bluuebunny/crossref_metadata_2025 huggingface dataset:
Characterization of viability and proliferation of alginate‐poly‐L‐lysine–alginate encapsulated myoblasts using flow cytometry
Authors: Thakur Ajit , Sengupta Ruchira , Matsui Hideto , Lillicrap David , Jones Kim , Hortelano Gonzalo
Published: May 2010

Genetically modified cells encapsulated in alginate‐poly‐L‐lysine‐alginate (APA) are being developed to deliver therapeutic products to treat a variety of diseases. The characterization of the encapsulated cells thus becomes paramount. This study reports a novel method to assess the viability, granularity and proliferation of encapsulated cells based on flow cytometry. The in vitro viability of encapsulated G8 murine myoblasts secreting canine FVIII (cFVIII) measured by flow cytometry was comparable to the traditional trypan blue exclusion method and both correlated with cFVIII secretion levels. In contrast, after implantation into mice, only viability measured by flow cytometry correlated with cFVIII secretion. Further, flow cytometry analysis of encapsulated cells maintained in vitro and in vivo revealed a greater fraction of granular cells compared to free cells, suggesting that encapsulation influences the morphology (cytoplasmic composition) of cells within APA microcapsules. Interestingly, the proliferation study showed that encapsulated cells proliferate faster, on average, and were more heterogeneous in vivo compared to in vitro culture conditions, suggesting that encapsulated cell proliferation is complex and environment‐dependent. In conclusion, we show that flow cytometry analysis allows for a more consistent and comprehensive examination of encapsulated cells to aid in the development of cell therapy protocols. © 2010 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2010.
Anti‐Proliferative Effects of Evodiamine on Human Thyroid Cancer Cell Line ARO
Authors: Chen Meng‐Ching , Yu Ching‐Han , Wang Shyi‐Wu , Pu Hsiao‐Fung , Kan Shu‐Fen , Lin Lie‐Chwen , Chi Chin‐Wen , Ho Lary Low‐Tone , Lee Chen‐Hsen , Wang Paulus S.
Published: May 2010

The incidence of thyroid cancer increases with age, and it is twice in women as common as in men. The undifferentiated thyroid cancer (UTC) is the most aggressive of all thyroid cancers. Unfortunately, there are almost no efficacious therapeutic modalities. It is important to develop some new effective therapies. Evodiamine is a chemical extracted from a kind of Chinese herb named Wu‐Chu‐Yu and has been demonstrated to be effective in preventing the growth of a variety of cancer cells. In the present study, the mechanism by which evodiamine inhibited the undifferentiated thyroid cancer cell line ARO was examined. Based on 3‐(4,5‐dimethylthiazol ‐2‐yle)2,5‐diphenyltetrazolium bromide (MTT) assay, cell proliferation rate was reduced dose‐dependently by evodiamine, but not by rutaecarpine. According to the flow cytometric analysis, evodiamine treatment resulted in G2/M arrest and DNA fragmentation in ARO cells. The G2/M arrest was accompanied with an increase of the expression of cdc25C, cyclin B1, and cdc2‐p161 protein, and it was also with a decrease of the expression of cdc2‐p15. Furthermore, by using the TUNEL assay, evodiamine‐induced apoptosis was observed at 48 h and extended to 72 h. Western blotting demonstrated that evodiamine treatment induced the activation of caspase‐8, caspase‐9, caspase‐3, and the cleavage of poly ADP‐ribose polymerase (PARP). These results suggested that evodiamine inhibited the growth of the ARO cells, arrested them at M phase, and induced apoptosis through caspases signaling. J. Cell. Biochem. 110: 1495–1503, 2010. © 2010 Wiley‐Liss, Inc.
On the stability of metal–aminoacid complexes in water based on water–ligand exchange reactions and electronic properties: Detailed study on iron–glycine hexacoordinated complexes
Authors: Mandado Marcos , Cordeiro M. Natália D. S.
Published: May 2010

Thermodynamic stability of metal–aminoacid complexes in water is discussed in terms of the Gibbs free energy of water–ligand exchange processes, and the electronic stabilizing factors thoroughly investigated by means of 1‐electron and 2‐electron density properties. Hexacoordinated complexes formed between iron cations and glycine molecules acting as monodentate or bidentate ligands have been chosen as targets for the current study. Results agree with experimental findings, and complexes formed with bidentate ligands are found to be more stable than those formed with monodentate ones. The larger the number of the coordinated glycine molecules the more stable is the complex. Fe(III) complexes are more stable than Fe(II) ones, but differences are small and the Fe3+/Fe2+exchange process appears to be energetically feasible for these complexes. Formation of the second glycine–iron interaction involving the amino nitrogen in the bidentate ligands is enthalpycally unfavorable but takes place due to the large entropy rise of the process. The larger stability of Fe(III) complexes is due however to the balance between energetic and solvation terms, which is favorable to these complexes. Electron density properties account satisfactorily for the electronic energy changes along the complex formation in terms of ligand–metal electron transfer and covalent bond orders. © 2010 Wiley Periodicals, Inc. J Comput Chem 2010
Theoretical studies on the reactions CH3SCH3 with OH, CF3, and CH3 radicals
Authors: Zhang Hui , Zhang Gui‐Ling , Liu Jing‐Yao , Sun Miao , Liu Bo , Li Ze‐Sheng
Published: May 2010

The multiple‐channel reactions OH + CH3SCH3 → products, CF3 + CH3SCH3 → products, and CH3 + CH3SCH3 → products are investigated by direct dynamics method. The optimized geometries, frequencies, and minimum energy path are all obtained at the MP2/6‐31+G(d,p) level, and energetic information is further refined by the MC‐QCISD (single‐point) method. The rate constants for eight reaction channels are calculated by the improved canonical variational transition state theory with small‐curvature tunneling contribution over the temperature range 200–3000 K. The total rate constants are in good agreement with the available experimental data and the three‐parameter expressions k1 = 4.73 × 10−16T1.89 exp(−662.45/T), k2 = 1.02 × 10−32T6.04 exp(933.36/T), k3 = 3.98 × 10−35T6.60 exp(660.58/T) (in unit of cm3 molecule−1 s−1) over the temperature range of 200–3000 K are given. Our calculations indicate that hydrogen abstraction channels are the major channels and the others are minor channels over the whole temperature range. © 2010 Wiley Periodicals, Inc. J Comput Chem, 2010
vmdICE: A plug‐in for rapid evaluation of molecular dynamics simulations using VMD
Authors: Knapp Bernhard , Lederer Nadja , Omasits Ulrich , Schreiner Wolfgang
Published: May 2010

Molecular dynamics (MD) is a powerful in silico method to investigate the interactions between biomolecules. It solves Newton's equations of motion for atoms over a specified period of time and yields a trajectory file, containing the different spatial arrangements of atoms during the simulation. The movements and energies of each single atom are recorded. For evaluating of these simulation trajectories with regard to biomedical implications, several methods are available. Three well‐known ones are the root mean square deviation (RMSD), the root mean square fluctuation (RMSF) and solvent accessible surface area (SASA). Herein, we present a novel plug‐in for the software “visual molecular dynamics” (VMD) that allows an interactive 3D representation of RMSD, RMSF, and SASA, directly on the molecule. On the one hand, our plug‐in is easy to handle for inexperienced users, and on the other hand, it provides a fast and flexible graphical impression of the spatial dynamics of a system for experts in the field. © 2010 Wiley Periodicals, Inc. J Comput Chem, 2010
Combat experience and the acquired capability for suicide
Authors: Bryan Craig J. , Cukrowicz Kelly C. , West Christopher L. , Morrow Chad E.
Published: May 2010

Rising suicide rates are an increasing concern among military personnel. The interpersonal‐psychological theory of suicide proposes that three necessary factors are needed to die by suicide: feelings that one does not belong with other people, feelings that one is a burden on others or society, and an acquired capability to overcome the fear and pain associated with suicide. The current study tests the theory's proposal that acquired capability may be particularly influenced by military experience, because combat exposure may cause habituation to fear of painful experiences such as suicide. Utilizing clinical and nonclinical samples of military personnel deployed to Iraq, results of the current study indicate that a greater range of combat experiences predicts acquired capability above and beyond depression and post‐traumatic stress disorder symptoms, previous suicidality, and other common risk factors for suicide. Combat experiences did not, however, predict perceived burdensomeness or thwarted belongingness. The authors discuss how combat experiences might serve as a mechanism for elevating suicide risk and implications for clinical interventions and suicide prevention efforts. © 2010 Wiley Periodicals, Inc. J Clin Psychol: 66:1–13, 2010.
Evaluation of antioxiodant drugs for the treatment of ochronotic alkaptonuria in an in vitro human cell model
Authors: Tinti Laura , Spreafico Adriano , Braconi Daniela , Millucci Lia , Bernardini Giulia , Chellini Federico , Cavallo Giovanni , Selvi Enrico , Galeazzi Mauro , Marcolongo Roberto , Gallagher James A. , Santucci Annalisa
Published: May 2010

Alkaptonuria (AKU) is a rare autosomal recessive disease, associated with deficiency of homogentisate 1,2‐dioxygenase activity in the liver. This leads to an accumulation of homogentisic acid (HGA) and its oxidized derivatives in polymerized form in connective tissues especially in joints. Currently, AKU lacks an appropriate therapy. Hence, we propose a new treatment for AKU using the antioxidant N‐acetylcysteine (NAC) administered in combinations with ascorbic acid (ASC) since it has been proven that NAC counteracts the side‐effects of ASC. We established an in vitro cell model using human articular primary chondrocytes challenged with an excess of HGA (0.33 mM). We used this experimental model to undertake pre‐clinical testing of potential antioxidative therapies for AKU, evaluating apoptosis, viability, proliferation, and metabolism of chondrocytes exposed to HGA and treated with NAC and ASC administered alone or in combination addition of both. NAC decreased apoptosis induced in chondrocytes by HGA, increased chondrocyte growth reduced by HGA, and partially restored proteoglycan release inhibited by HGA. A significantly improvement in efficacy was found with combined addition of the two antioxidants in comparison with NAC and ASC alone. Our novel in vitro AKU model allowed us to demonstrate the efficacy of the co‐administration of NAC and ASC to counteract the negative effects of HGA for the treatment of ochronotic arthropathy. J. Cell. Physiol. 225: 84–91, 2010. © 2010 Wiley‐Liss, Inc.
Dauricine induces apoptosis, inhibits proliferation and invasion through inhibiting NF‐κB signaling pathway in colon cancer cells
Authors: Yang Zhengfeng , Li Chenghai , Wang Xiu , Zhai Chunyan , Yi Zhengfang , Wang Lei , Liu Bisheng , Du Bing , Wu Huihui , Guo Xizhi , Liu Mingyao , Li Dali , Luo Jian
Published: May 2010

Dauricine, a bioactive component of Asiatic Moonseed Rhizome, has been widely used to treat a large number of inflammatory diseases in traditional Chinese medicine. In our study, we demonstrated that dauricine inhibited colon cancer cell proliferation and invasion, and induced apoptosis by suppressing nuclear factor‐kappaB (NF‐κB) activation in a dose‐ and time‐dependent manner. Addition of dauricine inhibited the phosphorylation and degradation of IκBα, and the phosphorylation and translocation of p65. Moreover, dauricine down‐regulated the expression of various NF‐κB‐regulated genes, including genes involved cell proliferation (cyclinD1, COX2, and c‐Myc), anti‐apoptosis (survivin, Bcl‐2, XIAP, and IAP1), invasion (MMP‐9 and ICAM‐1), and angiogenesis (VEGF). In athymic nu/nu mouse model, we further demonstrated that dauricine significantly suppressed colonic tumor growth. Taken together, our results demonstrated that dauricine inhibited colon cancer cell proliferation, invasion, and induced cell apoptosis by suppressing NF‐κB activity and the expression profile of its downstream genes. These findings provide evidence for a novel role of dauricine in preventing or treating colon cancer through modulation of NF‐κB singling pathway. J. Cell. Physiol. 225: 266–275, 2010. © 2010 Wiley‐Liss, Inc.
Potential of ultraviolet wide‐field imaging and multiphoton microscopy for analysis of dehydroergosterol in cellular membranes
Authors: Wüstner Daniel , Brewer Jonathan R. , Bagatolli Luis , Sage Daniel
Published: May 2010

Dehydroergosterol (DHE) is an intrinsically fluorescent sterol with absorption/emission in the ultraviolet (UV) region and biophysical properties similar to those of cholesterol. We compared the potential of UV‐sensitive low‐light‐level wide‐field (UV‐WF) imaging with that of multiphoton (MP) excitation microscopy to monitor DHE in living cells. Significantly reduced photobleaching in MP microscopy of DHE enabled us to acquire three‐dimensional z‐stacks of DHE‐stained cells and to obtain high‐resolution maps of DHE in surface ruffles, nanotubes, and the apical membrane of epithelial cells. We found that the lateral resolution of MP microscopy is ∼1.5‐fold higher than that of UV‐WF deconvolution microscopy, allowing for improved spatiotemporal analysis of plasma membrane sterol distribution. Surface intensity patterns of DHE with a diameter of 0.2 μm persisting over several minutes could be resolved by MP time‐lapse microscopy. Diffusion coefficients of 0.25‐μm‐diameter endocytic vesicles containing DHE were determined by MP spatiotemporal image correlation spectroscopy. The requirement of extremely high laser power for visualization of DHE by MP microscopy made this method less potent for multicolor applications with organelle markers like green fluorescent protein–tagged proteins. The signal‐to‐noise ratio obtainable by UV‐WF imaging could be significantly improved by pixelwise bleach rate fitting and calculation of an amplitude image from the decay model and by frame averaging after pixelwise bleaching correction of the image stacks. We conclude that UV‐WF imaging and MP microscopy of DHE provide complementary information regarding membrane distribution and intracellular targeting of sterols. Microsc. Res. Tech. 74:92‐108, 2011. © 2010 Wiley‐Liss, Inc.
Amino acids–nucleotides biomolecular recognition: from biological occurrence to affinity chromatography
Authors: Sousa F. , Cruz C. , Queiroz J. A.
Published: May 2010

In this review, the protein–DNA interactions are discussed considering different perspectives, and the biological occurrence of this interaction is explained at atomic level. The evaluation of the amino acid–nucleotide recognition has been investigated analysing datasets for predicting the association preferences and the geometry that favours the interaction.

Based on this knowledge, an affinity chromatographic method was developed also exploiting this biological favoured contact. In fact, the implementation of this technique brings the possibility to apply the concept of molecular interactions to the development of new purification methodologies. In addition, the integration of the information recovered by all the different perspectives can bring new insights about some biological mechanisms, though not totally clarified. Copyright © 2010 John Wiley & Sons, Ltd.
Adipose tissue distribution in children: Automated quantification using water and fat MRI
Authors: Kullberg Joel , Karlsson Ann‐Katrine , Stokland Eira , Svensson Pär‐Arne , Dahlgren Jovanna
Published: May 2010

Purpose:

To develop and validate a method for rapid acquisition and automated processing of magnetic resonance (MR) images for analysis of abdominal adipose tissue distribution in children.

Materials and Methods:

The study included 21 (10 girls, 11 boys) healthy 5‐year‐old children. Rapid water and fat MR imaging (6 sec) was performed using a 2‐point‐Dixon technique on a 1.5T MR scanner using an 8‐channel cardiac coil. An automated image processing algorithm was developed for automated segmentation of visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT), respectively. The results from the fully automated analysis were compared to those from a semiautomated analysis, performed by three operators, from the same images.

Results:

The automated analysis was seen to give results with strong correlation to the reference measurements (r ≥ 0.997); however, the SAT volume was underestimated by 9.4 ± 3.8%. The accuracy of the automated segmentation of VAT and SAT (TP: true positive, FP: false positive, mean ± SD, %) was TP: 83.6 ± 8.5, FP: 12.7 ± 6.8; and TP: 89.9 ± 3.6, FP: 0.7 ± 0.3, respectively.

Conclusion:

A method for rapid imaging and fully automated postprocessing of abdominal adipose tissue distribution is presented. The method allows robust and time‐efficient measurement of adipose tissue distribution in young children. J. Magn. Reson. Imaging 2010. © 2010 Wiley‐Liss, Inc.
Dynamic visualization of arachnoid adhesions in a patient with idiopathic syringomyelia using high‐resolution cine magnetic resonance imaging at 3T
Authors: Gottschalk Andreas , Schmitz Bernd , Mauer Uwe M. , Bornstedt Axel , Steinhoff Silke , Danz Burkhardt , Schlötzer Wiebke , Rasche Volker
Published: May 2010

A 39‐year‐old female patient with thoracic syringomyelia underwent routine magnetic resonance imaging (MRI) and 3 T MRI to investigate the value of retrospectively cardiac‐gated cine steady‐state free precession (SSFP) MRI in the preoperative and postoperative diagnosis of arachnoid membranes in the spinal subarachnoid space. Therefore, 3T MRI included sagittal and transverse retrospectively cardiac‐gated cine balanced fast‐field echo (balanced‐FFE) sequences both preoperatively and after microsurgical lysis of arachnoid adhesions and expansive duraplasty. Arachnoid membranes were detected and this result was correlated with intraoperative findings and the results of routine cardiac‐gated phase‐contrast cerebrospinal fluid (CSF) flow MRI. Retrospectively cardiac‐gated cine SSFP MRI enabled imaging of arachnoid membranes with high spatial resolution and sufficient contrast to delineate them from hyperintense CSF preoperatively and postoperatively. The images were largely unaffected by artifacts. Surgery confirmed the presence of arachnoid adhesions in the upper thoracic spine. Not all arachnoid membranes that were seen on cine balanced‐FFE sequences caused significant spinal CSF flow blockages in cardiac‐gated phase‐contrast CSF flow studies. In conclusion, retrospectively cardiac‐gated cine SSFP MRI may become a valuable tool for the preoperative detection of arachnoid adhesions and the postoperative evaluation of microsurgical adhesiolysis in patients with idiopathic syringomyelia. J. Magn. Reson. Imaging 2010. © 2010 Wiley‐Liss, Inc.
Reducing the impact of white matter lesions on automated measures of brain gray and white matter volumes
Authors: Chard Declan T. , Jackson Jonathan S. , Miller David H. , Wheeler‐Kingshott Claudia A.M.
Published: May 2010

Purpose:

To develop an automated lesion‐filling technique (LEAP; LEsion Automated Preprocessing) that would reduce lesion‐associated brain tissue segmentation bias (which is known to affect automated brain gray [GM] and white matter [WM] tissue segmentations in people who have multiple sclerosis), and a WM lesion simulation tool with which to test it.

Materials and Methods:

Simulated lesions with differing volumes and signal intensities were added to volumetric brain images from three healthy subjects and then automatically filled with values approximating normal WM. We tested the effects of simulated lesions and lesion‐filling correction with LEAP on SPM‐derived tissue volume estimates.

Results:

GM and WM tissue volume estimates were affected by the presence of WM lesions. With simulated lesion volumes of 15 mL at 70% of normal WM intensity, the effect was to increase GM fractional (relative to intracranial) volumes by ≈2.3%, and reduce WM fractions by ≈3.6%. Lesion filling reduced these errors to ≈0.1%.

Conclusion:

The effect of WM lesions on automated GM and WM volume measures may be considerable and thereby obscure real disease‐mediated volume changes. Lesion filling with values approximating normal WM enables more accurate GM and WM volume measures and should be applicable to structural scans independently of the software used for the segmentation. J. Magn. Reson. Imaging 2010. © 2010 Wiley‐Liss, Inc.
VUV photon‐induced ionization/dissociation of antipyrine and propyphenazone: mass spectrometric and theoretical insights
Authors: Deng Liulin , Zhang Lidong , Guo Huijun , Jia Liangyuan , Pan Yang , Yin Hao , Qi Fei
Published: May 2010

Two analgesic and anti‐inflammatory drugs, antipyrine and propyphenazone, were investigated with infrared laser desorption/tunable synchrotron vacuum ultraviolet (VUV) photoionization mass spectrometry (IR LD/VUV PIMS) and theoretical calculations. Mass spectra of the two drugs were measured at various photon energies. Fragment ions were gradually produced as photon energy increases. The structural assignment of the dominant fragment ions was supported by the results from a commercial electron impact time‐of‐flight mass spectrometer (EI‐TOF MS). Primary fragmentation pathways were established from experimental observations combining with theoretical calculations. Methyl radical elimination is a common fragmentation pathway for two analytes. However, for propyphenazone cation, isopropyl group elimination to form antipyrine cation is another competitive pathway. Copyright © 2010 John Wiley & Sons, Ltd.
Appearance of phagocytic microglia adjacent to motoneurons in spinal cord tissue from a presymptomatic transgenic rat model of amyotrophic lateral sclerosis
Authors: Sanagi Tomomi , Yuasa Shigeki , Nakamura Yasuko , Suzuki Eri , Aoki Masashi , Warita Hitoshi , Itoyama Yasuto , Uchino Shigeo , Kohsaka Shinichi , Ohsawa Keiko
Published: May 2010

Microglial activation occurs early during the pathogenesis of amyotrophic lateral sclerosis (ALS). Recent evidence indicates that the expression of mutant Cu2+/Zn2+ superoxide dismutase 1 (SOD1) in microglia contributes to the late disease progression of ALS. However, the mechanism by which microglia influence the neurodegenerative process and disease progression in ALS remains unclear. In this study, we revealed that activated microglia aggregated in the lumbar spinal cord of presymptomatic mutant SOD1H46R transgenic rats, an animal model of familial ALS. The aggregated microglia expressed a marker of proliferating cell, Ki67, and phagocytic marker proteins ED1 and major histocompatibility complex (MHC) class II. The motoneurons near the microglial aggregates showed weak choline acetyltransferase (ChAT) immunoreactivity and contained reduced granular endoplasmic reticulum and altered nucleus electron microscopically. Furthermore, immunopositive signals for tumor necrosis factor‐α (TNFα) and monocyte chemoattractant protein‐1 (MCP‐1) were localized in the aggregated microglia. These results suggest that the activated and aggregated microglia represent phagocytic features in response to early changes in motoneurons and possibly play an important role in ALS disease progression during the presymptomatic stage. © 2010 Wiley‐Liss, Inc.
Investigation of annexin A5 as a biomarker for Alzheimer's disease using neuronal cell culture and mouse model
Authors: Yamaguchi Mami , Kokai Yasuo , Imai Shin‐Ichi , Utsumi Kumiko , Matsumoto Kyoichi , Honda Hirohito , Mizue Yuka , Momma Masako , Maeda Tetsu , Toyomasu Shozo , Ito Yoichi M. , Kobayashi Seijyu , Hashimoto Eri , Saito Toshikazu , Sohma Hitoshi
Published: May 2010

Alzheimer's disease (AD) differs from other forms of dementia in its relation to amyloid β peptide (Aβ). Aβ, a proteolytic product of amyloid precursor proteins (APP), has a toxic effect on neuronal cells, which involves perturbation of their Ca2+ homeostasis. This effect implies that changes of protein expression in neuronal cells with calcium stress should provide a molecular marker for this disease. In the present study, we used the supernatant from a neuronal cell culture after incubation with or without Aβ and isolated a Ca2+‐dependent acidic phospholipid binding fraction to perform a proteomic study. Several unique proteins were identified after incubation with Aβ. We focused on annexin A5, among these proteins, because it binds both Ca2+ and lipids likely to be involved in calcium homeostasis. Tg2576 transgenic mice (AD model) overexpressing mutant human APP showed a significant increase of annexin A5 in the brain cortex but not in other organs, including liver, kidney, lung, and intestine. In human plasma samples, the level of annexin A5 was significantly increased in a proportion of AD patients compared with a control group (P < 0.0001 in the logistic regression analysis). From the receiver operating characteristic (ROC) curve with plasma annexin A5 concentrations, the mean area under the curve (AUC 0.898) suggests that annexin A5 is a favorable marker for AD. © 2010 Wiley‐Liss, Inc.
Bone morphogenetic proteins 2, 5, and 6 in combination stimulate osteoblasts but not osteoclasts in vitro
Authors: Wutzl Arno , Rauner Martina , Seemann Rudolf , Millesi Werner , Krepler Petra , Pietschmann Peter , Ewers Rolf
Published: May 2010

Bone regeneration is required for fracture healing. Various procedures have been used to promote osteogenesis with bone morphogenetic proteins (BMPs). We assessed the effects of BMP‐2, BMP‐5, and BMP‐6 in isolated and combined use on the generation of osteoblasts and osteoclasts by comparing the osteoclastic potency of each on osteoclasts of primary murine bone marrow cells. Subsequently, cells were stained for tartrate‐resistant acid phosphatase, and real time PCR analysis of receptor activator of NKκB ligand and osteoprotegerin was conducted. The same combination of BMPs was used to assess their potential to enhance osteoblasts, employing a mineralization assay and real‐time PCR analysis of collagen type‐1, runx2, and osterix. While BMP‐2 alone and the combination of BMP‐2 and BMP‐5 significantly enhanced osteoclastogenesis, BMP‐2, BMP‐5, and BMP‐6 in combination did not have additional effects. However, the combined use of BMP‐2, BMP‐5, and BMP‐6 had an additive effect on matrix mineralization and osterix expression in osteoblasts. Our study shows that the combination of BMP‐2, BMP‐5, and BMP‐6 stimulates osteoblasts but not osteoclastogenesis. Thus, the synergistic use of various BMPs might improve effective bone regeneration in the clinical setting. © 2010 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res
Temporal tissue patterns in bone healing of sheep
Authors: Vetter Andreas , Epari Devakara R. , Seidel Robin , Schell Hanna , Fratzl Peter , Duda Georg N. , Weinkamer Richard
Published: May 2010

Secondary fracture healing in long bones leads to the successive formation of intricate patterns of tissues in the newly formed callus. The main aim of this work was to quantitatively describe the topology of these tissue patterns at different stages of the healing process and to generate averaged images of tissue distribution. This averaging procedure was based on stained histological sections (2, 3, 6, and 9 weeks post‐operatively) of 64 sheep with a 3 mm tibial mid‐shaft osteotomy, stabilized either with a rigid or a semi‐rigid external fixator. Before averaging, histological images were sorted for topology according to six identified tissue patterns. The averaged images were obtained for both fixation types and the lateral and medial side separately. For each case, the result of the averaging procedure was a collection of six images characterizing quantitatively the progression of the healing process. In addition, quantified descriptions of the newly formed cartilage and the bone area fractions (BA/TA) of the bony callus are presented. For all cases, a linear increase in the BA/TA of the bony callus was observed. The slope was greatest in the case of the most rigid stabilization and lowest in the case of the least stiff. This topological description of the progression of bone healing will allow quantitative validation (or falsification) of current mechano‐biological theories. © 2010 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 28:1440–1447, 2010
The position and context of Middle Palaeolithic industries from the Ebbsfleet Valley, Kent, UK
Authors: Scott Beccy , Ashton Nick , Penkman Kirsty E. H. , Preece Richard C. , White Mark
Published: May 2010

The late Middle Pleistocene sites in the Ebbsfleet Valley, Kent, UK, have yielded archaeological assemblages critical to understanding the early Middle Palaeolithic of northwestern Europe. Despite a long history of research, the nature and context of these assemblages are still poorly understood. This paper clarifies the stratigraphic, environmental and archaeological records at Ebbsfleet. These reflect a cold–warm–cold sequence of climatic events, preserved within part of the Taplow/Mucking Formation of the Thames (Marine Isotope Stage (MIS) 8/7/6). Levallois artefacts are shown to be restricted to the lower part of the Ebbsfleet Channel sequence (Phases I and II) and are assigned to late MIS 8/early MIS 7. This material is associated with fauna indicative of open environments during both cool and temperate conditions. Handaxe assemblages are recorded from higher up the sequence (Phases III–V), but have been redeposited from higher terrace units nearby. No primary context archaeology is apparent during these later phases of aggradation. This may indicate that humans abandoned the site once available raw material became inaccessible, and may also reflect a decline in human presence in Britain during the latter part of MIS 7. Copyright © 2010 John Wiley & Sons, Ltd.
Did the Indo‐Asian summer monsoon decrease during the Holocene following insolation?
Authors: Tiwari Manish , Ramesh Rengaswamy , Bhushan Ravi , Sheshshayee Madavalm S. , Somayajulu Bammidipati L. K. , Jull A. J. Timothy , Burr George S.
Published: May 2010

A few studies from the western Arabian Sea indicate that the Indian summer (or southwest) monsoon (ISM), after attaining its maximum intensity at ca. 9 ka, declined during the Holocene, as did insolation. In contrast, earlier and later observations from both the eastern and the western Arabian Sea do not support this inference. Analysis of multiple proxies of productivity in a new sediment core from the western Arabian Sea fails to confirm the earlier, single‐proxy (e.g. abundance of Globigerina bulloides) based, inference of the Holocene weakening of ISM, following insolation. The reason for the observed decreasing trend in foraminiferal abundance – the basis for the earlier inference – could be the favouring of silicate rather than carbonate productivity by the increased ISM wind strength. Although ISM exhibits several multi‐millennial scale fluctuations, there is no evidence from several multi‐proxy data to conclude that it declined during the Holocene; this is consistent with the phase lag analysis of longer time series of monsoon proxies. Thus, on sub‐Milankovitch timescales, ISM did not follow insolation, highlighting the importance of internal feedbacks. A comparison with East Asian summer monsoon (EASM) records suggests that both ISM and EASM varied in unison, implying common forcing factors on such longer timescales. Copyright © 2010 John Wiley & Sons, Ltd.
Severe tumor budding is a risk factor for lateral lymph node metastasis in early rectal cancers
Authors: Homma Yoichiro , Hamano Takashi , Otsuki Yoshiro , Shimizu Shinichi , Kobayashi Hiroshi , Kobayashi Yasuyuki
Published: May 2010

Background

Lateral lymph node (LLN) metastasis sometimes occurs in patients with early rectal cancer that has invaded the submucosa (SM) and muscularis propria (MP). This study aims to identify the risk factor(s) for LLN metastasis in such patients.

Method

We retrospectively analyzed 65 patients with pathological SM or MP lower rectal adenocarcinoma, for whom radical resection had been performed at a single institution.

Results

We performed LLN dissection in 52 (80%) patients. The LLN dissection rates in the case of pathological SM and MP tumors were 65.6% and 94.4%, respectively, and the corresponding LLN metastasis rates were 6.9% and 11.1%. Severe tumor budding was found to be a risk factor for LLN metastasis (P = 0.002). Further, of six patients with LLN metastasis, four did not have coincident mesenteric lymph node metastasis.

Conclusion

In rectal cancer that has pathologically invaded SM and MP, LLN metastasis is not negligible. LLN dissection could lower the local recurrence rate of SM and MP rectal cancer. In case LLN dissection is not performed, patients with a high tumor budding grade should be administered adjuvant therapy. J. Surg. Oncol. 2010;102:230–234. © 2010 Wiley‐Liss, Inc.
Safety and efficacy of curative intent surgery for peri‐ampullary liver metastasis
Authors: de Jong Mechteld C. , Tsai Susan , Cameron John L. , Wolfgang Christopher L. , Hirose Kenzo , van Vledder Mark G. , Eckhauser Frederic , Herman Joseph M. , Edil Barish H. , Choti Michael A. , Schulick Richard D. , Pawlik Timothy M.
Published: May 2010

Introduction

The management of patients with peri‐ampullary liver metastasis remains controversial. We sought to assess the safety and efficacy of curative intent surgery for peri‐ampullary liver metastasis.

Methods

Between 1993 and 2009, 40 patients underwent curative intent surgery (resection and/or radiofrequency ablation (RFA)) for peri‐ampullary liver metastasis. Clinicopathologic and outcome data were collected and analyzed.

Results

Location of the primary tumor was pancreas head (n = 20), ampulla of Vater (n = 10), distal bile duct (n = 5), or duodenum (n = 5). Most patients (n = 27) presented with synchronous disease, while 13 patients presented with metachronous disease following a median disease‐free interval of 22 months. Most patients (n = 25) presented with hepatic metastasis from pancreaticobiliary origin (pancreatic or distal common bile duct) compared with 15 patients who had metastasis from an intestinal‐type primary (ampullary or duodenal). There were no differences in metastatic tumor number or size between these groups (P > 0.05). Post‐operative morbidity and mortality was 30% and 5% respectively. Overall 1‐ and 3‐year survival was 55% and 18%. Patients who underwent resection of liver metastasis from intestinal‐type tumors experienced a longer survival compared with patients who had pancreaticobiliary lesions (median: 13 months vs. 23 months; P = 0.05).

Conclusion

Curative intent surgery for peri‐ampullary liver metastasis was associated with post‐operative morbidity and a 5% mortality rate. Although the overall survival benefit was modest, patients with liver metastasis from intestinal‐type tumors experienced improved survival following resection of liver metastasis compared with pancreaticobiliary lesions. J. Surg. Oncol. 2010;102:256–263. © 2010 Wiley‐Liss, Inc.
Tumour budding, uPA and PAI‐1 are associated with aggressive behaviour in colon cancer
Authors: Märkl B. , Renk Prof. I. , Oruzio D.V. , Jähnig H. , Schenkirsch G. , Schöler C. , Ehret Prof. W. , Arnholdt H.M. , Anthuber M. , Spatz H.
Published: May 2010

Aims

The proteases PAI‐1 and uPA play a major role in extracellular matrix degradation, which facilitates tumour progression. Tumour budding is a histomorphological expression of enhanced tumour cell migration.

Materials and Methods

To investigate their prognostic value for and correlation with colon cancer, a prospective study was performed. We analysed tissue levels of uPA and PAI‐1 of 55 colon cancer tumours employing a commercially available enzyme‐linked immunosorbent assay (ELISA). Tumour budding was analysed on cytokeratin‐stained slides.

Results

There was a strong correlation between uPA and tumour budding (R = 0.440; P < 0.001). uPA levels were increased in high grade tumours, whereas PAI‐1 was elevated in cases with venous invasion (P = 0.004 and P = 0.028). PAI‐1 values and tumour budding are associated significantly with the occurrence of distant metastases (P < 0.001 and P = 0.034, respectively). Tumour budding was significantly associated with lymph node metastases (P = 0.034). Multivariate analysis revealed PAI‐1 and lymph node metastases to be independently predictive of distant metastases (P = 0.007 and P = 0.004, respectively).

Conclusions

The results of our study show that tumour budding and the plasmin/plasminogen system are related. PAI‐1 was independently predictive for the occurrence of distant metastasis. J. Surg. Oncol. 2010;102:235–241. © 2010 Wiley‐Liss, Inc.
Extraction, separation and quantitative structure–retention relationship modeling of essential oils in three herbs
Authors: Wei Yuhui , Xi Lili , Chen Dongxia , Wu Xin'an , Liu Huanxiang , Yao Xiaojun
Published: May 2010

The essential oils extracted from three kinds of herbs were separated by a 5% phenylmethyl silicone (DB‐5MS) bonded phase fused‐silica capillary column and identified by MS. Seventy‐four of the compounds identified were selected as origin data, and their chemical structure and gas chromatographic retention times (RT) were performed to build a quantitative structure–retention relationship model by genetic algorithm and multiple linear regressions analysis. The predictive ability of the model was verified by internal validation (leave‐one‐out, fivefold, cross‐validation andY‐scrambling). As for external validation, the model was also applied to predict the gas chromatographic RT of the 14 volatile compounds not used for model development from essential oil ofRadix angelicae sinensis. The applicability domain was checked by the leverage approach to verify prediction reliability. The results obtained using several validations indicated that the best quantitative structure–retention relationship model was robust and satisfactory, could provide a feasible and effective tool for predicting the gas chromatographic RT of volatile compounds and could be also applied to help in identifying the compound with the same gas chromatographic RT.
Expression of advanced glycation end‐product receptors in the cochlea
Authors: Hanusek Claudia , Setz Cristian , Radojevic Vesna , Brand Yves , Levano Soledad , Bodmer Daniel
Published: May 2010

Objectives/Hypothesis:

Advanced glycation end products (AGE) have recently been implicated in aging changes within different tissues of the body. The role of AGEs and their receptors in the mammalian inner ear is largely unknown. In this study we analyzed for the expression of two AGE receptors, namely RAGE and Ddost (AGE‐R1).

Study Design:

Controlled animal study. Controlled animal study. This study confirmed the expression of the AGEs receptors RAGE and Ddost (AGE‐R1) both on mRNA and protein level in the cochlea. Furthermore, we were able to localize these two receptors in the organ of Corti (OC).

Methods:

Expression of RAGE and Ddost (AGE‐R1) receptors in the rat cochlea were analyzed by reverse transcriptase‐polymerase chain reaction (RT‐PCR) and Western blot. Specific localization of these two AGE receptors was also obtained within the mouse OC using immunohistochemistry.

Results:

We detected RAGE and Ddost (AGE‐R1) at the mRNA and protein level in the OC, spiral ganglion, and stria vascularis. Moreover, RAGE and Ddost (AGE‐R1) could specifically be identified within the immature and mature OC.

Conclusions:

The AGE receptors RAGE and Ddost (AGE‐R1) could be identified within the cochlea. Different expression patterns of these two receptors were observed in the immature and mature inner ear. Laryngoscope, 120:1227–1232, 2010
Effectiveness of lubricant additives for copper‐alloy‐steel sliding contacts
Authors: Jisa Robin , Ristic Andjelka , Brenner Josef , Lebersorger Thomas , Ilo Sotiraq , Neumayer Hans , Franek Friedrich
Published: May 2010

A selection of additives and their performance and compatibility with a variety of copper alloys have been evaluated in an SRV test set‐up. The tests show a remarkable variation of tribological behaviour with a clear relation to both the type of lubricant / additive and the type of alloy. One ester‐based additive showed outstanding friction and wear reduction for some groups of copper alloys. In order to better understand the fundamental mechanisms, we applied a variety of surface analyses, such as 3D confocal white light microscopy, scanning electron microscopy and X‐ray photoelectron microscopy. Copyright © 2010 John Wiley & Sons, Ltd.
Usefulness 18F-FDG positron emission tomography/computed tomography for detecting recurrence of hepatocellular carcinoma in posttransplant patients
Authors: Kim Young-Kyu , Lee Kwang-Woong , Cho Seong Yeon , Han Sung-Sik , Kim Seong Hoon , Kim Seok-Ki , Park Sang-Jae
Published: May 2010

18F-fluoro-2-deoxy-D-glucose (18F-FDG) positron emission tomography (PET)/computed tomography (CT) has recently been shown to be able to predict a poor outcome after liver transplantation (LT) for patients with hepatocellular carcinoma (HCC). However, there are few reports on the usefulness of PET during follow-up after LT. In this study, we assessed the efficacy of 18F-FDG PET/CT for the detection of HCC recurrence after LT. From February 2005 to December 2008, out of 93 adult LT cases (91 living donors and 2 deceased donors), 10 patients who showed HCC recurrence and received 18F-FDG PET/CT during follow-up were included. The accuracy of 18F-FDG PET/CT was assessed with imaging and histological studies. The most common sites of recurrence were extrahepatic (60%). The most common extrahepatic sites were the lungs and bone (31.3% each). Among 4 patients with intrahepatic recurrence, 1 patient (25%) was positive according to 18F-FDG PET/CT. The detection rate of 18F-FDG PET/CT was 92.9% for extrahepatic metastases ≥ 1 cm and 0% for lesions < 1 cm. The detection rate of 18F-FDG PET/CT was 100% in bone and the lymph nodes, 60% in the lungs, and 0% in the brain. 18F-FDG PET/CT identified 2 lesions in bone that were not found in a bone scan. In conclusion, because of its limitations for small lesions, intrahepatic lesions, and brain lesions, 18F-FDG PET/CT is not suitable as a screening tool after LT. However, 18F-FDG PET/CT could provide additional information beyond that provided by conventional modalities, and it could contribute to the clinical management of HCC recurrence after LT, especially in patients with extrahepatic recurrence. Liver Transpl 16:767-772, 2010. © 2010 AASLD.
Iron Tris(dibenzoylmethane) Centered PCL and PCL‐b‐PLA Stars from Dual Functional Catalyst‐Initiators
Authors: Chen Jianbin , Gorczynski Jessica L. , Fraser Cassandra L.
Published: May 2010

Iron(III)‐centered star polymers of poly(ε‐caprolactone) (PCL) and copolymers with polylactide have been synthesized by a divergent ring‐opening polymerization initiated from an alcohol‐functionalized dibenzoylmethane (dbm) complex. This strategy provides a route to well‐defined polyester products with relatively high molecular weights and low PDIs. These reactions require no added catalyst. The resulting Fe‐centered star polymers and the associated polymeric ligands obtained upon Fe complex fragmentation in acid are responsive materials that could find application in delivery vectors, sensors, supported polymeric catalysts, and optical materials.

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Poly(ε‐caprolactone)‐graft‐poly(2‐(dimethylamino)ethyl methacrylate) Amphiphilic Copolymers Prepared via a Combination of ROP and ATRP: Synthesis, Characterization, and Self‐Assembly Behavior
Authors: Guo Shutao , Wang Weiwei , Deng Liandong , Xing Jinfeng , Dong Anjie
Published: May 2010

Poly(ε‐caprolactone)‐graft‐poly(2‐(dimethylamino) ethyl methacrylate) (PCL‐g‐PDMAEMAs), a kind of amphiphilic graft copolymer, was prepared by combination of ROP and ATRP. The FTIR, 1H NMR, and GPC results indicate that well‐defined polymers with controlled graft density and length of side chain were successfully synthesized. We prepared PCL‐g‐PDMAEMA nanoparticles by employing a nanoprecipitation technique. The pH‐ and thermosensitive properties of PCL‐g‐PDMAEMA nanoparticles were investigated by 1H NMR, TEM, and DLS. It was found that the nanoparticles with an average size of 120 nm presented core–shell structure in aqueous dispersion. Furthermore, the nanoparticles are sensitive to temperature in base while not in an acidic environment.

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Poly[ethylene‐co‐(methacrylic acid)] Healing Agents for Mendable Carbon Fiber Laminates
Authors: Meure Sam , Furman Scott , Khor Sarah
Published: May 2010

The first reported use of two‐dimensional mesh thermoplastic fibers in an epoxy matrix for mendable composites is presented, yielding 100% restoration of GIC, failure energy, and peak loads over repeated damage‐healing cycles. SEM imaging and EDS mapping showed different surface structures between CFRPp and CFRPf and confirmed strength recoveries were attained by delivery of EMAA to the fracture plane which enabled the fractured surfaces to rebind after heating to 150 °C for 30 min.

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Deformation of a singularity of type E8 and Mordell‐Weil lattices in characteristic 2
Authors: Ito Hiroyuki
Published: May 2010

In this paper, we study the Mordell‐Weil lattices of the family of elliptic surfaces which is arising from the E84 singularity, one of the ADE singularities in characteristic 2. And we construct a subfamily of the universal family of supersingular K 3 surfaces in characteristic 2 as an application (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Generalized implicit Euler method for hyperbolic functional differential equations
Authors: Czernous Wojciech
Published: May 2010

Nonlinear hyperbolic functional differential equations with initial boundary conditions are considered. Theorems on the convergence of difference schemes and error estimates of approximate solutions are presented. The proof of the stability of the difference functional problem is based on a comparison technique. Nonlinear estimates of the Perron type with respect to the functional variable for given functions are used. Numerical examples are given (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Injectable Biodegradable Poly(ethylene glycol)/RGD Peptide Hybrid Hydrogels for in vitro Chondrogenesis of Human Mesenchymal Stem Cells
Authors: Liu Shao Qiong , Tian Quan , Wang Lei , Hedrick James L. , Hui James Hoi Po , Yang Yi Yan , Ee Pui Lai Rachel
Published: May 2010

In this study, an injectable and biodegradable poly(ethylene glycol) (PEG)/arginine–glycine–aspartic (RGD) peptide hybrid hydrogel has been synthesized and used as a biomimetic scaffold for encapsulation of human mesenchymal stem cells (hMSCs). Tetrahydroxyl PEG was functionalized with acrylate, and then reacted with thiol‐containing peptide (RGD). Gelation occurred within 30 min with the addition of cells and PEG‐dithiol via Michael addition. The hydrogels synthesized with a peptide concentration of 1.0–5.0 mM achieved significantly greater cell viability when compared to the hydrogels without the RGD peptide. However, the effect of RGD on chondrogenesis was found to be dose‐dependent. Immunohistology studies demonstrated that hMSCs encapsulated in the hydrogel matrix with 1.0 mM RGD and TGF‐β3 showed enhanced positive staining for aggrecan and type II collagen as compared to that with 5.0 mM RGD and unmodified PEG hydrogels. RT‐PCR results further revealed that the cells in hydrogels with 1 mM RGD expressed significantly higher levels of type II collagen than those in PEG hydogels without RGD peptide. These findings have demonstrated that the PEG‐RGD hydrogels can be a promising scaffold to deliver hMSCs for cartilage repair.

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