bluuebunny/crossref_metadata_2025_split huggingface dataset:
Nuclear trafficking of FGFR1: A role for the transmembrane domain
Authors: Myers Jason M. , Martins Gabriel G. , Ostrowski Jacek , Stachowiak Michal K. 
Published: March 2003

AbstractSeveral members of the fibroblast growth factor (FGF) family lack signal peptide (SP) sequences and are present only in trace amounts outside the cell. However, these proteins contain nuclear localization signals (NLS) and accumulate in the cell nucleus. Our studies have shown that full length FGF receptor 1 (FGFR1) accumulates within the nuclear interior in parallel with FGF‐2. We tested the hypothesis that an atypical transmembrane domain (TM) plays a role in FGFR1 trafficking into the nuclear interior. With FGFR1 destined for constitutive fusion with the plasma membrane due to its SP, how the receptor may enter the nucleus is unclear. Sequence analysis identified that FGFR1 has an atypical TM containing short stretches of hydrophobic amino acids (a.a.) interrupted by polar a.a. The β‐sheet is the predicted conformation of the FGFR1 TM, in contrast to the α‐helical conformation of other single TM tyrosine kinase receptors, including FGFR4. Receptor trafficking in live cells was studied by confocal microscopy via C‐terminal FGFR1 fusions to enhanced green fluorescent protein (EGFP) and confirmed by subcellular fractionation and Western immunoblotting. Nuclear entry of FGFR1–EGFP was independent of karyokinessis, and was observed in rapidly proliferating human TE671 cells, in slower proliferating glioma SF763 and post‐mitotic bovine adrenal medullary cells (BAMC). In contrast, a chimeric FGFR1/R4‐EGFP, where the TM of FGFR1 was replaced with that of FGFR4, was associated with membranes (golgi‐ER, plasma, and nuclear), but was absent from the nucleus and cytosol. FGFR1Δ‐EGFP mutants, with hydrophobic TM a.a. replaced with polar a.a., showed reduced association with membranes and increased cytosolic/nuclear accumulation with an increase in TM hydrophilicity. FGFR1(TM−)‐EGFP (TM deleted), was detected in the golgi‐ER vesicles, cytosol, and nuclear interior; thus demonstrating that the FGFR1 TM does not function as a NLS. To test whether cytosolic FGFR1 provides a source of nuclear FGFR1, cells were transfected with FGFR1(SP−) (SP was deleted), resulting in cytosolic, non‐membrane, protein accumulation in the cytosol and the cell nucleus. Our results indicate that an unstable association with cellular membranes is responsible for the release of FGFR1 into the cytosol and cytosolic FGFR1 constitutes the source of the nuclear receptor. J. Cell. Biochem. 88: 1273–1291, 2003. © 2003 Wiley‐Liss, Inc.
The anatomy of the posterolateral aspect of the rabbit knee
Authors: Crum Joshua A. , Laprade Robert F. , Wentorf Fred A. 
Published: March 2003

AbstractThe purpose of this study was to determine the anatomy of the posterolateral aspect of the rabbit knee to serve as a basis for future in vitro and in vivo posterolateral knee biomechanical and injury studies. Twelve nonpaired fresh‐frozen New Zealand white rabbit knees were dissected to determine the anatomy of the posterolateral corner.The following main structures were consistently identified in the rabbit posterolateral knee: the gastrocnemius muscles, biceps femoris muscle, popliteus muscle and tendon, fibular collateral ligament, posterior capsule, ligament of Wrisberg, and posterior meniscotibial ligament. The fibular collateral ligament was within the joint capsule and attached to the femur at the lateral epicondyle and to the fibula at the midportion of the fibular head. The popliteus muscle attached to the medial edge of the posterior tibia and ascended proximally to give rise to the popliteus tendon, which inserted on the proximal aspect of the popliteal sulcus just anterior to the fibular collateral ligament. The biceps femoris had no attachment to the fibula and attached to the anterior compartment fascia of the leg.This study increased our understanding of these structures and their relationships to comparative anatomy in the human knee. This knowledge of the rabbit's posterolateral knee anatomy is important to understand for biomechanical and surgical studies which utilize the rabbit knee as a model for human posterolateral knee injuries. © 2003 Orthopaedic Research Society. Published by Elsevier Science Ltd. All rights reserved.
Pamidronate treatment improves bone mineral density in children with Menkes disease
Authors: Kanumakala S. , Boneh A. , Zacharin M. 
Published: March 2003

AbstractMenkes disease is a severe multisystem disorder due to defective bioavailability and transport of copper at the cellular level. Deficient activity of lysyl oxidase, a copper‐dependent enzyme, causes defective collagen cross‐linking leading to osteoporosis and pathological fractures in these children. The objective of the study was to evaluate the changes in bone mineral density following pamidronate treatment in children with Menkes disease. The study design was an open observational study of three children with Menkes disease and significant osteoporosis with or without pathological fractures, all of whom received pamidronate treatment for 1 year. There were 34–55% and 16–36% increases in lumbar spine bone mineral content and areal bone mineral density, respectively, following 1 year of treatment with pamidronate. There were no further fractures in two of the three children treated. No adverse effects of pamidronate treatment were noted. Pamidronate treatment was associated with an increase in bone mineral density and may be an effective treatment modality for the management of osteoporosis in children with Menkes disease.
Propranolol for reemergent posttraumatic stress disorder following an event of retraumatization: A case study
Authors: Taylor Fletcher , Cahill Larry 
Published: March 2003

AbstractThis case report concerns a 44‐year‐old woman who experienced 5 similar motor vehicle accidents, the last 3 causing severe PTSD episodes of over 6 months each, despite multiple pharmacotherapies. Following a 6th accident, severe PTSD symptoms reemergcd. Forty‐eight hours after this trauma, propranolol (60 mg) orally, twice a day (1.75 mg/kg/day) was begun, and the PTSD symptoms were rapidly and markedly reduced. The Clinician‐Administered PTSD Rating Scale score was reduced from an initial 86 to 56 by 11 days posttrauma. To our knowledge, this is the first report of the effects of propranolol treatment on reemergent PTSD symptoms. Propranolol may be particularly efficacious in the prevention of initial or reemergent PTSD symptoms.
Identification of novel mutations in the NPC1 gene in German patients with Niemann–Pick C disease
Authors: Kaminski W. E. , Klünemann H. H. , Ibach B. , Aslanidis C. , Klein H. E. , Schmitz G. 
Published: March 2003

AbstractNiemann–Pick disease type C (NPC) is an inherited neuro degenerative disorder associated with intracellular cholesterol trafficking defects. Mutations in two distinct genes, NPC1 and HE1, have recently been shown to cause this disease. We have analysed the NPC1 gene in five German patients with NPC from four unrelated families. We identified a total of five novel mutations in the coding region of the NPC1 gene (G231V, D874V, I642M, I1094T and R116stop). All affected individuals displayed compound heterozygosity. The mutated alleles were transmitted by the nonaffected parents with the exception of one patient, in whom a de novo mutation (G231V) had occurred. Interestingly, the G231V/P237S NPC1 genotype in this individual is associated with an early‐onset form of NPC. In contrast, we found that the D874V/D948N genotype, observed in another NPC patient, is characterized by a late onset of clinical symptoms that presents with a pronounced white‐matter disease. Our results will contribute to defining the association between the clinical phenotypes and the genetic abnormalities in Niemann–Pick C disease.
Fear activation and habituation patterns as early process predictors of response to prolonged exposure treatment in PTSD
Authors: van Minnen Agnes , Hagenaars Muriel 
Published: March 2003

AbstractImproved (n = 21) and nonimproved (n = 13) PTSD patients (a mixed trauma population) were compared for fear activation and habituation patterns during and between the 1st and 2nd prolonged exposure sessions. Drop‐outs (n = 11) were also evaluated. Nonimproved patients had significantly higher ratings of anxiety at the start of the first exposure session. Improved patients showed more within‐session habituation during the self‐exposures at home and more between‐session habituation. Even after controlling for initial PTSD and depression symptom severity, habituation between the first and second exposure sessions was significantly related to treatment outcome. Patients who dropped out of the treatment were found not to differ from completers on fear activation and within‐session habituation during the first exposure session.
Ceramide Glycanase Activities in Human Cancer Cells
Authors: Basu Manju , Kelly Patrick , O'Donnell Peter , Miguel Maria , Bradley Mathew , Sonnino Sandro , Banerjee Sipra , Basu Subhash 
Published: March 2003

Ceramide glycanase (CGase) activities have been detected in different human tumor cells (colon, carcinoma Colo-205; neuroblastoma, IMR-32; breast cancer lines, SKBr3 and MCF7). However, the level of enzymatic activity is lower in these cells compared to that present in other mammalian tissues reported before (Basu, M., Kelly, P., Girzadas, M. A., Li, Z., and Basu, S. Methods Enzymol. (in press)). The majority of CGase activity was found in the 100,000g soluble supernatant fraction isolated from all these cell lines and tissues. Using the soluble enzyme, the requirement for optimum CGase activity was found to be consistent with previous observations found for rat and rabbit tissues (Basu, M., Dastgheib, S., Girzadas, M. A., O'Donnell, P. H., Westervelt, C. W., Li, Z., Inokuchi, J. I., and Basu, S. (1998) Acta Pol. Biochim. 42:327). The CGase activities from both Colo-205 and IMR-32 cells are optimum at a protein to detergent ratio of one. All the mammalian CGases, including human cancer cells, show an optimum pH between 5.5 and 5.8 in sodium acetate buffer. The CGase activities from cancer cells are found to be cation-independent; however, mercury, zinc, and copper ions seem to inhibit the enzyme activity substantially in both tumor cells lines. The mercury ion inhibition of CGase activities from all different sources indicates a possible structural homology in the CGase proteins.
Radiolabeled substrates, labeled at the sphingosine double bond or at the 3-position of sphingosine without modifying double bond of sphingosine were used in this investigation. Both were active substrates with all enzyme preparations isolated from different cancer cells (apparent Km, 500 μM for nLcOse5[3H-DT]Cer and 350 μM for GgOse4[sph-3-3H]Cer with Colo-205 enzyme). Structural analogues of ceramide and sphingosine (L-PPMP, L-PDMP, alkylamines, and Tamoxifen) inhibited cancer cell CGase activities in vitro.
Gonadal Steroids Differentially Regulate the Messenger Ribonucleic Acid Expression of Pituitary Orexin Type 1 Receptors and Adrenal Orexin Type 2 Receptors
Authors: Jöhren Olaf , Brüggemann Norbert , Dendorfer Andreas , Dominiak Peter 
Published: March 2003

Abstract
Hypothalamic prepro-orexin as well as pituitary and adrenal orexin receptors are gender-specifically expressed. To assess the regulation by gonadal steroids, we investigated the effect of 17β-estradiol in female and of testosterone in male rats on prepro-orexin and orexin receptor mRNA expression. Rats were either sham-operated or gonadectomized and subsequently treated with placebo, 17β-estradiol, or testosterone for 21 d. Tissue mRNA levels of prepro-orexin, orexin type-1 (OX1), and orexin type-2 (OX2) receptors were measured using quantitative real-time RT-PCR.
In female rats, pituitary OX1 receptor mRNA levels were increased 12-fold after ovariectomy compared with sham- operated rats. The increase of pituitary OX1 receptor mRNA was inhibited by treatment with 17β-estradiol. Adrenal mRNA levels of OX2 receptors in ovariectomized rats were increased 2-fold compared with sham-operated rats and were also reduced by treatment with 17β-estradiol. In male rats, orchidectomy increased the mRNA levels of pituitary OX1 receptors compared with sham-operated rats. In contrast, adrenal OX2 receptor mRNA was reduced after orchidectomy. Testosterone treatment reversed the effect of orchidectomy on pituitary OX1 and adrenal OX2 receptors. In the hypothalamus, no differences were found in the mRNA levels of prepro-orexin, OX1, and OX2 receptors between sham-operated, placebo-treated, and steroid-treated female or male rats.
Our results indicate that gonadal steroids differentially regulate pituitary OX1 receptors and adrenal OX2 receptors in male and female rats and may contribute to specific sex- dependent neuroendocrine and endocrine actions of orexins.
Bone morphogenetic protein‐2 enhances osterix gene expression in chondrocytes
Authors: Yagi K. , Tsuji K. , Nifuji A. , Shinomiya K. , Nakashima K. , deCrombrugghe B. , Noda Masaki 
Published: March 2003

AbstractOsterix is a recently identified zinc‐finger‐containing transcription factor, which is required for skeletogenesis as no bone formation was observed in osterix‐deficient mice. Osterix was first cloned as a gene whose expression was enhanced by BMP in C2C12 cells. As BMP induces ectopic bone formation in vivo via a pathway reminiscent to endochondral bone formation, BMP may also regulate osterix gene expression in chondrocytes. However, no information was available regarding the BMP actions on osterix gene expression in chondrocytes. We therefore examined the effects of BMP‐2 on osterix gene expression in chondrocytes in culture. RT‐PCR analysis indicated that osterix mRNA was expressed in the primary cultures of chondrocytes derived from mouse rib cartilage. The treatment with BMP‐2 enhanced the levels of osterix transcripts within 24 h and the enhancement was still observed at 48 h based on RT‐PCR analysis. This BMP effect was specific to this cytokine, as TGF‐β did not alter osterix gene expression. BMP effects on the osterix mRNA levels were also confirmed by Northern blot analysis. The enhancing effect of BMP on osterix gene expression was observed in a dose‐dependent manner starting at 200 ng/ml. The BMP enhancement of the osterix gene expression in chondrocytes was blocked in the presence of a protein synthesis inhibitor, cycloheximide, while it was still observed in the presence of 5,6‐dichloro‐1‐β D‐ribofuranosylbenzimidazol (DRB) suggesting the involvement of post‐transcriptional events, which require new protein synthesis. These results indicated that osterix gene is expressed in the primary cultures of chondrocytes and its expression is under the control of BMP‐2. J. Cell. Biochem. 88: 1077–1083, 2003. © 2003 Wiley‐Liss, Inc.
Retention and Degradation of N-Glycoproteins in the Rough Endoplasmic Reticulum
Authors: Duvet Sandrine , Dubuisson Jean , Ermonval Myriam , Cacan René , Verbert André 
Published: March 2003

Recent studies have shown that newly synthesized proteins and glycoproteins are submitted to a quality control mechanism in the rough endoplasmic reticulum (ER). In this report we present two models: One model will illustrate a transient retention in rough ER leading to a further degradation of glycoproteins in the cytosol, (soluble alkaline phosphatase expressed in Man-P-Dol deficient CHO cells lines). The second model will illustrate a strict retention of glycoproteins in rough ER without degradation nor recycling through the Golgi (E1, E2 glycoproteins of Hepatitis C virus in stably transfected UHCV-11.4 cells and in infected Hep G2 cells).
In both cases, oligomannoside structures are markers of these phenomena, either as free soluble released oligomannosides in the case of degradation, or as N-linked oligomannosides for strict retention in rough ER.
Paraventricular Nucleus Administration of Calcitonin Gene-Related Peptide Inhibits Food Intake and Stimulates the Hypothalamo-Pituitary-Adrenal Axis
Authors: Dhillo Waljit S. , Small Caroline J. , Jethwa Preeti H. , Russell Sabina H. , Gardiner James V. , Bewick Gavin A. , Seth Asha , Murphy Kevin G. , Ghatei Mohammad A. , Bloom Stephen R. 
Published: March 2003

Abstract
Calcitonin gene-related protein (CGRP) inhibits food intake and stimulates the hypothalamo-pituitary-adrenal (HPA) axis after intracerebroventricular injection in rats. However, the hypothalamic site and mechanism of action are unknown. We investigated the effects of intraparaventricular nucleus administration (iPVN) of CGRP on food intake and the HPA axis in rats and the effect of CGRP on the release of hypothalamic neuropeptides in vitro. In addition, we investigated the effects of food deprivation on hypothalamic CGRP expression. CGRP dose-dependently reduced food intake in the first hour after iPVN injection in fasted male rats (saline, 5.1 ± 0.8 g; 0.3 nmol CGRP, 1.1 ± 0.5 g; P &lt; 0.001 vs. saline). iPVN injection of CGRP8–37 (a CGRP1 receptor antagonist) alone had no effect on food intake. However, the reduction in food intake by iPVN CGRP was attenuated by prior administration of CGRP8–37 [CGRP8–37 (10 nmol)/CGRP (0.3 nmol), 3.0 ± 0.8 g; P &lt; 0.05 vs. 0.3 nmol CGRP]. CGRP (100 nm) stimulated the release of α-melanocyte stimulating hormone, cocaine- and amphetamine-related transcript, corticotropin-releasing hormone, and arginine vasopressin from hypothalamic explants to 127 ± 19%, 148 ± 10%, 158 ± 17%, and 198 ± 21% of basal levels, respectively (P &lt; 0.05 vs. basal), but did not alter the release of either neuropeptide Y or agouti-related protein. Hypothalamic CGRP mRNA levels in 24-h fasted rats were increased to 130 ± 8% of control levels [CGRP mRNA (arbitrary units), 4.75 ± 0.4; controls, 3.65 ± 0.34; P &lt; 0.05]. Our data suggest that CGRP administered to the PVN inhibits food intake and stimulates the HPA axis.
Suppressive role of endogenous regucalcin in the enhancement of nitric oxide synthase activity in liver cytosol of normal and regucalcin transgenic rats
Authors: Yamaguchi Masayoshi , Takahashi Hiroko , Tsurusaki Yoshinori 
Published: March 2003

AbstractThe suppressive role of endogenous regucalcin, which is a regulatory protein of calcium signaling, in the enhancement of nitric oxide (NO) synthase activity in the liver cytosol of rats was investigated. The enzyme activity was measured in a reaction mixture containing either vehicle or calcium chloride (1–20 μM) in the absence or presence of regucalcin (0.1, 0.25, or 0.5 μM). NO synthase activity was significantly increased by the addition of calcium (5–20 μM). This increase was completely abolished in the presence of trifluoperazine (TFP; 10–50 μM), an antagonist of Ca2+/calmodulin. The addition of regucalcin (0.1–0.5 μM) caused a significant fall in the calcium‐increased enzyme activity. The effect of regucalcin (0.25 μM) in decreasing NO synthase activity was seen in the presence of ethylene glycol bis‐(2‐aminoethylether) N,N,N′,N′‐tetraacetic acid (EGTA, 1 mM) or TFP (20 μM), indicating that regucalcin acts independent on Ca2+/calmodulin. NO synthase activity was significantly raised in the presence of anti‐regucalcin monoclonal antibody (10–50 ng/ml) in the reaction mixture. The effect of the antibody (50 ng/ml) or calcium (10 μM) in elevating NO synthase activity in the liver cytosol of normal rats was not seen in the liver cytosol obtained from regucalcin transgenic rats. Moreover, the increase in NO synthase activity in the liver cytosol of normal rats induced by a single intraperitoneal administration of calcium (5.0 mg/100 g body weight) was significantly enhanced in the presence of anti‐regucalcin monoclonal antibody (50 ng/ml) in the reaction mixture. The administration of calcium caused a significant increase in regucalcin level in the liver cytosol of normal rats. The present study demonstrated that endogenous regucalcin plays a suppressive role in the enhancement of NO synthase activity in the liver cytosol of rats. J. Cell. Biochem. 88: 1226–1234, 2003. © 2003 Wiley‐Liss, Inc.
Progression of chondrogenesis in C3H10T1/2 cells is associated with prolonged and tight regulation of ERK1/2
Authors: Seghatoleslami M. Reza , Roman‐Blas Jorge A. , Rainville Anne M. , Modaressi Rozbeh , Danielson Keith G. , Tuan Rocky S. 
Published: March 2003

AbstractClose contact of mesenchymal cells in vivo and also in super dense micromass cultures in vitro results in cellular condensation and alteration of existing cellular signaling required for initiation and progression of chondrogenesis. To investigate chondrogenesis related changes in the activity of ubiquitous cell signaling mediated by mitogen‐activated protein kinases (MAP kinase), we have compared the effect of cell seeding of pluripotent C3H10T1/2 mesenchymal cells as monolayers (non‐chondrogenic culture) or high density micromass cultures (chondrogenic) on the regulation and phosphorylation state of extracellular signal‐regulated kinase 1 and 2 (ERK1/2) and also on regulation of ERK1/2 nuclear targets, namely, activation protein‐1 (AP‐1) and serum response factor (SRF). Increasing cell density resulted in reduced DNA binding as well as activity of AP‐1. SRF activity, on the other hand, was up‐regulated in confluent monolayer cultures but like AP‐1 was inhibited in micromass cultures. Low levels of PD 98059 (5 μM), a specific inhibitor of ERK1/2, resulted in delayed induction of AP‐1 and SRF activity whereas higher concentrations of this inhibitor (10–50 μM) conferred an opposite effect. Increasing concentrations of the PD 98059 inhibitor in long term monolayer or micromass cultures (2.5 day) resulted in differential regulation of c‐Fos and c‐Jun protein levels as well as total expression and phosphorylation levels of ERK1/2. PD 98059 treatment of C3H10T1/2 micromass cultures also resulted in up‐regulation of type IIB collagen and Sox9 gene expression. While high expression of aggrecan and type IIB collagen genes were dependent on BMP‐2 signaling, ERK inhibition of BMP‐2 treated micromass cultures resulted in reduced activity of both genes. Our findings show that the activity of ERK1/2 in chondrogenic cultures of C3H10T1/2 cells is tightly controlled and can cross interact with other signaling activities mediated by BMP‐2 to positively regulate chondrogensis. J. Cell. Biochem. 88: 1129–1144, 2003. Published 2003 Wiley‐Liss, Inc.