bluuebunny/crossref_metadata_2025_split huggingface dataset:
Flow cytometric sorting of neuronal and glial nuclei from central nervous system tissue
Authors: Okada Seiji , Saiwai Hirokazu , Kumamaru Hiromi , Kubota Kensuke , Harada Akihito , Yamaguchi Masahiro , Iwamoto Yukihide , Ohkawa Yasuyuki 
Published: August 2010

AbstractDue to the complex cellular heterogeneity of the central nervous system (CNS), it is relatively difficult to reliably obtain molecular descriptions with cell‐type specificity. In particular, comparative analysis of epigenetic regulation or molecular profiles is hampered by the lack of adequate methodology for selective purification of defined cell populations from CNS tissue. Here, we developed a direct purification strategy of neural nuclei from CNS tissue based on fluorescence‐activated cell sorting (FACS). We successfully fractionated nuclei from complex tissues such as brain, spinal cord, liver, kidney, and skeletal muscle extruded mechanically or chemically, and fractionated nuclei were structurally maintained and contained nucleoproteins and nuclear DNA/RNA. We collected sufficient numbers of nuclei from neurons and oligodendrocytes using FACS with immunolabeling for nucleoproteins or from genetically labeled transgenic mice. In addition, the use of Fab fragments isolated from papain antibody digests, which effectively enriched the specialized cell populations, significantly enhanced the immunolabeling efficacy. This methodology can be applied to a wide variety of heterogeneous tissues and is crucial for understanding the cell‐specific information about chromatin dynamics, nucleoproteins, protein–DNA/RNA interactions, and transcriptomes retained in the nucleus, such as non‐coding RNAs. J. Cell. Physiol. 226: 552–558, 2011. © 2010 Wiley‐Liss, Inc.
Characterization of GFP‐tagged GnRH‐containing terminalis neurons in transgenic zebrafish
Authors: Wang Xiaokai , Huang Luoxiu , Li Yue , Li Xinle , Li Ping , Ray John , Li Lei 
Published: August 2010

AbstractThe terminalis nerve (TN) has been described in all vertebrate species, in which it plays important roles in animal behavior and physiology. In teleost fish, the TN is located in the olfactory bulb and in its nerve tract. Here, we report a study on the characterization of the TN cell development, axon projection and physiology in zebrafish (Danio rerio). We have generated several lines of transgenic zebrafish [Tg (GnRH‐3::GFP)] that express GFP in the TN cells. The transgenes are expressed under the transcriptional control of the zebrafish GnRH‐3 promoter. During development, the first GFP‐positive TN cell was identified at approximately 34 h post‐fertilization (hpf). By 38 hpf, several clusters of TN cells were identified in the olfactory bulb and olfactory nerve tract. In the olfactory bulb, the TN cells projected axons caudally. In the forebrain, some of the TN axons extended caudally, but most crossed the midline of the brain at the commissural anterior. In the midbrain, some of the TN axons extended dorsally towards the tectum, whereas other axons extended caudally, or extended ventrally to the optic nerve where they entered the neural retina. We also examined the cell membrane property of the TN cells. Using patch‐clamp techniques, we recorded spontaneous and evoked action potentials from isolated TN cells. We examined the expression of glutamate receptors in the TN cells. The data shed light on the mechanisms of TN function in the nervous system and in the regulation of animal physiology. J. Cell. Physiol. 226: 608–615, 2011. © 2010 Wiley‐Liss, Inc.
Involvement of β1‐integrin via PIP complex and FAK/paxillin in dexamethasone‐induced human mesenchymal stem cells migration
Authors: Yun Seung Pil , Ryu Jung Min , Han Ho Jae 
Published: August 2010

AbstractAlthough glucocorticoids strongly affect numerous biological processes including cell growth, development, and homeostasis, their effects on migration of human mesenchymal stem cells (hMSCs) are unclear. Therefore, we investigated the role of dexamethasone (DEX) and its related signaling pathways on migration of hMSCs. We found that DEX, at 10−8 to 10−6 M, significantly increased migration after a 24 h incubation, and DEX (10−6 M) increased migration at >12 h. Moreover, DEX (10−6 M) increased the level of glucocorticoid receptor (GR)‐α mRNA and protein expression, but not GR‐β mRNA. The increases in DEX‐induced migration were inhibited by the GR antagonist mifepristone (10−7 M). In addition, DEX increased integrin‐linked kinase (ILK) and α‐parvin expression but did not change PINCH‐1/2 expression in lysate. DEX also increased formations of complex with ILK and α‐parvin, and ILK and PINCH‐1/2 as shown by immunoprecipitation, which were all inhibited by mifepristone. DEX‐induced migration was blocked by ILK and α‐parvin small interfering(si)RNAs. In addition, DEX increased focal adhesion kinase (FAK) and paxillin expression, which were attenuated by ILK and α‐parvin siRNAs. DEX‐induced cell migration was inhibited by FAK/paxillin siRNAs. DEX also increased β1‐integrin expression, which was blocked by FAK/paxillin siRNAs. In addition, DEX‐induced cell migration was inhibited by β1‐integrin siRNA. Downregulation of ILK, α‐parvin, FAK/paxillin and β1‐integrin expression by siRNAs decreased DEX‐induced filamentous(F)‐actin organization and migration of hMSCs. In conclusion, DEX partially stimulates hMSC migration by the expression of β1‐integrin through formation of a PINCH‐1/2/ILK/α‐parvin complex (PIP complex), and FAK and paxillin expression. J. Cell. Physiol. 226: 683–692, 2011. © 2010 Wiley‐Liss, Inc.
Exercise training reduces insulin resistance and upregulates the mTOR/p70S6k pathway in cardiac muscle of diet‐induced obesity rats
Authors: Medeiros Cleber , Frederico Marisa J. , da Luz Gabrielle , Pauli José R. , Silva Adelino S.R. , Pinho Ricardo A. , Velloso Lício A. , Ropelle Eduardo R. , De Souza Cláudio T. 
Published: August 2010

AbstractObesity and insulin resistance are rapidly expanding public health problems. These disturbances are related to many diseases, including heart pathology. Acting through the Akt/mTOR pathway, insulin has numerous and important physiological functions, such as the induction of growth and survival of many cell types and cardiac hypertrophy. However, obesity and insulin resistance can alter mTOR/p70S6k. Exercise training is known to induce this pathway, but never in the heart of diet‐induced obesity subjects. To evaluate the effect of exercise training on mTOR/p70S6k in the heart of obese Wistar rats, we analyzed the effects of 12 weeks of swimming on obese rats, induced by a high‐fat diet. Exercise training reduced epididymal fat, fasting serum insulin and plasma glucose disappearance. Western blot analyses showed that exercise training increased the ability of insulin to phosphorylate intracellular molecules such as Akt (2.3‐fold) and Foxo1 (1.7‐fold). Moreover, reduced activities and expressions of proteins, induced by the high‐fat diet in rats, such as phospho‐JNK (1.9‐fold), NF‐kB (1.6‐fold) and PTP‐1B (1.5‐fold), were observed. Finally, exercise training increased the activities of the transduction pathways of insulin‐dependent protein synthesis, as shown by increases in Raptor phosphorylation (1.7‐fold), p70S6k phosphorylation (1.9‐fold), and 4E‐BP1 phosphorylation (1.4‐fold) and a reduction in atrogin‐1 expression (2.1‐fold). Results demonstrate a pivotal regulatory role of exercise training on the Akt/mTOR pathway, in turn, promoting protein synthesis and antagonizing protein degradation. J. Cell. Physiol. 226: 666–674, 2011. © 2010 Wiley‐Liss, Inc.
Resveratrol induces apoptosis of human nasopharyngeal carcinoma cells via activation of multiple apoptotic pathways
Authors: Huang Tsung‐Teng , Lin Hung‐Chi , Chen Chang‐Chieh , Lu Chia‐Chen , Wei Chia‐Fong , Wu Ting‐Shu , Liu Fu‐Guo , Lai Hsin‐Chih 
Published: August 2010

AbstractResveratrol, a naturally occurring dietary compound with chemopreventive properties has been reported to trigger a variety of cancer cell types to apoptosis. Whether resveratrol shows any activity on human nasopharyngeal carcinoma (NPC) cells remained to be determined. The aim of this study was to investigate the effect and mechanism of resveratrol on human NPC cells. Treatment of resveratrol resulted in significant decrease in cell viability of NPC cell lines in a dose‐ and time‐dependent manner. A dose‐dependent apoptotic cell death was also measured by flow cytometery analysis. Molecular mechanistic studies of apoptosis unraveled resveratrol treatment resulted in a significant loss of mitochondrial transmembrane potential, release of cytochrome c, enhanced expression of Fas ligand (FasL), and suppression of glucose‐regulated protein 78 kDa (GRP78). These were followed by activation of caspases‐9, ‐8, ‐4, and ‐3, subsequently leading to DNA fragmentation and cell apoptosis. Furthermore, up‐regulation of proapoptotic Bax and down‐regulation of antiapoptotic Bcl‐2 protein were also observed. Taken together, resveratrol induces apoptosis in human NPC cells through regulation of multiple apoptotic pathways, including death receptor, mitochondria, and endoplasmic reticulum (ER) stress. Resveratrol can be developed as an effective compound for human NPC treatment. J. Cell. Physiol. 226: 720–728, 2011. © 2010 Wiley‐Liss, Inc.
Dexamethasone modulates osteogenesis and adipogenesis with regulation of osterix expression in rat calvaria‐derived cells
Authors: Mikami Yoshikazu , Lee Mio , Irie Seiko , Honda Masaki J. 
Published: August 2010

AbstractOsteoblasts and adipocytes originate from common mesenchymal progenitor cells and although a number of compounds can induce osteoblastic and adipogenic differentiation from progenitor cells, the underlying mechanisms have not been elucidated. The present study examined the synergistic effects of dexamethasone (Dex) and bone morphogenetic protein (BMP)‐2 on the differentiation of clonal mesenchymal progenitor cells isolated from rat calvaria into osteoblasts and adipocytes, as well as the effects of the timing of treatment. Cells were cultured for various periods of time in the presence of Dex and/or BMP‐2. When cells were treated with Dex + BMP‐2 during the early phase of differentiation, they differentiated into adipocytes. However, when cells were treated with Dex + BMP‐2 during the late phase of differentiation, a synergistic effect on in vitro matrix mineralization was observed. To examine differences between the early and late phases of differentiation, ALP activity was measured in the presence of BMP‐2. ALP activity increased markedly on Day 9, corresponding to the onset of the synergistic effect of Dex. Dex treatment inhibited osterix (OSX) expression in cells committed to adipogenic differentiation, but not in cells committed to osteogenic differentiation following BMP‐2 treatment. The isoform2 OSX promoter region was found to be involved in the effects of Dex on cells during the early phase of differentiation. Furthermore, cells stably expressing OSX (isoform2) formed mineralized nodules even though they had been treated with Dex + BMP‐2 during the early phase of differentiation. It appears that Dex modulates osteogenesis and adipogenesis in mesenchymal stem cells by regulating OSX expression. J. Cell. Physiol. 226: 739–748, 2011. © 2010 Wiley‐Liss, Inc.
Regulation of UDP‐glucose dehydrogenase is sufficient to modulate hyaluronan production and release, control sulfated GAG synthesis, and promote chondrogenesis
Authors: Clarkin Claire E. , Allen Steve , Kuiper Nikki J. , Wheeler Benjamin T. , Wheeler‐Jones Caroline P. , Pitsillides Andrew A. 
Published: August 2010

AbstractGlycosaminoglycans (GAGs) are critical for extracellular matrix (ECM) integrity in cartilage but mechanisms regulating their synthesis are not defined. UDP‐glucose dehydrogenase (UGDH) catalyses UDP‐glucose oxidation to UDP‐glucuronic acid, an essential monosaccharide in many GAGs. Our previous studies in articular surface (AS) cells from embryonic joints have established pivotal roles for mitogen‐activated protein kinases (MAPK) in synthesis of the unsulfated GAG, hyaluronan (HA). We investigated the functional significance of UGDH in GAG production and chondrogenesis, and determined roles for MEK–ERK and p38MAPK pathways in regulating UGDH expression and function. Inhibitors of MEK and p38MAPK reduced UGDH protein in AS cells. Treatment with TGF‐β (archetypal growth factor) increased UGDH expression, sulfated (s)‐GAG/HA release and pericellular matrix formation in a p38MAPK‐dependent manner. Retroviral overexpression of UGDH augmented HA/sGAG release and pericellular matrix elaboration, which were blocked by inhibiting MEK but not p38MAPK. UGDH overexpression increased cartilage nodule size in bone marrow culture, promoted chondrogenesis in limb bud micromass culture and selectively suppressed medium HA levels and modified GAG sulfation, as assessed by FACE analysis. Our data provide evidence that: (i) TGF‐β regulates UGDH expression via p38MAPK to modulate sGAG/HA secretion, (ii) MEK–ERK, but not p38MAPK facilitates UGDH‐induced HA and sGAG release, and (iii) increased UGDH expression promotes chondrogenesis directly and differential modifies GAG levels and sulfation. These results indicate a more diverse role for UGDH in the support of selective GAG production than previously described. Factors regulating UGDH may provide novel candidates for restoring ECM integrity in degenerative cartilage diseases, such as osteoarthritis.Arthritis Research Campaign. J. Cell. Physiol. 226: 749–761, 2011. © 2010 Wiley‐Liss, Inc.
Induction of small G protein RhoB by non‐genotoxic stress inhibits apoptosis and activates NF‐κB
Authors: Li Yi‐Dong , Liu Ya‐Ping , Cao Dong‐Mei , Yan Ya‐Min , Hou You‐Na , Zhao Jie‐Ying , Yang Rui , Xia Zhao‐Fan , Lu Jian 
Published: August 2010

AbstractIt has been reported by us and other groups that the expression of small GTP binding protein RhoB can be induced by genotoxic stressors and glucocorticoid (GC), a stress hormone that plays a key role in stress response. Until now stress‐induced genes that confer cytoprotection under stressed conditions are largely unknown. In this study, we investigated the effects and mechanism of non‐genotoxic stressors, including scalding in vivo and heat stress in vitro on the expression of RhoB. We found for the first time that both scalding, which could induce typical neuroendocrine responses of acute stress and cellular heat stress significantly increased the expression of RhoB at mRNA and protein levels. Moreover, in vitro experiments in human lung epithelial cells (A549) showed that induction of RhoB by heat stress was in a glucocorticoid receptor (GR)‐independent manner and through multiple pathways including stabilization of RhoB mRNA and activation of p38 MAPK. Further experiments demonstrated that up‐regulation of RhoB significantly inhibited heat stress‐induced apoptosis and elevated transcriptional activity of NF‐κB, but did not affect the expression of Hsp70 in A549 cells. In conclusion, we showed for the first time that RhoB was up‐regulated by scalding in vivo and heat stress in vitro and played an important cytoprotective role during heat stress‐induced apoptotic cell death. J. Cell. Physiol. 226: 729–738, 2011. © 2010 Wiley‐Liss, Inc.
PLGA based drug delivery systems (DDS) for the sustained release of insulin: insight into the protein/polyester interactions and the insulin release behavior
Authors: Lassalle Verónica , Ferreira María Luján 
Published: August 2010

AbstractBACKGROUND: Drug delivery systems (DDS) were designed using insulin as model drug and poly (lactic–co‐glycolic) copolymers (PLGA) as polymeric matrix. The carriers were synthesized by direct self‐assembly of the insulin and the polyester under mild conditions.RESULTS: The kind and level of association between the protein and the polymer were studied using computational methods (combined MM2/PM3) and spectroscopic tools (Fourier transform infrared (FTIR), energy dispersive X‐ray (EDX) and X‐ray fluorescence spectroscopy (XFS)). The effect of the number average molecular weight (Mn) of the copolymer on the association efficiency (AE) drug–polymer as well as on the release profile has been explored. Mathematical models were used to predict the insulin release kinetic and mechanism.CONCLUSIONS: Satisfactory protein/PLGA association efficiencies (between 77 and 99%) were registered depending on the Mn of the PLGA. Hydrophobic and hydrophilic interactions were detected between the protein and the polymeric network by computational analysis. In vitro release studies demonstrated that copolyesters of about 8600 and 1500 Da were suitable for the gradual release of insulin while PLGA oligomers of average molecular weight between 700 and 800 Da were unsuitable as DDS. The insulin release kinetics fits well with the Korsmeyer model, following the anomalous transport mechanism. Copyright © 2010 Society of Chemical Industry
Solvent‐free synthesis of glycerol carbonate and glycidol from 3‐chloro‐1,2‐propanediol and potassium (hydrogen) carbonate
Authors: Gómez‐Jiménez‐Aberasturi Olga , Ochoa‐Gómez José R. , Pesquera‐Rodríguez Amaia , Ramírez‐López Camilo , Alonso‐Vicario Ainhoa , Torrecilla‐Soria Jesús 
Published: August 2010

AbstractBACKGROUND: An indirect solvent‐free synthetic approach for obtaining glycerol carbonate and glycidol from glycerol and CO2 through their more reactive and easily synthesizable derivatives 3‐chloro‐1,2‐propanediol (HAL) and potassium (hydrogen) carbonate has been studied.RESULTS: The reaction is fast with source of carbonation and temperature having a strong influence on the results. A yield of 80% glycerol carbonate together with a simultaneous substantial production of glycidol (0.56 mol mol−1 glycerol carbonate) are obtained using K2CO3 as the carbonation source at 80 °C, a reaction time of 30 min and a 3:1 HAL/K2CO3 molar ratio. A lower yield of glycerol carbonate (60%) is obtained from KHCO3 after 50 min with the other experimental conditions remaining unchanged. In this case, glycidol formation is zero or insignificant. Glycerol is also obtained in high yields, although in much lower amounts from KHCO3 (∼0.59 mol mol−1 glycerol carbonate independent of operating conditions) than from K2CO3 (0.84–1.1 mol mol−1 glycerol carbonate, depending on experimental conditions).CONCLUSIONS: The proposed synthetic strategy overcomes the currently difficult direct reaction between glycerol and CO2, leading to the simultaneous synthesis of two valuable chemicals: glycerol carbonate and glycidol. However, glycerol is also obtained in substantial amounts thus decreasing the overall yield of the process. Thus, methods for preventing its formation must be developed for industrial feasibility. Copyright © 2010 Society of Chemical Industry
Differences in Brachypelma albopilosa (theraphosidae) hemolymph proteome between subadult and adult females
Authors: Trabalon Marie , Carapito Christine , Voinot Florian , Martrette Jean‐Marc , Van Dorsselaer Alain , Gilbert Caroline , Bertile Fabrice 
Published: August 2010

AbstractThe changes in the hemolymph proteome of mygalomorph Brachypelma albopilosa females were examined for the first time in relation to their developmental stage (subadult and adult period). Seven distinct subunits of hemocyanin (a, b, c, d, e, f, and g chains), as well as actin were clearly identified and their sequence partly characterized using a combination of one‐ and two‐dimensional gel electrophoresis and mass spectrometry. The different structures determined along with possible post‐translational modifications may reflect a role of hemocyanin in molting, immunity, and reproduction. In addition, despite no precise identification, additional peptide sequences from eight protein bands (four bands >200 kDa and four bands in the 95–200 kDa mass range) were determined. As reported in other spider species, the putative corresponding structures are the coagulogen protein and/or lipoproteins (HDL‐1, HDL‐2, VHDL) for which quantitative differences between adult and subadult individuals could be related to the molting process and/or cuticle lipid and protein composition according to the developmental stage. J. Exp. Zool. 313A:651–659, 2010. © 2010 Wiley‐Liss, Inc.
Effectiveness of an inpatient smoking cessation program
Authors: Gadomski Anne M. , Gavett Jessie , Krupa Nicole , Tallman Nancy , Jenkins Paul 
Published: August 2010

AbstractBACKGROUND:Inpatient smoking cessation may increase the success of quitting smoking post‐hospital discharge.METHODS:Using a quasiexperimental study design, use of cessation methods, mortality, self‐reported abstinence, and quit status 6 months post–hospital discharge were measured to assess the effectiveness of an inpatient smoking cessation program. Subjects were interviewed by telephone 6 months post‐hospital discharge. Outcomes for patients who were seen by the inpatient smoking cessation counselor were compared to consecutive patients who were not seen by the counselor. Electronic medical records (EMRs) and administrative data were used to construct baseline measures, comorbidity covariates, pharmaceutical use rates during hospitalization, readmission, and mortality outcomes. Multivariate methods included logistic regression and survival analysis.RESULTS:At baseline, the study groups varied by mean age, length of stay (LOS), comorbidity index, cardiovascular diagnosis, and acuity. At 6 months post–hospital discharge, the intent to treat estimate for point prevalence abstinence was 16% in the intervention group compared to 10% in the comparison group (P = 0.02) while self‐reported quit status in the intervention group was 44% vs. 30% in the comparison group (P = 0.00). The intervention group used more nicotine replacement therapy (NRT) than the comparison group both in‐hospital and following discharge. Crude post–hospital discharge mortality was significantly less in the intervention group (0.02) than in the comparison group (0.04). A multivariate survival model, controlling for baseline imbalances, showed a significantly reduced mortality in the intervention group (hazard ratio [HR] = 0.37; P = 0.04).CONCLUSIONS:Inpatient smoking cessation programs effectively improve quit outcomes, NRT use, and mortality post–hospital discharge. Journal of Hospital Medicine 2011. © 2010 Society of Hospital Medicine.
Cognitive error as the most frequent contributory factor in cases of medical injury: A study on verdict's judgment among closed claims in Japan
Authors: Tokuda Yasuharu , Kishida Naoki , Konishi Ryota , Koizumi Shunzo 
Published: August 2010

AbstractBACKGROUND:Cognitive errors in the course of clinical decision‐making are prevalent in many cases of medical injury. We used information on verdict's judgment from closed claims files to determine the important cognitive factors associated with cases of medical injury.METHODS:Data were collected from claims closed between 2001 to 2005 at district courts in Tokyo and Osaka, Japan. In each case, we recorded all the contributory cognitive, systemic, and patient‐related factors judged in the verdicts to be causally related to the medical injury. We also analyzed the association between cognitive factors and cases involving paid compensation using a multivariable logistic regression model.RESULTS:Among 274 cases (mean age 49 years old; 45% women), there were 122 (45%) deaths and 67 (24%) major injuries (incomplete recovery within a year). In 103 cases (38%), the verdicts ordered hospitals to pay compensation (median; 8,000,000 Japanese Yen). An error in judgment (199/274, 73%) and failure of vigilance (177/274, 65%) were the most prevalent causative cognitive factors, and error in judgment was also significantly associated with paid compensation (odds ratio, 1.9; 95% confidence interval [CI], 1.0‐3.4). Systemic causative factors including poor teamwork (11/274, 4%) and technology failure (5/274, 2%) were less common.CONCLUSIONS:The closed claims analysis based on verdict's judgment showed that cognitive errors were common in cases of medical injury, with an error in judgment being most prevalent and closely associated with compensation payment. Reduction of this type of error is required to produce safer healthcare. Journal of Hospital Medicine 2010. © 2010 Society of Hospital Medicine.