bluuebunny/crossref_metadata_2025 huggingface dataset:
P–T evolution of the Precambrian Metamorphic Complex, NW Iran: a study of metapelitic rocks
Authors: Saki A. , Moazzen M. , Oberhänsli R.
Published: May 2010

The Mahneshan Metamorphic Complex (MMC) is one of the Precambrian terrains exposed in the northwest of Iran. The MMC underwent two main phases of deformation (D1and D2) and at least two metamorphic events (M1and M2). Critical metamorphic mineral assemblages in the metapelitic rocks testify to regional metamorphism under amphibolite‐facies conditions. The dominant metamorphic mineral assemblage in metapelitic rocks (M1) is muscovite, biotite I, Garnet I, staurolite, Andalusite I and sillimanite. Peak metamorphism took place at 600–620°C and ∼7 kbar, corresponding to a depth ofca. 24 km. This was followed by decompression during exhumation of the crustal rocks up to the surface. The decrease of temperature and pressure during exhumation produced retrograde metamorphic assemblages (M2). Secondary phases such as garnet II biotite II, Andalusite II constrain the temperature and pressure of M2retrograde metamorphism to 520–560°C and 2.5–3.5 kbar, respectively. The geothermal gradient obtained for the peak of metamorphism is 33°C km−1, which indicates that peak metamorphism was of Barrovian type and occurred under medium‐pressure conditions. The MMC followed a ‘clockwise’ P–T path during metamorphism, consistent with thermal relaxation following tectonic thickening. The bulk chemistry of the MMC metapelites shows that their protoliths were deposited at an active continental margin. Together with the presence of palaeo‐suture zones and ophiolitic rocks around the high‐grade metamorphic rocks of the MMC, these features suggest that the Iranian Precambrian basement formed by an island‐arc type cratonization. Copyright © 2010 John Wiley & Sons, Ltd.
CHFR hypermethylation and EGFR mutation are mutually exclusive and exhibit contrastive clinical backgrounds and outcomes in non‐small cell lung cancer
Authors: Koga Takaomi , Takeshita Masafumi , Yano Tokujiro , Maehara Yoshihiko , Sueishi Katsuo
Published: May 2010

Aberrant promoter methylation of the checkpoint gene with forkhead‐associated domain and ring finger (CHFR) gene is frequently detected in human cancer. We previously demonstrated that diminished CHFR expression was significantly correlated with both poor prognosis and heavy smoking in nonsmall cell lung cancer (NSCLC). Conversely, epidermal growth receptor (EGFR) mutation is detected in NSCLC among those who have never smoked or smoked lightly. To address the frequency of CHFR hypermethylation as well as differences in the distributions and clinicopathologic backgrounds against EGFR mutation in NSCLC, we investigated a large group of 208 NSCLC patients, including 165 with adenocarcinoma (ADC), 40 with squamous cell carcinoma and three others. We found that CHFR hypermethylation and EGFR mutation are mutually exclusive and have contrastive clinicopathologic backgrounds in NSCLC. Methylation‐specific polymerase chain reaction (MSP) and direct DNA sequencing were performed to detect CHFR hypermethylation and EGFR mutation, respectively. CHFR hypermethylation was found in 29 cases (14%) (16 ADC (8%), 12 SCC (6%) and one adenosquamous carcinoma), while EGFR mutation was detected in 48 (23%) cases, all of which were ADC. CHFR hypermethylation and EGFR mutation were mutually exclusive (p = 0.004). NSCLC with altered CHFR was significantly correlated with smoking history, poor differentiation, lymphatic invasion, and poor prognosis; this contrasted sharply with EGFR mutation, which had statistically better clinical outcomes. Our results demonstrate that CHFR loss might be critical for the tumorigenesis of NSCLC in patients with a history of smoking and induces tumors of a more malignant phenotype than the EGFR mutation. Thus, CHFR alteration should be considered a therapeutic target against NSCLC in patients with poor prognoses.
Molecular signatures classify astrocytic gliomas by IDH1 mutation status
Authors: Toedt Grischa , Barbus Sebastian , Wolter Marietta , Felsberg Jörg , Tews Björn , Blond Frederic , Sabel Michael C. , Hofmann Stefanie , Becker Natalia , Hartmann Christian , Ohgaki Hiroko , von Deimling Andreas , Wiestler Otmar D. , Hahn Meinhard , Lichter Peter , Reifenberger Guido , Radlwimmer Bernhard
Published: May 2010

To identify novel glioma‐associated pathomechanisms and molecular markers, we performed an array‐based comparative genomic hybridization analysis of 131 diffuse astrocytic gliomas, including 87 primary glioblastomas (pGBIV), 13 secondary glioblastomas (sGBIV), 19 anaplastic astrocytomas (AAIII) and 12 diffuse astrocytomas (AII). All tumors were additionally screened for IDH1 and IDH2 mutations. Expression profiling was performed for 74 tumors (42 pGBIV, 11 sGBIV, 13 AAIII, 8 AII). Unsupervised and supervised bioinformatic analyses revealed distinct genomic and expression profiles separating pGBIV from the other entities. Classifier expression signatures were strongly associated with the IDH1 gene mutation status. Within pGBIV, the rare subtype of IDH1 mutant tumors shared expression profiles with IDH1 mutant sGBIV and was associated with longer overall survival compared with IDH1 wild‐type tumors. In patients with IDH1 wild‐type pGBIV, PDGFRA gain or amplification as well as 19q gain were associated with patient outcome. Array‐CGH analysis additionally revealed homozygous deletions of the FGFR2 gene at 10q26.13 in 2 pGBIV, with reduced FGFR2 mRNA levels being frequent in pGBIV and linked to poor outcome. In conclusion, we report that diffuse astrocytic gliomas can be separated into 2 major molecular groups with distinct genomic and mRNA profiles as well as IDH1 gene mutation status. In addition, our results suggest FGFR2 as a novel glioma‐associated candidate tumor suppressor gene on the long arm of chromosome 10.
Long‐term follow‐up of patients with resected pancreatic cancer following vaccination against mutant K‐ras
Authors: Wedén Synne , Klemp Marianne , Gladhaug Ivar P. , Møller Mona , Eriksen Jon Amund , Gaudernack Gustav , Buanes Trond
Published: May 2010

K‐ras mutations are frequently found in adenocarcinomas of the pancreas and can elicit mutation‐specific immune responses. Targeting the immune system against mutant Ras may thus influence the clinical course of the disease. Twenty‐three patients who were vaccinated after surgical resection for pancreatic adenocarcinoma (22 pancreaticoduodenectomies, one distal resection), in two previous Phase I/II clinical trials, were followed for more than 10 years with respect to long‐term immunological T‐cell reactivity and survival. The vaccine was composed of long synthetic mutant ras peptides designed mainly to elicit T‐helper responses. Seventeen of 20 evaluable patients (85%) responded immunologically to the vaccine. Median survival for all patients was 27.5 months and 28 months for immune responders. The 5‐year survival was 22% and 29%, respectively. Strikingly, 10‐year survival was 20% (four patients out of 20 evaluable) versus zero (0/87) in a cohort of nonvaccinated patient treated in the same period. Three patients mounted a memory response up to 9 years after vaccination. The present observation of long‐term immune response together with 10‐year survival following surgical resection indicates that K‐ras vaccination may consolidate the effect of surgery and represent an adjuvant treatment option for the future.
A tritherapy combination of a fusion protein vaccine with immune‐modulating doses of sequential chemotherapies in an optimized regimen completely eradicates large tumors in mice
Authors: Song Xinxin , Guo Wenzhong , Cui Jianfeng , Qian Xinlai , Yi Linan , Chang Mengjiao , Cai Qiliang , Zhao Qingzheng
Published: May 2010

Tumor‐induced immunosuppression plays a critical role in both impeding tumor‐specific immune responses and limiting the effects of cancer immunotherapy. Analyses of regulatory cells recruited during the growth of the E7‐expressing tumor, TC‐1, revealed a high percentage of regulatory T cells (Tregs) as well as myeloid‐derived suppressor cells (MDSCs) in spleens and tumors. In this study, we proposed that treatment with immune‐modulating doses of cyclophosphamide (CTX) and all‐trans retinoic acid (ATRA) would result in a beneficial tumor microenvironment with the suppression of Tregs and MDSCs and, thus, enhance the effect of a human papillomavirus protein vaccine. Our results showed that CTX preconditioning and persistent ATRA treatment along with the vaccine achieved long‐term survival and induced long‐term memory responses. However, the effect of the antitumor response sharply declined when the tritherapy was initiated after the optimal therapeutic time. The more intensive regimen could rescue the effect of the tritherapy accompanied by the decreased percentage of Tregs and MDSCs in spleens and tumors. Besides, a favorable host environment was created by the reduced secretion of interleukin‐10 and 6 and vascular endothelial growth factor (VEGF) in the tumor niche and decreased the expression of phosphorylation‐signal transducer and activator of transcription 3 of TC‐1 tumors. Our data shed light on the immune‐modulating doses of sequential chemoimmunotherapeutic strategy targeting not only the tumor but also its microenvironment, which suggests a potential clinical benefit for the immunotherapy of HPV‐associated malignancies.
miR‐137 targets Cdc42 expression, induces cell cycle G1 arrest and inhibits invasion in colorectal cancer cells
Authors: Liu Ming , Lang Nan , Qiu Meng , Xu Feng , Li Qiu , Tang Qiulin , Chen Ji , Chen Xi , Zhang Siyuan , Liu Zhen , Zhou Jitao , Zhu Yajie , Deng Yu , Zheng Yi , Bi Feng
Published: May 2010

miRNAs have emerged as post‐transcriptional regulators that are critically involved in the pathogenesis of a number of human cancers. Cdc42, one of the best characterized members of the Rho GTPase family, is found to be up‐regulated in several types of human tumors and has been implicated in cancer initiation and progression. In the present study, we have identified miR‐137 as a potential regulator of Cdc42 expression. A bioinformatics search revealed a putative target‐site for miR‐137 within the Cdc42 3′ UTR at nt 792–798, which is highly conserved across different species. Expression of miR‐137 in colorectal cancer cell lines was found inversely correlated with Cdc42 expression. miR‐137 could significantly suppress Cdc42 3′ UTR luciferase‐reporter activity, and this effect was not detectable when the putative 3′ UTR target‐site was mutated. Consistent with the results of the reporter assay, ectopic expression of miR‐137 reduced both mRNA and protein expression levels of Cdc42 and mimicked the effect of Cdc42 knockdown in inhibiting proliferation, inducing G1 cell cycle arrest, and blocking invasion of the colorectal cancer cells, whereas anti‐miR‐137 expression led to the opposite effect. Furthermore, expression of miR‐137 suppressed the immediate downstream effector of Cdc42, PAK signaling. Our results suggest that miR‐137 may have a tumor suppressor function by directly targeting Cdc42 to inhibit the proliferation and invasion activities of colorectal cancer cells. They raise an interesting possibility that Cdc42 activity and function can be controlled by miRNAs in addition to the classic regulators such as guanine nucleotide exchange factors and GTPase‐activating proteins.
Fabrication and characterization of chitosan‐gelatin blend nanofibers for skin tissue engineering
Authors: Dhandayuthapani Brahatheeswaran , Krishnan Uma Maheswari , Sethuraman Swaminathan
Published: May 2010

Tissue engineering scaffolds produced by electrospinning feature a structural similarity to the natural extracellular matrix. Polymer blending is one of the effective methods to provide new and desirable biocomposites for tissue engineering applications. In this study chitosan was blended with gelatin and the effect of processing parameters of electrospinning and the solution properties of the polymer on the morphology of the fibers obtained were investigated. The morphology of the electrospun chitosan, gelatin and the chitosan‐gelatin blend were characterized using a scanning electron microscope (SEM). The miscibility of the blend was determined using a SEM, and differential scanning calorimetry (DSC) Fourier transform Infrared spectrometer (FTIR). Further the tensile properties of the blend nanofibers were studied and compared with chitosan and gelatin fibers. In this study we have been able to electrospin defect‐free chitosan, gelatin and chitosan‐gelatin blend nanofibers with smooth morphology and diameter ranging from 120 to 200 nm, 100 to 150 nm, and 120–220 nm, respectively by optimizing the process and solution parameters. Chitosan and gelatin formed completely miscible blends as evidenced from DSC and FTIR measurements. The tensile strength of the chitosan‐gelatin blend nanofibers (37.91 ± 4.42 MPa) was significantly higher than the gelatin nanofibers (7.23 ± 1.15 MPa) (p < 0.05) and comparable with that of normal human skin. Thus the novel chitosan‐gelatin blend nanofiber system has potential application in skin regeneration. © 2010 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2010.
CRTAM: A molecule involved in epithelial cell adhesion
Authors: Garay Erika , Patiño‐López Genaro , Islas Socorro , Alarcón Lourdes , Canche‐Pool Elsy , Valle‐Rios Ricardo , Medina‐Contreras Oscar , Granados Giovana , Chávez‐Munguía Bibiana , Juaristi Eusebio , Ortiz‐Navarrete Vianney , González‐Mariscal Lorenza
Published: May 2010

Class I‐restricted T cell associated molecule (CRTAM) is a member of the immunoglobulin superfamily that complies with the structural characteristics of the JAM family of proteins and is phylogenetically more closely related to nectin‐like proteins. Here we demonstrate for the first time, that CRTAM is expressed in epithelial cells along the lateral membrane and is important for early cell–cell contacts and cell–substrate interactions. CRTAM is sensitive to intermediate filament disruption and treatment of monolayers with soluble CRTAM enhances cell–cell dissociation and lowers transepithelial electrical resistance. Incubation of newly plated cells with anti‐CRTAM antibody decreases the formation of cell aggregates and promotes cell detachment. Co‐cultures of epithelial cells and fibroblasts that lack CRTAM expression and in vitro binding assays, demonstrate the participation of CRTAM in homotypic and heterotypic trans‐interactions. Hence we conclude that CRTAM is a molecule involved in epithelial cell adhesion. J. Cell. Biochem. 111: 111–122, 2010. © 2010 Wiley‐Liss, Inc.
Endothelial cells regulate cardiomyocyte development from embryonic stem cells
Authors: Chen Kang , Bai Hao , Arzigian Melanie , Gao Yong‐Xing , Bao Jing , Wu Wen‐Shu , Shen Wei‐Feng , Wu Liqun , Wang Zack Z.
Published: May 2010

The molecules and environment that direct pluripotent stem cell differentiation into cardiomyocytes are largely unknown. Here, we determined a critical role of receptor tyrosine kinase, EphB4, in regulating cardiomyocyte generation from embryonic stem (ES) cells through endothelial cells. The number of spontaneous contracting cardiomyocytes, and the expression of cardiac‐specific genes, including α‐MHC and MLC‐2V, was significantly decreased in EphB4‐null ES cells. EphB4 was expressed in endothelial cells underneath contracting cardiomyocytes, but not in cardiomyocytes. Angiogenic inhibitors, including endostatin and angiostatin, inhibited endothelial cell differentiation and diminished cardiomyogenesis in ES cells. Generation of functional cardiomyocytes and the expression of cardiac‐specific genes were significantly enhanced by co‐culture of ES cells with human endothelial cells. Furthermore, the defects of cardiomyocyte differentiation in EphB4‐deficient ES cells were rescued by human endothelial cells. For the first time, our study demonstrated that endothelial cells play an essential role in facilitating cardiomyocyte differentiation from pluripotent stem cells. EphB4 signaling is a critical component of the endothelial niche to regulate regeneration of cardiomyocytes. J. Cell. Biochem. 111: 29–39, 2010. © 2010 Wiley‐Liss, Inc.
Inhibition of LPS‐induced C/EBPδ by trichostatin a has a positive effect on LPS‐induced cyclooxygenase 2 expression in RAW264.7 cells
Authors: Liu Yi‐Wen , Wang Shao‐An , Hsu Tsung‐Yi , Chen Tsu‐An , Chang Wen‐Chang , Hung Jan‐Jong
Published: May 2010

Cyclooxygenase 2 (COX‐2) is an important inflammatory factor. Previous studies have indicated that COX‐2 is induced with lipopolysaccharide (LPS) treatment. Here, we found that an inhibitor of histone deacetylase (HDAC), trichostatin A (TSA), cannot repress LPS‐induced COX‐2 but it increased the COX‐2 level in RAW264.7 cells. We found no significant difference in NF‐κB activation and ERK1/2 phosphorylation, but LPS‐induced C/EBPδ expression was completely abolished after TSA treatment of LPS‐treated cells. Interesting, reporter assay of C/EBPδ promoter revealed that Sp1‐binding site is important. Although there was no alteration in c‐Jun levels, but the phosphorylation of c‐Jun at its C‐terminus was increased dramatically. A DNA‐associated protein assay (DAPA) and chromatin immunoprecipitation assay (ChIP) indicated that c‐Jun was recruited via Sp1 to the promoter of C/EBPδ after LPS treatment; this recruitment of c‐Jun was repressed by TSA. C/EBPδ inhibition by TSA resulted in increased binding of C/EBPα and C/EBPβ to the COX‐2 promoter. Therefore, TSA has a positive effect on LPS‐induced COX‐2 since it decreases the C/EBPδ level by reducing c‐Jun recruitment by Sp1 to the C/EBPδ promoter, resulting in increased the recruitment of C/EBPα and C/EBPβ to the COX‐2 promoter. J. Cell. Biochem. 110: 1430–1438, 2010. © 2010 Wiley‐Liss, Inc.
Role of annexin A6 isoforms in catecholamine secretion by PC12 cells: Distinct influence on calcium response
Authors: Podszywalow‐Bartnicka Paulina , Kosiorek Michalina , Piwocka Katarzyna , Sikora Ewa , Zablocki Krzysztof , Pikula Slawomir
Published: May 2010

Noradrenaline and adrenaline are secreted by adrenal medulla chromaffin cells via exocytosis. Exocytosis of catecholamines occurs after cell stimulation with various endogenous activators such as nicotine or after depolarization of the plasma membrane and is regulated by calcium ions. Cytosolic [Ca2+] increases in response to cell excitation and triggers a signal‐initiated secretion. Annexins are known to participate in the regulation of membrane dynamics and are also considered to be involved in vesicular trafficking. Some experimental evidence suggests that annexins may participate in Ca2+‐regulated catecholamine secretion. In this report the effect of annexin A6 (AnxA6) isoforms 1 and 2 on catecholamine secretion has been described. Overexpression of AnxA6 isoforms and AnxA6 knock‐down in PC12 cells were accompanied by almost complete inhibition or a 20% enhancement of dopamine secretion, respectively. AnxA6‐1 and AnxA6‐2 overexpression reduced Δ[Ca2+]c upon depolarization by 32% and 58%, respectively, while AnxA6 knock‐down increased Δ[Ca2+]c by 44%. The mechanism of AnxA6 action on Ca2+ signalling is not well understood. Experimental evidence suggests that two AnxA6 isoforms interact with different targets engaged in regulation of calcium homeostasis in PC12 cells. J. Cell. Biochem. 111: 168–178, 2010. © 2010 Wiley‐Liss, Inc.
EGR1, EGR2, and EGR3 activate the expression of their coregulator NAB2 establishing a negative feedback loop in cells of neuroectodermal and epithelial origin
Authors: Kumbrink Joerg , Kirsch Kathrin H. , Johnson Judith P.
Published: May 2010

The inducible zinc finger transcription factors EGR1, EGR2, and EGR3 regulate the expression of numerous genes involved in differentiation, growth, and response to extracellular signals. Their activity is modulated in part through NAB2 which is induced by the same stimuli. In melanoma and carcinoma cells EGR1 activates NAB2 expression. In T lymphocytes EGR2 and EGR3 have been shown to inhibit NAB2 expression. Therefore, we investigated the influence of EGR2 and EGR3 on NAB2 expression in melanoma and carcinoma cells. Here, we show that like EGR1, EGR2 and EGR3 induced NAB2 expression in these cells. EGR1 and EGR3 act in concert on the NAB2 promoter and are more potent activators of NAB2 transcription than EGR2. EGR1‐, EGR2‐, and EGR3‐induced NAB2 promoter activity is mediated through similar cis‐regulatory elements and the activation by each EGR is repressed by NAB2. Kinetic studies suggest that induction of EGR1 leads to low NAB2 expression, while EGR2 and EGR3 are necessary for maximal and sustained expression. As already shown for EGR1, reduction of EGR2 or EGR3 expression by siRNAs reduced endogenous NAB2 levels. Depletion of EGR3 also resulted in a reduction of EGR2 levels confirming EGR2 as a target gene of EGR3. Our results suggest that in many cells of neuroectodermal and epithelial origin EGR1, EGR2, and EGR3 activate NAB2 transcription which is in turn repressed by NAB2, thus establishing a negative feedback loop. This points to a complex relationship between the EGR factors and NAB2 expression likely depending on the cellular context. J. Cell. Biochem. 111: 207–217, 2010. © 2010 Wiley‐Liss, Inc.
RNA‐mediated gene silencing of FUT1 and FUT2 influences expression and activities of bovine and human fucosylated nucleolin and inhibits cell adhesion and proliferation
Authors: Palumberi Domenico , Aldi Silvia , Ermini Leonardo , Ziche Marina , Finetti Federica , Donnini Sandra , Rosati Floriana
Published: May 2010

In a previous article, we demonstrated the existence of fucosyl‐containingO‐glycans forms of nucleolin in bovine post‐capillary venular endothelial cells (CVEC) and malignant cultured human A431 cells. The tool for this discovery was an antibody found to interact strongly and exclusively with nucleolin in total protein extracts. The antibody was originally raised against a mollusc glycoprotein and was demonstrated to be directed against itsO‐glycans, recently found to belong prevalently to the blood group H‐antigen type with fucose linked in alpha1, 2 to galactose. Here, we show that si‐RNA induced down‐regulation of the expression of FUT1 and FUT2, the fucosyltransferases required for the biosynthesis of the terminal glycan motif Fucα‐2‐Galβ‐R, reduced expression of the fucosylated nucleolin glycoforms and their exposure at the cell surface in CVEC. Treatment of the cells with FUT1/2 siRNA also reduced their ability to bind and internalize endostatin and their adhesion efficiency and inhibited cell growth. Expression of FUT1, FUT2, and FUT6 was also analyzed in serum‐stimulated versus serum‐starved cells and in cells treated with FUT1 and FUT2 siRNA. A reduced expression of fucosylated nucleolin and inhibition of cell growth by suppressing FUT1/2 expression was also tested and shown to be exhibited in human A431 cells. J. Cell. Biochem. 111: 229–238, 2010. © 2010 Wiley‐Liss, Inc.
Gab1 mediates hepatocyte growth factor‐stimulated mitogenicity and morphogenesis in multipotent myeloid cells
Authors: Felici Angelina , Giubellino Alessio , Bottaro Donald P.
Published: May 2010

Hepatocyte growth factor (HGF)‐stimulated mitogenesis, motogenesis and morphogenesis in various cell types begins with activation of the Met receptor tyrosine kinase and the recruitment of intracellular adaptors and kinase substrates. The adapter protein Gab1 is a critical effector and substrate of activated Met, mediating morphogenesis, among other activities, in epithelial cells. To define its role downstream of Met in hematopoietic cells, Gab1 was expressed in the HGF‐responsive, Gab1‐negative murine myeloid cell line 32D. Interestingly, the adhesion and motility of Gab1‐expressing cells were significantly greater than parental cells, independent of growth factor treatment. Downstream of activated Met, Gab1 expression was specifically associated with rapid Shp‐2 recruitment and activation, increased mitogenic potency, suppression of GATA‐1 expression and concomitant upregulation of GATA‐2 transcription. In addition to enhanced proliferation, continuous culture of Gab1‐expressing 32D cells in HGF resulted in cell attachment, filopodia extension and phenotypic changes suggestive of monocytic differentiation. Our results suggest that in myeloid cells, Gab1 is likely to enhance HGF mitogenicity by coupling Met to Shp‐2 and GATA‐2 expression, thereby potentially contributing to normal myeloid differentiation as well as oncogenic transformation. J. Cell. Biochem. 111: 310–321, 2010. © 2010 Wiley‐Liss, Inc.
Opposite expression of securin and γ‐H2AX regulates baicalein‐induced cancer cell death
Authors: Jiang Ren‐Huei , Su Wen‐Chi , Liu Huei‐Fang , Huang Hou‐Syun , Chao Jui‐I
Published: May 2010

Securin and γ‐H2AX have been shown to regulate cell survival and genomic stability. However, it is still unknown how the expression and regulation of these proteins is altered following treatment with baicalein, a natural flavonoid extracted from the Scutellaria baicalensis root. In the present study, we investigate the possible roles of securin and γ‐H2AX in baicalein‐induced cancer cell death. Baicalein reduced cell viability in a variety of human cancer cell lines, including bladder, cervical, colon, and lung cancer cells. Interestingly, baicalein treatment (40–80 µM for 24 h) markedly inhibited securin expression, while the levels of γ‐H2AX were elevated. Abnormal spindle formation and chromosomal segregation were induced by baicalein. Furthermore, wild type HCT116 cancer cells had a higher incidence of cytotoxicity and apoptosis than securin‐null HCT116 cells following treatment with baicalein. In contrast, baicalein increased the levels of γ‐H2AX to a similar extent in both cell types. Transfection with H2AX siRNA further increased baicalein‐induced cell death. Additionally, blockade of the AKT pathway by treatment with wortmannin or AKT shRNA lowered the levels of γ‐H2AX and enhanced cytotoxicity in baicalein‐treated cells. Taken together, our findings suggest that the opposing effects of baicalein on securin and γ‐H2AX levels may be involved in the regulation of cell viability and genomic stability by this compound. J. Cell. Biochem. 111: 274–283, 2010. © 2010 Wiley‐Liss, Inc.
About the synthesis of N‐1‐substituted 3‐aminopyrroles. A comparison
Authors: Almerico Anna Maria , Cirrincione Girolamo , Aiello Enrico , Dattolo Gaetano
Published: May 2010

A general method for the preparation of 3‐amino‐1‐methylpyrroles in excellent yields is reported. The key step involves the N‐methylation of the nitro derivatives 2, under phase transfer catalysis conditions.
Studies on pyrrolidinones. Synthesis of 5‐(5‐oxo‐2‐pyrrolidinyl)‐1,2,4‐triazole‐3‐thione derivatives
Authors: Rigo Benoít , Couturier Daniel
Published: May 2010

Starting from readily available pyroglutamic esters 1, some new 1,2,4‐triazole‐3‐thione derivatives, bonded to a pyrrolidinone ring were synthesized and are characterized by their spectral data.
10th international symposium on the synthesis and applications of isotopes and isotopically labelled compounds—whole‐body autoradiography (WBA)‐drug discovery and development Session 3: Monday, June 15, 2009
Authors: Whitby Brian , Lordi Alfred
Published: May 2010

The use of QWBA and MARG in studying tissue distribution of receptor binding proteins was discussed. Copyright © 2010 John Wiley & Sons, Ltd.
Radiosynthesis of the HIV integrase inhibitor [18F]MK‐0518 (Isentress)
Authors: Li Wenping , Thompson Wayne , Fisher Thorsten , Wai John S. , Hazuda Daria , Burns H. Donald , Hamill Terence G.
Published: May 2010

The human immunodeficiency virus integrase inhibitor, [18F]MK‐0518, was prepared via a three‐step, one‐pot radiosynthesis. [18F]4‐Fluorobenzylamine was produced from the fluorination of 4‐cyano‐N,N,N‐trimethylammonium triflate with [18F]fluoride and reduction with borane methylsulfide complex in 50–68% radiochemical yield. The final step, the coupling of [18F]4‐fluorobenzylamine with an ester coupling partner, achieved an overall uncorrected radiochemical yield after HPLC purification of ∼2%, based on the starting [18F]fluoride. In a typical run, the total synthesis time was about 90 min and gave 0.37–1.74 GBq (10–47 mCi) of [18F]MK‐0518. The radiochemical purity of [18F]MK‐0518 was>98% and the specific activity was 243–1275 Ci/mmol (EOS, n=4). A convenient three‐step, one‐pot radiosynthesis of [18F]MK‐0518 via [18F]4‐fluorobenzylamine has been developed, giving sufficient quantities of [18F]MK‐0518 for animal positron emission tomography studies. Copyright © 2010 John Wiley & Sons, Ltd.
MALDI linear TOF mass spectrometry of PEGylated (glyco)proteins
Authors: Seyfried Birgit K. , Siekmann Jürgen , Belgacem Omar , Wenzel Ryan J. , Turecek Peter L. , Allmaier Günter
Published: May 2010

PEGylation of proteins is a fast growing field in biotechnology and pharmaceutical sciences owing to its ability to prolong the serum half‐life time of recombinant proteins. Matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI TOF MS) has been shown to be a powerful tool in the analysis of several PEGylated small proteins. Here we present data obtained with a standard secondary electron multiplier (SEM) and a high mass (HM) detector combined with a MALDI linear TOF MS system for the detection of PEGylated (glyco)proteins in the range of 60–600 kDa. Examples of MALDI TOF MS of small (interferon α2a), middle (human serum albumin (HSA)) and high molecular mass proteins (coagulation factor VIII and von Willebrand factor (vWF), both heavily glycosylated proteins) are presented. The particular challenge for the analysis was the heterogeneity of the (glyco)proteins in the high molecular weight range in combination with additional PEGylation, which even introduced more heterogeneity and was more challenging for interpretation. Nevertheless, the performance of MALDI linear TOF MS with both detector systems in terms molecular weight and heterogeneity determination depending on the m/z range was superior to the other methods. Although the SEM was able to obtain information about protein PEGylation in the mass range up to 100 kDa (e.g. PEGylated HSA), the HM system was crucial for detection of HM ions (e.g. PEGylated recombinant vWF), which was impossible with the standard SEM. Copyright © 2010 John Wiley & Sons, Ltd.
L‐Valine assisted distinction between the stereo‐isomers of D‐hexoses by positive ion ESI tandem mass spectrometry
Authors: Li Gang , Huang Zhongbin , Fu Chuan , Xu Pengxiang , Liu Yan , Zhao Yu‐fen
Published: May 2010

The binary mixtures of 7 hexoses and 20 amino acids were investigated by electrospray ionization ion trap mass spectrometry (ESI‐ITMS). The adduct ions of the amino acid and the hexose were detected for 12 amino acids but not for the other 8 amino acids which are basic acidic amino acids and amides. The ions of amino acid–hexose complexes were further investigated by tandem mass spectrometry (MS/MS), and some of them just split easily into two parts whereas the others gave rich fragmentation, such as the complex ions of isoleucine, phenylalanie, tyrosine, and valine. We found that hexoses could be complexed by two molecules of valine but only by one molecule of the other amino acids. Among seven kinds of valine–hexose complexes coordinated by potassium ion, the MS2 spectra of the ion at m/z 453 yielded unambiguous differentiation. And the fragmentation ions are sensitive to the stereochemical differences at the carbon‐4 of hexoses in the complexes, as proved by the MS2. Copyright © 2010 John Wiley & Sons, Ltd.
Differentiation of Boc‐protected α,δ‐/δ,α‐ and β,δ‐/δ,β‐hybrid peptide positional isomers by electrospray ionization tandem mass spectrometry
Authors: Raju G. , Ramesh V. , Srinivas R. , Sharma G. V. M. , Shoban Babu B.
Published: May 2010

Two new series of Boc‐N‐α,δ‐/δ,α‐ and β,δ‐/δ,β‐hybrid peptides containing repeats of L‐Ala‐δ5‐Caa/δ5‐Caa‐L‐Ala and β3‐Caa‐δ5‐Caa/δ5‐Caa‐β3‐Caa (L‐Ala = L‐alanine, Caa = C‐linked carbo amino acid derived from D‐xylose) have been differentiated by both positive and negative ion electrospray ionization (ESI) ion trap tandem mass spectrometry (MS/MS). MSn spectra of protonated isomeric peptides produce characteristic fragmentation involving the peptide backbone, the Boc‐group, and the side chain. The dipeptide positional isomers are differentiated by the collision‐induced dissociation (CID) of the protonated peptides. The loss of 2‐methylprop‐1‐ene is more pronounced for Boc‐NH‐L‐Ala‐δ‐Caa‐OCH3 (1), whereas it is totally absent for its positional isomer Boc‐NH‐δ‐Caa‐L‐Ala‐OCH3 (7), instead it shows significant loss of t‐butanol. On the other hand, second isomeric pair shows significant loss of t‐butanol and loss of acetone for Boc‐NH‐δ‐Caa‐β‐Caa‐OCH3 (18), whereas these are insignificant for its positional isomer Boc‐NH‐β‐Caa‐δ‐Caa‐OCH3 (13). The tetra‐ and hexapeptide positional isomers also show significant differences in MS2 and MS3 CID spectra. It is observed that ‘b’ ions are abundant when oxazolone structures are formed through five‐membered cyclic transition state and cyclization process for larger ‘b’ ions led to its insignificant abundance. However, b1+ ion is formed in case of δ,α‐dipeptide that may have a six‐membered substituted piperidone ion structure. Furthermore, ESI negative ion MS/MS has also been found to be useful for differentiating these isomeric peptide acids. Thus, the results of MS/MS of pairs of di‐, tetra‐, and hexapeptide positional isomers provide peptide sequencing information and distinguish the positional isomers. Copyright © 2010 John Wiley & Sons, Ltd.
Spectroscopy study of mural paintings from the Pyrenean Church of Saint Eulàlia of Unha
Authors: Clark Robin J. H. , Hark Richard R. , Salvadó Nati , Butí Salvador , Pradell Trinitat
Published: May 2010

A series of wall paintings dating from the Romanesque period to the 16th century were uncovered during restoration projects at the church of Santa Eulàlia of Unha in the Val d'Aran. This study centres on the use of complementary spectroscopic techniques to examine comprehensively the pigments, binders and other materials associated with these mural paintings. These include Raman microscopy, far and mid‐Fourier transform infrared (FTIR) spectroscopy both with synchrotron radiation (SR), as well as optical microscopy and scanning electron microscopy (SEM‐EDS), and build upon earlier SR‐X‐ray diffraction studies. Traditional materials such as hematite, goethite, vermilion, red lead, carbon black and calcite were identified, but the artists' palette varied with the period. The blue pigment identified in the Romanesque frescoes was determined to be aerinite, a rare Fe(II)/Fe(III)‐containing aluminosilicate found locally in the Pyrenees region. Electronic, Raman and far‐SR‐FTIR spectra of this pigment are presented for the first time in the context of a work of art. The presence of some nitrates, sulphates and oxalates found in the mural paintings is commented upon. Copyright © 2010 John Wiley & Sons, Ltd.
Model for estimating the threshold mechanical stability of structural cartilage grafts used in rhinoplasty
Authors: Zemek Allison , Garg Rohit , Wong Brian J. F.
Published: May 2010

Objectives/Hypothesis:

Characterizing the mechanical properties of structural cartilage grafts used in rhinoplasty is valuable because softer engineered tissues are more time‐ and cost‐efficient to manufacture. The aim of this study is to quantitatively identify the threshold mechanical stability (e.g., Young's modulus) of columellar, L‐strut, and alar cartilage replacement grafts.

Study Design:

Descriptive, focus group survey.

Methods:

Ten mechanical phantoms of identical size (5 × 20 × 2.3 mm) and varying stiffness (0.360 to 0.85 MPa in 0.05 MPa increments) were made from urethane. A focus group of experienced rhinoplasty surgeons (n = 25, 5 to 30 years in practice) were asked to arrange the phantoms in order of increasing stiffness. Then, they were asked to identify the minimum acceptable stiffness that would still result in favorable surgical outcomes for three clinical applications: columellar, L‐strut, and lateral crural replacement grafts. Available surgeons were tested again after 1 week to evaluate intra‐rater consistency.

Results:

For each surgeon, the threshold stiffness for each clinical application differed from the threshold values derived by logistic regression by no more than 0.05 MPa (accuracy to within 10%). Specific thresholds were 0.56, 0.59, and 0.49 MPa for columellar, L‐strut, and alar grafts, respectively. For comparison, human nasal septal cartilage is approximately 0.8 MPa.

Conclusions:

There was little inter‐ and intra‐rater variation of the identified threshold values for adequate graft stiffness. The identified threshold values will be useful for the design of tissue‐engineered or semisynthetic cartilage grafts for use in structural nasal surgery.
Under the microscope: Assessing surgical aptitude of otolaryngology residency applicants
Authors: Carlson Matthew L. , Archibald David J. , Sorom Abraham J. , Moore Eric J.
Published: May 2010

Objectives/Hypothesis:

Application to otolaryngology residency is a highly competitive process. Programs identify the best candidates by evaluating academic performance in medical school, board scores, research experience, performance during an interview, and letters of recommendation. Unfortunately, none of these metrics completely assess an applicant's capacity to learn and perform surgical skills. We describe a direct assessment of an applicant's ability for rapid surgical skill acquisition, manual dexterity, and response to stress that can be performed during the interview process.

Study Design:

A retrospective study at an academic otolaryngology residency program.

Methods:

After orientation, applicants were seated at a microsurgical training station and allotted 20 minutes to suture an incision using 10‐0 nylon suture on a latex practice card. Their performance was graded using a 1‐to‐5 scoring system for the following categories: microscope use, respect for tissue, instrument handling, knot tying and suture control, skills acquisition, and attitude toward the exercise. Applicants were given some instruction and assessed on their ability to incorporate what they had learned into their technique.

Results:

The average total applicant score was 23.2, standard deviation (SD) 3.6 (maximum 30); 13.4% of applicants scored <1 SD below the mean, 66.1% scored within 1 SD of the mean, and 20.5% scored >1 SD above the mean.

Conclusions:

The value of applicant screening tests in predicting surgical competency is controversial. We describe a direct assessment tool that may prove useful in identifying outliers, both high and low, to aid in final applicant ranking.
Teasing out the best molecular marker in the AKT/mTOR pathway in head and neck squamous cell cancer patients
Authors: Clark Cheryl , Shah Shivang , Herman‐Ferdinandez Lilantha , Ekshyyan Oleksandr , Abreo Fleurette , Rong Xiaohua , McLarty Jerry , Lurie Aubrey , Milligan Edward J. , Nathan Cherie‐Ann O.
Published: May 2010

Objectives/Hypothesis:

No reliable molecular biomarker is currently available for clinical application in the management of head and neck cancer patients. The AKT/mTOR pathway is activated in 90% to 100% of head and neck squamous cell cancer (HNSCC) and could be promising biomarkers closely linked to cancer incidence.

Study Design:

Retrospective study of HNSCC and non‐cancer patients.

Methods:

Oral mucosa from noncancer patients were compared to HNSCC tumors and junctional zone mucosa. The candidate biomarkers mTOR, AKT, 4EBP1, and S6 kinase, signaling components upstream and downstream of mTOR that appear dysregulated in HNSCC, were evaluated using immunohistochemistry (IHC) and Western blot analysis.

Results:

Expression of phosphorylated AKT and phosphorylated mTOR were significantly higher in cancer patient tumors compared to noncancer oral mucosa samples (P = .004 and P = .026, respectively) by Western blot analysis. Expression of p‐mTOR and p‐4EBP1 were higher in patient junctional zones compared to tumors (p = 0.017 and p = 0.022, respectively) and no difference in p‐AKT or p‐S6 expression in HNSCC patients' junctional zone compared to tumors. IHC‐demonstrated p‐mTOR expression was 81.9% sensitive and 100% specific in differentiating cancer from noncancer mucosa, whereas p‐4EBP1 expression by IHC was only 50.0% sensitive and 95.5% specific in differentiating normal mucosa from HNSCC (P < .01).

Conclusions:

Phosphorylated mTOR appears to be a reliable biomarker by both Western blot analysis (P = .026) and IHC in human head and neck cancer (P < .001). Moreover, phosphorylated AKT, which is immediately upstream of mTOR, is a potential biomarker that should be further studied. Clinical trials with mTOR inhibitors are being evaluated for HNSCC, and selecting patients that are likely to respond to these inhibitors requires identifying and validating predictive biomarkers of response.
Perioperative medication errors in otolaryngology
Authors: Rosenwasser Rebecca , Winterstein Almut G. , Rosenberg Amy F. , Rosenberg Eric I. , Antonelli Patrick J.
Published: May 2010

Objectives/Hypothesis:

Medication errors are a common cause of poor clinical outcomes. Information on perioperative medication errors is scarce. This study was aimed at identifying the nature, cause, and potential remedies for medication errors in otolaryngologic surgery.

Study Design:

Prospective and descriptive.

Methods:

Clinicians were incentivized for reporting possible medication errors that occurred from the preoperative through the first postoperative clinic visit over a 2‐month period. Each report was investigated by an expert panel to determine validity, preventability, contributing factors, and potential preventative measures. A random sample of procedures and clinic visits were monitored for compliance with safe medication practices and information flow.

Results:

From 589 surgeries, 20 medication errors were reported (two preoperative, four operative, five during hospital admission, two in transition between services, four during discharge, and three postoperative). Errors included wrong dose (seven), omitted dose (six), wrong drug (five), wrong site (two), and unnecessary drug (one). Causes included failure to consider weight‐based dosing, use accurate drug references, calculate the total medication supply needed, verify the administration site, consider pertinent patient information (e.g., allergies), reconcile medications upon transfers, and document medication histories. Use of preprinted order forms was flawed, and discharge instructions were insufficient to guide patients postoperatively.

Conclusions:

Failure to adhere to safe medication‐use practices occurred throughout perioperative care. Improvement in medication documentation, following established safe practices, integration of patient information in prescribing decisions, and use of clinical decision support systems appear necessary to prevent perioperative medication errors in otolaryngology.
Prospective clinical phase II study of two new indwelling voice prostheses (Provox Vega 22.5 and 20 Fr) and a novel anterograde insertion device (Provox Smart Inserter)
Authors: Hilgers Frans J. M. , Ackerstaff Annemieke H. , Jacobi Irene , Balm Alfons J. M. , Tan I. Bing , van den Brekel Michiel W. M.
Published: May 2010

Objectives/Hypothesis:

To assess device life of the new Provox Vega 22.5 and 20 Fr prostheses, to establish whether the optimized airflow characteristics of these devices materialize in better voice characteristics in comparison to Provox2, and to assess the feasibility of voice prosthesis replacement with the new Smart Inserter.

Study Design:

Prospective clinical phase II study.

Methods:

Prospective clinical phase II study including two patient cohorts (Provox Vega 22.5 Fr: N = 33; Provox Vega 20 Fr: N = 26). Device‐life assessment, subjective data collection with study‐specific structured questionnaires, perceptual analysis (Comparative Mean Opinion Score), and objective voice analysis (standard Dutch text, maximum phonation time, and dynamic range).

Results:

Device life of both prostheses appears comparable to Provox2. Several voice characteristics were better in Vega 22.5, which coincides well with patients' preference. Voicing with Vega 20 appears comparable to Provox2. Use of Smart Inserter showed a short learning curve, and was considered an improvement by the clinicians.

Conclusions:

The new Provox Vega 22.5 and 20 Fr prostheses have a device life comparable to Provox2, whereas voicing is better than (with the 22.5Fr version) or comparable (with the 20 Fr version) to Provox2. The Smart Inserter, after a short learning curve, turns out to be a further improvement of the anterograde replacement of these indwelling devices.
Multiple nontrivial solutions for resonant Neumann problems
Authors: Filippakis Michael , Papageorgiou Nikolaos S.
Published: May 2010

We consider semilinear second order elliptic Neumann problems, which are resonant both at infinity (with respect to an eigenvalue λk, k ≥ 1) and at zero (with respect to the principal eigenvalue λ0 = 0). Using techniques from Morse theory, combined with variational methods, we are able to show that the problem has at least four nontrivial smooth solutions (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Dietary‐feeding of grape seed extract prevents azoxymethane‐induced colonic aberrant crypt foci formation in fischer 344 rats
Authors: Velmurugan Balaiya , Singh Rana P. , Agarwal Rajesh , Agarwal Chapla
Published: May 2010

Chemoprevention by dietary agents/supplements has emerged as a novel approach to control various malignancies, including colorectal cancer (CRC). This study assessed dietary grape seed extract (GSE) effectiveness in preventing azoxymethane (AOM)‐induced aberrant crypt foci (ACF) formation and associated mechanisms in Fischer 344 rats. Six‐week‐old rats were injected with AOM, and fed control diet or the one supplemented with 0.25% or 0.5% (w/w) GSE in pre‐ and post‐AOM or only post‐AOM experimental protocols. At 16 wk of age, rats were sacrificed and colons were evaluated for ACF formation followed by cell proliferation, apoptosis, and molecular analyses by immunohistochemistry. GSE‐feeding caused strong chemopreventive efficacy against AOM‐induced ACF formation in terms of up to 60% (P < 0.001) reduction in number of ACF and 66% (P < 0.001) reduction in crypt multiplicity. Mechanistic studies showed that GSE‐feeding inhibited AOM‐induced cell proliferation but enhanced apoptosis in colon including ACF, together with a strong decrease in cyclin D1, COX‐2, iNOS, and survivin levels. Additional studies showed that GSE‐feeding also decreased AOM‐caused increase in β‐catenin and NF‐κB levels in colon tissues. Compared to control animals, GSE alone treatment did not show any considerable change in these biological and molecular events in colon, and was nontoxic. Together, these findings show the chemopreventive efficacy of GSE against the early steps of colon carcinogenesis in rats via likely targeting of β‐catenin and NF‐κB signaling, and suggest its potential usefulness for the prevention of human CRC. © 2010 Wiley‐Liss, Inc.
Preparation of gel‐silica/ammonium polyphosphate core‐shell flame retardant and properties of polyurethane composites
Authors: Ni Jianxiong , Chen Lijuan , Zhao Kuiming , Hu Yuan , Song Lei
Published: May 2010

A novel intumescent gel‐silica/ammonium polyphosphate core‐shell flame retardant (MCAPP), which contains silicon, phosphorus, and nitrogen, has been prepared by in situ polymerization. The structure of MCAPP was characterized by Fourier transform infrared (FTIR) and X‐ray photoelectron spectroscopy (XPS). The properties of MCAPP were investigated by water solubility, hydrophilicity, and morphological determination. The flame retardancy and thermal stability of polyurethane (PU) composite with MCAPP were evaluated by limiting oxygen index (LOI), UL‐94 test, cone calorimetry, and thermogravimetric analysis (TGA). The results showed that MCAPP could decrease the heat release rate (HRR) and increase the thermal stability of PU materials greatly. Finally, water‐resistant properties of PU/FR composites were also studied. Copyright © 2010 John Wiley & Sons, Ltd.