bluuebunny/crossref_metadata_2025_split huggingface dataset:
Proximal cis-Acting Elements, Including Steroidogenic Factor 1, Mediate the Efficiency of a Distal Enhancer in the Promoter of the Rat Gonadotropin-Releasing Hormone Receptor Gene
Authors: Pincas Hanna , Amoyel Karine , Counis Raymond , Laverrière Jean-Noël 
Published: January 2014

AbstractThe gonadotrope-specific and regulated expression of the GnRH receptor (GnRH-R) gene is dependent on multiple transcription factors that interact with the noncanonical GnRH-R activating sequence (GRAS), the activator protein-1 (AP-1) element, and the steroidogenic factor-1 (SF-1) binding site. However, these three elements are not sufficient to mediate the complete cell-specific expression of the rat GnRH-R gene. In the present study, we demonstrate, by transient transfection in gonadotrope-derived αT3–1 and LβT2 cell lines, the existence of a distal enhancer [GnRH-R- specific enhancer (GnSE)] that is highly active in the context of the GnRH-R gene promoter. We show that the GnSE activity (–1,135/–753) is mediated through a functional interaction with a proximal region (–275/–226) that includes the SF-1 response element. Regions of similar length containing either the AP-1 or GRAS elements are less active or inactive. Transfection assays using an artificial promoter containing two SF-1 elements fused to a minimal PRL promoter indicate that SF-1 is crucial in this interaction. In addition, by altering the promoter with deletion and block- replacement mutations, we have identified the active elements of GnSE within two distinct sequences at positions –983/–962 and –871/–862. Sequence analysis and electrophoretic mobility shift experiments suggest that GnSE response elements interact, in these two regions, with GATA- and LIM-related factors, respectively. Altogether, these data establish the importance of the GnSE in the GnRH-R gene expression and reveal a novel role for SF-1 as a mediator of enhancer activity, a mechanism that might regulate other SF-1 target genes.
A Single Pitx1 Binding Site Is Essential for Activity of the LHβ Promoter in Transgenic Mice
Authors: Quirk Christine C. , Lozada Kristen L. , Keri Ruth A. , Nilson John H. 
Published: January 2014

AbstractReproduction depends on regulated expression of the LHβ gene. Tandem copies of regulatory elements that bind early growth response protein 1 (Egr-1) and steroidogenic factor 1 (SF-1) are located in the proximal region of the LHβ promoter and make essential contributions to its activity as well as mediate responsiveness to GnRH. Located between these tandem elements is a single site capable of binding the homeodomain protein Pitx1. From studies that employ overexpression paradigms performed in heterologous cell lines, it appears that Egr-1, SF-1, and Pitx1 interact cooperatively through a mechanism that does not require the binding of Pitx1 to its site. Since the physiological ramifications of these overexpression studies remain unclear, we reassessed the requirement for a Pitx1 element in the promoter of the LHβ gene using homologous cell lines and transgenic mice, both of which obviate the need for overexpression of transcription factors. Our analysis indicated a striking requirement for the Pitx1 regulatory element. When assayed by transient transfection using a gonadotrope-derived cell line (LβT2), an LHβ promoter construct harboring a mutant Pitx1 element displayed attenuated transcriptional activity but retained responsiveness to GnRH. In contrast, analysis of wild-type and mutant expression vectors in transgenic mice indicated that LHβ promoter activity is completely dependent on the presence of a functional Pitx1 binding site. Indeed, the dependence on an intact Pitx1 binding site in transgenic mice is so strict that responsiveness to GnRH is also lost, suggesting that the mutant promoter is inactive. Collectively, our data reinforce the concept that activity of the LHβ promoter is determined, in part, through highly cooperative interactions between SF-1, Egr-1, and Pitx1. While Egr-1 can be regarded as a key downstream effector of GnRH, and Pitx1 as a critical partner that activates SF-1, our data firmly establish that the Pitx1 element plays a vital role in permitting these functions to occur in vivo.
Cloning and Characterization of a Rat Ortholog of MMP-23 (Matrix Metalloproteinase-23), a Unique Type of Membrane-Anchored Matrix Metalloproteinase and Conditioned Switching of Its Expression during the Ovarian Follicular Development
Authors: Ohnishi Junji , Ohnishi Eriko , Jin Mulan , Hirano Wakako , Nakane Dai , Matsui Hitoshi , Kimura Atsushi , Sawa Hirofumi , Nakayama Kazuhisa , Shibuya Hiroshi , Nagashima Kazuo , Takahashi Takayuki 
Published: January 2014

AbstractIn our attempt to study the role of matrix metalloproteinases (MMPs) in the process of mammalian ovulation, we isolated a rat ortholog of the recently reported human MMP-23 from gonadotropin-primed immature rat ovaries. Transient expression of epitope-tagged rat and human MMP-23 in COS-1 cells revealed that they were synthesized as a membrane-anchored glycoprotein with type II topology. Indirect immunofluorescent analysis showed that subcellular localization of MMP-23 was predominantly in the perinuclear regions. The transfected human MMP-23 protein was processed endogenously to the soluble form in COS-1 cells. However, cotransfection of MMP-23 with the mouse furin cDNA did not enhance this processing, indicating that furin may not be involved in this event. Notably, in situ hybridization analysis revealed a dramatic switching of MMP-23 mRNA localization from granulosa cells to theca-externa/fibroblasts and ovarian surface epithelium during the follicular development. In serum-free primary culture of rat granulosa cells, a drastic diminution of MMP-23 mRNA expression was observed in response to FSH action between 24 h and 48 h of culture. The observed effect of FSH on MMP-23 expression was mimicked by treatment of granulosa cells with forskolin or 8-bromo (Br)-cAMP. In contrast, MMP-23 mRNA levels increased in theca-interstitial cells regardless of the presence of LH in the culture. However, treatment of theca-interstitial cells with forskolin or 8-Br-cAMP markedly reduced the expression of MMP-23 with a concomitant increase in progesterone production. These results indicate that the MMP-23 gene is spatially and temporally regulated in a cell type-specific manner in ovary via the cAMP signaling pathway.
Phosphorylation of the Receptor for PTH and PTHrP Is Required for Internalization and Regulates Receptor Signaling
Authors: Tawfeek Hesham A. W. , Qian Fang , Abou-Samra Abdul B. 
Published: January 2014

Abstract
We have previously shown that agonist-dependent phosphorylation of the PTH/PTHrP receptor occurs on its carboxyl-terminal tail. Using site- directed mutagenesis, phosphopeptide mapping, and direct sequencing of cyanogen bromide-cleaved fragments of phosphoreceptors, we report here that PTH-dependent phosphorylation occurs on the serine residues at positions 491, 492, 493, 495, 501, and 504, and that the serine residue at position 489 is required for phosphorylation. When these seven sites were mutated to alanine residues, the mutant receptor was no longer phosphorylated after PTH stimulation. The phosphorylation-deficient receptor, stably expressed in LLCPK-1 cells, was impaired in PTH-dependent internalization and showed an increased sensitivity to PTH stimulation; the EC50 for PTH-stimulated cAMP accumulation was decreased by 7-fold. Furthermore, PTH stimulation of the phosphorylation-deficient PTH/PTHrP receptor caused a sustained elevation in intracellular cAMP levels. These data indicate that agonist-dependent phosphorylation of the PTH/PTHrP receptor plays an important role in receptor function.
The mRNA Structure Has Potent Regulatory Effects on Type 2 Iodothyronine Deiodinase Expression
Authors: Gereben Balázs , Kollár Anna , Harney John W. , Larsen P. Reed 
Published: January 2014

AbstractType 2 deiodinase (D2) is a selenoenzyme catalyzing the activation of T4 to T3. D2 activity/mRNA ratios are often low, suggesting that there is significant posttranscriptional regulation. The D2 mRNA in higher vertebrates is more than 6 kb, containing long 5′ and 3′ untranslated regions (UTRs). The D2 5′UTRs are greater than 600 nucleotides and contain 3–5 short open reading frames. These full-length 5′UTRs reduce the D2 translation efficiency approximately 5-fold. The inhibition by human D2 5′UTR is localized to a region containing the first short open reading frame encoding a tripeptide—MKG. This inhibition was abolished by mutating the AUG start codon and weakened by modification of the essential purine of the Kozak consensus. Deletion of the 3.7-kb 3′UTR of the chicken D2 mRNA increased D2 activity approximately 3.8-fold due to an increase in D2 mRNA half-life. In addition, alternatively spliced D2 mRNA transcripts similar in size to the major 6- to 7-kb D2 mRNAs but not encoding an active enzyme are present in both human and chicken tissues. Our results indicate that a number of factors reduce the D2 protein levels. These mechanisms, together with the short half-life of the protein, ensure limited expression of this key regulator of T4 activation.
Frequency of Impaired Fibroblast Growth Factor Receptor 1 Signaling as a Cause of Normosmic Idiopathic Hypogonadotropic Hypogonadism
Authors: Raivio Taneli , Sidis Yisrael , Plummer Lacey , Chen Huaibin , Ma Jinghong , Mukherjee Abir , Jacobson-Dickman Elka , Quinton Richard , Van Vliet Guy , Lavoie Helene , Hughes Virginia A. , Dwyer Andrew , Hayes Frances J. , Xu Shuyun , Sparks Susan , Kaiser Ursula B. , Mohammadi Moosa , Pitteloud Nelly 
Published: January 2014

ABSTRACT

Context
FGFR1 mutations have been identified in about 10% of patients with Kallmann syndrome. Recently cases of idiopathic hypogonadotropic hypogonadism (IHH) with a normal sense of smell (nIHH) have been reported.

Aims
The objective of the study was to define the frequency of FGFR1 mutations in a large cohort of nIHH, delineate the spectrum of reproductive phenotypes, assess functionality of the FGFR1 mutant alleles in vitro, and investigate genotype-phenotype relationships.

Design
FGFR1 sequencing of 134 well-characterized nIHH patients (112 men and 22 women) and 270 healthy controls was performed. The impact of the identified mutations on FGFR1 function was assessed using structural prediction and in vitro studies.

Results
Nine nIHH subjects (five males and four females; 7%) harbor a heterozygous mutation in FGFR1 and exhibit a wide spectrum of pubertal development, ranging from absent puberty to reversal of IHH in both sexes. All mutations impair receptor function. The Y99C, Y228D, and I239T mutants impair the tertiary folding, resulting in incomplete glycosylation and reduced cell surface expression. The R250Q mutant reduces receptor affinity for FGF. The K618N, A671P, and Q680X mutants impair tyrosine kinase activity. However, the degree of functional impairment of the mutant receptors did not always correlate with the reproductive phenotype, and variable expressivity of the disease was noted within family members carrying the same FGFR1 mutation. These discrepancies were partially explained by additional mutations in known IHH loci.

Conclusions
Loss-of-function mutations in FGFR1 underlie 7% of nIHH with different degrees of impairment in vitro. These mutations act in concert with other gene defects in several cases, consistent with oligogenicity.
Melatonin Sensitizes Human Myometrial Cells to Oxytocin in a Protein Kinase Cα/Extracellularly Regulated Kinase-Dependent Manner
Authors: Sharkey James T. , Cable Casey , Olcese James 
Published: January 2014

abstract
Context: Studies have shown that labor occurs primarily in the night/morning hours. Recently, we identified the human myometrium as a target for melatonin (MEL), the neuroendocrine output signal coding for circadian night.
Objective: The purpose of this study was to determine the signaling pathway underlying the effects of MEL on contractility and the contractile machinery in immortalized human myometrial cells.
Design: To ascertain the signaling pathway of MEL leading to its effects on myometrial contractility in vitro, we performed gel retraction assays with cells exposed to iodo-MEL (I-MEL) with or without oxytocin and the Rho kinase inhibitor Y27632. I-MEL effects on inositol trisphosphate (IP3)/diacylglycerol (DAG)/protein kinase C (PKC) signaling were also investigated. Additionally, we assayed for caldesmon phosphorylation and ERK1/2 activation.
Results: I-MEL was found to activate PKCα via the phospholipase C/IP3/DAG signaling pathway, which was confirmed by PKC enzyme assay. I-MEL did not affect myosin light chain phosphatase activity, and its effects on contractility were insensitive to Rho kinase inhibition. I-MEL did increase phosphorylation of ERK1/2 and caldesmon, which was inhibited by the MAPK kinase inhibitor PD98059 or the PKC inhibitor C1.
Conclusions: MEL sensitizes myometrial cells to subsequent procontractile signals in vitro through activation of the phospholipase C/IP3/DAG signaling pathway, resulting in specific activation of PKCα and ERK1/2, thereby phosphorylating caldesmon, which increases actin availability for myosin binding and cross-bridging. In vivo, this sensitization would provide a mechanism for the increased nocturnal uterine contractility and labor that has been observed in late-term human pregnancy.
Novel Activating Mutations of the Calcium-Sensing Receptor: The Calcilytic NPS-2143 Mitigates Excessive Signal Transduction of Mutant Receptors
Authors: Letz Saskia , Rus Ramona , Haag Christine , Dörr Helmuth-Günther , Schnabel Dirk , Möhlig Matthias , Schulze Egbert , Frank-Raue Karin , Raue Friedhelm , Mayr Bernhard , Schöfl Christof 
Published: January 2014

Abstract
Context and Objective: Activating mutations in the calcium-sensing receptor (CaSR) gene cause autosomal dominant hypocalcemia (ADH). The aims of the present study were the functional characterization of novel mutations of the CaSR found in patients, the comparison of in vitro receptor function with clinical parameters, and the effect of the allosteric calcilytic NPS-2143 on the signaling of mutant receptors as a potential new treatment for ADH patients.
Methods: Wild-type and mutant CaSR (T151R, P221L, E767Q, G830S, and A844T) were expressed in human embryonic kidney cells (HEK 293T). Receptor signaling was studied by measuring intracellular free calcium in response to different concentrations of extracellular calcium ([Ca2+]o) in the presence or absence of NPS-2143.
Results: All ADH patients had lowered serum calcium ranging from 1.7 to 2.0 mm and inadequate immunoreactive PTH and urinary calcium excretion. In vitro testing of CaSR mutations from these patients revealed exaggerated [Ca2+]o-induced cytosolic Ca2+ responses with EC50 values for [Ca2+]o ranging from 1.56 to 3.15 mm, which was lower than for the wild-type receptor (4.27 mm). The calcilytic NPS-2143 diminished the responsiveness to [Ca2+]o in the CaSR mutants T151R, E767Q, G830S, and A844T. The mutant P221L, however, was only responsive when coexpressed with the wild-type CaSR.
Conclusion: Calcilytics might offer medical treatment for patients with autosomal dominant hypocalcemia caused by calcilytic-sensitive CaSR mutants.
Rheological Characterization of Starch Gelatinization: Effects of plasticizer/starch ratio and clay content in corn-starch/glycerol thermoplastic starch (TPS)
Authors: Lara Sandra , Salcedo Felipe 
Published: January 2014

ABSTRACTThis work presents a systematic rheological study of the gelatinization process of corn-starch plasticized with glycerol, showing the effects of the glycerol/starch ratio, water/starch ratio, and clay (montmorillonite) content. Gelatinization temperatures at different heating rates in rheological oscillatory temperature-sweep experiments were determined for different corn-starch/glycerol/clay formulations. The influences of the different formulation variables on the gelatinization processes and on the gel properties are analyzed. Some hypotheses postulating how the different intermolecular interactions present in the composites are responsible for these effects are discussed.
Identity Fusion and Status of the Evaluator as Moderators of Self-Enhancement and Self-Verification at the Group Level of Self-Description
Authors: Besta Tomasz , Kaźmierczak Maria , Błażek Magdalena 
Published: January 2014

Abstract
We examined the influence of identity fusion and status of evaluator on willingness to fight for one’s group after group-descriptive or not group-descriptive feedback. The valence of evaluative information was varied as well: feedback either support negative group-stereotype (negative but descriptive condition) or contradict negative group-stereotype (positive but not descriptive condition). In two studies (N=208 & N=119) we partially replicated previous findings on self-verification. Individuals fused with one’s group were more prone than non fused to fight for group members after receiving, challenging, not group-describing feedback, but only when evaluator’s status was high. When the evaluator’ status was low, fused participants (vs. non fused) engaged in self-enhancement strategies: that is they endorse radical behavior after negative, even if group-describing, feedback. We discuss the importance of identity fusion and evaluator’s status for determining when self-enhancement and self-verification will and will not be evoke.
Repetitive thought as a moderator of the impact of control deprivation on emotional and cognitive functioning
Authors: Jarmakowski-Kostrzanowski Tomasz 
Published: January 2014

Abstract
The present research explores the role of repetitive thought (RT) in developing control deprivation deficits. The two main RT theories lead to diverging predictions. The response style theory suggests that RT in reaction to distress leads to negative effects in terms of emotional and cognitive functioning. However, the theory of Marin and Tesser and its elaboration by Watkins, suggest that the effects of RT depend on its form and that individuals who are not depression-prone usually adopt the constructive form of RT that leads to positive effects. To test which of these predictions is true for control deprivation situation, two experimental studies were conducted. Participants after control deprivation were induced RT or distraction, followed by the measurement of their emotional and cognitive functioning. The results suggest that repetitive thought reduces both emotional and cognitive helplessness deficits and has no effect in no control deprivation condition. This supports the theory of Martin and Tesser and its elaboration by Watkins.
Detection of Adolescent Idiopathic Scoliosis among Nepalese Children Through the School Screening Program
Authors: Pokharel Rohit Kumar , Lakhey Rajesh Bahadur , Kafle Dinesh , Shah Lakahn Lall 
Published: January 2014

Introduction: Idiopathic Adolescent Scoliosis (IAS) is the commonest type of scoliosis without any known cause. There has been studies in different countries and population on IAS; and its prevalence rate varies for different population. There has been no reported study done in Nepal on prevalence of adolescent idiopathic scoliosis (AIS). The aim of this study was to detect and evaluate the prevalence of idiopathic adolescent scoliosis in school children in different parts of Nepal. Methods: This was a descriptive cross sectional study conducted at five schools from different geographical parts of Nepal, from January 2010 to December 2010. Scoliosis screening was done in 5 schools from different geographical locations of Nepal. Children (from grade 6 to 10; age more than 10 years) with positive Adam’s forward bending test and scoliometer angle of 5 degrees or more were subjected for radiographic evaluation of the spine. The criterion for the diagnosis of scoliosis was detection of Cobb’s angle of more than 10 degrees. Evaluation of scoliosis according to the age, gender, ethnic group, spinal region and magnitude of curve was done. Results: Out of 1094 children screened, 31 (2.83%) were referred for radiological evaluation; and 12 of them had lateral spinal curvature of more than 10 degrees. Hence the prevalence of scoliosis was 1.09%. The girl to boy ratio of distribution of scoliosis was 3:1 for larger curves of more than 20 degrees. Maximum number of scoliosis cases was seen in girls of 13 years, and the right sided thoracic scoliosis was most common. There is no ethnic predominance of scoliosis. Conclusion: Prevalence of idiopathic scoliosis among school children who need treatment in Nepal is low; however, school screening program has its epidemiological value. DOI: http://dx.doi.org/10.3126/noaj.v3i2.9514 NOAJ July-December 2013, Vol 3, Issue 2, 14-19
Development and characterization of 25 EST‐SSR markers in Pinus sylvestris var. mongolica (Pinaceae)
Authors: Fang Pan , Niu Shihui , Yuan Huwei , Li Zhexin , Zhang Yuncheng , Yuan Lu , Li Wei 
Published: January 2014

• Premise of the study: A set of novel expressed sequence tag (EST) microsatellite markers was developed in Pinus sylvestris var. mongolica to promote further genetic studies in this species.• Methods and Results: One hundred seventy‐five EST–simple sequence repeat (SSR) primers were designed and synthesized for 31,653 isotigs based on P. tabuliformis EST sequences. The primer pairs were used to identify 25 polymorphic loci in 48 individuals. The number of alleles ranged from two to eight with observed and expected heterozygosity values of 0.0435 to 0.8125 and 0.0430 to 0.7820, respectively.• Conclusions: These new polymorphic EST‐SSR markers will be useful for assessing genetic diversity, molecular breeding and genetic improvement, and conservation of P. sylvestris var. mongolica.
Detecting super‐thin clouds with polarized sunlight
Authors: Sun Wenbo , Videen Gorden , Mishchenko Michael I. 
Published: January 2014

AbstractWe report a novel method for detecting cloud particles in the atmosphere. Solar radiation backscattered from clouds is studied with both satellite data and a radiative transfer model. A distinct feature is found in the angle of linear polarization of solar radiation that is backscattered from clouds. The dominant backscattered electric field from the clear‐sky Earth‐atmosphere system is nearly parallel to the Earth surface. However, when clouds are present, this electric field can rotate significantly away from the parallel direction. Model results demonstrate that this polarization feature can be used to detect super‐thin cirrus clouds having an optical depth of only ~0.06 and super‐thin liquid water clouds having an optical depth of only ~0.01. Such clouds are too thin to be sensed using any current passive satellite instruments.