bluuebunny/crossref_metadata_2025_split huggingface dataset:
A cell‐based Dkk1 binding assay reveals roles for extracellular domains of LRP5 in Dkk1 interaction and highlights differences between wild‐type and the high bone mass mutant LRP5(G171V)
Authors: Murrills Richard J. , Matteo Jeanne J. , Bhat Bheem M. , Coleburn Valerie E. , Allen Kristina M. , Chen Wei , Damagnez Veronique , Bhat Ramesh A. , Bex Frederick J. , Bodine Peter V.N. 
Published: September 2009

AbstractDkk1 is a secreted antagonist of the LRP5‐mediated Wnt signaling pathway that plays a pivotal role in bone biology. Because there are no well‐documented LRP5‐based assays of Dkk1 binding, we developed a cell‐based assay of Dkk1/LRP5 binding using radioactive 125I‐Dkk1. In contrast to LRP6, transfection of LRP5 alone into 293A cells resulted in a low level of specific binding that was unsuitable for routine assay. However, co‐transfection of LRP5 with the chaperone protein MesD (which itself does not bind Dkk1) or Kremen‐2 (a known Dkk1 receptor), or both, resulted in a marked enhancement of specific binding that was sufficient for evaluation of Dkk1 antagonists. LRP5 fragments comprising the third and fourth β‐propellers plus the ligand binding domain, or the first β‐propeller, each inhibited Dkk1 binding, with mean IC50s of 10 and 196 nM, respectively. The extracellular domain of Kremen‐2 (“soluble Kremen”) was a weaker antagonist (mean IC50 806 nM). We also found that cells transfected with a high bone mass mutation LRP5(G171V) had a subtly reduced level of Dkk1 binding, compared to wild type LRP5‐transfected cells, and no enhancement of binding by MesD. We conclude that (1) LRP5‐transfected cells do not offer a suitable cell‐based Dkk1 binding assay, unless co‐transfected with either MesD, Kremen‐2, or both; (2) soluble fragments of LRP5 containing either the third and fourth β‐propellers plus the ligand binding domain, or the first β‐propeller, antagonize Dkk1 binding; and (3) a high bone mass mutant LRP5(G171V), has subtly reduced Dkk1 binding, and, in contrast to LRP5, no enhancement of binding with MesD. J. Cell. Biochem. 108: 1066–1075, 2009. © 2009 Wiley‐Liss, Inc.
Polyamine effects on cell function: Possible central role of plasma membrane PI(4,5)P2
Authors: Coburn Ronald F. 
Published: September 2009

AbstractThe polyamines spermidine, spermine, and putrescine are intimately involved in and required for cell growth and proliferation. There are also multiple effects of polyamines on other cellular processes that seem not to be a result of changes in protein expression. It is a daunting task to classify and understand cellular effects of endogenous polyamines. There has been no central hypothesis how these effects can occur or how spermine and spermidine could be targeted to various signal transduction cascades. However, now there is evidence that multiple effects of endogenous polyamines on different cellular processes may involve plasma membrane PI(4,5)P2 and recent evidence of how polyamines could be targeted to specific cellular functions. J. Cell. Physiol. 221: 544–551, 2009. © 2009 Wiley‐Liss, Inc.
The mammalian anti‐proliferative BTG/Tob protein family
Authors: Winkler G. Sebastiaan 
Published: September 2009

AbstractThe mammalian BTG/Tob family comprises six proteins (BTG1, BTG2/PC3/Tis21, BTG3/ANA, BTG4/PC3B, Tob1/Tob and Tob2), which regulate cell cycle progression in a variety of cell types. They are characterised by the conserved N‐terminal domain spanning 104–106 amino acids. Recent biochemical and structural data indicate that the conserved BTG domain is a protein–protein interaction module, which is capable of binding to DNA‐binding transcription factors as well as the paralogues CNOT7 (human Caf1/Caf1a) and CNOT8 (human Pop2/Calif/Caf1b), two deadenylase subunits of the Ccr4‐Not complex. Consistent with this finding, several members of the BTG/Tob family are shown to be implicated in transcription in the nucleus and cytoplasmic mRNA deadenylation and turnover. The C‐terminal regions are less conserved and appear to mediate protein–protein interactions that are unique to each family member. The human and mouse BTG/Tob proteins will be the focus of this review and structural aspects of BTG/Tob interactions with components of the Ccr4‐Not complex, and the role of the BTG/Tob proteins in the regulation of gene expression, tumourigenesis and cancer will be discussed. J. Cell. Physiol. 222:66–72, 2010. © 2009 Wiley‐Liss, Inc.
Runx2, p53, and pRB status as diagnostic parameters for deregulation of osteoblast growth and differentiation in a new pre‐chemotherapeutic osteosarcoma cell line (OS1)
Authors: Pereira Barry P. , Zhou Yefang , Gupta Anurag , Leong David T. , Aung Khin Zarchi , Ling Ling , Pho Robert W.H. , Galindo Mario , Salto‐Tellez Manuel , Stein Gary S. , Cool Simon M. , van Wijnen Andre J. , Nathan Saminathan S. 
Published: September 2009

AbstractOsteosarcomas are the most prevalent primary bone tumors found in pediatric patients. To understand their molecular etiology, cell culture models are used to define disease mechanisms under controlled conditions. Many osteosarcoma cell lines (e.g., SAOS‐2, U2OS, MG63) are derived from Caucasian patients. However, patients exhibit individual and ethnic differences in their responsiveness to irradiation and chemotherapy. This motivated the establishment of osteosarcoma cell lines (OS1, OS2, OS3) from three ethnically Chinese patients. OS1 cells, derived from a pre‐chemotherapeutic tumor in the femur of a 6‐year‐old female, were examined for molecular markers characteristic for osteoblasts, stem cells, and cell cycle control by immunohistochemistry, reverse transcriptase‐PCR, Western blotting and flow cytometry. OS1 have aberrant G‐banded karyotypes, possibly reflecting chromosomal abnormalities related to p53 deficiency. OS1 had ossification profiles similar to human fetal osteoblasts rather than SAOS‐2 which ossifies ab initio (P < 0.05). Absence of p53 correlates with increased Runx2 expression, while the slow proliferation of OS1 cells is perhaps attenuated by pRB retention. OS1 express mesenchymal stem cell markers (CD44, CD105) and differ in relative expression of CD29, CD63, and CD71 to SAOS‐2. (P < 0.05). Cell cycle synchronization with nocodazole did not affect Runx2 and CDK1 levels but decreased cyclin‐E and increased cyclin‐A (P < 0.05). Xenotransplantion of OS1 in SCID mice yields spontaneous tumors that were larger and grew faster than SAOS‐2 transplants. Hence, OS1 is a new osteosarcoma cell culture model derived from a pre‐chemotherapeutic ethnic Chinese patient, for mechanistic studies and development of therapeutic strategies to counteract metastasis and deregulation of mesenchymal development. J. Cell. Physiol. 221: 778–788, 2009. © 2009 Wiley‐Liss, Inc.
Activation of nuclear factor kappa B in mammary epithelium promotes milk loss during mammary development and infection
Authors: Connelly Linda , Barham Whitney , Pigg Rachel , Saint‐Jean Leshana , Sherrill Taylor , Cheng Dong‐Sheng , Chodosh Lewis A. , Blackwell Timothy S. , Yull Fiona E. 
Published: September 2009

AbstractWe investigated whether nuclear factor kappa B (NF‐κB), which exhibits a regulated pattern of activity during murine mammary gland development, plays an important role during lactation and involution, when milk production ceases and the gland undergoes apoptosis and re‐modeling. We generated a doxycycline inducible transgenic mouse model to activate NF‐κB specifically in the mammary epithelium through expression of a constitutively active form of IKK2, the upstream kinase in the classical NF‐κB signaling cascade. We found that activation of NF‐κB during involution resulted in a more rapid reduction in milk levels and increased cleavage of caspase‐3, an indicator of apoptosis. We also found that activation of NF‐κB during lactation with no additional involution signals had a similar effect. The observation that NF‐κB is a key regulator of milk production led us to investigate the role of NF‐κB during mastitis, an infection of the mammary gland in which milk loss is observed. Mammary gland injection of E. coli LPS resulted in activation of NF‐κB and milk loss during lactation. This milk loss was decreased by selective inhibition of NF‐κB in mammary epithelium. Together, our data reveal that activation of NF‐κB leads to milk clearance in the lactating mammary gland. Therefore, targeting of NF‐κB signaling may prove therapeutic during mastitis in humans and could be beneficial for the dairy industry, where such infections have a major economic impact. J. Cell. Physiol. 222:73–81, 2010. © 2009 Wiley‐Liss, Inc.
Induction of Th17 differentiation by corneal epithelial‐derived cytokines
Authors: Zheng Xiaofen , Bian Fang , Ma Ping , De Paiva Cintia S. , Stern Michael , Pflugfelder Stephen C. , Li De‐Quan 
Published: September 2009

AbstractThis study was to explore a potential role of epithelium‐derived cytokines in Th17 differentiation. Th17 induction was evaluated by murine CD4+T cells treated with different combinations of five inducing cytokines, or conditioned media of human corneal epithelial cells (HCECs) exposed to a variety of stimuli. Th17 differentiation was determined by measuring Th17 associated molecules, IL‐17A, IL‐17F, IL‐22, CCL‐20, and STAT3 at mRNA and protein levels, and numbers of IL‐17‐producing T cells by real‐time PCR, and cytokine immunobead and ELISPOT assays, respectively. IL‐23 was the strongest inducer for expanding Th17 cells in the presence of TGF‐β1 + IL‐6; and IL‐1β was the strongest Th17 amplifier in the presence of TGF‐β1 + IL‐6 + IL‐23. These inducing cytokines were found to be significantly stimulated in HCECs challenged by hyperosmotic media (450 mOsM), microbial components (polyI:C, flagellin, R837, and other TLR ligands) and TNF‐α. Interestingly, when incubated with conditioned media of HCECs irritated by polyI:C or TNF‐α, CD4+T cells displayed increased mRNA levels of IL‐17A, IL‐17F, IL‐22, CCL‐20, and STAT3, increased IL‐17 protein in the supernatant, and increased numbers of IL‐17‐producing T cells (Th17 cells). These findings demonstrate for the first time that Th17 differentiation can be promoted by cytokines produced by corneal epithelium that are exposed to hyperosmotic, microbial, and inflammatory stimuli. J. Cell. Physiol. 222:95–102, 2010. © 2009 Wiley‐Liss, Inc.
Variations of the origin of renal arteries in the fetus identified on power Doppler and 3D sonography
Authors: Degani Shimon , Leibovitz Zvi , Shapiro Israel , Ohel Gonen 
Published: September 2009

AbstractPurpose.To investigate anatomic variations of renal arteries (RAs) using 2‐dimensional (2D)/3D power Doppler sonography (PDUS) in normal fetuses and in fetuses with renal anomalies.Methods.The origin, direction, and distribution of RAs were studied in 120 fetuses without renal malformations using 2D/3D PDUS. Studies were performed between 14 and 17 weeks of gestation age on 64 male and 56 female fetuses. PDUS of the RAs was also performed in 12 fetuses with renal anomalies.Results.In 117/120 fetuses, a single RA was found to originate from each side of the abdominal aorta. The origin of the right RA from the abdominal aorta was superior to, at the same level as, and inferior to that of the left RA in 47% (55/117), 25.5% (30/117), and 27.5% (32/117) of the cases, respectively. An accessory right RA was found in 3 cases. Bifurcation of the right RA was found in 2 cases. There were no variations of the origin of renal vessels in 69% of the cases (83/120), whereas the remaining 31% (37/120) had 1 or several variation patterns. Variations in renal vessels were found in 8 of the 12 fetuses with renal malformations: accessory artery in 3 cases, origin of the right RA from the iliac artery in horseshoe kidney and in 2 cases with pelvic kidney, and bifurcation in 2 of the fetuses with double collecting system.Conclusions.Variations in origin and distribution of fetal RAs are frequently observed in normal pregnancies. The majority of renal malformations are associated with such vascular variations. © 2009 Wiley Periodicals, Inc. J Clin Ultrasound, 2010
Evaluation of the hepatic artery anastomosis by intraoperative sonography with high‐frequency transducer in right‐lobe graft living donor liver transplantation
Authors: Mun Han Song , Kim Kyoung Won , Song Gi‐Won , Ahn Chul‐Soo , Kim So Yeon , Hwang Shin , Lee Sung Gyu 
Published: September 2009

AbstractObjectiveTo describe the usefulness of intraoperative ultrasonography (IOUS) with high‐frequency transducer in living donor liver transplantation (LDLT) using right‐lobe graft (RLG).MethodThis retrospective study was approved by our institutional review board. We performed IOUS in 22 patients (17 men and 5 women, aged 51 ± 9.0 years) during LDLT with RLG using a Sequoia 512 scanner with an 8–12‐MHz linear transducer. Hepatic artery (HA) anastomosis was identified on gray‐scale US, and the diameter and percentage of stenosis of the anastomosis were measured. The HA was evaluated to detect thrombus or dissection in the region of anastomosis. Doppler study of the graft HA was also performed. Patients were divided into those with and without abnormalities, including thrombosis, dissection, and abnormal Doppler parameters (peak systolic velocity < 30 cm/s or > 2 m/s, resistance index < 0.5, and systolic acceleration time > 80 msec).ResultOn gray‐scale and Doppler IOUS study, abnormalities were found in 10 of 22 patients. Diagnoses were anastomotic stenosis (n = 2), celiac stenosis (n = 1), compromise of HA inflow due to systemic hypotension (n = 1), HA thrombosis (n = 2), and HA dissection (n = 4). Re‐anastomoses were done in 3 case (2 stenoses and 1 thrombosis). Uneventful postoperative recovery occurred in the other 7 patients without re‐anastomosis.ConclusionIOUS with high‐frequency transducer is a useful method to make an early diagnosis of HA complications of LDLT with RLG. © 2009 Wiley Periodicals, Inc. J Clin Ultrasound, 2010
Trumpet maneuver in the sonographic diagnosis of an external laryngocele
Authors: Conkbayir Isik , Keyik Bahri , Hekimoglu Koray , Cifci Egemen , Yanik Bahar 
Published: September 2009

AbstractLaryngocele is an uncommon disease defined as an abnormal dilatation or herniation of the laryngeal saccule, forming an air‐filled cavity. Radiologic evaluation of a suspected laryngocele generally includes plain radiographs, CT, and MRI. There are very few reports in the literature on sonographic findings of laryngocele. Trumpet maneuver has been reported in CT examination for the diagnosis of laryngocele. However, to the best of our knowledge, the use of trumpet maneuver in the sonographic diagnosis of laryngocele has not been previously described. This report presents the sonographic findings of an external laryngocele while performing the trumpet maneuver. © 2009 Wiley Periodicals, Inc. J Clin Ultrasound, 2010
Deep thiopental anesthesia alters steady‐state glucose homeostasis but not the neurochemical profile of rat cortex
Authors: Lei Hongxia , Duarte Joao M.N. , Mlynarik Vladimir , Python Agathe , Gruetter Rolf 
Published: September 2009

AbstractBarbiturates are regularly used as an anesthetic for animal experimentation and clinical procedures and are frequently provided with solubilizing compounds, such as ethanol and propylene glycol, which have been reported to affect brain function and, in the case of 1H NMR experiments, originate undesired resonances in spectra affecting the quantification. As an alternative, thiopental can be administrated without any solubilizing agents. The aim of the study was to investigate the effect of deep thiopental anesthesia on the neurochemical profile consisting of 19 metabolites and on glucose transport kinetics in vivo in rat cortex compared with α‐chloralose using localized 1H NMR spectroscopy. Thiopental was devoid of effects on the neurochemical profile, except for the elevated glucose at a given plasma glucose level resulting from thiopental‐induced depression of glucose consumption at isoelectrical condition. Over the entire range of plasma glucose levels, steady‐state glucose concentrations were increased on average by 48% ± 8%, implying that an effect of deep thiopental anesthesia on the transport rate relative to cerebral glucose consumption ratio was increased by 47% ± 8% compared with light α‐chloralose‐anesthetized rats. We conclude that the thiopental‐induced isoelectrical condition in rat cortex significantly affected glucose contents by depressing brain metabolism, which remained substantial at isoelectricity. © 2009 Wiley‐Liss, Inc.
KATP channel openers protect mesencephalic neurons against MPP+‐induced cytotoxicity via inhibition of ROS production
Authors: Xie Juan , Duan Lei , Qian Xia , Huang Xu , Ding Jianhua , Hu Gang 
Published: September 2009

AbstractOpening of ATP‐sensitive potassium (KATP) channels has been demonstrated to exert significant neuroprotection in in vivo and in vitro models of Parkinson's disease (PD), but the exact mechanism remains unclear. In the present study, various KATP channel openers (KCOs) sensitive to diverse KATP subunits were used to clarify the protective role of KATP channel opening in 1‐methyl‐4‐phenylpyridinium (MPP+)‐induced oxidative stress injury in mouse primary cultured mesencephalic neurons. The results showed that pretreatment with nonselective KCO pinacidil (Pin) or diazoxide (Dia), a KCO sensitive to Kir6.2/SUR1 KATP channels, protected mesencephalic neurons, especially dopaminergic neurons, against MPP+‐induced injury in a concentration‐dependent manner. However, cromakalim (Cro), an opener of Kir6.1/SUR2 but not Kir6.2/SUR1 KATP channels, failed to protect against MPP+‐induced cytotoxicity. Furthermore, Pin and Dia but not Cro significantly suppressed the elevation of reactive oxygen species (ROS) triggered by MPP+ and prevented the loss of mitochondrial member potential (ΔΨm) and the release of mitochondrial cyotchrome c. Consequently, opening of KATP channels expressed in neurons could protect primary mesencephalic neurons against MPP+‐induced cytotoxicity via inhibiting ROS overproduction and subsequently ameliorating mitochondrial function. © 2009 Wiley‐Liss, Inc.
Dispersive liquid–liquid microextraction using extraction solvent lighter than water
Authors: Farajzadeh Mir Ali , Seyedi Seyed Esmaeil , Shalamzari Mohammad Safi , Bamorowat Mehdi 
Published: September 2009

AbstractFor the first time a dispersive liquid–liquid microextraction method on the basis of an extraction solvent lighter than water was presented in this study. Three organophosphorus pesticides (OPPs) were selected as model compounds and the proposed method was carried out for their preconcentration from water samples. In this extraction method, a mixture of cyclohexane (extraction solvent) and acetone (disperser) is rapidly injected into the aqueous sample in a special vessel (see experimental section) by syringe. Thereby, a cloudy solution is formed. In this step, the OPPs are extracted into the fine droplets of cyclohexane dispersed into aqueous phase. After centrifuging the fine droplets of cyclohexane are collected on the upper of the extraction vessel. The upper phase (0.40 μL) is injected into the gas chromatograph (GC) for separation. Analytes were detected by a flame ionization detector (FID) (for high concentrations) or MS (for low concentrations). Some important parameters, such as the kind of extraction and dispersive solvents and volume of them, extraction time, temperature, and salt amount were investigated. Under the optimum conditions, the enrichment factors (EFs) ranged from 100 to 150 and extraction recoveries varied between 68 and 105%, both of which are relatively high over those of published methods. The linear ranges were wide (10–100 000 μg/L for GC‐FID and 0.01–1 μg/L for GC‐MS) and LODs were low (3–4 μg/L for GC‐FID and 0.003 μg/L for GC‐MS). The RSDs for 100.0 μg/L of each OPP in water were in the range of 5.3–7.8% (n = 5).
HPLC with polysaccharide chiral stationary phase in polar‐organic phase mode: Application to the asymmetric epoxidation of allylic alcohols
Authors: Morante‐Zarcero Sonia , del Hierro Isabel , Fajardo Mariano , Sierra Isabel 
Published: September 2009

AbstractA simple and rapid HPLC method using a polysaccharide‐based chiral stationary phase (Chiralpak AD‐H) in polar‐organic phase mode has been developed for direct resolution of glycidyl nitrobenzoate (GNB) and 2‐methyl glycidyl nitrobenzoate (MGNB) enantiomers. ACN and methanol were used as mobile phase and the effects of the addition of ethanol and 2‐propanol as organic modifier in the mobile phase, flow rate and the column temperature were tested. The optimized conditions were: methanol/ethanol (80:20) at a flow rate of 0.9 mL/min and 40°C. Analysis time was ⪇13 min and the chiral resolution was ⪈2. The method was validated and resulted to be selective, precise and accurate. The method was found to be linear in 2–300 μg/mL range (R2 >0.999) with an LOD nearly 0.5 μg/mL for four enantiomers. GNB and MGNB enantiomers were obtained by asymmetric epoxidation of allyl alcohol and 2‐methyl allyl alcohol, respectively, using chiral titanium–tartrate complexes as catalyst and dichloromethane as solvent after in situ derivatization of the intermediate glycidols derivatives. The quite simple and rapid validated method was applied successfully for direct determination of the enantiomeric excess (⪈90%) and yield obtained in real samples of asymmetric epoxidation of allylic alcohols without further purification, workup or solvent removal. The method provides a useful and value‐added tool for controlling the enantiomeric purity of the synthesized epoxides.
Uncertainty in the determination of glucose in aqueous solutions by high‐performance liquid chromatography with evaporative light scattering detection
Authors: Nogueiro Estevinho Berta , Ferraz Amélia , Rocha Fernando , Santos Lúcia , Alves Arminda 
Published: September 2009

AbstractThe determination of glucose and other carbohydrates is the most widespread chemical analysis that is performed within the industries of food, beverage, forage, biomass, pulp and paper, pharmaceuticals among others. Besides that, sugar refineries need to control their products, by‐products and effluents, and furthermore, glucose in the sucrose refining process, is considered an impurity, which shall be controlled. Being HPLC the most currently instrumental technique used for glucose analysis, the evaporative light scattering detector (ELSD) offers advantages (sensitivity, possibility for operating in gradient mode) over the also used refractive index detector. In this work, an HPLC‐ELSD methodology was optimised and validated, aiming the estimate of the uncertainty associated with the results at low levels of concentration of glucose to be measured. Linearity of the response was obtained in the range of glucose concentrations from 20 to 300 mg/L, with an analysis time of 10 min. The global uncertainty was estimated accordingly to the bottom‐up approach used by Eurachem. It was 13% on average for concentrations from 100 to 300 mg/L. For lower concentrations, uncertainty increased significantly up to 30% in the vicinity of the LOD of the method.
Lysozyme in wine: A risk evaluation for consumers allergic to hen's egg
Authors: Weber Patrick , Kratzin Hartmut , Brockow Knut , Ring Johannes , Steinhart Hans , Paschke Angelika 
Published: September 2009

AbstractLysozyme used in wine production could present a risk for consumers allergic to hen's egg. Thus, precautionary labeling of lysozyme on wines has been adopted within the European Community by updating Annex IIIa by Directive 2007/68/EC on November 27, 2007. Since no scientific data is known about the actual amounts and risks of lysozyme in wines, various in vitro efforts and skin prick tests were applied in this study to evaluate the presence of lysozyme in wines and the reactivity of those residues in allergic individuals and to fulfill the claim of updating Annex IIIa announced in Directive 2003/89/EC. Depending on the wine's color (red or white wine) and fining with bentonite, which is known as an important step to remove unstable proteins mainly from white wines, diverse results were obtained concerning the amounts of lysozyme in finished wines and their in vitro and in vivo reactivity in humans allergic to hen's egg.