bluuebunny/crossref_metadata_2025_split huggingface dataset:
Centrifugal methods and devices for rapid in‐gel digestion of proteins
Authors: Lazarev Alexander V. , Rejtar Tomas , Dai Shujia , Karger Barry L. 
Published: March 2009

AbstractModern proteomic research frequently relies upon separation of proteins in a polyacrylamide gel matrix followed by in‐gel enzymatic digestion and extraction of peptides for subsequent analysis by MS. In this work, we propose a novel semi‐automated method of mechanical processing of gel bands by passing these bands through a specially designed centrifugal device termed a Gel Shredder prior to digestion and extraction of peptides. Such a device allows integrated washing, destaining and shredding of gel bands into uniform blocks of controlled size, approximately 150–300 μm, prior to the enzymatic digestion and extraction of peptides. Shredding into uniform blocks increases the surface area of the gel pieces and promotes improved gel rehydration, allowing improved diffusion of the proteolytic enzymes and solvent into the gel lattice. We demonstrate that the new method substantially reduces the time spent on tedious manual handling of gel bands, while minimizing the risk of sample contamination. The performance of the Gel Shredder has been compared with a conventional in‐gel digestion protocol using several standard proteins and a complex proteomic sample in terms of relative quantitation by either MALDI‐TOF/TOF or nanoLC‐ESI IT‐Fourier transformation ion cyclotron resonance MS. It is shown that significant time savings and improved peptide recovery can be obtained for many proteins using the Gel Shredder compared with the traditional in‐gel digestion protocol.
Hexokinase inhibitor screening based on adenosine 5′‐diphosphate determination by electrophoretically mediated microanalysis
Authors: Wang Tongdan , Kang Jingwu 
Published: March 2009

AbstractA CE‐based method for hexokinase inhibitor screening was developed in the present paper. In this method, hexokinase activity was assayed via electrophoretically mediated microanalysis (EMMA), which combines on‐column hexokinase‐mediated reaction and measurement of produced adenosine 5′‐diphosphate (ADP) via electrophoretical separation and UV detection. Enzyme inhibition can be read out directly from the reduced peak area of ADP in comparison with a reference electropherogram obtained in the absence of any inhibitor. Conditions for on‐column enzyme reaction and separation of adenosine 5′‐triphosphate (ATP) and ADP were optimized. The optimal buffer composition for enzymatic reaction was 25 mM HEPES buffer (pH 7.5) containing 5 mM MgCl2, whereas the optimal buffer composition for separation was 100 mM Tris‐phosphate buffer (pH 5.5) containing 0.02% (m/v) hexadimethrine bromide (HDB). Fortunately, discontinuous buffer system can be adapted easily in the EMMA method. The time for separation was reduced dramatically to less than 3 min by reversing the direction of EOF via dynamically coating the capillary wall with the cationic polyelectrolyte HDB. Moreover, the peak tailing of ATP was also reduced by HDB coating. The Z′ factor as high as 0.98 was obtained, indicating a high quality of the screening data. The present method is simple, robust and cost‐effective.
Exposure of HepG2 cells to low levels of PAH‐containing extracts from contaminated soils results in unpredictable genotoxic stress responses
Authors: Mattsson Åse , Lundstedt Staffan , Stenius Ulla 
Published: March 2009

AbstractContaminated soil is a serious environmental problem, constituting a risk to humans and the environment. Polycyclic aromatic hydrocarbons (PAHs) are often present at contaminated sites. However, risk levels are difficult to estimate because of the complexity of contaminants present. Here, we compare cellular effects of extracts from contaminated soils collected at six industrial settings in Sweden. Chemical analysis showed that all soils contained complex mixtures of PAHs and oxy‐PAHs. Western blotting and immunocytochemistry were used to investigate DNA damage signaling in HepG2 cells exposed to extracts from these soils. The effects on phosphorylated Mdm2, p53, Erk, H2AX, 53BP1, and Chk2, cell cycle regulating proteins (cyclin D1 and p21), and cell proliferation were compared. We found that most soil extracts induced phosphorylation of Mdm2 at the 2A10 epitope at low concentrations. This is in line with previous studies suggesting that this endpoint reflects readily repaired DNA‐damage. However, we found concentration‐ and time‐dependent γH2AX and 53BP1 responses that were sustained for 48 hr. These endpoints may reflect the presence of different types of persistent DNA‐damage. High concentrations of soil extracts decreased cyclin D1 and increased p21 response, indicating cell cycle arrest. Phosphorylation of Mdm2 at Ser166, which attenuates the p53 response and is induced by many tumor promoters, was induced in a time‐dependent manner and was associated with Erk phosphorylation. Taken together, the PAH extracts elicited unpredictable signaling responses that differed between samples. More polar compounds, i.e., oxy‐PAHs, also contributed to the complexity. Environ. Mol. Mutagen. 2009. © 2009 Wiley‐Liss, Inc.
Factors related to motivation for change in adolescents with eating disorders
Authors: Zaitsoff Shannon L. , Taylor Andrew 
Published: March 2009

AbstractObjectiveTo examine the association between motivation for change and eating disorder symptom severity, the quality of the parent–adolescent relationship and depressive symptom severity in adolescents with eating disorders.MethodFifty‐four female adolescents with eating disorders (mean age = 15.8 years, SD = 1.6) participated in this research prior to beginning outpatient treatment. Participants completed the Motivational Stages of Change for Adolescents Recovering from an Eating Disorder, the Eating Disorders Inventory‐2, the Relations with Parents scale from the Behavioral Assessment System for Children and Adolescents Self‐report and the Child Depression Inventory.ResultsGreater motivation for change was associated with less body dissatisfaction, more adaptive parent–adolescent relationships and fewer depressive symptoms. The association between motivation for change and adolescents' perception of their relationship with their parents remained significant even after controlling for age, body dissatisfaction and depressive symptoms.ConclusionInvolvement of families in treatment for adolescents with eating disorders may serve to enhance their motivation to recover. Copyright © 2009 John Wiley & Sons, Ltd and Eating Disorders Association.
The prevalence, incidence and development of eating disorders in finnish adolescents—a two‐step 3‐year follow‐up Study
Authors: Isomaa Rasmus , Isomaa Anna‐Lisa , Marttunen Mauri , Kaltiala‐Heino Riittakerttu , Björkqvist Kaj 
Published: March 2009

AbstractObjectiveTo investigate the prevalence, incidence and development of eating disorders and subclinical eating pathology.MethodA two‐step three‐year follow‐up study on eating disorders in adolescence (N = 595) was conducted in western Finland. A screening questionnaire followed by a semi‐structured interview was used to determine the prevalence, incidence and development of eating disorders.ResultsThe lifetime prevalence rates for females age 18 were 2.6% for anorexia nervosa (AN), 0.4 for bulimia nervosa (BN), 7.7% for AN‐NOS, 1.3% for BN‐NOS and 8.5% for subclinical eating disorders. No prevalent case of DSM‐IV eating disorders was found among the male participants. The incidence rate of any eating disorder in females age 15–18 was 1641 per 100 000 person‐years.ConclusionEating disorders are relatively common in female adolescents. As many as one in five adolescent females are or have been struggling with eating disorder related issues during their teenage years. Copyright © 2009 John Wiley & Sons, Ltd and Eating Disorders Association.
Perfectionism and eating attitudes in portuguese students: A longitudinal study
Authors: Soares Maria João , Macedo António , Bos Sandra Carvalho , Marques Mariana , Maia Berta , Pereira Ana Telma , Gomes Ana , Valente José , Pato Michele , Azevedo Maria Helena 
Published: March 2009

AbstractAimTo investigate the role of perfectionism in the development of disordered eating behaviours.Method382 female university students completed the Hewitt & Flett MPS and the EAT‐40 at baseline, and 1 year after (T1) and 206 2 years later (T2).ResultsPerfectionism at baseline was significantly associated with long‐term abnormal eating attitudes/behaviours. Self‐Oriented Perfectionism (SOP) and Socially Prescribed Perfectionism (SPP) were significant predictors of disordered eating behaviours. Regression analysis revealed that SOP at baseline was predictive of Diet Concerns and overall eating disturbance (EAT total score), at T1 and T2. SPP was a significant predictor of Social Pressure to Eat at T1 and T2 and of Bulimic Behaviours only at T1.ConclusionOur findings contribute to a more clear understanding of the association between perfectionism and eating disorders. SOP and SPP were prospectively associated with abnormal eating attitudes/behaviours and SOP was found to be predictive of diet concerns and overall eating disturbance. Copyright © 2009 John Wiley & Sons, Ltd and Eating Disorders Association.
Integrative high‐resolution microarray analysis of human myeloma cell lines reveals deregulated miRNA expression associated with allelic imbalances and gene expression profiles
Authors: Lionetti Marta , Agnelli Luca , Mosca Laura , Fabris Sonia , Andronache Adrian , Todoerti Katia , Ronchetti Domenica , Deliliers Giorgio Lambertenghi , Neri Antonino 
Published: March 2009

AbstractIt is thought that altered microRNA (miRNA) expression due to various mechanisms plays a critical role in the pathogenesis of most human cancers. Notably, about half of the known miRNAs are intragenic and frequently coexpressed with their host genes. To date there is little evidence concerning miRNA expression in multiple myeloma (MM). In an attempt to provide insights into miRNA deregulation in MM, we profiled global miRNA expression in a panel of molecularly well‐characterized human myeloma cell lines (HMCLs) using high‐resolution microarrays, and then used integrative analyses to identify altered patterns correlated with DNA copy number (CN) or gene expression profiles. We identified 16 miRNAs mapped to chromosomal regions frequently involved in numerical imbalances in MM, whose expression significantly correlated with the CN of the corresponding miRNA genes; among these, miR‐22 expression was also affected by chromosome arm 17p loss in a representative panel of primary MM tumors. The expression of 32 intronic miRNAs significantly correlated with that of their host transcripts, some of which were highly deregulated in MM patients. The expression of some of the miRNAs was validated by quantitative RT‐PCR. Finally, a number of the identified miRNAs have previously been reported to play important roles in tumorigenesis. Overall, our data highlight that genomic alterations may significantly affect miRNA expression in HMCLs and demonstrate a frequent coexpression of intronic miRNAs with their host genes that may have a pathogenetic relevance in plasma cell transformation. © 2009 Wiley‐Liss, Inc.
Stat3 inhibition activates tumor macrophages and abrogates glioma growth in mice
Authors: Zhang Leying , Alizadeh Darya , Van Handel Michelle , Kortylewski Marcin , Yu Hua , Badie Behnam 
Published: March 2009

AbstractAs the main effector‐cell population of the central nervous system, microglia (MG) are considered to play an important immunoregulatory function in a number of pathological conditions such as inflammation, trauma, degenerative disease, and brain tumors. Recent studies, however, have suggested that the anti‐neoplastic function of MG may be suppressed in malignant brain tumors. Considering the proposed suppressive role of signal transducers and activators of transcription 3 (Stat3) in antitumor immunity, we evaluated the role of Stat3 inhibition on MG and macrophage (MP) activation and tumor growth in a murine glioma model. N9 MG cells were exposed to GL261 glioma conditioned medium (GL261‐CM) and evaluated for Stat3 activity and cytokine expression. Furthermore, the role of Stat3 inhibition on MG and MP activation was studied both in vitro and in vivo. Finally, the effect of Stat3 inhibition on tumor growth was assessed in intracranial GL261 gliomas. GL261‐CM increased Stat3 activity in N9 cells in vitro and resulted in overexpression of IL‐10 and IL‐6, and downregulation of IL1‐β, a pro‐inflammatory cytokine. Inhibition of Stat3 by CPA‐7 or siRNA reversed glioma‐induced cytokine expression profile in N9 cells. Furthermore, inactivation of Stat3 in intracranial GL261 tumors by siRNA resulted in MG/MP activation and tumor growth inhibition. Glioma‐induced MG and MP suppression may be mediated thorough Stat3. Inhibition of Stat3 function in tumor MG/MP may result in their activation and can potentially be used as an adjunct immunotherapy approach for gliomas. © 2009 Wiley‐Liss, Inc.
Enhancement of neuronal outward delayed rectifier K+ current by human monocyte‐derived macrophages
Authors: Hu Dehui , Liu Jianuo , Xiong Huangui 
Published: March 2009

AbstractMacrophages are critical cells in mediating the pathology of neurodegenerative disorders and enhancement of neuronal outward potassium (K+) current has implicated in neuronal apoptosis. To understand how activated macrophages induce neuronal dysfunction and injury, we studied the effects of lipopolysaccharide (LPS)‐stimulated human monocytes‐derived macrophage (MDM) on neuronal outward delayed rectifier K+ current (IK) and resultant change on neuronal viability in primary rat hippocampal neuronal culture. Bath application of LPS‐stimulated MDM‐conditioned media (MCM) enhanced neuronal IK in a concentration‐dependentmanner, whereas non‐stimulated MCM failed to alter neuronal IK. The enhancement of neuronal IK was repeated in a macrophage‐neuronal co‐culture system. The link of stimulated MCM (MCM(+))‐associated enhancement of IK to MCM(+)‐induced neuronal injury, as detected by PI/DAPI (propidium iodide/4′,6‐diamidino‐2‐phenylindol) staining and MTT assay, was demonstrated by experimental results showing that addition of IK blocker tetraethylammonium to the culture protected hippocampal neurons from MCM(+)‐associated challenge. Further investigation revealed elevated levels of Kv 1.3 and Kv 1.5 channel expression in hippocampal neurons after addition of MCM(+) to the culture. These results suggest that during brain inflammation macrophages, through their capacity of releasing bioactive molecules, induce neuronal injury by enhancing neuronal IK and that modulation of Kv channels is a new approach to neuroprotection. © 2009 Wiley‐Liss, Inc.
Differential expression of sPLA2 following spinal cord injury and a functional role for sPLA2‐IIA in mediating oligodendrocyte death
Authors: Titsworth W. Lee , Cheng Xiaoxin , Ke Yan , Deng Lingxiao , Burckardt Kenneth A. , Pendleton Chris , Liu Nai‐Kui , Shao Hui , Cao Qi‐Lin , Xu Xiao‐Ming 
Published: March 2009

AbstractAfter the initial mechanical insult of spinal cord injury (SCI), secondary mediators propagate a massive loss of oligodendrocytes. We previously showed that following SCI both the total phospholipase activity and cytosolic PLA2‐IVα protein expression increased. However, the expression of secreted isoforms of PLA2 (sPLA2) and their possible roles in oligodendrocyte death following SCI remained unclear. Here we report that mRNAs extracted 15 min, 4 h, 1 day, or 1 month after cervical SCI show marked upregulation of sPLA2‐IIA and IIE at 4 h after injury. In contrast, SCI induced down regulation of sPLA2‐X, and no change in sPLA2‐IB, IIC, V, and XIIA expression. At the lesion site, sPLA2‐IIA and IIE expression were localized to oligodendrocytes. Recombinant human sPLA2‐IIA (0.01, 0.1, or 2 μM) induced a dose‐dependent cytotoxicity in differentiated adult oligodendrocyte precursor cells but not primary astrocytes or Schwann cells in vitro. Most importantly, pretreatment with S3319, a sPLA2‐IIA inhibitor, before a 30 min H2O2 injury (1 or 10 mM) significantly reduced oligodendrocyte cell death at 48 h. Similarly, pretreatment with S3319 before injury with IL‐1β and TNFα prevented cell death and loss of oligodendrocyte processes at 72 h. Collectively, these findings suggest that sPLA2‐IIA and IIE are increased following SCI, that increased sPLA2‐IIA can be cytotoxic to oligodendrocytes, and that in vitro blockade of sPLA2 can create sparing of oligodendrocytes in two distinct injury models. Therefore, sPLA2‐IIA may be an important mediator of oligodendrocyte death and a novel target for therapeutic intervention following SCI. © 2009 Wiley‐Liss, Inc.
IL‐12 p40 homodimer, the so‐called biologically inactive molecule, induces nitric oxide synthase in microglia via IL‐12Rβ1
Authors: Jana Malabendu , Dasgupta Subhajit , Pal Utpal , Pahan Kalipada 
Published: March 2009

AbstractEarlier we have demonstrated that IL‐12 p40 homodimer (p402) induces the expression of inducible nitric oxide synthase (iNOS) in microglia. This study was undertaken to investigate underlying mechanisms required for IL‐12 p402‐ and IL‐12 p70‐induced expression of iNOS in microglia. IL‐12 p402 alone induced the activation of both extracellular signal‐regulated kinase (ERK) and p38 mitogen‐activated protein kinase (MAPK). Interestingly, the ERK pathway coupled p402 to iNOS expression via C/EBPβ, but not NF‐κB, whereas the p38 pathway relayed the signal from p402 to iNOS expression via both NF‐κB and C/EBPβ. Furthermore, by using microglia from IL‐12Rβ1 (−/−) and IL‐12Rβ2 (−/−) mice or siRNA against IL‐12Rβ1 and IL‐12Rβ2, we demonstrate that p402 induced the expression of iNOS in microglia via IL‐12Rβ1–(ERK+p38)–(NF‐κB +C/EBPβ) pathway. In contrast, both IL‐12Rβ1 and IL‐12Rβ2 were involved for IL‐12 p70‐induced microglial expression of iNOS. Although IL‐12Rβ1 coupled p70 to NF‐κB and C/EBPβ, IL‐12Rβ2 was responsible for p70‐mediated activation of GAS. This study delineates a new role of IL‐12Rβ1 and IL‐12Rβ2 for the expression of iNOS and production of NO in microglia that may participate in the pathogenesis of neuroinflammatory diseases. © 2009 Wiley‐Liss, Inc.
Sp proteins and Runx2 mediate regulation of matrix gla protein (MGP) expression by parathyroid hormone
Authors: Suttamanatwong Supaporn , Jensen Eric D. , Schilling Jody , Franceschi Renny T. , Carlson Ann E. , Mansky Kim C. , Gopalakrishnan Rajaram 
Published: March 2009

AbstractAs part of its catabolic action in bone, parathyroid hormone (PTH) inhibits extracellular matrix mineralization. We previously showed that PTH dose‐dependently induces matrix gla protein (MGP) expression in osteoblasts and this induction is at least partially responsible for PTH‐mediated inhibition of mineralization. Recently, we identified PKA and ERK/MAPK as the key signaling pathways involved in PTH regulation of MGP expression. The goal of this study was to further characterize the mechanism by which PTH stimulates expression of MGP. Deletion analysis of the murine Mgp gene promoter identified a PTH‐responsive region between −173 bp and−49 bp. Using gel‐mobility shift assays we found that Sp1/Sp3, and Runx2 bind to distinct sites within this region. Mutation of either the Sp or the Runx2 site reduced MGP induction by PTH, while mutation of both sites completely abolished PTH responsiveness. Overexpression of Runx2 or Sp1 activated the Mgp reporter, while Sp3 was a dose‐dependent repressor of Sp1 and PTH‐induced MGP expression. Collectively, these data show that PTH regulates MGP gene transcription in osteoblasts through altered activities of Sp and Runx2 transcription factors. J. Cell. Biochem. 107: 284–292, 2009. © 2009 Wiley‐Liss, Inc.
Suppression of PAI‐1 expression through inhibition of the EGFR‐mediated signaling cascade in rat kidney fibroblast by ascofuranone
Authors: Cho Hyun‐Ji , Kang Jeong‐Han , Kim Teoan , Park Kwang‐Kyun , Kim Cheorl‐Ho , Lee In‐Seon , Min Kwan‐Sik , Magae Junji , Nakajima Hiroo , Bae Young‐Seuk , Chang Young‐Chae 
Published: March 2009

AbstractFibrosis in glomerulosclerosis causes progressive loss of renal function. Transforming growth factor (TGF)‐β, one of the major profibrotic cytokines, induces the synthesis of plasminogen activator inhibitor (PAI)‐1, a factor that plays a crucial role in the development of fibrosis. Here, we found that an isoprenoid antibiotic, ascofuranone, suppresses expression of profibrotic factors including matrix proteins and PAI‐1 induced by TGF‐β in renal fibroblasts. Ascofuranone selectively inhibits phosphorylation of epidermal growth factor receptor (EGFR), and downstream kinases such as Raf‐1, MEK‐1/2, and ERK‐1/2. PAI‐1 transcription also is suppressed by treatment with kinase inhibitors for MEK‐1/2 or EGFR, and with small interfering RNA for EGFR. Ascofuranone inhibits cellular metalloproteinase activity, and an inhibitor of metalloproteinases suppresses EGFR phosphorylation and PAI‐1 transcription. These results suggest that ascofuranone suppresses expression of profibrotic factors through the inhibition of an EGFR‐dependent signal transduction pathway activated by metalloproteinases. J. Cell. Biochem. 107: 335–344, 2009. © 2009 Wiley‐Liss, Inc.
Iodothyronine deiodinases and thyroid hormone receptors regulation during flatfish (Solea senegalensis) metamorphosis
Authors: Isorna Esther , Obregon Maria Jesus , Calvo Rosa Maria , Vázquez Rosa , Pendón Carlos , Falcón Jack , Muñoz‐Cueto José Antonio 
Published: March 2009

AbstractThyroid hormone‐induced metamorphosis seems to represent an ancestral feature of chrordates (urochordates, cephalochordates and vertebrates), but also of nonchordate animals. Although thyroid hormones and thyroid hormone receptor profiles during metamorphosis have been analyzed in different vertebrate taxa, including fish, developmental expression and activity of type 2 (dio2, D2) and type 3 (dio3, D3) iodothyronine deiodinases, two key enzymes in anuran metamorphosis, remain unknown in any fish species. The aim of this work was to investigate the development of thyroid hormone system during the metamorphosis of a flatfish species, the Senegalese sole, focusing on the deiodinases developmental profile. We have cloned sole D2 and D3 and analyzed several parameters of thyroid hormones system in pre‐, early‐, middle‐, and late‐metamorphic larvae. Both deiodinases contain in their catalytic centers an UGA triplet encoding for a selenocystein (Sec) residue as expected. Left eye migration and rotation in body position were associated with a significant increase in both thyroid hormones and thyroid hormone receptors at the middle–late metamorphic stages. Although dio2 expression slightly increased during metamorphosis, D2 activity augmentation was much more significant. Sole dio3 expression declined only slightly, whereas the D3 activity clearly decreased at mid‐late metamorphic period. This developmental profile of deiodinases sustained the rise of thyroid hormones levels observed during sole metamorphosis. No clear cut daily rhythms were observed in the parameters analyzed although it seemed that thyroid hormone system was more active during daytime, in particular at late metamorphic stages. These developmental changes point out the importance not only of thyroid hormones and their receptors but also of dio2 and dio3 in mediating flatfish metamorphosis, as it has been described in amphibians. J. Exp. Zool. (Mol. Dev. Evol.) 312B:231–246, 2009. © 2009 Wiley‐Liss, Inc.