bluuebunny/crossref_metadata_2025 huggingface dataset:
Multilineage differentiation potential of cells isolated from the human amniotic membrane
Authors: Díaz‐Prado Silvia , Muiños‐López Emma , Hermida‐Gómez Tamara , Rendal‐Vázquez Maria Esther , Fuentes‐Boquete Isaac , de Toro Francisco J. , Blanco Francisco J.
Published: July 2010

The human amniotic membrane (HAM) contains two cell types from different embryological origins. Human amnion epithelial cells (hAECs) are derived from the embryonic ectoderm, while human amnion mesenchymal stromal cells (hAMSCs) are derived from the embryonic mesoderm. In this study, we localized, isolated, quantified and phenotypically characterized HAM‐derived cells and analysed their in vitro differentiation potential towards mesodermal cell lineages. Human amnion‐derived cells were isolated and characterized by flow cytometry. Immunohistochemistry and quantitative real‐time reverse transcription‐polymerase chain reaction studies were performed for the analysis of multipotentiality. Immunophenotypic characterization of both cell types demonstrated the presence of the common, well‐defined human mesenchymal stem cell (MSC) markers (CD90, CD44, CD73, CD166, CD105, CD29), as well as the embryonic stem‐cell markers SSEA‐4 and STRO‐1. Phenotypes of both cell populations were maintained from passages P0 to P9. The assessment of multilineage potential demonstrated that the hAMSCs showed greater adipogenic and chondrogenic potential. Both populations had the ability to retain their capacity for differentiation during culture passages from P0 to P4. Our data demonstrate the successful localization and isolation of hAMSCs and hAECs from the HAM. Both cell populations possessed similar immunophenotype. However, they differed in cell yield and multipotential for differentiation into the major mesodermal lineages. Our functional differentiation studies demonstrated that hAMSCs possess a much greater mesodermal differentiation capacity than hAECs. These considerations will be important for use of these cells for cell therapy. J. Cell. Biochem. 111: 846–857, 2010. © 2010 Wiley‐Liss, Inc.
Lancemaside A inhibits lipopolysaccharide‐induced inflammation by targeting LPS/TLR4 complex
Authors: Joh Eun‐Ha , Kim Dong‐Hyun
Published: July 2010

In our previous study, lancemaside A isolated from Codonopsis lanceolata (family Campanulaceae) ameliorated colitis in mice. In this study, the anti‐inflammatory effects of lancemaside A was investigated in lipopolysaccharide (LPS)‐stimulated mice and their peritoneal macrophage cells. Lancemaside A suppressed the production of pro‐inflammatory cytokines, TNF‐α and IL‐1β, in vitro and in vivo. Lancemaside A also down‐regulated inducible nitric oxide synthase (iNOS) and cyclooxygenase‐2 (COX‐2), as well as the inflammatory mediators, nitric oxide (NO), and PGE2. Lancemaside A also inhibited the expression of IL‐1 receptor‐associated kinase‐4 (IRAK‐4), the phosphorylation of IKK‐β and IκB‐α, the nuclear translocation of NF‐κB and the activation of mitogen‐activated protein kinases in LPS‐stimulated peritoneal macrophages. Furthermore, lancemaisde A inhibited the interaction between LPS and TLR4, as well as IRAK‐4 expression in peritoneal macrophages. Based on these findings, lancemaside A expressed anti‐inflammatory effects by regulating both the binding of LPS to TLR4 on macrophages. J. Cell. Biochem. 111: 865–871, 2010. © 2010 Wiley‐Liss, Inc.
Teratomas from pluripotent stem cells: A clinical hurdle
Authors: Fong Chui‐Yee , Gauthaman Kalamegam , Bongso Ariff
Published: July 2010

Although basic research on human embryonic stem cells (hESCs) at the laboratory bench has progressed with enviable speed there has been little head way in terms of its clinical application. A look at the Internet however shows several stem cell clinics worldwide offering direct transplantation of undifferentiated hESCs to patients for the cure of a variety of diseases before bona fide evidence‐based results can be demonstrated from large controlled studies. This raises concern because reliable protocols have to be first developed to resolve the three major hurdles delaying clinical trials such as inadequate cell numbers, immunorejection and tumorigenesis. Cell expansion methods using bioreactors, rotary culture and mitotic agents have now been developed to generate stem cell derivatives in large numbers. The problem of immunorejection can now be overcome with the development of indirect and direct reprogramming protocols to personalize tissues to patients (human induced pluripotent stem cells, hiPSCs; nuclear transfer stem cells, NTSCs; induced neuronal cells, iN). However, hESC, hiPSC, and NTSCs being pluripotent have the disadvantage of teratoma formation in vivo which has to be carefully addressed so as to provide safe stem cell based therapies to the patient. This review addresses the issue of tumorigenesis and discusses approaches by which this concern may be overcome and at the same time emphasizes the need to concurrently explore alternative stem cell sources that do not confer the disadvantages of pluripotency but are widely multipotent so as to yield safe desirable tissues for clinical application as soon as possible. J. Cell. Biochem. 111: 769–781, 2010. © 2010 Wiley‐Liss, Inc.
Development and evaluation of a novel nano‐scale vector for siRNA
Authors: Jing Guang‐Jun , Fu Zuo‐Gen , Dan Bing , Lin Li‐Rong , Yang Tian‐Ci , Shi Song‐Lin
Published: July 2010

To synthesize a lipid‐cationic polymer (LCP) containing brassidic acid side chain and to investigate its transfection efficiency and characteristics as a siRNA gene vector. The LCP was chemically synthesized and its nucleic acid binding capacity was determined by gel electrophoresis. HeLa‐EGFP and TH1080‐EGFP cell lines were transfected with siRNA against enhanced green fluorescent protein (EGFP) gene using a LCP to investigate the transfection efficiency. An MTT assay was performed to evaluate the cellular toxicity of the LCP vector. Its degradability and stability under acidic conditions were also investigated. The LCP vector possessed high DNA binding capacity. More than 73% of the cellular fluorescence was inhibited by the LCP‐mediated transfection of siRNA against EGFP gene, indicating that vector had high transfection efficiency. Cellular viability was about 95% at the optimum transfection efficiency of LCP, suggesting that the cellular toxicity of LCP was very low. The LCP was also observed to be degradable; moreover, it could be easily stored at normal temperature. A gene vector used for the transfection of siRNA was successfully fabricated from synthesized LCP. Its numerous excellent properties entitle values for further scientific research. J. Cell. Biochem. 111: 881–888, 2010. © 2010 Wiley‐Liss, Inc.
Elevated disgust proneness in schizophrenia
Authors: Ille Rottraut , Schöny Michaela , Kapfhammer Hans‐Peter , Schienle Anne
Published: July 2010

Schizophrenia is a mental disorder hardly investigated with regard to habitual disgust. We compared disgust propensity, trait anger, trait anxiety and disgust sensitivity in 69 patients with schizophrenia, 68 depressive patients and 70 mentally healthy controls. Patients with schizophrenia reported more pronounced overall disgust propensity than healthy individuals. Whereas food‐related disgust was crucial for schizophrenia, depressed patients experienced elevated death‐related disgust. Females reported greater disgust proneness than males. Both patient groups showed higher trait anxiety and trait anger than controls. Depressive patients additionally reported elevated anger expression and even higher anger suppression than the schizophrenia group. We found a positive relationship between disgust proneness and disgust sensitivity in controls and patients. The importance of disorder‐specific disgust domains is discussed. © 2010 Wiley Periodicals, Inc. J Clin Psychol 66:1–11, 2010.
Removal of 4‐chlorophenol from contaminated water using coconut shell waste pretreated with chemical agents
Authors: Kurniawan Tonni Agustiono , Waihung Lo , Repo Eveliina , Sillanpää Mika ET
Published: July 2010

BACKGROUND: At concentrations higher than 1 mg L−1, 4‐chlorophenol (4‐CP) is very toxic to living organisms, and if ingested beyond the permitted concentration it causes health disorders such as cancer and mutation. This laboratory study investigates treatment of contaminated water laden with 4‐CP using coconut shell charcoal (CSC) waste. Batch studies were conducted to study the effects of dose, pH, and equilibrium time on 4‐CP removal. To improve 4‐CP removal, surface modification of the adsorbent with TiO2, HNO3, and/or NaOH was undertaken.

RESULTS: At an initial 4‐CP concentration of 25 mg L−1 under optimized conditions (dose 13.5 g L−1, pH 2.0; agitation speed 150 rpm and 50 min equilibrium time), the NaOH‐treated CSC demonstrated a greater removal of 4‐CP (71%) than those oxidized with HNO3 (40%) and/or coated with TiO2 (52%). The adsorption capacity of the NaOH‐treated CSC (54.65 mg g−1) was higher than those treated with HNO3 (23.13 mg g−1) or coated with TiO2 (48.42 mg g−1).

CONCLUSION: Although treatment results using the NaOH‐treated CSC alone were promising, the treated effluents were still unable to meet the required limit of less than 1 mg L−1. Therefore, subsequent treatments are still required to complement the removal of 4‐CP from the wastewater. Copyright © 2010 Society of Chemical Industry
A fragmentation study of the Rhodamine 610 cation using visible‐laser desorption ionization
Authors: Yang C. Y. , Hu X. K. , Loboda A. V. , Lipson R. H.
Published: July 2010

The fragmentation of a potential visible matrix‐assisted laser desorption ionization: Rhodamine 610 was studied under 532 nm visible irradiation, as a function of anion counter ion. It was found that at a fixed fluence, the chloride salt produced fewer fragments than those formed with ClO4− or BF4−. Evidence presented suggests that the degree of fragmentation is inversely proportional to the strength of the contact ion pair in the solid state; that is, more energy is deposited into the radical cation which can lead to fragmentation when less energy is required to separate the ion pair. Similar results were found for salts of Rhodamine 6G. Copyright © 2010 John Wiley & Sons, Ltd.
Identification and quantification of 30 antipsychotics in blood using LC‐MS/MS
Authors: Saar Eva , Gerostamoulos Dimitri , Drummer Olaf H. , Beyer Jochen
Published: July 2010

Over the last decade, the prescription rates of antipsychotic (AP) drugs have increased worldwide. Studies have shown that the risk of sudden cardiac death is threefold higher among patients treated with APs. To investigate the presence of APs in postmortem cases, a liquid chromatography (LC)‐MS/MS method was developed using only 0.1 ml of blood sample with 10 µl of internal standard (IS) (haloperidol‐d4, 1 µg/ml). After the addition of 0.2 ml of Trizma buffer, the blood sample was extracted using liquid–liquid extraction (LLE) with 1 ml of 1‐chlorobutane for 5 min on a shaker at 1500 rpm. After centrifugation at 12 000 rpm for 1 min, the separated solvent layer was transferred to an autosampler vial and evaporated to dryness under N2. The residue was reconstituted in 0.05 ml acetonitrile containing 0.1% formic acid, vortexed for 30 s and an additional 0.45 ml of 50 mmol/l ammonium formate pH 3.5 was added and the sample vortexed; 0.1 ml of the final extract was injected into a Shimadzu Prominence HPLC system, with detection of drugs achieved using an Applied Biosystems 3200 Q‐TRAP® LC‐MS/MS system equipped with a Turbo V ion source [electron spray ionization (ESI), multiple reaction monitoring (MRM) mode]. The method has been validated according to international guidelines and was found to be selective for all tested compounds. Calibration was satisfactory for all drugs, except olanzapine, from subtherapeutic to toxic concentrations. The lower limits of quantifications (LLOQs) corresponded to the lowest concentrations used for the calibration curves. With the exception of the lowest concentrations of bromperidol, buspirone and perphenazine, accuracy data were within the acceptance interval of ± 15% ( ± 20% at LLOQ) of the nominal values for all drugs. The method has been proven to be useful for the routine analysis of APs in postmortem blood samples. Copyright © 2010 John Wiley & Sons, Ltd.
Animal urine as painting materials in African rock art revealed by cluster ToF‐SIMS mass spectrometry imaging
Authors: Mazel Vincent , Richardin Pascale , Touboul David , Brunelle Alain , Richard Caroline , Laval Eric , Walter Philippe , Laprévote Olivier
Published: July 2010

The rock art site at the village of Songo in Mali is a very important Dogon ritual place where, since the end of the nineteenth century until today, takes place the ceremony of circumcision. During these ceremonies, paintings are performed on the walls of the shelter with mainly three colors: red, black and white. Ethnological literature mentions the use of animal urine of different species such as birds, lizards or snakes as a white pigment. Urine of these animals is mainly composed of uric acid or urate salts. In this article, time‐of‐flight secondary ion mass spectrometry (ToF‐SIMS) is used to compare uric acid, snake urine and a sample of a white pigment of a Dogon painting coming from the rock art site of Songo. ToF‐SIMS measurements in both positive and negative ion modes on reference compounds and snake urine proved useful for the study of uric acid and urate salts. This method enables to identify unambiguously these compounds owing to the detection in negative ion mode of the ion corresponding to the deprotonated molecule ([M − H]− at m/z 167.01) and its fragment ions. Moreover, the mass spectra obtained in positive ion mode permit to differentiate uric acid and urate salts on the basis of specific ions. Applying this method to the Dogon white pigments sample, we show that the sample is entirely composed of uric acid. This proves for the first time, that animal urine was used as a pigment by the Dogon. The presence of uric acid instead of urate salts as normally expected in animal urine could be explained by the preparation of the pigment for its application on the stone. Copyright © 2010 John Wiley & Sons, Ltd.
How long do you benefit from vacation? A closer look at the fade‐out of vacation effects
Authors: Kühnel Jana , Sonnentag Sabine
Published: July 2010

This study adds to research on the beneficial effects of vacation to employees' well‐being and on the fade‐out of these effects. One hundred and thirty‐one teachers completed questionnaires one time before and three times after vacationing. Results indicated that teachers' work engagement significantly increased and teachers' burnout significantly decreased after vacation. However, these beneficial effects faded out within one month. Applying hierarchical regression analyses, we investigated the fade‐out of vacation effects in detail. In line with the Job Demands‐Resources model, job demands after vacation sped up the fade‐out of beneficial effects. Additionally, leisure time relaxation experiences after vacation delayed the fade‐out of beneficial effects. We conclude that reducing job demands and ensuring leisure time relaxation can prolong relief from vacation. Copyright © 2010 John Wiley & Sons, Ltd.
Climatological aspects of explosive cyclones in the Mediterranean
Authors: Kouroutzoglou J. , Flocas H. A. , Keay K. , Simmonds I. , Hatzaki M.
Published: July 2010

An objective climatology of explosive cyclones is performed over a 40‐year period (1962–2001), for the Mediterranean region with the aid of the Melbourne University automatic cyclone finding and tracking scheme. The major temporal and spatial characteristics of Mediterranean explosive cyclones are investigated, including frequency, movement, generation and dissipation, scale, depth, and intensity. It was verified that the explosive cyclogenesis in the Mediterranean is a rather rare phenomenon that occurs mainly from November to March. Explosive cyclones preferentially form along the northern Mediterranean coast, with maximum density in the Ligurian and Aegean Seas. The cyclogenesis mechanism that predominates in their development is characterized by an interaction of a short and an open long wave in a pre‐existing development (referred to as the KF mechanism). There are distinct differences between Western Mediterranean and Eastern Mediterranean explosive cyclones. Although explosive cyclones tend to form in the Western Mediterranean, their scale and depth is greater in the Eastern Mediterranean. Explosive cyclones more likely move from the main cyclogenetic areas in the Western Mediterranean along specific eastward paths over southern Italy and the Ionian Sea where they dissipate or they generate in Eastern Mediterranean and move north‐eastwards to the Black Sea or eastwards to Cyprus and Middle East. Copyright © 2010 Royal Meteorological Society
Temperature‐induced phase transformations in Na2WO4 and Na2MoO4 crystals
Authors: Lima C. Luz , Saraiva G. D. , Freire P. T. C. , Maczka M. , Paraguassu W. , de Sousa F. F. , Mendes Filho J.
Published: July 2010

Temperature‐dependent Raman studies on Na2WO4and Na2MoO4 crystals were performed in order to obtain information on structural changes induced by temperature evolution. The stability of the cubic phase of Na2WO4and Na2MoO4crystals was assessed and our results indicate that this phase is stable in the 8–823 K and 15–773 K ranges for Na2WO4 and Na2MoO4, respectively. The crystal of Na2WO4 shows a phase transition occurring at 833 K. Na2MoO4undergoes a sequence of three phase transitions, which were observed at 783–803, 823–913 and 943–950 K. In both crystals, a strong first‐order phase transition occurs as indicated by the behavior of the Raman modes: the Raman bands split and new bands appear below 100 cm−1. These transitions are connected with tilting and/or rotations of the WO4and MoO4 tetrahedra, which leads to a disorder at the WO4and MoO4 sites. Copyright © 2010 John Wiley & Sons, Ltd.
An international taxi cab evaluation: Comparing Madrid with London, New York, and Paris
Authors: Skok Walter , Martinez Juan Antonio
Published: July 2010

Taxi cabs are a key feature of daily life in Madrid, which is one of the most important European cities. An exploratory study was undertaken in order to evaluate the quality of Madrid's taxi cab industry.

This paper analyses the current situation by using the Taxi Cab Management Model (TCMM) which identifies the key processes for managing and operating a taxi cab service in any international city. The model provides a benchmark for measuring performance against other capital cities in developed nations. By comparing the results of this analysis with similar case studies of London, New York and Paris, areas of best practice are identified, as well as possible operational improvements.

The study concludes that the Madrid taxi cab industry is ahead of cities like London, New York and Paris in terms of IT integration within the industry, but that some key issues need to be reviewed e.g. the potential for a more centralised fleet management system. One of the best features of the Madrid taxi cab industry was found to be the test for candidate drivers. It is well designed in terms of its user interface (candidates do not need to be computer literate), it has a large bank of questions (up to 6000) that are used randomly to generate examinations, and it offers a highly efficient way to assess candidates. Copyright © 2010 John Wiley & Sons, Ltd.
Stretching‐Controlled Micromolding Process with Etched Metal Surfaces as Templates Towards Mass‐Producing Superhydrophobic Polymer Films
Authors: Feng Jie , Lin Feiyun , Zhong Mingqiang
Published: July 2010

A novel technique is described that uses stretching‐controlled thermal micromolding with etched metal surfaces as templates for the mass‐production of superhydrophobic polymer films. First, the metal surface is etched and then used as a template to thermally replica‐mold the polymer (e.g., polyethylene). The resulting film surfaces exhibited stable superhydrophobicity with water contact angles >150° and sliding angles ≈7°. SEM imaging demonstrates that the microstructure on the superhydrophobic surface is formed by stretching from the microholes of the template during separation. This technique can be easily combined with melt‐flow casting for manufacturing superhydrophobic polymer surfaces on a large scale.

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Focal conduction block in a case of tarsal tunnel syndrome
Authors: Almeida Diogo F. , Scremin Luciano , Zúniga Sérgio F. , Oh Shin J.
Published: July 2010

We report a case of tarsal tunnel syndrome (TTS) with focal conduction block across the tarsal tunnel (TT). A 46‐year‐old woman had pain in the left foot, sensory loss on the plantar surface, and positive Tinel sign over the TT. TTS was confirmed by magnetic resonance imaging (MRI) scan and surgery. Motor nerve conduction studies showed focal conduction block across the TT. Conduction block has rarely been reported in TTS. In this case, conduction block provides evidence for focal demyelination as the primary pathological process in TTS. Muscle Nerve, 2010
Distributed low‐frequency functional electrical stimulation delays muscle fatigue compared to conventional stimulation
Authors: Malešević Nebojša M. , Popović Lana Z. , Schwirtlich Laszlo , Popović Dejan B.
Published: July 2010

We present a low‐frequency stimulation method via multi‐pad electrodes for delaying muscle fatigue. We compared two protocols for muscle activation of the quadriceps in paraplegics. One protocol involved a large cathode at 30 HZ(HPR, high pulse‐rate), and the other involved four smaller cathodes at 16 HZ(LPR, low pulse‐rate). The treatment included 30‐min daily sessions for 20 days. One leg was treated with the HPR protocol and the other with the LPR protocol. Knee‐joint torque was measured before and after therapy to assess the time interval before the knee‐joint torque decreased to 70% of the initial value. The HPR therapy provided greater increases in muscle endurance and force in prolonged training. Yet the LPR stimulation produced less muscle fatigue compared to the HPR stimulation. The results suggest that HPR is the favored protocol for training, and LPR is better suited for prolonged stimulation. Muscle Nerve, 2010
IgM MGUS anti‐MAG neuropathy with predominant muscle weakness and extensive muscle atrophy
Authors: Kawagashira Yuichi , Kondo Naohide , Atsuta Naoki , Iijima Masahiro , Koike Haruki , Katsuno Masahisa , Tanaka Fumiaki , Kusunoki Susumu , Sobue Gen
Published: July 2010

We report a patient with anti–myelin‐associated glycoprotein (MAG) neuropathy, predominantly exhibiting severe motor symptoms, accompanied by extensive muscle atrophy mimicking Charcot–Marie–Tooth disease. Nerve conduction studies revealed mild retardation of motor conduction velocities and significant prolongation of distal latency. Sural nerve biopsy revealed widely spaced myelin and positive staining of myelinated fibers with an IgM antibody. Predominant motor symptoms with muscle atrophy can be one of the clinical manifestations of anti‐MAG neuropathy. Muscle Nerve, 2010
Effects of intravesical instillation of ATP on rat bladder primary afferent activity and its relationship with capsaicin‐sensitivity
Authors: Aizawa Naoki , Igawa Yasuhiko , Andersson Karl‐Erik , Iijima Kazuyoshi , Nishizawa Osamu , Wyndaele Jean‐Jacques
Published: July 2010

Aims

Previous studies have suggested that ATP has a role in mechano‐afferent transduction, at least partly mediated by nerves other than capsaicin (Cap)‐sensitive nerves. We investigated the direct effect of ATP on single fiber activity (SFA) of the primary bladder afferent nerves and its relationship with Cap‐sensitivity.

Methods

Female Sprague–Dawley rats were used. On the basis of conduction velocities, SFA was grouped as Aδ‐ or C‐fibers. First, SFA and intravesical pressure were measured during filling as the baseline. Then, Cap (10−5 M) was instilled intravesically and the SFA response was monitored. To desensitize Cap‐sensitive nerves, resiniferatoxin (RTX) (10−6 M) was administered intravesically, and then Cap was again administered to confirm the desensitization effect. Thereafter, ATP (10−3 M) or its vehicle was instilled intravesically and another filling cycles recorded.

Results

Thirty‐two single afferent fibers were discriminated. Aδ‐fibers did not respond to intravesical instillation of Cap, RTX or ATP. Based on Cap‐sensitivity, C‐fibers could be divided into two subtypes: Cap‐sensitive (n = 8) and Cap‐insensitive (n = 16). In the Cap‐sensitive C‐fibers, the response to bladder filling mostly disappeared after RTX desensitization, no matter if the bladder was filled with vehicle or ATP. On the other hand, in the Cap‐insensitive C‐fibers, even after RTX‐treatment, the response was preserved and more enhanced when the bladder was filled with ATP compared to vehicle.

Conclusions

Mechanosensitive bladder afferents can be classified as (1) Aδ‐fibers, (2) Cap‐insensitive, and (3) Cap‐sensitive C‐fibers. The activation of the bladder afferents induced by intravesical application of ATP is mediated mainly through Cap‐insensitive C‐fibers. Neurourol. Urodyn. 30:163–168, 2011. © 2010 Wiley‐Liss, Inc.
The closed mind: ‘Experience’ and ‘cognition’ aspects of openness to experience and need for closure as psychological bases for right–wing attitudes
Authors: Onraet Emma , Van Hiel Alain , Roets Arne , Cornelis Ilse
Published: July 2010

Openness to Experience and Need for Closure (NFC) are dispositional variables related to social–cultural right–wing attitudes. The present study investigated their joint effects. Factor analysis revealed an ‘experiential’ dimension with high loading openness items, and a ‘cognition’ dimension with high loadings for most NFC items and about a quarter of the openness item set. The experiential openness items were weakly related to right–wing attitudes, demonstrating little predictive value. Conversely, the cognitive openness and NFC items were powerful predictors of right–wing attitudes, and also played an important role in integrative models, both as a predictor of authoritarianism–based racism and as a mediator of age related increments in right–wing attitudes. It is concluded that right–wing attitudes should be primarily understood in terms of (motivated) cognition, and to a smaller extent in terms of experiential openness. The distinction between ‘experiential’ and ‘cognitive’ openness is critically assessed, and it is asserted that because cognition is a multifaceted construct openness contains more than one cognitive dimension. Copyright © 2010 John Wiley & Sons, Ltd.
Cytoskeletal scaffolding proteins interact with Lynch‐Syndrome associated mismatch repair protein MLH1
Authors: Brieger Angela , Adryan Boris , Wolpert Fabian , Passmann Sandra , Zeuzem Stefan , Trojan Jörg
Published: July 2010

The involvement of MLH1 in several mismatch repair‐independent cellular processes has been reported. In an attempt to gain further insight into the protein's cellular functions, we screened for novel interacting partners of MLH1 utilizing a bacterial two‐hybrid system. Numerous unknown interacting proteins were identified, suggesting novel biological roles of MLH1. The network of MLH1 and its partner proteins involves a multitude of cellular processes. Integration of our data with the “General Repository for Interaction Datasets” highlighted that MLH1 exhibits relationships to three interacting pairs of proteins involved in cytoskeletal and filament organization: Thymosin β 4 and Actin γ, Cathepsin B and Annexin A2 as well as Spectrin α and Desmin. Coimmunoprecipitation and colocalization experiments validated the interaction of MLH1 with these proteins. Differential mRNA levels of many of the identified proteins, detected by microarray analysis comparing MLH1‐deficient and ‐proficient cell lines, support the assumed interplay of MLH1 and the identified candidate proteins. By siRNA knock down of MLH1, we demonstrated the functional impact of MLH1–Actin interaction on filament organization and propose that dysregulation of MLH1 plays an essential role in cytoskeleton dynamics. Our data suggest novel roles of MLH1 in cellular organization and colorectal cancerogenesis.
Upregulation of plasma C9 protein in gastric cancer patients
Authors: Chong Poh‐Kuan , Lee Huiyin , Loh Marie Chiew Shia , Choong Lee‐Yee , Lin Qingsong , So Jimmy Bok Yan , Lim Khong Hee , Soo Ross Andrew , Yong Wei Peng , Chan Siew Pang , Smoot Duane T. , Ashktorab Hassan , Yeoh Khay Guan , Lim Yoon Pin
Published: July 2010

Gastric cancer is one of the leading causes of cancer‐related deaths worldwide. Current biomarkers used in the clinic do not have sufficient sensitivity for gastric cancer detection. To discover new and better biomarkers, protein profiling on plasma samples from 25 normal, 15 early‐stage and 21 late‐stage cancer was performed using an iTRAQ‐LC‐MS/MS approach. The level of C9 protein was found to be significantly higher in gastric cancer compared with normal subjects. Immunoblotting data revealed a congruent trend with iTRAQ results. The discriminatory power of C9 between normal and cancer states was not due to inter‐patient variations and was independent from gastritis and Helicobacter pylori status of the patients. C9 overexpression could also be detected in a panel of gastric cancer cell lines and their conditioned media compared with normal cells, implying that higher C9 levels in plasma of cancer patients could be attributed to the presence of gastric tumor. A subsequent blind test study on a total of 119 plasma samples showed that the sensitivity of C9 could be as high as 90% at a specificity of 74%. Hence, C9 is a potentially useful biomarker for gastric cancer detection.
Illuminating the molecular basis of diabetic arteriopathy: A proteomic comparison of aortic tissue from diabetic and healthy rats
Authors: Jüllig Mia , Chen Xiuyin , Middleditch Martin J. , Vazhoor Greeshma , Hickey Anthony J. , Gong Deming , Lu Jun , Zhang Shaoping , Phillips Anthony R. J. , Cooper Garth J. S.
Published: July 2010

Arterial disease is a major diabetic complication, yet the component molecular mechanisms of diabetic arteriopathy remain poorly understood. In order to identify major proteins/pathways implicated in diabetic arteriopathy, we studied the effect of 16‐wk untreated streptozotocin‐induced diabetes on the rat aortic proteome. Specific protein levels in isolated aortas were compared in six discrete, pair‐wise (streptozotocin‐diabetic and non‐diabetic age‐matched controls) experiments in which individual proteins were identified and quantified by iTRAQ combined with LC‐MS/MS. A total of 398 unique non‐redundant proteins were identified in at least one experiment and 208 were detected in three or more. Between‐group comparisons revealed significant changes or trends towards changes in relative abundance of 51 proteins (25 increased, 26 decreased). Differences in levels of selected proteins were supported by Western blotting and/or enzyme assays. The most prominent diabetes‐associated changes were in groups of proteins linked to oxidative stress responses and the structure/function of myofibrils and microfilaments. Indexes of mitochondrial content were measurably lower in aortic tissue from diabetic animals. Functional cluster analysis also showed decreased levels of glycolytic enzymes and mitochondrial electron transport system‐complex components. These findings newly implicate several proteins/functional pathways in the pathogenesis of arteriosclerosis/diabetic arteriopathy.
Large pore gels to separate mega protein complexes larger than 10 MDa by blue native electrophoresis: Isolation of putative respiratory strings or patches
Authors: Strecker Valentina , Wumaier Zibiernisha , Wittig Ilka , Schägger Hermann
Published: July 2010

Here, we expand the application of blue native electrophoresis to the separation of mega protein complexes larger than 10 MDa by introducing novel large pore acrylamide gels. We tailored the bis‐acrylamide cross‐linker amounts relative to the acrylamide monomer to enlarge the pore size of acrylamide gels and to obtain elastic and sufficiently stable gels. The novel gel types were then used to search for suprastructures of mitochondrial respiratory supercomplexes, the hypothetical respiratory strings, or patches. We identified 4–8 MDa assemblies that contain respiratory complexes I, III, and IV and most likely represent dimers, trimers, and tetramers of respiratory supercomplexes. We also isolated multimeric respiratory supercomplexes with apparent masses of 35–45 MDa, the presumed core pieces of respiratory strings or patches. Electron microscopic investigations will be required to clarify whether the isolated assemblies of complexes are ordered and specific, as predicted for respiratory strings and patches in the mitochondrial membrane.
The stromal‐vascular fraction of adipose tissue contributes to major differences between subcutaneous and visceral fat depots
Authors: Peinado Juan R. , Jimenez‐Gomez Yolanda , Pulido Marina R. , Ortega‐Bellido Maria , Diaz‐Lopez Cesar , Padillo Francisco J. , Lopez‐Miranda Jose , Vazquez‐Martínez Rafael , Malagón María M.
Published: July 2010

Adipose tissue represents a complex tissue both in terms of its cellular composition, as it includes mature adipocytes and the various cell types comprising the stromal‐vascular fraction (SVF), and in relation to the distinct biochemical, morphological and functional characteristics according to its anatomical location. Herein, we have characterized the proteomic profile of both mature adipocyte and SVF from human visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT) fat depots in order to unveil differences in the expression of proteins which may underlie the distinct association of VAT and SAT to several pathologies. Specifically, 24 proteins were observed to be differentially expressed between SAT SVF versus VAT SVF from lean individuals. Immunoblotting and RT‐PCR analysis confirmed the differential regulation of the nuclear envelope proteins lamin A/C, the membrane‐cytoskeletal linker ezrin and the enzyme involved in retinoic acid production, aldehyde dehydrogenase 1A2, in the two fat depots. In sum, the observation that proteins with important cell functions are differentially distributed between VAT and SAT and their characterization as components of SVF or mature adipocytes pave the way for future research on the molecular basis underlying diverse adipose tissue‐related pathologies such as metabolic syndrome or lipodystrophy.
Hydrogen‐bonded nucleophile effects in ANS: the reactions of 1‐chloro and 1‐fluoro‐2,4‐dinitrobenzene with 2‐guanidinobenzimidazole, 1‐(2‐aminoethyl)piperidine andN‐(3‐aminopropyl)morpholine in aprotic solvents
Authors: Alvaro Cecilia E. Silvana , Ayala Alicia D. , Nudelman Norma S.
Published: July 2010

The kinetics of the reactions of 2,4‐dinitrofluorobenzene (DNFB) and 2,4‐dinitrochlorobenzene (DNClB) with 2‐guanidinobenzimidazole (2‐GB) at 40 ± 0.2 °C in dimethylsulphoxide (DMSO), toluene, and in toluene–DMSO mixtures, and with 1‐(2‐aminoethyl)piperidine (2‐AEPip) andN‐(3‐aminopropyl)morpholine (3‐APMo) in toluene at 25 ± 0.2 °C were studied under pseudo first‐order conditions. For the reactions of 2‐GB carried out in pure DMSO, the second‐order rate coefficients were independent of the amine concentration. In contrast, the reactions of 2‐GB with DNFB in toluene, showed a kinetic behaviour consistent with a base‐catalysed decomposition of the zwitterionic intermediate. These results suggest an intramolecular H‐bonding of 2‐GB in toluene, which is not present in DMSO. To confirm this interpretation the reactions were studied in DMSO–toluene mixtures. Small amounts of DMSO produce significant increase in rate that is not expected on the basis of the classical effect of a dipolar aprotic medium; the effect is consistent with the formation of a nucleophile/co‐solventmixed aggregate. For the reactions of 3‐APMo with both substrates in toluene, the second‐order rate coefficients,kA, show a linear dependence on the [amine]. 3‐APMo is able to form a six‐membered ring by an intramolecular H‐bond which prevents the formation of self‐aggregates. In contrast, a third order was observed in the reactions with 2‐AEPip: these results can be interpreted as a H‐bonded homo‐aggregate of the amine acting as a better nucleophile than the monomer. Most of these results can be well explained within the frame of the ‘dimer nucleophile’ mechanism. Copyright © 2010 John Wiley & Sons, Ltd.
Paracrine factors produced by bone marrow stromal cells induce apoptosis and neuroendocrine differentiation in prostate cancer cells
Authors: Zhang Chu , Soori Mehrnoosh , Miles Fayth L. , Sikes Robert A. , Carson Daniel D. , Chung Leland W.K. , Farach‐Carson Mary C.
Published: July 2010

BACKGROUND

Preferential bony metastasis of human prostate cancer (PCa) cells contributes to disease mortality and morbidity. Local factors in bone stromal extracellular matrix microenvironment affect tumor growth through paracrine interactions between tumor and stromal cells.

METHODS

Using co‐culture and medium transfer, we used several methods to assess interactions between PCa and bone stromal cells using three PCa cell lines: PC3, LNCaP, and the LNCaP derivative, C4‐2B.

RESULTS

Co‐culture of LNCaP and C4‐2B cells with bone marrow stromal cell lines, HS27a and HS5, decreased cell number, as did culture with conditioned medium (CM) harvested from these two cell lines suggesting a soluble paracrine factor was responsible. PC3 cell growth was unaffected. CM harvested from bone stromal cell lines triggered apoptosis in LNCaP and C4‐2B cell lines, but not in PC3 cells. Surviving C4‐2B cells grown in bone stromal cell CM over several days were growth arrested, suggesting presence of a growth inhibitor. Apoptosis induced by CM was dose‐dependent. Flow cytometry demonstrated that over a 5‐day culture period in stromal cell CM, LNCaP, and C4‐2B cell lines, but not PC3 cells, underwent greater apoptosis than parallel cultures in SF medium. The LNCaP and C4‐2B cells showed morphology and biomarker expression consistent with transdifferentiation towards a neuroendocrine phenotype after exposure to stromal cell CM.

CONCLUSIONS

The reactive bone stromal microenvironment initially is hostile to PCa cells producing widespread apoptosis. Activation of transdifferentiation in a subset of apoptotic resistant cells may support phenotypic adaptation during disease progression in bone, eventually favoring lethal disease. Prostate 71: 157–167, 2011. © 2010 Wiley‐Liss, Inc.
Micro‐ and nano‐electromechanical resonators based on SiC and group III‐nitrides for sensor applications
Authors: Brueckner K. , Niebelschuetz F. , Tonisch K. , Foerster Ch. , Cimalla V. , Stephan R. , Pezoldt J. , Stauden T. , Ambacher O. , Hein M. A.
Published: July 2010

Wide‐bandgap semiconductors represent an attractive option to meet the increasing demands of micro‐ and nano‐electromechanical systems (MEMS/NEMS) by offering new functionalities, high stability, biocompatibility and the potential for miniaturization and integration. Here, we report on resonant MEMS and NEMS devices with functional layers of SiC, AlN and AlGaN/GaN heterostructures on different substrates, which have been investigated and analysed in the course of an interdisciplinary research focus programme of the German Research Foundation (DFG). The specific deposition and etching technologies necessary for the three‐dimensional micro‐structuring are explained. Further, the implementation of appropriate electromechanical transduction schemes is discussed. In case of SiC and AlN resonators, actuation and sensing was achieved by a magnetomotive scheme. A piezoelectric coupling scheme where the counter electrode is formed by the two‐dimensional electron gas at the interface of the III/V heterostructure was realized for the AlGaN/GaN resonators.

Thus, flexural and longitudinal vibration modes were excited and characterized using electrical and optical techniques. The measured key parameters of resonant frequency and quality factor are related to geometry, material and environmental parameters using analytical and finite element (FE) models. Finally, potential sensor applications are experimentally investigated.
Influence of the dielectric substrate on the field emitted by a subwavelength slit in a metal film
Authors: Nikitin A. Yu. , García‐Vidal F. J. , Martín‐Moreno L.
Published: July 2010

The field emitted by a localized source in the metal surface bounding to a half‐media with arbitrary dielectric permittivity is studied. The far field at the surface is composed of both the surface plasmon polariton and the algebraically decaying Norton wave contributions. The crossover distance at which the Norton wave overtakes the surface plasmon, depends upon the refractive index of the bounding media. Substrates with high dielectric permittivity bring the crossover substantially closer to the source.

magnified image

The field emitted by a subwavelength slit. Far away from the source the field presents contributions from both the surface plasmon and the Norton wave.

(© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Near‐field studies of arrays of chirped subwavelength apertures
Authors: Neshev Dragomir N. , Minovich Alexander , Dieing Thomas , Hattori Haroldo T. , McKerracher Ian , Tan Hark Hoe , Jagadish Chennupati , Kivshar Yuri S.
Published: July 2010

We study light transmission through a metal film perforated with an array of square apertures of varying size. We combine near‐field scanning optical microscopy with confocal microscopy to identify surface plasmon modes and propagating hole modes in the different size apertures. We also perform spectral measurements to reveal the complex characteristics of light transmission through the chirped subwavelength apertures. (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
The effect of shave‐off scanning at a fabricated section by focused ion beam
Authors: Morita Y. , Kuroda A. , Iwata T. , Owari M.
Published: July 2010

Three‐dimensional microanalysis for a minute structure that consists of organic compounds and polymers is important to develop and to practically use the method to analyze a micronanometer order area. However, it is difficult because of the insulating nature of the material, need of structural analysis and a dose limit not to destruct surface organic structure (static limit). We developed a novel three‐dimensional microanalysis method by means of focused ion beam (FIB) for section processing (shave‐off scanning) and ToF‐SIMS for mapping method. For the purpose of realization of three‐dimensional microanalysis and a chemical and structural analysis of the organic matter, a polymer destruction of fabricated section by shave‐off scanning was examined. To determine spectrum changing, polystyrene was fabricated as a function of dose time by shave‐off scanning, and these samples were analyzed by ToF‐SIMS. As a result, a flat surface was made along with shave‐off scanning time, and a surface organic structure fabricated by FIB was detected with a certain rate. Copyright © 2010 John Wiley & Sons, Ltd.
An AFM contribution to the understanding of surface effects caused by ageing and cleaning on acrylic glass. The Shadows by Lourdes Castro, a case study
Authors: de Sá M. H. , Ferreira Joana L. , Melo M. J. , Ramos Ana M.
Published: July 2010

Atomic Force Microscopy (AFM) was used to study the ageing and cleaning effects on the surface of acrylic glass sheets (PMMA) like the ones used by the Portuguese artist Lourdes Castro. Measurements were done in transparent (colourless and blue) samples. The samples were aged in a SolarBox; cleaning and polishing products, similar to the ones applied by the artist, were used. From the AFM images the craze/scratch phenomena in the pristine samples surface was confirmed. The experiments led to an increase of the surface roughness, more pronounced for the polished samples due to the presence of granular features. These modifications are due to the interaction of the acrylic sheets with the polishing products, which seems to be stronger in the coloured samples. These findings are useful to define the best strategy to prolong the longevity of such materials, in particular the works of art created by Lourdes Castro. Copyright © 2010 John Wiley & Sons, Ltd.
Additional requirements for process assessment in safety–critical software and systems domain
Authors: Johansson Mika , Nevalainen Risto
Published: July 2010

Certification of safety–critical software is a multi‐disciplinary topic. Process assessment is an essential part of that, but is not enough for software certification. Certification also employs several other method families, such as inspections and reviews, independent V&V, conformance with selected reference standard(s) and use of selected measurements and analyses. Process assessment directly supports qualification of safety–critical applications but is less relevant for certification of platforms and environments. Anyway, qualification and certification are closely related. Certification as a whole supports qualification and makes it more effective. It is possible to adapt and evolve process assessment so that it supports both qualification and certification. Typical process assessment is done for improvement purpose. In qualification and certification it is not so relevant as conformance and management of risks. In this article we discuss the possibilities to develop process assessment to achieve that goal. In most cases assessment is a combination of several approaches. Copyright © 2010 John Wiley & Sons, Ltd.
Lessons learned from an ISO/IEC 15504 SPI programme in a company
Authors: Mas Antonia , Fluxà Bartomeu , Amengual Esperança
Published: July 2010

This paper describes a first‐hand experience in Software Process Improvement within a Spanish Company which, since 2002, has been involved in an improvement programme led by our research group. We discuss the experience and the results of this improvement programme, as well as the lessons learned to deal with new future improvement initiatives in other companies of our environment. Copyright © 2010 John Wiley & Sons, Ltd.
UML‐based regression testing for OO software
Authors: Mansour Nashat , Takkoush Husam , Nehme Ali
Published: July 2010

In software maintenance, a system has to be regression tested after modifying it. The goal of regression testing is to ensure that modifications have not adversely affected the system. Regression test selection determines a subset of test cases, from the initial test suite, which concentrates on the parts of the system affected by the modification. Previous techniques have been mainly code‐based and several of them have addressed procedural programs. When working with large and complex object‐oriented systems, source code‐based regression testing is usually costly. This paper proposes a programming‐language‐independent technique for regression test selection for object‐oriented software based on Unified Modeling Language (UML 2.0) design diagrams. These diagrams are: the newly introduced interaction overview diagram, class diagrams, and sequence diagrams. We assume a test suite that contains both unit and system test cases. Based on the software changes reflected in the class and the interaction overview diagrams, our proposed technique selects test cases in phases. In the first phase, we select both unit and system test cases that directly traverse the changed methods and their calling methods. For the second phase, we present algorithms for detecting system level changes in the interaction overview diagram. If the change is at the action level, which is represented by a sequence diagram, only the test cases that execute changed methods will be selected. We apply our proposed technique to a few object‐oriented subject applications and evaluate its precision and inclusiveness in addition to the number of selected tests; the results demonstrate the advantages of the technique. Copyright © 2010 John Wiley & Sons, Ltd.