaling cascades in granulosa cells (GCs) and cumulus cell-oocyte complexes (COCs) in culture, indicating that this kinase might impact follicular development and COC expansion in vivo. Because Mapk14 knockout mice are embryonic lethal, we generated GC specific Mapk14 knockout mice (Mapk14gc−/−) by mating Mapk14fl/fl and Cyp19a1-Cre mice. Unexpectedly, the Mapk14gc−/− female mice were fertile. Analyses of gene expression patterns showed that amphiregulin (Areg) and epiregulin (Ereg), two key regulators of ovulation and COC expansion, were up-regulated in the GCs but down-regulated in cumulus cells of the mutant mice in vivo. COCs from the mutant mice expanded and expressed matrix-related genes, if cultured with AREG, but not when cultured with forskolin or PGE2, the latter being a key factor regulating MAPK14 activity in cumulus cells. Conversely, when GCs from the Mapk14gc−/− mice were cultured with forskolin, they produced more Areg and Ereg mRNA than did wild-type GCs. These results indicate that disruption of Mapk14 selectively alters the expression of Areg and other genes in each cell type. Greater AREG and EREG produced by the GCs appears to by-pass and compensate for the critical need for MAPK14 signaling and induction of Areg/Ereg (and hence matrix genes) by PGE2 in cumulus cells of the mutant mice. In conclusion, although MAPK14 is not overtly essential for preovulatory follicle development or events associated with ovulation and luteinization in vivo, it does impact gene expression profiles.
Developmental Programming: Differential Effects of Prenatal Testosterone Excess on Insulin Target Tissues
Authors: Nada Shadia E. , Thompson Robert C. , Padmanabhan Vasantha
Published: January 2014

Polycystic ovarian syndrome (PCOS) is the leading cause of infertility in reproductive-aged women with the majority manifesting insulin resistance. To delineate the causes of insulin resistance in women with PCOS, we determined changes in the mRNA expression of insulin receptor (IR) isoforms and members of its signaling pathway in tissues of adult control (n = 7) and prenatal testosterone (T)-treated (n = 6) sheep (100 mg/kg twice a week from d 30-90 of gestation), the reproductive/metabolic characteristics of which are similar to women with PCOS. Findings revealed that prenatal T excess reduced (P < 0.05) expression of IR-B isoform (only isoform detected), insulin receptor substrate-2 (IRS-2), protein kinase B (AKt), peroxisome proliferator-activated receptor-γ (PPARγ), hormone-sensitive lipase (HSL), and mammalian target of rapamycin (mTOR) but increased expression of rapamycin-insensitive companion of mTOR (rictor), and eukaryotic initiation factor 4E (eIF4E) in the liver. Prenatal T excess increased (P < 0.05) the IR-A to IR-B isoform ratio and expression of IRS-1, glycogen synthase kinase-3α and -β (GSK-3α and -β), and rictor while reducing ERK1 in muscle. In the adipose tissue, prenatal T excess increased the expression of IRS-2, phosphatidylinositol 3-kinase (PI3K), PPARγ, and mTOR mRNAs. These findings provide evidence that prenatal T excess modulates in a tissue-specific manner the expression levels of several genes involved in mediating insulin action. These changes are consistent with the hypothesis that prenatal T excess disrupts the insulin sensitivity of peripheral tissues, with liver and muscle being insulin resistant and adipose tissue insulin sensitive.
Implication of Oxygen-Regulated Protein 150 (ORP150) in Apoptosis Induced by Proteasome Inhibitors in Human Thyroid Cancer Cells
Authors: Gao Yan-Yan , Liu Bao-Qin , Du Zhen-Xian , Zhang Hai-Yan , Niu Xiao-Fang , Wang Hua-Qin
Published: January 2014

Context

The inhibition of the 26S proteasome may lead to endoplasmic reticulum stress, which has been shown to be implicated in the antitumoral effects of proteasome inhibitors. Oxygen-regulated protein 150 (ORP150) is an inducible endoplasmic reticulum chaperone that is up-regulated after numerous cellular insults and has a cytoprotective role for the maintenance of cellular viability.

Objective

The purpose of this study was to determine the involvement of ORP150 in cytotoxicity of thyroid cancer cells mediated by proteasome inhibition.

Design

The effects of proteasome inhibition on the expression of ORP150 were analyzed using real-time RT-PCR and Western blot. To ascertain the effect of ORP150, cells were transfected with ORP150 plasmid or small interfering RNA (siRNA) against ORP150, apoptotic cells, and induction of CCAAT/enhancer-binding protein homologous transcription factor (CHOP) mediated by proteasome inhibition were investigated.

Results

ORP150 was induced in thyroid cancer cells after proteasome inhibition. Suppression of activating transcription factor 4 expression by siRNA inhibited the up-regulation of ORP150 mediated by proteasome inhibitors. siRNA for ORP150 stimulated MG132-mediated apoptosis and induction of CHOP, a transcription factor with apoptosis-inducing activity. In contrast, ORP150-overexpressing cells demonstrated less susceptibility to MG132-induced apoptosis and displayed less up-regulation of CHOP. In addition, the sensitizing effect of small interfering ORP150 on apoptosis was suppressed by siRNA for CHOP.

Conclusions

These results suggest that up-regulation of ORP150 in thyroid cancer cells inhibits MG132-induced apoptosis via suppression of CHOP induction, thereby decreasing the potential antitumor activity of MG132.
CXCL9 and CXCL11 Chemokines Modulation by Peroxisome Proliferator-Activated Receptor-α Agonists Secretion in Graves’ and Normal Thyrocytes
Authors: Antonelli Alessandro , Ferrari Silvia Martina , Frascerra Silvia , Pupilli Cinzia , Mancusi Caterina , Metelli Maria Rita , Orlando Claudio , Ferrannini Ele , Fallahi Poupak
Published: January 2014

Context

Peroxisome proliferator-activated receptor (PPAR)-α has been shown to exert immunomodulatory effects in autoimmune disorders. However, until now, no data were present in the literature about the effect of PPARα activation on CXCL9 and CXCL11 chemokines in general or on secretion of these chemokines in thyroid cells.

Objective and Design

The presence of PPARα and PPARγ has been evaluated by RT-PCR in Graves’ disease (GD) and control cells in primary culture. Furthermore, we have tested the role of PPARα and PPARγ activation on CXCL9 and CXCL11 secretion in GD and control cells after stimulation of these chemokines secretion with IFNγ and TNFα.

Results

This study shows the presence of PPARα and PPARγ in GD and control cells. A potent dose-dependent inhibition by PPARα-agonists was observed on the cytokines-stimulated secretion of CXCL9 and CXCL11 in GD and control cells. The potency of the PPARα agonists used was maximum on the secretion of CXCL9, reaching about 90% of inhibition by fenofibrate and 85% by ciprofibrate. The relative potency of the compounds was different with each chemokine; for example, gemfibrozil exerted a 55% inhibition on CXCL11, whereas it had a weaker activity on CXCL9 (40% inhibition). PPARα agonists were stronger (ANOVA, P < 0.001) inhibitors of CXCL9 and CXCL11 secretion in thyrocytes than PPARγ agonists.

Conclusions

Our study shows the presence of PPARα in GD and control thyrocytes. PPARα activators are potent inhibitors of the secretion of CXCL9 and CXCL11, suggesting that PPARα may be involved in the modulation of the immune response in the thyroid.
Regulation of Placental Growth by Aldosterone and Cortisol
Authors: Moser Carine , Khankin Eliyahu V. , Schüller Simone , Escher Geneviève , Frey Brigitte M. , Portmann C.-Bettina , Baumann Marc U. , Lehmann Andrea D. , Surbek Daniel , Karumanchi S. Ananth , Frey Felix J. , Mohaupt Markus G.
Published: January 2014

During pregnancy, trophoblasts grow to adapt the feto-maternal unit to fetal requirements. Aldosterone and cortisol levels increase, the latter being inactivated by a healthy placenta. By contrast, preeclamptic placental growth is reduced while aldosterone levels are low and placental cortisol tissue levels are high due to improper deactivation. Aldosterone acts as a growth factor in many tissues, whereas cortisol inhibits growth. We hypothesized that in preeclampsia low aldosterone and enhanced cortisol availability might mutually affect placental growth and function. Proliferation of cultured human trophoblasts was time- and dose-dependently increased with aldosterone (P < 0.04 to P < 0.0001) and inhibited by spironolactone and glucocorticoids (P < 0.01). Mineralo- and glucocorticoid receptor expression and activation upon agonist stimulation was verified by visualization of nuclear translocation of the receptors. Functional aldosterone deficiency simulated in pregnant mice by spironolactone treatment (15 μg/g body weight/day) led to a reduced fetal umbilical blood flow (P < 0.05). In rat (P < 0.05; R2 = 0.2055) and human (X2 = 3.85; P = 0.0249) pregnancy, placental size was positively related to plasma aldosterone. Autocrine production of these steroid hormones was excluded functionally and via the absence of specific enzymatic transcripts for CYP11B2 and CYP11B1. In conclusion, activation of mineralocorticoid receptors by maternal aldosterone appears to be required for trophoblast growth and a normal feto-placental function. Thus, low aldosterone levels and enhanced cortisol availability may be one explanation for the reduced placental size in preeclampsia and related disorders.
Loss of the C Terminus of Melanocortin Receptor 2 (MC2R) Results in Impaired Cell Surface Expression and ACTH Insensitivity
Authors: Hirsch Andrea , Meimaridou Eirini , Fernandez-Cancio Monica , Pandey Amit V. , Clemente María , Audi Laura , Clark Adrian J. L. , Flück Christa E.
Published: January 2014

Objective

Mutations in melanocortin receptor 2 (MC2R) and its related melanocortin receptor accessory protein (MRAP) cause familial glucocorticoid deficiency. We identified a novel MC2R mutation, K289fs. This unique mutation in the C terminus of MC2R is located in the intracellular part of the protein for which the exact function is unknown.

Setting

A 6-wk-old boy presented with severe hypoglycemia, unmeasurable cortisol, and grossly elevated ACTH but normal electrolytes. Genetic analysis revealed homozygote K289fs mutation in MC2R. His parents and siblings were heterozygous but phenotypically normal.

Intervention and Results

The role of the C terminus of MC2R was studied in two cell systems. Because the K289fs mutant changes the last eight amino acids of the protein and leads to protein elongation, wild-type MC2R and C-terminally mutated constructs were tested for activity to respond to ACTH in an OS3 cell-based reporter assay. Wild-type and alanine-substituted constructs responded normally to ACTH. By contrast K289fs and M290X had a total loss of activity. Cell surface assays and confocal localization studies revealed that K289fs and M290X receptors were not found at the cell surface, indicating that their transport from the endoplasmic reticulum to the cell membrane is disrupted. Interestingly, coimmunoprecipitation experiments showed no alteration in the interaction of mutant MC2R with MRAP, suggesting that interaction between these two proteins does not guarantee normal localization.

Conclusions

Loss of the C terminus of MC2R impairs cell surface expression and ACTH sensitivity but does not disrupt interaction of MC2R with MRAP. These findings highlight the extreme sensitivity of MC2R to structural disruption.
Depression in Women with Spontaneous 46, XX Primary Ovarian Insufficiency
Authors: Schmidt Peter J. , Luff Jamie A. , Haq Nazli A. , Vanderhoof Vien H. , Koziol Deloris E. , Calis Karim A. , Rubinow David R. , Nelson Lawrence M.
Published: January 2014

Context

A high prevalence of depressive symptoms is observed in women with primary ovarian insufficiency (POI) compared with women in whom the menopause is normally timed. Indeed, studies suggest that depression and/or its pharmacological treatment contribute to the onset of POI.

Objectives

We characterize the prevalence of psychiatric disorders and the timing of onset of clinically significant depression relative to both the diagnosis of POI and the onset of menstrual irregularity in women with POI.

Design and Setting

We conducted a cross-sectional clinic-based study at the National Institutes of Health Clinical Research Center.

Patients

A total of 174 women with spontaneous 46, XX POI and 100 women with Turner syndrome participated in the study.

Main Outcome Measures

The structured clinical interview for DSM-IV was performed.

Results

Lifetime histories of depression in POI exceeded rates of depression reported in women with Turner syndrome and community-based samples of women (P < 0.001). The onset of depression frequently preceded the diagnosis of POI but occurred after the onset of menstrual irregularity. Analyses standardizing the periods of risk for depression showed that similar numbers of depressions occurred before and after these events.

Conclusions

POI is associated with an increased lifetime risk for major depression. Attention to the presence of depression in POI should become an important part of the care for these women. The onset of depression frequently occurs after signs of altered ovarian function but before the diagnosis of POI. Thus, in some women the association between POI and depression suggests an overlapping pathophysiology rather than a causal relationship.
S1P1 and VEGFR-2 Form a Signaling Complex with Extracellularly Regulated Kinase 1/2 and Protein Kinase C-α Regulating ML-1 Thyroid Carcinoma Cell Migration
Authors: Bergelin Nina , Löf Christoffer , Balthasar Sonja , Kalhori Veronica , Törnquist Kid
Published: January 2014

Sphingosine 1-phosphate (S1P) and vascular endothelial growth factor receptor 2 (VEGFR-2) signaling have been shown to integrate in many biological processes. The follicular thyroid carcinoma cell line ML-1 expresses VEGFR-2 and secretes substantial amounts of both vascular endothelial growth factor (VEGF)-A and VEGF-C. ML-1 cells also express S1P-receptors (S1P1–3,5). S1P is able to phosphorylate VEGFR-2, and inhibiting VEGFR-2 attenuates S1P-induced migration and down-regulates S1P1 expression in ML-1 cells. In the present study, we focused on the interactions between S1P1 and VEGFR-2. We show that S1P receptors form complexes with VEGFR-2 and that the S1P1/VEGFR-2 complex associates with protein kinase C (PKC)-α and ERK1/2. Furthermore, the complex evokes bidirectional signaling since the S1P-induced ERK1/2 phosphorylation is sensitive to VEGFR-2 kinase inhibition and VEGF-A-induced ERK1/2 phosphorylation is sensitive to pertussis toxin treatment as well as S1P1 small interfering RNA (siRNA) treatment. Both S1P- and VEGF-A-induced haptotaxis is sensitive to pertussis toxin treatment and S1P1 siRNA treatment. Phosphorylation of ERK1/2 evoked by both VEGF-A and the S1P1 agonist SEW-2871 is inhibited by PKC-α and PKC-βI siRNA. We hypothesize that VEGFR-2 forms a signaling complex with S1P1, evoking bidirectional signaling regulating both ERK1/2 phosphorylation and haptotaxis of ML-1 cells.