bluuebunny/crossref_metadata_2025 huggingface dataset:
Capillary electrophoresis–electrochemiluminescence detection method for the analysis of ibandronate in drug formulations and human urine
Authors: Huang Yu‐Shan , Chen Shun‐Niang , Whang Chen‐Wen
Published: July 2011

A simple, rapid and sensitive CE method coupled with electrochemiluminescence (ECL) detection for direct analysis of ibandronate (IBAN) has been developed. Using a buffer solution of 20 mM sodium phosphate (pH 9.0) and a voltage of 13.5 kV, separation of IBAN in a 30‐cm length capillary was achieved in 3 min. ECL detection was performed with an indium tin oxide working electrode bias at 1.6 V (versus a Pt wire reference) in a 200‐mM sodium phosphate buffer (pH 8.0) containing 3.5 mM Ru(bpy)32+ (where bpy=2,2′‐bipyridyl). Derivatization of IBAN prior to CE‐ECL analysis was not needed. Linear correlation (r=0.9992, n=7) between ECL intensity and analyte concentration was obtained in the range of 0.25–50 μM IBAN. The LOD of IBAN in water was 0.08 μM. The developed method was applied to the analysis of IBAN in a drug formulation and human urine sample. SPE using magnetic Fe3O4@Al2O3 nanoparticles as the extraction phase was employed to pretreat the urine sample before CE‐ECL analysis. The linear range was 0.2–12.0 μM IBAN in human urine (r=0.9974, n=6). The LOD of IBAN in urine was 0.06 μM. Total analysis time including sample preparation was <1 h.
The alluvial history and sedimentary record of the Priiskovoe site and its place in the Paleolithic prehistory of Siberia
Authors: Buvit Ian , Terry Karisa , Kolosov Viktor K. , Konstantinov Mikhail V.
Published: July 2011

The Priiskovoe site in the Transbaikal Region of Siberia has long been considered one of the most ancient examples of the area's Middle Paleolithic. Priiskovoe and many other regional Paleolithic sites are part of a sequence of alluvial sediments found on or close to valley floor environments. We present several key pieces of evidence to suggest that Priiskovoe and its chipped‐stone assemblage are Upper Paleolithic in age. We examine the depositional history of the Selenga and Chikoi rivers and assess Priiskovoe's stratigraphy, geomorphology, and geochronology to argue that the Paleolithic component is dated around 26,000 14C yr B.P. within a thick buried soil ascribed to the Karginsk Interstadial. We then analyze the lithic material to conclude that the artifacts are characteristically Upper Paleolithic in terms of regional raw material selection, tool frequencies, and core reduction. Although several sites in the Transbaikal are potentially Middle Paleolithic contenders, Priiskovoe is best considered part of Siberia's Upper Paleolithic tradition. © 2011 Wiley Periodicals, Inc.
A 3.51 Ga bimodal volcanics‐BIF‐ultramafic succession from Singhbhum Craton: implications for Palaeoarchaean geodynamic processes from the oldest greenstone succession of the Indian subcontinent
Authors: Mukhopadhyay Joydip , Ghosh Gautam , Zimmermann Udo , Guha Susmita , Mukherjee Titir
Published: July 2011

The Palaeoarchaean greenstone belt of the southern Iron Ore Group (SIOG) (3.51 Ga) in the Singhbhum Craton, eastern India, includes low‐strained and low‐greenstone grade bimodal volcanics, Banded Iron Formation (BIF) and chromiferous ultramafics as enclaves within tonalite–trondjhemite–granodiorite (TTG) granitoids, collectively referred to as the Singhbhum Granite (3.4 Ga to 3.1 Ga). The succession comprises, from base to top, a lower unit of massive and pillowed basalt conformably overlain by dacitic lava and pyroclastics which in turn is overlain by a major BIF unit. The ultramafics are juxtaposed with the volcanics‐BIF succession along a thrust fault. The lithological association of pillow lava, subaqueous dacitic lava and pyroclastic rocks and BIF collectively, suggests that the entire succession was deposited in a deep‐marine depositional setting. The ash‐poor dacitic volcanic rock succession with evidences of a transition from suppressed‐volatile deep‐water lava flow and pyroclastics to more evolved mass‐flow deposits with increasing trend of subaqueous flow transformation, records a transition from a deep‐water low‐height volcanic chain to a shallower subaqueous eruption in an aggradational volcanic chain. Geochemical proxies from the bimodal volcanics and ultramafics showing enrichment of La/Nb, Th/Nb, Th/La, Ba/La, Pb/Ce, depletion in Nb–Ta relative to neighbouring REE, together with tectonic discrimination criteria using Nb, Y, Zr, Ti compositions, suggest an extending oceanic arc–forearc geodynamic setting similar to many of the Phanerozoic supra‐subduction zone ophiolites where ophiolite development in the extending upper plate in a relatively short time span is facilitated by slab rollback processes. The positive Eu‐anomaly together with high Y/Ho values from the BIFs also suggests their deposition in close proximity to spreading centres that might have developed over a rifted arc. The bimodal volcanic rock‐BIF‐ultramafic succession of the SIOG with evidence of a convergent margin geodynamic setting is an important example for Palaeoarchaean plate tectonic processes operating on Earth. Copyright © 2011 John Wiley & Sons, Ltd.
Surrogates, meta‐analysis and cost‐effectiveness modelling: a combined analytic approach
Authors: Hawkins Neil , Richardson Gerry , Sutton Alex J , Cooper Nicola J , Griffiths Chris , Rogers Anne , Bower Peter
Published: July 2011

Estimates of cost‐effectiveness analyses are typically obtained either directly from ‘trial’ based analyses or indirectly via surrogate endpoints in ‘model’ based analyses. Data from clinical trials that include both surrogate and final endpoints can be used in a joint analysis that combines these two approaches. This joint approach allows the inclusion of information regarding the effects of treatment on surrogate endpoints while relaxing the strong assumption of ‘conditional independence’ associated with indirect model‐based analyses. An example cost‐effectiveness analysis of Chronic Disease Self‐Management Programme is used to compare the different approaches. It is shown that despite using a common data set, the different analytic approaches produce differing estimates of the cost‐effectiveness of the intervention and the value of future research. The paper concludes by discussing the selection of the appropriate analytic approach. Copyright © 2011 John Wiley & Sons, Ltd.
Connective tissue growth factor autocriny in human hepatocellular carcinoma: Oncogenic role and regulation by epidermal growth factor receptor/yes-associated protein–mediated activation
Authors: Urtasun Raquel , Latasa Maria U. , Demartis Maria I. , Balzani Stella , Goñi Saioa , Garcia-Irigoyen Oihane , Elizalde Maria , Azcona Maria , Pascale Rosa M. , Feo Francesco , Bioulac-Sage Paulette , Balabaud Charles , Muntané Jordi , Prieto Jesus , Berasain Carmen , Avila Matias A.
Published: July 2011

The identification of molecular mechanisms involved in the maintenance of the transformed phenotype of hepatocellular carcinoma (HCC) cells is essential for the elucidation of therapeutic strategies. Here, we show that human HCC cells display an autocrine loop mediated by connective tissue growth factor (CTGF) that promotes DNA synthesis and cell survival. Expression of CTGF was stimulated by epidermal growth factor receptor (EGFR) ligands and was dependent on the expression of the transcriptional coactivator, Yes-associated protein (YAP). We identified elements in the

CTGF

gene proximal promoter that bound YAP-enclosing complexes and were responsible for basal and EGFR-stimulated CTGF expression. We also demonstrate that YAP expression can be up-regulated through EGFR activation not only in HCC cells, but also in primary human hepatocytes. CTGF contributed to HCC cell dedifferentiation, expression of inflammation-related genes involved in carcinogenesis, resistance toward doxorubicin, and

in vivo

HCC cell growth. Importantly, CTGF down-regulated tumor necrosis factor–related apoptosis-inducing ligand (TRAIL) receptor 2 expression and was involved in the reduced sensitivity of these cells toward TRAIL-mediated apoptosis.

Conclusion:

We have identified autocrine CTGF as a novel determinant of HCC cells' neoplastic behavior. Expression of CTGF can be stimulated through the EGFR-signaling system in HCC cells in a novel cross-talk with the oncoprotein YAP. Moreover, to our knowledge, this is the first study that identifies a signaling mechanism triggering YAP gene expression in healthy and transformed liver parenchymal cells. (Hepatology 2011)
Development of squamous neoplasia in esophageal iodine‐unstained lesions and the alcohol and aldehyde dehydrogenase genotypes of Japanese alcoholic men
Authors: Yokoyama Akira , Hirota Teruyuki , Omori Tai , Yokoyama Tetsuji , Kawakubo Hirofumi , Matsui Toshifumi , Mizukami Takeshi , Mori Shuka , Sugiura Hitoshi , Maruyama Katsuya
Published: July 2011

We investigated the development of esophageal neoplasia in biopsy specimens of the distinct iodine‐unstained lesions (DIULs) ≥5 mm detected in 280 of 2,115 Japanese alcoholic men who underwent screening by esophageal iodine staining. Low‐grade intraepithelial neoplasia (LGIN) was diagnosed in 155 of them, high‐grade intraepithelial neoplasia (HGIN) in 57, and invasive SCC in 35. The size of the DIULs increased with the degree of neoplasia. Most LGINs were flat and were missed before iodine staining. Some DIULs became a light pink color (PC) about 2 min after staining, and 2.6, 56.1 and 96.0% of the LGIN, HGIN and invasive SCC lesions, respectively, were PC‐sign‐positive. Multiple DIULs of any size markedly increased the risk of LGIN [adjusted OR (95%CI) = 10.1 (7.12–14.5)], HGIN [27.9 (14.6–53.4)] and invasive SCC [21.6 (10.1–46.4)], and were strongly associated with the presence vs. absence of DIULs ≥ 5 mm [13.3 (9.21–19.1)], inactive heterozygous aldehyde dehydrogenase‐2 (ALDH2*1/*2) vs. ALDH2*1/*1 [2.60 (1.79–3.78)], and less‐active alcohol dehydrogenase‐1B (ADH1B*1/*1) vs. ADH1B*2 allele [2.61 (1.87–3.64)]. The combination of ALDH2*1/*2 and ADH1B*1/*1 synergistically increased the risk of LGIN [4.53 (2.17–9.47)], HGIN [10.4 (4.34–24.7)] and invasive SCC [21.7 (7.96–59.3)]. Esophageal neoplasia developed at earlier ages in those with ALDH2*1/*2. Biopsy‐proven HGIN was diagnosed as invasive SCC in 15 (39.5%) of 38 patients after endoscopic mucosectomy or surgery. In conclusion, large size, non‐flat appearance, positive PC sign and multiplicity of DIULs and ALDH2*1/*2 and ADH1B*1/*1 were associated with development of esophageal neoplasia in Japanese alcoholics. Biopsy‐proven HGIN should be totally resected for both diagnostic and therapeutic purposes.
Vaccination‐induced functional competence of circulating human tumor‐specific CD8 T‐cells
Authors: Baumgaertner Petra , Jandus Camilla , Rivals Jean‐Paul , Derré Laurent , Lövgren Tanja , Baitsch Lukas , Guillaume Philippe , Luescher Immanuel F. , Berthod Gregoire , Matter Maurice , Rufer Nathalie , Michielin Olivier , Speiser Daniel E.
Published: July 2011

T‐cells specific for foreign (e.g., viral) antigens can give rise to strong protective immune responses, whereas self/tumor antigen‐specific T‐cells are thought to be less powerful. However, synthetic T‐cell vaccines composed of Melan‐A/MART‐1 peptide, CpG and IFA can induce high frequencies of tumor‐specific CD8 T‐cells in PBMC of melanoma patients. Here we analyzed the functionality of these T‐cells directly ex vivo, by multiparameter flow cytometry. The production of multiple cytokines (IFNγ, TNFα, IL‐2) and upregulation of LAMP‐1 (CD107a) by tumor (Melan‐A/MART‐1) specific T‐cells was comparable to virus (EBV‐BMLF1) specific CD8 T‐cells. Furthermore, phosphorylation of STAT1, STAT5 and ERK1/2, and expression of CD3 zeta chain were similar in tumor‐ and virus‐specific T‐cells, demonstrating functional signaling pathways. Interestingly, high frequencies of functionally competent T‐cells were induced irrespective of patient's age or gender. Finally, CD8 T‐cell function correlated with disease‐free survival. However, this result is preliminary since the study was a Phase I clinical trial. We conclude that human tumor‐specific CD8 T‐cells can reach functional competence in vivo, encouraging further development and Phase III trials assessing the clinical efficacy of robust vaccination strategies.
Ten‐year survival among HIV‐1‐infected subjects with AIDS or non‐AIDS‐defining malignancies
Authors: Spagnuolo Vincenzo , Galli Laura , Salpietro Stefania , Gianotti Nicola , Guffanti Monica , Cossarini Francesca , Bigoloni Alba , Cinque Paola , Bossolasco Simona , Travi Giovanna , Fumagalli Luca , Lazzarin Adriano , Castagna Antonella
Published: July 2011

Few data are available regarding the 10‐year survival among subjects with HIV and cancer. The aim of this study was to evaluate the 10‐year survival of HIV‐infected subjects with AIDS‐defining malignancies (ADM) or non‐AIDS‐defining malignancies (NADM). This was a single center, retrospective, observational study of subjects with HIV infection and a subsequent cancer diagnosis; the data were collected from January 1991 to April 2010. Malignancies were divided into ADM or NADM on the basis of the Centre of Diseases Control‐1993 classification. Survival curves were estimated using Kaplan–Meyer method and compared by the log‐rank test. Six hundred and fifteen (9.5%) of the 6,495 subjects recorded in the San Raffaele Infectious Diseases Database developed a malignancy: 431 (70%) an ADM and 184 (30%) a NADM. In the case of ADM, survival was more favorable when cancer was diagnosed during post‐highly active antiretroviral therapy (HAART) era (10‐year survival: 43.2% ± 4.4%) than when diagnosed during the pre‐HAART era (10‐year survival: 16.4% ± 2.7%; log‐rank test: p < 0.001). The same was true in the case of NADM (10‐year survival: 44.7% ± 5.5% vs. 33.3 ± 9.6%; log‐rank test: p = 0.03). An evaluation of survival probability by cancer type showed higher survival rates during the post‐HAART era in the case of non‐Hodgkin lymphoma (10‐year survival: 42.1% ± 5.3% vs. 11.4% ± 3.3%; log‐rank test: p = <0.001), Kaposi's sarcoma (10‐year survival: 44.0% ± 8.4% vs. 23.5% ± 3.9%; log‐rank test: p < 0.001) and Hodgkin's disease (10‐year survival: 49.5% ± 14.5% vs. 40.0% ± 12.7%; log‐rank test: p = 0.005). Despite the better cancer prognosis during the post‐HAART era, the 10‐year survival of HIV‐infected subjects with an ADM or NADM is poor.
N‐acetyl‐cysteine protects chicken growth plate chondrocytes from T‐2 toxin‐induced oxidative stress
Authors: He Shao‐jun , Hou Jia‐fa , Dai Yu‐yi , Zhou Zhen‐lei , Deng Yi‐feng
Published: July 2011

T‐2 toxin is now considered to be related to bone malformation such as incomplete ossification, absence of bones and fused bones. In this study, primary cultures of chicken tibial growth plate chondrocytes (GPCs) were treated with various concentrations of T‐2 toxin (5, 50, and 500 n m) in the absence and presence of N‐acetyl‐cysteine (NAC) to investigate the effects of the antioxidant NAC on T‐2 toxin‐induced toxicity. Our results showed that T‐2 toxin markedly decreased cell viability, alkaline phosphatase activity and glutathione content (P < 0.05). In addition, T‐2 toxin significantly increased reactive oxygen species levels and malondialdehyde in a dose‐dependent manner. However, the T‐2 toxin‐induced cytotoxicity was reversed, in part, by the antioxidant NAC (P < 0.05). These results suggest that T‐2 toxin inhibits the proliferation and differentiation of GPCs in vitro by altering cellular homeostasis and NAC can protect GPCs against T‐2 toxin cytotoxicity by reducing the T‐2 toxin‐induced oxidative stress. Copyright © 2011 John Wiley & Sons, Ltd.
Accelerated neovascularization and endothelialization of vascular grafts promoted by covalently bound laminin type 1
Authors: Williams Stuart K. , Kleinert Leigh B. , Patula‐Steinbrenner Vangie
Published: July 2011

Development of a small diameter (<6 mm) synthetic vascular graft with clinically acceptable patency must overcome the inherent thrombogenicity of polymers and the development of neointimal thickening. Establishment of an endothelial cell lining on the lumenal surface has been hypothesized as a mechanism to improve the function of vascular grafts. The major aim of this study is to evaluate the use of laminin type 1, covalently bound to all surfaces of expanded polytetrafluoroethylene (ePTFE) grafts, on neovascularization of the interstices and lumenal surface endothelialization. One millimeter i.d. vascular grafts were surface modified through covalent attachment of laminin type 1. Grafts were subsequently implanted as interpositional aortic grafts in rats. Following 5‐weeks implantation, the grafts were explanted and morphologically evaluated using scanning electron microscopy and light microscopy. Scanning electron microscopy identified an extensive coverage of antithrombogenic cells on the lumenal flow surface of laminin type 1 modified grafts. Histological evaluation confirmed the presence of endothelial cells on the midgraft lumenal surface of laminin 1 modified grafts. Extensive neovascularization of the interstices of the laminin‐modified grafts occurred as compared with control grafts. We conclude that surface modification using laminin type 1 accelerates both the neovascularization and endothelialization of porous ePTFE vascular grafts. © 2011 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 99A: 67–73, 2011.
Osteopontin functionalization of hydroxyapatite nanoparticles in a PDLLA matrix promotes bone formation
Authors: Jensen T. , Baas J. , Dolathshahi‐Pirouz A. , Jacobsen T. , Singh G. , Nygaard J. V. , Foss M. , Bechtold J. , Bünger C. , Besenbacher F. , Søballe K.
Published: July 2011

We studied the osteoconductive tissue response of hydroxyapatite (HA) nanoparticles functionalized with osteopontin (OPN) in a matrix of poly‐D,L‐lactic‐acid (PDLLA). In a canine endosseus 0.75‐mm gap implant model, we tested the osteointegrative impact of the OPN functionalized composite as an implant coating, and a non‐functionalized composite was used as reference control. During the four weeks of observation, the OPN functionalized composite coating significantly increased the formation of new bone in the porosities of the implant, but no differences were observed in the gap. The study provides evidence of its potential use either alone or in combination with other osteoconductive compounds. © 2011 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 99A: 94–101, 2011.
Implant‐assisted meniscal repair in vivo using a chondrocyte‐seeded flexible PLGA scaffold
Authors: Yoo Jeong Joon , Bichara David A. , Zhao Xing , Randolph Mark A. , Gill Thomas J.
Published: July 2011

A cell‐based engineered construct can be used for healing of intractable meniscal lesions. Our aims were to assess the culture conditions (static versus dynamic oscillation) and the healing capacity of the chondrocyte‐seeded flexible implants in a heterotopic mouse model. Swine articular chondrocytes were labeled with PKH 26 or DiI dye and seeded onto a flexible PLGA scaffold using dynamic oscillating conditions for 24 h. Half of cell‐seeded scaffolds were cultured in the same dynamic conditions, while the remaining scaffolds were cultured statically. After 7 days, scaffolds were placed between swine meniscal discs and were implanted subcutaneously in nude mice for 6 weeks. Additional constructs for assessing in vivo cell tracking were implanted for 12 weeks. Live/dead assays demonstrated labeled chondrocytes attached throughout the scaffold in both culture conditions. DNA measurements showed no significant difference between the culture conditions. A continuous fibro‐cartilaginous healing tissue was observed between meniscal discs in all 12 dynamically cultured constructs and 9 of 11 statically cultured ones. There was no evidence of meniscal healing using acellular scaffold as well as in meniscal constructs lacking an implant. Both PKH 26‐ and DiI‐labeled cells were identified along the healing interface. We conclude the chondrocyte‐seeded flexible PLGA implants induce healing of meniscal discs in nude mice. Culture conditions after seeding have no apparent effects on healing. © 2011 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 99A: 102–108, 2011.
Interfacial interactions of apolipoprotein AI and high density lipoprotein: Overlooked phenomena in blood‐material contact
Authors: Cornelius Rena M. , Macri Joseph , Brash John L.
Published: July 2011

Apolipoprotein AI (apo AI) is the major protein component of high density lipoprotein (HDL), and represents ∼1% of the total protein content of plasma. In previous work, apo AI was identified as a major component of the protein layers adsorbed from plasma to biomaterials having a wide range of surface properties. Notwithstanding such indications of the major contribution of lipoprotein interactions, these phenomena have been largely overlooked in the blood‐contacting biomaterials area. In this communication, detailed quantitative data on the adsorption of apo AI to typical “biomedical” segmented polyurethane (PU) are reported. Using radiolabeled apo AI, adsorption levels from buffer and plasma were found to be ∼0.5 and ∼0.2 μg/cm2, respectively. Albumin adsorption from plasma was comparable at about ∼0.17 μg/cm2. Since, it is unknown how much of the adsorbed apo AI is associated with HDL versus how much is in the free state, the corresponding molar quantities cannot be determined with certainty. However, if it is assumed that all of the adsorbed apo AI is free, the molar quantities adsorbed from plasma were in the ratio apo AI:albumin = 2.77, compared with 0.00063 in plasma. Immunoblot data showed similar trends with respect both to the variation in adsorbed quantity with plasma concentration and to the relative adsorbed quantities of apo AI and albumin. These data show unequivocally the very strong surface activity of apo AI and suggest that a key future focus for blood compatibility research should be lipoprotein interactions. © 2011 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 99A: 109–115, 2011.
PEGylated silica nanoparticles encapsulating multiple magnetite nanocrystals for high‐performance microscopic magnetic resonance angiography
Authors: Wu Si‐Han , Lin Chien‐Yuan , Hung Yann , Chen Wei , Chang Chen , Mou Chung‐Yuan
Published: July 2011

A novel magnetic resonance (MR) angiographic method, 3DΔR2‐mMRA (three dimensional and ΔR2 based microscopy magnetic resonance angiography), is developed as a clinical diagnosis for depicting the function and structure of cerebral small vessels. However, the visibility of microvasculatures and the precision of cerebral blood volume calculation greatly rely on the transverse relaxivity and intravascular half‐life of contrast agent, respectively. In this work, we report a blood pool contrast agent named H‐Fe3O4@SiO2–PEG where multiple Fe3O4 nanocrystals are encapsulated in a thin silica shell to enhance the T2‐relaxivity (r2 = 342.8 mM−1 s−1) and poly(ethylene glycol) (PEG) is employed to reduce opsonization and prolong circulation time of nanoparticles. Utilization of the newly developed H‐Fe3O4@SiO2–PEG with a novel MR angiographic methodology, a high‐resolution MR image of rat cerebral microvasculatures is successfully obtained. © 2011 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2011.
The structure of intersession experience in psychotherapy and its relation to the therapeutic alliance
Authors: Hartmann Armin , Orlinsky David , Zeeck Almut
Published: July 2011

During the days‐long or week‐long intervals between their therapy sessions, patients typically recollect, reflect on, practice, and imaginatively elaborate on experiences they had during sessions with their therapists. These “intersession experiences” (IE) have been studied for some time with the Intersession Experience Questionnaire (IEQ) in Germany and the United States. This study aims to compare the factor structure of the English and the German versions of the IEQ and to explore the relation of IE to the therapeutic alliance, as a key process variable usually associated with outcome. Factor analyses on a total of 7,550 IEQ reports from 769 patients in outpatient, day hospital, and inpatient treatment showed almost identical factor structures in both countries and across different therapy settings. Significant associations were found between certain IE factors and therapeutic alliance, varying in strength (0.27≤r2≤0.66) by treatment duration and country. Theoretical, clinical and research implications of the findings are discussed. © 2011 Wiley Periodicals, Inc. J Clin Psychol 67:1–20, 2011.
Regulation of cell cycle components during exposure to anoxia or dehydration stress in the wood frog, Rana sylvatica
Authors: Roufayel Rabih , Biggar Kyle K. , Storey Kenneth B.
Published: July 2011

The wood frog (Rana sylvatica) exhibits a well‐developed natural anoxia and dehydration tolerance. The degree of stress tolerance depends on numerous biochemical adaptations, including stress‐induced hypometabolism that helps to preserve long‐term viability by reducing ATP demand. We hypothesized that the mechanisms involved in cell cycle control could act to aid in the establishment of the hypometabolic state required for stress survival. Selected proteins involved in the proliferation of cells were evaluated using immunoblotting in liver and skeletal muscle of wood frogs comparing controls with animals subjected to either 24‐hr anoxia exposure under a nitrogen gas atmosphere or dehydration to 40% of total body water lost (all at 5°C). Levels of cyclins (type A, B, D, and E) decreased significantly under both stresses in liver and skeletal muscle. Similar reductions were seen for Cyclin‐dependant kinases (Cdk) types 2, 4, and 6 in both liver and skeletal muscle; however, an increase in the relative amount of phosphorylated inactive p‐Cdk (Thr14/Tyr15) was observed in liver under both stresses. Levels of positive regulators of Cdk activity (Cdc25 type A and C) were significantly reduced in both tissues under both stresses, whereas negative regulators of Cdk activity (p16INK4a and p27KIP1) increased significantly in liver under both anoxia and dehydration stress (but not in muscle). This study provides the first report of differential regulation of cell cycle components in an anoxia and dehydration tolerant vertebrate, the wood frog, suggesting that cell cycle suppression is an active part of stress resistance and life extension in hypometabolic states. J. Exp. Zool. 315:487–494, 2011. © 2011 Wiley‐Liss, Inc.
The position of the AUG start codon in MFG‐based γ‐retroviral vectors has a dramatic effect on translation‐dependent protein expression
Authors: Frankel Timothy L. , Zhang Ling , Burns William R. , Zheng Zhili , Morgan Richard A.
Published: July 2011

Background

In the past three decades, much advancement has been made in γ‐retroviral vector mediated gene transfer. One widely used vector design is based on the MFG vector, which uses the Moloney murine leukemia virus (MoMLV) transcriptional unit with extended packaging signals and insertion of the native MoMLV envelope splice acceptor region immediate 5′ to the gene of interest inserted at an NcoI restriction site, which contains a translation start codon. Little is known about the impact of variations in start codon location within MFG‐based vectors on protein expression.

Methods

To evaluate variation in start condo placement, a gene encoding a T‐cell receptor (TCR) was cloned into an MFG‐based vector and site‐directed mutagenesis was used to move the gene away from the splice acceptor, as well as alter the frame with respect to the upstream start codon. Kozak consensus sequences were also added to the gene in an attempt to improve translation.

Results

Protein expression as measured by TCR surface expression and biological activity was substantially reduced when the gene was placed downstream and out‐of‐frame with the NcoI start codon. Expression was reestablished by mutation of the upstream start site, although at a reduced level. These findings were repeated with two other genes, a dominant negative TGFβRII and the reporter protein dEGFP.

Conclusions

These finding emphasize the scanning rule for translation initiation and stress the importance of cloning genes of interest into or near the native NcoI start site of MFG‐based retroviral vectors. Copyright © 2011 John Wiley & Sons, Ltd.
An improved procedure for the three‐component synthesis of benzo[g]chromene derivatives using basic ionic liquid
Authors: Yu Yi , Guo Hongyun , Li Xiaojun
Published: July 2011

A basic ionic liquid, 1‐butyl‐3‐methyl imidazolium hydroxide ([bmim]OH), efficiently promotes a one‐pot, three‐component condensation of aromatic aldehydes, malononitrile or ethyl cyanoacetate, and 2‐hydroxy‐1,4‐naphthoquinone to produce 4‐aryl‐5,10‐dihydro‐4H‐benzo[g]chromene‐5,10‐dione derivatives in high yields at room temperature. This reaction does not involve any hazardous organic solvent and toxic catalyst. The ionic liquid is recovered and recycled for subsequent reactions. Moreover, this protocol has the advantages of easy work‐up, short reaction time, high yields, and environmentally benign procedure compared with the reported methods. J. Heterocyclic Chem., (2011).
Pelvic and hindlimb myology of the basal archosaur Poposaurus gracilis (archosauria: Poposauroidea)
Authors: Schachner Emma R. , Manning, Phillip L. , Dodson Peter
Published: July 2011

The discovery of a largely complete and well preserved specimen of Poposaurus gracilis has provided the opportunity to generate the first phylogenetically based reconstruction of pelvic and hindlimb musculature of an extinct nondinosaurian archosaur. As in dinosaurs, multiple lineages of basal archosaurs convergently evolved parasagittally erect limbs. However, in contrast to the laterally projecting acetabulum, or “buttress erect” hip morphology of ornithodirans, basal archosaurs evolved a very different, ventrally projecting acetabulum, or “pillar erect” hip. Reconstruction of the pelvic and hindlimb musculotendinous system in a bipedal suchian archosaur clarifies how the anatomical transformations associated with the evolution of bipedalism in basal archosaurs differed from that of bipedal dinosaurs and birds. This reconstruction is based on the direct examination of the osteology and myology of phylogenetically relevant extant taxa in conjunction with osteological correlates from the skeleton of P. gracilis. This data set includes a series of inferences (presence/absence of a structure, number of components, and origin/insertion sites) regarding 26 individual muscles or muscle groups, three pelvic ligaments, and two connective tissue structures in the pelvis, hindlimb, and pes of P. gracilis. These data provide a foundation for subsequent examination of variation in myological orientation and function based on pelvic and hindlimb morphology, across the basal archosaur lineage leading to extant crocodilians. J. Morphol., 2011. © 2011 Wiley‐Liss, Inc.
Liquid chromatography–tandem mass spectroscopic method for the determination of zerumbone in human plasma and its application to pharmacokinetics
Authors: Eid Eltayeb E. M. , Bustamam Abdul Ahmad , Rasedee A. , Suliman Fakhr Eldin O. , Sukari Mohd A. , Fatah Safa A.
Published: July 2011

A rapid, sensitive, specific and selective LC–MS/MS method for the determination of zerumbone (ZER) in human plasma using 2,4‐diamino‐6‐(4‐methoxyphenyl)‐1,3,5‐triazine (DMTZ) as an internal standard (IS) has been developed and validated. ZER was chromatographed on C8 column using a mobile phase of acetonitrile/water (80:20, v/v) at a flow rate of 0.25 ml min−1. Quantitation was achieved using ESI+ interface, employing multiple reaction monitoring (MRM) mode at m/z 219 > 81 and 218 > 134 for ZER and IS, respectively. The calibration standards were linear over a range of 5–3000 ng ml−1 (r2 = 0.9994) with an LLOQ of 5 ng ml−1 (RSD %; 11.4% and bias%; 9.5%). Intra‐ and inter‐day precision of ZER assay ranged from 0.18 to 3.56% with accuracy (bias) that varied between − 5.09 and 4.3%, demonstrating good precision and accuracy. Recoveries of ZER and the IS from human plasma were above 85%. The developed method was validated for the determination of ZER in rat plasma. Linearity, stability of ZER and the ME on rat plasma were discussed. The applicability of the developed method was demonstrated by measuring ZER in rat plasma samples following intravenous and intraperitoneal administration of ZER prepared in hydroxypropyl‐β‐cyclodextrin (HPβCD) and sodium carboxymethyl cellulose (CMC), respectively, in 20 mg kg−1 and this study indicated a clear significant difference (p < 0.05) in pharmacokinetic parameters of ZER in ZER/HPβCD complex compared with ZER in CMC preparation. Copyright © 2011 John Wiley & Sons, Ltd.
High throughput identification of components from traditional Chinese medicine herbs by utilizing graphene or graphene oxide as MALDI‐TOF‐MS matrix
Authors: Liu Yang , Liu Junyan , Yin Peng , Gao Mingxia , Deng Chunhui , Zhang Xiangmin
Published: July 2011

In this work, graphene or graphene oxide was utilized, for the first time, to identify small molecular components from traditional Chinese medicine (TCM) herbs, by acting as matrix of matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF‐MS). Due to the large surface area of graphene or graphene oxide, the analytes were trapped tightly to the matrix, which avoids the contamination of the ion source and vacuum system. Besides, their excellent electronic, thermal and mechanical properties make them desired matrices for MALDI‐TOF‐MS. Stable analysis was achieved with no background inference even at the concentration of 100 nM. Moreover, the limit of detection (LOD) could be greatly lowered by utilizing graphene or graphene oxide as a pre‐enrichment adsorbent. In summary, the promoted MALDI‐TOF‐MS methodology was demonstrated to be simple, sensitive, fast, cost effective and, most importantly, high throughput. Copyright © 2011 John Wiley & Sons, Ltd.
NR2B phosphorylation at tyrosine 1472 in spinal dorsal horn contributed to N‐methyl‐D‐aspartate‐induced pain hypersensitivity in mice
Authors: Li Shuai , Cao Jing , Yang Xian , Suo Zhan‐Wei , Shi Lei , Liu Yan‐Ni , Yang Hong‐Bin , Hu Xiao‐Dong
Published: July 2011

Calcium influx via N‐methyl‐D‐aspartate (NMDA)‐subtype glutamate receptors (NMDARs) regulates the intracellular trafficking of NMDARs, leading to long‐lasting modification of NMDAR‐mediated synaptic transmission that is involved in development, learning, and synaptic plasticity. The present study investigated the contribution of such NMDAR‐dependent synaptic trafficking in spinal dorsal horn to the induction of pain hypersensitivity. Our data showed that direct activation of NMDARs by intrathecal NMDA application elicited pronounced mechanical allodynia in intact mice, which was concurrent with a specific increase in the abundance of NMDAR subunits NR1 and NR2B at the postsynaptic density (PSD)‐enriched fraction. Selective inhibition of NR2B‐containing NMDARs (NR2BR) by ifenprodil dose dependently attenuated the mechanical allodynia in NMDA‐injected mice, suggesting the importance of NR2BR synaptic accumulation in NMDA‐induced pain sensitization. The NR2BR redistribution at synapses after NMDA challenge was associated with a significant increase in NR2B phosphorylation at Tyr1472, a catalytic site by Src family protein tyrosine kinases (SFKs) that has been shown to prevent NR2B endocytosis. Intrathecal injection of a specific SFKs inhibitor, PP2, to block NR2B tyrosine phosphorylation eliminated NMDA‐induced NR2BR synaptic expression and also attenuated the mechanical allodynia. These data suggested that activation of spinal NMDARs was able to accumulate NR2BR at synapses via SFK signaling, which might exaggerate NMDAR‐dependent nociceptive transmission and contribute to NMDA‐induced nociceptive behavioral hyperresponsiveness. © 2011 Wiley‐Liss, Inc.
Differential activity of regions of the psoas major and quadratus lumborum during submaximal isometric trunk efforts
Authors: Park Rachel J. , Tsao Henry , Cresswell Andrew G. , Hodges Paul W.
Published: July 2011

Controversy exists regarding the function of psoas major (PM) and quadratus lumborum (QL) at the lumbar spine. The functions of discrete regions of PM and QL were studied during trunk loading tasks. Twelve healthy participants performed isometric trunk loading tasks in various directions in upright sitting. Fine‐wire electromyography (EMG) electrodes were inserted under ultrasound guidance into PM fascicles arising from the transverse process (PM‐t) and vertebral body (PM‐v) and the anterior (QL‐a) and posterior (QL‐p) layers of QL on the right side. Although right PM‐t and PM‐v were both active during right lateral‐flexion trunk efforts, their activity was opposite in the sagittal plane, with greater PM‐t towards extension and PM‐v towards flexion. QL‐a and QL‐p were similarly active, though QL‐p was active to a greater percentage of MVC during right trunk lateral‐flexion efforts. Activity of QL‐p was modulated with respiratory phase during the loading tasks with trunk efforts towards the right lateral‐flexion/flexion and right lateral‐flexion directions. These findings provide novel understanding of the unique activation of discrete regions of PM and QL. These differences must be considered in future EMG studies to better understand the function of these deeply situated trunk muscles in the control of the lumbar spine. © 2011 Orthopaedic Research Society Published by Wiley Periodicals, Inc. J Orthop Res 30:311–318, 2012
Anthocyanin composition and extractability in berry skin and wine of Vitis vinifera L. cv. Aglianico
Authors: Manfra Michele , Nisco Mauro De , Bolognese Adele , Nuzzo Vitale , Sofo Adriano , Scopa Antonio , Santi Luca , Tenore Gian Carlo , Novellino Ettore
Published: July 2011

BACKGROUND: The present article reports the anthocyanin content in the berry skin and wine of the Italian red grape cultivar Aglianico (clone VCR11 grafted onto 1103 Paulsen), one of the most ancient vines and famous for its deep‐red colour. Anthocyanins were extracted from frozen berry skin in an acidified methanol solution. The extraction mixtures, monitored for 120 h, were analysed by high‐performance liquid chromatography.

RESULTS: The extraction from berry skin of delphinidin, petunidin and malvidin appeared to be a time‐independent process, whereas the concentration of peonidin increased linearly with time. Peonidin‐O‐acetyl‐glucoside was transferred from skin more slowly than petunidin‐O‐acetyl‐glucoside and malvidin‐O‐acetyl‐glucoside. The anthocyanin composition of the resulting wine showed that the total anthocyanin content was about one‐tenth of the corresponding berry skin content. The ratio acetyl/coumaroyl anthocyanins in the wine was sharply higher than the value in berry skin (0.85 and 0.10, respectively), indicating an enrichment of acetyl derivatives in the wine.

CONCLUSION: Levels of single anthocyanins in wine were not always correlated with those detected in grapes, as they were affected by winemaking. The high values of some anthocyanins in Aglianico wine could ameliorate its quality, increasing the chromatic properties, aging stability and product acceptance. Copyright © 2011 Society of Chemical Industry
Modulation of post‐hatch growth and immunity through in ovo supplemented nutrients in broiler chickens
Authors: Bakyaraj Sanniyasi , Bhanja Subrat K , Majumdar Samir , Dash Banabihari
Published: July 2011

BACKGROUND: Early post‐hatch growth and immunity were assessed through in ovo supplementation of nutrients: amino acids (AA), trace elements (TE), fatty acids and vitamins (FAV) grouped under humoral immunity (HI) or cell‐mediated immunity (CMI) on the 18th day of incubation at the broad end of the egg using a 25 mm needle.

RESULTS: Hatchability in AA groups was better than TE and FAV groups. CMI groups had better hatchability than HI groups. AA and TE groups had higher chick‐to‐egg weight ratio (P < 0.01) than the FAV group. At 3 weeks of age, a higher body weight (P < 0.01) was recorded in AA for CMI, TE for HI and FAV for HI groups. FAV‐injected chicks had a higher bursa weight at hatch, but TE chicks had higher thymus weight at the 3rd week of age. Humoral immune response was not different in in ovo injected chicks compared to control. CMI was higher (P < 0.01) in AA for CMI, TE for CMI and FAV for CMI or HI nutrient‐injected chicks.

CONCLUSIONS: In ovo injection of AA for CMI and TE for HI may accelerate growth of broiler chickens. In ovo injection of AA, TE or FAV may modulate CMI in chicks. Copyright © 2011 Society of Chemical Industry
Performance evaluation of a rapid‐scanning quadrupole mass spectrometer in the comprehensive two‐dimensional gas chromatography analysis of pesticides in water
Authors: Purcaro Giorgia , Quinto Tranchida Peter , Conte Lanfranco , Obiedzińska Agnieszka , Dugo Paola , Dugo Giovanni , Mondello Luigi
Published: July 2011

The performance of a novel rapid‐scanning (20 000 amu/s) quadrupole mass spectrometer (qMS) has been evaluated in the comprehensive 2‐D gas chromatography (GC×GC) analysis of pesticides contained in water. Analyte extraction was performed by using direct solid‐phase microextraction (SPME). The MS system was operated using a rather wide m/z 50–450 mass range and a 33 Hz spectral production rate, a frequency which was found sufficient for reliable quantification. The qMS performance was evaluated considering: (i) number of data points per peak, (ii) mass spectral quality, (iii) extent of peak skewing, and (iv) consistency of retention times. Seven‐point calibration curves (external calibration) were constructed for 28 pesticides over the limit of quantification range of 100 μg/L (1, 5, 10, 25, 50, and 100 μg/L). The solid‐phase microextraction‐GC×GC‐qMS method was validated by calculating limits of detection and quantification, intraday peak area precision, accuracy, and intraday retention‐time precision. A series of tap water samples were subjected to analysis, fortunately giving negative results.
Self‐assembled highly ordered ethane‐bridged periodic mesoporous organosilica and its application in HPLC
Authors: Huang Lili , Lu Juan , Di Bin , Feng Fang , Su Mengxiang , Yan Fang
Published: July 2011

Monodisperse spherical periodic mesoporous organosilicas (PMOs) with ethane integrated in the framework were synthesized and their application as stationary phase for chromatographic separation is demonstrated. The ethane‐PMOs were prepared by condensation of 1,2‐bis(triethoxysilyl)ethane (BTSE) in basic condition using octadecyltrimethylammonium chloride (C18TMACl) as template and ethanol as co‐solvent. The morphology and mesoporous structure of ethane‐PMOs were controlled under different concentrations of sodium hydroxide (NaOH) and EtOH. The results of scanning electron microscopy (SEM), transmission electron microscopy (TEM), powder X‐ray diffraction (XRD), nitrogen sorption measurement, Fourier transform infrared spectroscopy (FT‐IR) and elemental analysis showed that ethane‐PMOs have spherical morphology, uniform particle distribution, highly ordered pore structure, high surface area and narrow pore‐size distribution. The column packed with these materials exhibits good permeability, high chemical stability and good selectivity of mixtures of aromatic hydrocarbons in normal phase high‐performance liquid chromatography (HPLC).
Capacitively coupled contactless conductivity detection as an alternative detection mode in CE for the analysis of kanamycin sulphate and its related substances
Authors: El‐Attug Mohamed N. , Adams Erwin , Hoogmartens Jos , Van Schepdael Ann
Published: July 2011

A method was developed to determine simultaneously kanamycin, its related substances and sulphate in kanamycin sulphate using capacitively coupled contactless conductivity detection. Kanamycin is an aminoglycoside antibiotic that lacks a strong UV‐absorbing chromophore. Due to its physicochemical properties, CE in combination with capacitively coupled contactless conductivity detection was chosen. The separation method uses a BGE composed of 40 mM 2‐(N‐morpholino)ethanesulphonic acid monohydrate and 40 mM L‐histidine, pH 6.35. A 0.6 mM N‐cetyltrimethyl ammonium bromide (CTAB) solution was added as electroosmotic flow modifier in a concentration below the critical micellar concentration (CMC). Ammonium acetate 50 mg/L was used as internal standard. In total, 30 kV was applied in reverse polarity on a fused‐silica capillary (65/41 cm; 75 μm id). The optimized separation was obtained in less than 6 min with good linearity (R2=0.9999) for kanamycin. It shows a good precision expressed as RSD on the relative peak areas equal to 0.3 and 1.1% for intra‐day and inter‐day precision, respectively. The LOD and LOQ are 0.7 and 2.3 mg/L, respectively. Similarly, for sulphate, a good linearity (R2=0.9996) and precision (RSD 0.4 and 0.6% for intra‐day and inter‐day, respectively) were obtained.
Simultaneous quantification of three iridoids in rat plasma after oral administration of Zhi‐zi‐chi decoction using LC‐MS
Authors: Long Zhi‐Min , Bi Kai‐Shun , Huo Yan‐Shuang , Yan Xiao‐Yu , Zhao Xu , Chen Xiao‐Hui
Published: July 2011

A sensitive and efficient liquid chromatography–mass spectrometry (LC‐MS) method was developed and validated for the simultaneous determination of geniposide, 6α‐hydroxygeniposide, and genipin gentiobioside in rat plasma. After the addition of internal standard (I.S.) salidroside and acidification (formic acid, 0.1%), plasma samples were carried out by protein precipitation with acetonitrile and separated on a Kromasil C18 column (200 mm×4.6 mm, 5 μm) within a runtime of 15.0 min. The linear ranges were 2–250 ng/mL for both 6α‐hydroxygeniposide and genipin gentiobioside and 2–2000 ng/mL for geniposide, respectively. The lower limit of quantification (LLOQ) was 2 ng/mL for all the analytes. The validated method was successfully applied to the pharmacokinetics study of geniposide, 6α‐hydroxygeniposide, and genipin gentiobioside in rats after oral administration of Zhi‐zi‐chi decoction.
Simultaneous determination of three naturally occurring estrogens in environmental waters by high‐performance liquid chromatography
Authors: Wang Chengjun , Xu Chunmei , Chen Fan , Tang Xuejiao
Published: July 2011

A simple, sensitive and accurate reversed‐phase high‐performance liquid chromatographic (HPLC) method for simultaneous determination of three naturally occurring estrogenic steroids including estrone (E1), 17β‐estradiol (E2) and estriol (E3) in environmental water samples was developed. Analytes were extracted with ethyl acetate solvents and preconcentrated prior to HPLC analysis. Separations were accomplished in <20 min using a reversed‐phase C18 column (4.6×250 mm id, 5 μm) with a gradient elution of mobile phase containing 3.0 mM ammonium acetate/acetonitrile mixtures (flow rate, 1.0 mL/min). UV light absorption responses at 205 nm were linear over a wide concentration range from 100 000 μg/L to the detection limits of 0.96 μg/L E1, 0.64 μg/L E2 and 0.78 μg/L E3. Quantitation was carried out by the peak area method. The relative standard deviation for the analysis of three estrogens was <3.0%. This method was applied for the simultaneous determination of estrogens in environmental water samples collected in Zhejiang, China. The higher concentrations of both E2 and E3 were found in Tang River and West Lake waters, and E1 was detected in lake water only. All three estrogens were below the detection limits in rain waters.