bluuebunny/crossref_metadata_2025 huggingface dataset:
BMP growth factor signaling in a biomechanical context
Authors: Kopf Jessica , Paarmann Pia , Hiepen Christian , Horbelt Daniel , Knaus Petra
Published: October 2013

Bone Morphogenetic Proteins (BMPs) are members of the transforming growth factor‐β superfamily of secreted polypeptide growth factors and are important regulators in a multitude of cellular processes. To ensure the precise and balanced propagation of their pleiotropic signaling responses, BMPs and their corresponding signaling pathways are subject to tight control. A large variety of regulatory mechanisms throughout different biological levels combines into a complex network and provides the basis for physiological BMP function. This regulatory network not only includes biochemical factors but also mechanical cues. Both BMP signaling and mechanotransduction pathways are tightly interconnected and represent an elaborate signaling network active during development but also during organ homeostasis. Moreover, its dysregulation is associated with a number of human pathologies. A more detailed understanding of this crosstalk in respect to molecular interactions will be indispensable in the future, in particular to understand BMP‐related diseases as well as with regard to an efficient clinical application of BMP ligands. © 2013 BioFactors, 40(2):171–187, 2014
A principal component analysis of the dynamics of subdomains and binding sites in human serum albumin
Authors: Paris Guillaume , Ramseyer Christophe , Enescu Mironel
Published: October 2013

The conformational dynamics of human serum albumin (HSA) was investigated by principal component analysis (PCA) applied to three molecular dynamics trajectories of 200 ns each. The overlap of the essential subspaces spanned by the first 10 principal components (PC) of different trajectories was about 0.3 showing that the PCA based on a trajectory length of 200 ns is not completely convergent for this protein. The contributions of the relative motion of subdomains and of the subdomains (internal) distortion to the first 10 PCs were found to be comparable. Based on the distribution of the first 3 PC, 10 protein conformers are identified showing relative root mean square deviations (RMSD) between 2.3 and 4.6 Å. The main PCs are found to be delocalized over the whole protein structure indicating that the motions of different protein subdomains are coupled. This coupling is considered as being related to the allosteric effects observed upon ligand binding to HSA. On the other hand, the first PC of one of the three trajectories describes a conformational transition of the protein domain I that is close to that experimentally observed upon myristate binding. This is a theoretical support for the older hypothesis stating that changes of the protein onformation favorable to binding can precede the ligand complexation. A detailed all atoms PCA performed on the primary Sites 1 and 2 confirms the multiconformational character of the HSA binding sites as well as the significant coupling of their motions. © 2013 Wiley Periodicals, Inc. Biopolymers 101: 561–572, 2014.
Limited proteolysis of natively unfolded protein 4E‐BP1 in the presence of trifluoroethanol
Authors: Hackl Ellen V.
Published: October 2013

Natively unfolded (intrinsically disordered (ID) proteins) have been attracting an increasing attention due to their involvement in many regulatory processes. Natively unfolded proteins can fold upon binding to their metabolic partners. Coupled folding and binding events usually involve only relatively short motifs (binding motifs). These binding motifs which are able to fold should have an increased propensity to form a secondary structure. The aim of the present work was to probe the conformation of the intrinsically disordered protein 4E‐BP1 in the native and partly folded states by limited proteolysis and to reveal regions with a high propensity to form an ordered structure. Trifuoroethanol (TFE) in low concentrations (up to 15 vol%) was applied to increase the helical population of protein regions with a high intrinsic propensity to fold. When forming helical structures, these regions lose mobility and become more protected from proteases than random/unfolded protein regions. Limited proteolysis followed by mass spectrometry analysis allows identification of the regions with decreased mobility in TFE solutions. Trypsin and V8 proteases were used to perform limited proteolysis of the 4E‐BP1 protein in buffer and in solutions with low TFE concentrations at 37°C and at elevated temperatures (42 and 50°C). Comparison of the results obtained with the previously established 4E‐BP1 structure and the binding motif illustrates the ability of limited proteolysis in the presence of a folding assistant (TFE) to map the regions with high and low propensities to form a secondary structure revealing potential binding motifs inside the intrinsically disordered protein. © 2013 Wiley Periodicals, Inc. Biopolymers 101: 591–602, 2014.
Development and validation of a rapid and high‐sensitivity liquid chromatography–tandem mass spectrometry assay for the determination of neostigmine in small‐volume beagle dog plasma and its application to a pharmacokinetic study
Authors: Cao Di , Li Wenxue , Zhao Xin , Ye Xiaolan , Sun Fanlu , Li Jinying , Song Fenyun , Fan Guorong
Published: October 2013

A simple, rapid and high sensitive liquid chromatography–tandem mass spectrometry (LC‐MS/MS) method for the determination of neostigmine in small‐volume beagle dog plasma was developed to assess the plasma pharmacokinetics of neostigmine. After protein precipitation in a Sirocco 96‐well filtration plate, the filtrate was directly injected into the LC‐MS/MS system. The analytes were separated on a Hanbon Hedera CN column (100 × 4.6 mm, 5 µm) with a mobile phase composed of methanol–water (60:40, v/v) and the water containing 0.01% formic acid at a flow rate of 0.6mL/min, with a split ratio of 1:1 flowing 300 μL into the mass spectrometer. The run time was 3 min. Detection was accomplished by electrospray ionization source in multiple reactions monitoring mode with the precursor‐to‐product ion transitions m/z 223.0 → 72.0 and 306.0 → 140.0 for neostigmine and anisodamine (internal standard), respectively. The method was sensitive with a lower limit of quantitation of 0.1 ng/mL, and good linearity in the range 0.1–100ng/mL for neostigmine (r ≥ 0.998). All the validation data, such as accuracy, intra‐run and inter‐run precision, were within the required limits. The method was successfully applied to pharmacokinetic study of neostigmine methylsulfate injection in beagle dogs. Copyright © 2013 John Wiley & Sons, Ltd.
In vitro evaluation of potential complexation between bovine insulin and bovine serum albumin
Authors: Al‐Domi Hayder , Alzweiri Muhammed , Hamdan Imad , Jaradat Ziad
Published: October 2013

The objective of this study was to examine the possible binding of bovine insulin (BI) with bovine serum albumin (BSA) to form a new potential diabetogenic irreversible complex protein. Several preparations of BSA and BI were prepared. Both capillary electrophoresis and spectrophotometric analysis were undertaken to test the possibility of complexation between BI and BSA. HPLC was used to test whether the potential complex of BI and BSA is reversible or irreversible. The optimum deviation between the real and calculated absorbances was observed at a BI/BSA ratio of 2. Moreover, the migration time of BI decreased substantially with increasing ratio of BI to BSA until it became almost constant at equal molar ratio of BI/BSA. While the majority of the 2:1 BI–BSA sample detached during the HPLC analysis, which confirms the reversible character of BI–BSA binding, the HPLC chromatogram also emphasizes the formation of an irreversible complexation between the two proteins. This study provides evidence of the formation of reversible and irreversible new BI–BSA complexes under physiological conditions. This highlights the importance of examining the possible diabetogenicity of BI–BSA complex in genetically susceptible people. Copyright © 2013 John Wiley & Sons, Ltd.
HPLC‐Q‐TOF‐MS/MS for analysis of major chemical constituents of Yinchen–Zhizi herb pair extract
Authors: Fu Zhiwen , Ling Yun , Li Zhixiong , Chen Mingcang , Sun Zhaolin , Huang Chenggang
Published: October 2013

The Yinchen–Zhizi herb pair (YZHP) consists of Herba Artemisiae Scopariae (Yinchen in Chinese) and Fructus Gardeniae (Zhizi in Chinese), and is mainly used to treat icteric hepatitis, itching skin and eczema. However, the bioactive constituents responsible for the pharmacological effects of YZHP are still unclear to date. In this work, a rapid and sensitive method was established to comprehensively study the constituents in YZHP extract by HPLC‐Q‐TOF MS/MS. The analysis was performed on an HPLC system equipped with an Agilent poroshell 120 EC‐C18 column (100 × 2.1 mm, 2.7 mm) working in a gradient elution program coupled to quadrupole‐time‐of‐flight mass spectrometry operating in the negative ion mode. As a result, a total of 46 compounds including 17 from Herba Artemisiae Scopariae and 36 from Fructus Gardeniae were detected and tentatively identified in YZHP extract by comparing the retention time and mass spectrometry and retrieving the reference literature. More importantly, a series of constituents, such as many iridoid glycosides, were reported for the first time in this formula. The HPLC‐Q‐TOF MS/MS method was developed and utilized successfully to identify the major constituents in YZHP extract and would be helpful for further metabolism and pharmacology research on YZHP. Copyright © 2013 John Wiley & Sons, Ltd.
Simultaneous determination of four components in Baige capsule by HPLC: application to pharmacokinetics and tissue distribution of normal and middle cerebral artery occlusion rats
Authors: Wang Yuanji , Wang Nan , Wu Lei , Lu Wen , Zhang Yanmin
Published: October 2013

A sensitive and specific high‐performance liquid chromatography (HPLC) method was developed and applied to the pharmacokinetics for simultaneous identification and quantification of four components – puerarin, daidzein, imperatorin and isoimperatorin – in the plasma and tissues of normal and middle cerebral artery occlusion rats after oral administration of Baige capsule. Ferulic was used as the internal standard. The extraction procedure was composed of two independent steps. The plasma was prepared by liquid–liquid extraction with light petroleum–acetic ether (1:1, v/v) first and then protein was precipitated with methanol. The tissue samples were weighted and homogenated with normal saline, then the homogenate was prepared by liquid–liquid extraction and protein precipitation. The parameters of pharmacokinetics were calculated using DAS 2.1.1 software. The calibration curves of all four components in the plasma and tissue homogenates were in good linearity in the measured range with R2 ≥ 0.9921. The relative standard deviation of the intra‐ and inter‐day accuracy at different levels was less than ±18.0%. In conclusion, the established method was a simple and effective one to simultaneously detect all four components in the plasma and tissues of rats, and was successfully applied in the pharmacokinetics of Baige capsule. Copyright © 2013 John Wiley & Sons, Ltd.
Prediction processes during multiple object tracking (MOT): involvement of dorsal and ventral premotor cortices
Authors: Atmaca Silke , Stadler Waltraud , Keitel Anne , Ott Derek V. M. , Lepsien Jöran , Prinz Wolfgang
Published: October 2013

Background

The multiple object tracking (MOT) paradigm is a cognitive task that requires parallel tracking of several identical, moving objects following nongoal‐directed, arbitrary motion trajectories.

Aims

The current study aimed to investigate the employment of prediction processes during MOT. As an indicator for the involvement of prediction processes, we targeted the human premotor cortex (PM). The PM has been repeatedly implicated to serve the internal modeling of future actions and action effects, as well as purely perceptual events, by means of predictive feedforward functions.

Materials and methods

Using functional magnetic resonance imaging (fMRI), BOLD activations recorded during MOT were contrasted with those recorded during the execution of a cognitive control task that used an identical stimulus display and demanded similar attentional load. A particular effort was made to identify and exclude previously found activation in the PM‐adjacent frontal eye fields (FEF).

Results

We replicated prior results, revealing occipitotemporal, parietal, and frontal areas to be engaged in MOT.

Discussion

The activation in frontal areas is interpreted to originate from dorsal and ventral premotor cortices. The results are discussed in light of our assumption that MOT engages prediction processes.

Conclusion

We propose that our results provide first clues that MOT does not only involve visuospatial perception and attention processes, but prediction processes as well.
Enhancing the Credibility of Reporting on Corporate Social Responsibility in Europe
Authors: De Beelde Ignace , Tuybens Sanne
Published: October 2013

This paper studies variables that have an effect on the choice of whether or not reports on corporate social responsibility are assured and who is selected to provide assurance. Such assurance reports increase the credibility of environmental and social reporting. In a European sample we observe some differences from previous research. Overall, industry effects seem small, but size and media visibility have a significant impact on assuring CSR reports. It seems that companies differentiate less between auditors and consultants. Copyright © 2013 John Wiley & Sons, Ltd and ERP Environment
It's Not All in Your Head (or at Least Your Brain): Association of Traumatic Brain Lesion Presence and Location with Performance on Measures of Response Bias in Forensic Evaluation
Authors: McBride Willie F. , Crighton Adam H. , Wygant Dustin B. , Granacher Robert P.
Published: October 2013

This study examined the relationship between lesion presence and localization and performance on measures of cognitive response bias, specifically in individuals purporting to have a traumatic brain injury. Ninety‐two participants, all of whom were involved in workers' compensation or personal injury litigation, were administered an extensive neuropsychological battery, including neuroimaging (magnetic resonance imaging and computed tomography), at a neuropsychiatric clinic in Lexington, KY. Those with evidence of intracranial injury on neuroimaging findings were placed in the head injury lesion litigation group and were coded based on the anatomical location and type of intracranial injury. Results demonstrated no significant relationships between lesion location and performance on performance validity tests (PVTs), as well as the Response Bias Scale of the Minnesota Multiphasic Personality Inventory‐2 Restructured Form. Given the lack of research concerning lesions and performance validity tests, this study addresses important questions about the validity of PVTs as specific measures of response bias in patients who have structural changes secondary to traumatic brain injury. Copyright © 2013 John Wiley & Sons, Ltd.
Investigation of vinegar production using a novel shaken repeated batch culture system
Authors: Schlepütz Tino , Büchs Jochen
Published: October 2013

Nowadays, bioprocesses are developed or optimized on small scale. Also, vinegar industry is motivated to reinvestigate the established repeated batch fermentation process. As yet, there is no small‐scale culture system for optimizing fermentation conditions for repeated batch bioprocesses. Thus, the aim of this study is to propose a new shaken culture system for parallel repeated batch vinegar fermentation. A new operation mode — the flushing repeated batch — was developed. Parallel repeated batch vinegar production could be established in shaken overflow vessels in a completely automated operation with only one pump per vessel. This flushing repeated batch was first theoretically investigated and then empirically tested. The ethanol concentration was online monitored during repeated batch fermentation by semiconductor gas sensors. It was shown that the switch from one ethanol substrate quality to different ethanol substrate qualities resulted in prolonged lag phases and durations of the first batches. In the subsequent batches the length of the fermentations decreased considerably. This decrease in the respective lag phases indicates an adaptation of the acetic acid bacteria mixed culture to the specific ethanol substrate quality. Consequently, flushing repeated batch fermentations on small scale are valuable for screening fermentation conditions and, thereby, improving industrial‐scale bioprocesses such as vinegar production in terms of process robustness, stability, and productivity. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:1158–1168, 2013
Protein misfolding and aggregation research: Some thoughts on improving quality and utility
Authors: Murphy Regina M. , Roberts Christopher J.
Published: October 2013

Once misfolded and aggregated proteins were as interesting as yesterday's trash, just a bothersome byproduct of productive activities. Today, they attract sustained interest from both basic researchers and practicing engineers. In the burgeoning biopharmaceutical industry, protein misfolding and aggregation pose significant challenges to the economic manufacture of safe and effective protein products. In the clinic, protein aggregates are believed to be pathological agents in a number of serious neurodegenerative disorders, such as Alzheimer's and Parkinson's. Over the past few years, the quantity of research into biotechnological aspects of protein misfolding and aggregation has skyrocketed. However, the quality of the published work is quite variable. In this brief opinion piece, we describe what we believe are some key features of high‐quality publications in protein aggregation. We focus on experimental studies that may also have a kinetic modeling component. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:1109–1115, 2013
Enveloped virus inactivation using neutral arginine solutions and applications in therapeutic protein purification processes
Authors: McCue Justin T. , Selvitelli Keith , Cecchini Doug , Brown Rhonda
Published: October 2013

For the manufacturing of recombinant protein therapeutics produced from mammalian cell culture, demonstrating the capacity of the purification process to effectively clear infectious viruses is a regulatory requirement. At least two process steps, using different mechanisms of virus removal and/or inactivation, should be validated in support of the regulatory approval process. For example, exposure of the product stream to low pH, detergents or solvent/detergent combinations is commonly incorporated in protein purification processes for the inactivation of lipid‐enveloped viruses. However, some proteins have limited stability at low pH or in the presence of the detergents, and alternative techniques for achieving the inactivation of enveloped viruses would be beneficial. We present here an alternative and novel approach for the rapid inactivation of enveloped viruses using pH‐neutral buffer solutions containing arginine. The implementation of this approach in a monoclonal antibody or Fc‐fusion protein purification process is described and illustrated with several different therapeutic proteins. The use of the neutral pH arginine solution was able to effectively inactivate two enveloped model viruses, with no measurable effect on the product quality of the investigated proteins. Thus, the use of pH‐neutral arginine containing buffer solutions provides an alternative means of virus inactivation where other forms of virus inactivation, such as low pH and/or solvent/detergent treatments are not possible or undesirable due to protein stability limitations. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 30:108–112, 2014
Development of a scale down cell culture model using multivariate analysis as a qualification tool
Authors: Tsang Valerie Liu , Wang Angela X. , Yusuf‐Makagiansar Helena , Ryll Thomas
Published: October 2013

In characterizing a cell culture process to support regulatory activities such as process validation and Quality by Design, developing a representative scale down model for design space definition is of great importance. The manufacturing bioreactor should ideally reproduce bench scale performance with respect to all measurable parameters. However, due to intrinsic geometric differences between scales, process performance at manufacturing scale often varies from bench scale performance, typically exhibiting differences in parameters such as cell growth, protein productivity, and/or dissolved carbon dioxide concentration. Here, we describe a case study in which a bench scale cell culture process model is developed to mimic historical manufacturing scale performance for a late stage CHO‐based monoclonal antibody program. Using multivariate analysis (MVA) as primary data analysis tool in addition to traditional univariate analysis techniques to identify gaps between scales, process adjustments were implemented at bench scale resulting in an improved scale down cell culture process model. Finally we propose an approach for small scale model qualification including three main aspects: MVA, comparison of key physiological rates, and comparison of product quality attributes. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 30:152–160, 2014
A novel approach to achieving modular retrovirus clearance for a parvovirus filter
Authors: Stuckey Juliana , Strauss Daniel , Venkiteshwaran Adith , Gao Jinxin , Luo Wen , Quertinmont Michelle , O'Donnell Sean , Chen Dayue
Published: October 2013

Viral filtration is routinely incorporated into the downstream purification processes for the production of biologics produced in mammalian cell cultures (MCC) to remove potential viral contaminants. In recent years, the use of retentive filters designed for retaining parvovirus (∼20 nm) has become an industry standard in a conscious effort to further improve product safety. Since retentive filters remove viruses primarily by the size exclusion mechanism, it is expected that filters designed for parvovirus removal can effectively clear larger viruses such as retroviruses (∼100 nm). In an attempt to reduce the number of viral clearance studies, we have taken a novel approach to demonstrate the feasibility of claiming modular retrovirus clearance for Asahi Planova 20N filters. Porcine parvovirus (PPV) and xenotropic murine leukemia virus (XMuLV) were co‐spiked into six different feedstreams and then subjected to laboratory scale Planova 20N filtration. Our results indicate that Planova 20N filters consistently retain retroviruses and no retrovirus has ever been detected in the filtrates even when significant PPV breakthrough is observed. Based on the data from multiple in‐house viral validation studies and the results from the co‐spiking experiments, we have successfully claimed a modular retrovirus clearance of greater than 6 log10 reduction factors (LRF) to support clinical trial applications in both USA and Europe. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 30:79–85, 2014
Engineering stem cell fate with biochemical and biomechanical properties of microcarriers
Authors: Sart Sébastien , Agathos Spiros N. , Li Yan
Published: October 2013

Microcarriers have been widely used for various biotechnology applications because of their high scale‐up potential, high reproducibility in regulating cellular behavior, and well‐documented compliance with current Good Manufacturing Practices (cGMP). Recently, microcarriers have been emerging as a novel approach for stem cell expansion and differentiation, enabling potential scale‐up of stem cell‐derived products in large bioreactors. This review summarizes recent advances of using microcarriers in mesenchymal stem cell (MSC) and pluripotent stem cell (PSC) cultures. From the reported data, efficient expansion and differentiation of stem cells on microcarriers rely on their ability to modulate cell shape (i.e. round or spreading) and cell organization (i.e. aggregate size). Nonetheless, current screening of microcarriers remains empirical, and accurate understanding of how stem cells interact with microcarriers still remains unknown. This review suggests that accurate characterization of biochemical and biomechanical properties of microcarriers is required to fully exploit their potential in regulating stem cell fate decision. Due to the variety of microcarriers, such detailed analyses should lead to the rational design of application‐specific microcarriers, enabling the exploitation of reproducible effects for large scale biomedical applications. © 2013 American Institute of Chemical EngineersBiotechnol. Prog., 29:1354–1366, 2013
The effect of antiapoptosis genes on clarification performance
Authors: Potty Ajish S. R. , Xenopoulos Alex , Patel Sonal , Prentice Holly , DiLeo Anthony
Published: October 2013

Optimal bioreactor harvest time is typically determined based on maximizing product titer without compromising product quality. We suggest that ease of downstream purification should also be considered during harvest. In this view, we studied the effect of antiapoptosis genes on downstream performance. Our hypothesis was that more robust cells would exhibit less cell lysis and thus generate lower levels of cell debris and host‐cell contaminants. We focused on the clarification unit operation, measuring postclarification turbidity and host‐cell protein (HCP) concentration as a function of bioreactor harvest time/cell viability. In order to mimic primary clarification using disk‐stack centrifugation, a scale‐down model consisting of a rotating disk (to simulate shear in the inlet feed zone of the centrifuge) and a swinging‐bucket lab centrifuge was used. Our data suggest that in the absence of shear during primary clarification (typical of depth filters), a 20–50% reduction in HCP levels and 50–65% lower postcentrifugation turbidity was observed for cells with antiapoptosis genes compared to control cells. However, on exposing the cells to shear levels typical in a disk‐stack centrifuge, the reduction in HCP was 10–15% while no difference in postcentrifugation turbidity was observed. The maximum benefit of antiapoptosis genes is, therefore, realized using clarification options that involve low shear, <1 × 106 W/m3 and minimal damage to the cells. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 30:100–107, 2014
Symptom burden and splenomegaly in patients with myelofibrosis in the United States: a retrospective medical record review
Authors: Mitra Debanjali , Kaye James A. , Piecoro Lance T. , Brown Jennifer , Reith Kelly , Mughal Tariq I. , Sarlis Nicholas J.
Published: October 2013

Myelofibrosis (MF) is a clonal hematopoietic malignancy characterized by constitutional and localized symptoms, progressive splenomegaly, bone marrow fibrosis, and cytopenias. Although MF is well studied, few studies exist regarding its symptomatic burden in routine clinical practice. This study aimed to characterize symptoms and other clinical features of MF among patients in the United States. We conducted a retrospective medical record review of adult patients with an MF diagnosis between 1 January 2005 and 31 March 2010, stratified by the presence of palpable splenomegaly. Eligible patients had 12 months or more of follow‐up after diagnosis (or after detection of splenomegaly, if present) unless death occurred. Demographic and clinical characteristics, MF‐related symptoms, and treatments were reported by treating physicians. We report on 180 MF patients: 102 with splenomegaly, 78 without. Median age was 66 years, 63% were male, and 82% had intermediate‐2 or high‐risk MF (International Prognostic Scoring System). Fatigue was reported by ~85% of patients; weight loss, night sweats, and fever (any grade) were each reported by 50% or more of patients. Generalized abdominal pain, left subcostal pain, and early satiety occurred more frequently among patients with splenomegaly. Multiple symptoms were reported by 95% of patients. Common comorbidities were hypertension, diabetes, and chronic pulmonary disease. Symptoms are common in MF patients, regardless of the presence of palpable splenomegaly. Careful assessment of symptom burden is an important aspect of the clinical evaluation of patients with MF.
Differential impact of the expression of the androgen receptor by age in estrogen receptor–positive breast cancer
Authors: Tokunaga Eriko , Hisamatsu Yuichi , Taketani Kenji , Yamashita Nami , Akiyoshi Sayuri , Okada Satoko , Tanaka Kimihiro , Saeki Hiroshi , Oki Eiji , Aishima Shinichi , Oda Yoshinao , Morita Masaru , Maehara Yoshihiko
Published: October 2013

We evaluated the expression of the androgen receptor (AR) to determine its significance in breast cancer. AR expression levels were analyzed in 250 invasive breast cancers by immunohistochemistry and any association with the clinicopathological features was evaluated. AR expression was higher in estrogen receptor (ER)‐positive cases than in ER‐negative cases (P < 0.0001). AR expression was associated with ER level, and it increased with age in ER‐positive cases. The cut‐off value was determined to be 75% (Cancer Res. 2009;69:6131–6140), and AR expression was considered to be high in 155 (62%) cases. High AR expression significantly correlated with lower nuclear grade (P < 0.0001), ER and progesterone receptor (PR) positivity (P < 0.0001 and P = 0.0022), HER2 negativity (P = 0.0113), lower Ki67 index (P < 0.0001) and a longer disease‐free survival (DFS) and distant metastasis‐free survival (DMFS) (P = 0.0003 and 0.0107). This association between a high AR expression and a good DFS and DMFS was significant for ER‐positive tumors (P < 0.0001 and P = 0.0018); however, no association existed between AR expression and prognosis for ER‐negative tumors. In patients ≤51 years old, a high AR expression level significantly correlated with a better prognosis, but this was not significant in patients who were 50 or younger. Multivariate Cox hazard analyses revealed AR expression to be independently associated with a good prognosis in overall patients (HR 0.46, P = 0.0052) and in the ER‐positive cohort (HR 0.34, P = 0.0009). AR expression is associated with a less aggressive phenotype and a good prognosis in patients with ER‐positive breast cancer. This is considered to be a specific phenomenon for postmenopausal breast cancer patients.
Ipilimumab and radiation therapy for melanoma brain metastases
Authors: Silk Ann W. , Bassetti Michael F. , West Brady T. , Tsien Christina I. , Lao Christopher D.
Published: October 2013

Ipilimumab, an antibody that enhances T‐cell activation, may augment immunogenicity of tumor cells that are injured by radiation therapy. We hypothesized that patients with melanoma brain metastasis treated with both ipilimumab and radiotherapy would have improved overall survival, and that the sequence of treatments may affect disease control in the brain. We analyzed the clinical and radiographic records of melanoma patients with brain metastases who were treated with whole brain radiation therapy or stereotactic radiosurgery between 2005 and 2012. The hazard ratios for survival were estimated to assess outcomes as a function of ipilimumab use and radiation type. Seventy patients were identified, 33 of whom received ipilimumab and 37 who did not. The patients who received ipilimumab had a censored median survival of 18.3 months (95% confidence interval 8.1–25.5), compared with 5.3 months (95% confidence interval 4.0–7.6) for patients who did not receive ipilimumab. Ipilimumab and stereotactic radiosurgery were each significant predictors of improved overall survival (hazard ratio = 0.43 and 0.45, with P = 0.005 and 0.008, respectively). Four of 10 evaluable patients (40.0%) who received ipilimumab prior to radiotherapy demonstrated a partial response to radiotherapy, compared with two of 22 evaluable patients (9.1%) who did not receive ipilimumab. Ipilimumab is associated with a significantly reduced risk of death in patients with melanoma brain metastases who underwent radiotherapy, and this finding supports the need for multimodality therapy to optimize patient outcomes. Prospective studies are needed and are underway.
Human motion reconstruction from sparse 3D motion sensors using kernel CCA‐based regression
Authors: Kim Jongmin , Seol Yeongho , Lee Jehee
Published: October 2013

This paper presents a real‐time performance animation system that reproduces full‐body character animation based on sparse three‐dimensional (3D) motion sensors on a performer. Producing faithful character animation from this setting is a mathematically ill‐posed problem, because input data from the sensors are not sufficient to determine the full degrees of freedom of a character. Given the input data from 3D motion sensors, we select similar poses from a motion database and build an online local model that transforms the low‐dimensional input signal into a high‐dimensional character pose. A regression method based on kernel canonical correlation analysis (CCA) is employed, because it effectively handles a wide variety of motions. Examples show that various human motions are naturally reproduced by the proposed method. Copyright © 2013 John Wiley & Sons, Ltd.
Cortisone induces insulin resistance in C2C12 myotubes through activation of 11beta‐hydroxysteroid dehydrogenase 1 and autocrinal regulation
Authors: Park Seung Yeon , Bae Ji Hyun , Cho Young Sik
Published: October 2013

The enzyme 11β‐hydroxysteroid dehydrogenase 1 (11β‐HSD1) is known to catalyse inactive glucocorticoids into active forms, and its dysregulation in adipose and muscle tissues has been implicated in the development of metabolic syndrome. To delineate the molecular mechanism by which active cortisol has an antagonizing effect against insulin, we optimized the metabolic production of cortisol and its biological functions in myotubes (C2C12). Myotubes supplemented with cortisone actively catalysed its conversion into cortisol, which in turn abolished phosphorylation of Akt in response to insulin treatment. This led to diminished uptake of insulin‐induced glucose. This was corroborated by the application of 11β‐HSD1 inhibitor glycyrrhetinic acid and a glucocorticoid receptor antagonist RU‐486, which reversed completely the antagonizing effects of cortisol on insulin action. Therefore, development of specific inhibitors targeting 11β‐HSD1 might be a promising way to improve impaired insulin‐stimulated glucose uptake. Copyright © 2013 John Wiley & Sons, Ltd.
Synthesis and Characterization of Cell‐Permeable Caged Phosphates that Can Be Photolyzed by Visible Light or 800 nm Two‐Photon Photolysis
Authors: Herbivo Cyril , Omran Ziad , Revol Julia , Javot Hélène , Specht Alexandre
Published: October 2013

We report the synthesis and photolytic properties of caged inorganic phosphates (Pi compounds) based on the 2‐(4′‐{bis[2‐(2‐methoxyethoxy)ethyl]amino}‐4‐nitro‐[1,1′‐biphenyl]‐3‐yl)propan‐1‐ol (EANBP) and 7‐(diethylamino)coumarin‐4‐yl]methyl (DEACM) protecting groups. The EANBP‐Pi showed unprecedented photolysis efficiency at 405 nm, with 95 % release of free phosphate and a quantum yield of 0.28. Thanks to the high two‐photon sensitivity of the EANBP chromophore, Pi release through two‐photon photolysis is also possible, with an action cross section of 20.5 GM at 800 nm. Two bioactivatable acetoxymethyl protection groups were added to the “caged‐Pi” compounds. The resulting triesters of phosphoric acid were able to diffuse through the cellular membranes of plant cells. Once inside a cell, the cleavage of these biocleavable motifs by intracellular esterases allows intracellular accumulation of EANBP‐Pi. Bis(AM)‐EANBP‐Pi therefore represents a very attractive tool for study of the Pi signal transduction cascade in living cells.
Therapeutic effects of hepatocyte growth factor‐overexpressing human umbilical cord blood‐derived mesenchymal stem cells on liver fibrosis in rats
Authors: Seo Kyoung‐Won , Sohn Suh‐Young , Bhang Dong‐Ha , Nam Myeong‐Jin , Lee Hee‐Woo , Youn Hwa‐Young
Published: October 2013

Fibrosis is a common end stage for a variety of liver diseases, including most chronic liver diseases, and results from an imbalance between collagen deposition and degradation. Mesenchymal stem cells (MSCs) have the ability to migrate into fibrotic livers and differentiate into hepatocytes. Hepatocyte growth factor (HGF) has potent anti‐apoptotic and mitogenic effects on hepatocytes during liver injury and plays an essential role in the development and regeneration of the liver. In this study, human HGF‐overexpressing human umbilical cord blood‐derived MSCs (hHGF‐HUCB‐MSCs) were prepared using the pMEX Expression System, and the upregulation of hHGF expression was confirmed by RT‐PCR and ELISA. HGF expressed by hHGF‐HUCB‐MSCs exerted a stimulatory effect on hepatocyte proliferation in vitro. hHGF‐HUCB‐MSCs were transplanted to investigate the therapeutic effects of these cells on carbon tetrachloride (CCL4)‐induced liver fibrosis in a rat model. After 4 weeks of cell treatment once per week with 2 × 106 cells, biochemical analysis of the serum and histopathological analysis of the liver tissue were performed. The results of the biochemical analysis of the serum show that the hHGF‐HUCB‐MSC‐treated group had higher levels of alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase, indicating the improvement of liver function. Histopathology showed that the hHGF‐HUCB‐MSC‐treated group had reduction in the density of collagen fibres. Thus hHGF‐HUCB‐MSCs can enhance liver regeneration and could be useful for the treatment of patients with liver fibrosis or cirrhosis.
Expression of embryonic stem cell markers and osteogenic differentiation potential in cells derived from periodontal granulation tissue
Authors: Ronay Valerie , Belibasakis Georgios N. , Attin Thomas , Schmidlin Patrick R. , Bostanci Nagihan
Published: October 2013

The aim of this study was to identify if cells obtained from periodontal granulation tissue possess embryonic stem cell properties and osteogenic capacities in vitro. Periodontal granulation tissue was removed from one furcation and one infrabony defect (FGTC/IGTC‐furcation/infrabony defect derived granulation tissue cells) of six patients. The extracted tissues were treated with collagenase/dispase solution, cultured and passaged twice, while a fraction of them was bacteriologically analyzed. Upon reaching confluence, total RNA was extracted, followed by cDNA synthesis and real‐time PCR analysis. Gene expression levels of collagen type I, alkaline phosphatase (ALP), and the embryonic stem cell markers Nanog, Oct‐4, Rex‐1 and Sox‐2 were measured, calibrated against the housekeeping gene GAPDH. Further, osteogenic differentiation was induced. Mineralized matrix formation was confirmed by von Kossa staining, and ALP activity was measured colorimetrically. The total bacterial load amounted to 9.4 ± 14.6 × 106 counts/mg of tissue for IGTC, and 11.1 ± 6.1 × 106 counts/of tissue for FGTC. Among the embryonic stem cell markers (FGTC/IGTC), Nanog was most highly expressed (3.48 ± 1.2/5.85 ± 5.7), followed by Oct‐4 (1.79 ± 0.69/2.85 ± 2.5), Sox‐2 (0.66 ± 0.3/1.26 ± 1.4) and Rex‐1 (0.06 ± 0.0/0.04 ± 0.0). The osteogenic differentiation process was positive in both FGTC and IGTC, judged by increased von Kossa staining, and elevated ALP activity and gene expression. This study provides evidence that infected periodontal granulation tissue harbors cells expressing embryonic stem cell markers, and exhibiting osteogenic capacities when in culture in vitro.
Development of interspecies nuclear transfer embryos reconstructed with argali (Ovis ammon) somatic cells and sheep ooplasm
Authors: Pan Xiaoyan , Zhang Yanli , Guo Zhiqin , Wang Feng
Published: October 2013

Interspecies nuclear transfer has already achieved success in several species, which shows great potential in recovery and conservation of endangered animals. The study was conducted to establish an efficient system for in vitro argali (Ovis ammon)‐sheep embryo reconstruction via interspecies somatic cell nuclear transfer (iSCNT). The competence of domestic sheep cytoplasts to reprogram the adult argali fibroblast nuclei was evaluated, and the effects of enucleation methods and donor cell passage and cell state on the in vitro development of argali‐sheep cloned embryos were also examined. Sheep oocytes could support argali and sheep fibroblast cell nuclei transfer and develop to blastocysts in vitro. Oocytes matured for 21–23 h and enucleated by chemically assisted enucleation (CAE) had a higher enucleation rate than blind enucleation (BE), but the development rate of iSCNT embryos was the same (P > 0.05). Moreover, passage numbers of fibroblast cells <10, as well as the cell cycle stages did not affect the development rate of iSCNT reconstructed embryos. Thus sheep cytoplasm successfully supports argali nucleus development to blastocyst stage after optimising the nuclear transfer procedure, which indicates that iSCNT can be used to conserve endangered argali in the near future.
Role of microRNA‐27a in myoblast differentiation
Authors: Chen Xiaoling , Huang Zhiqing , Chen Daiwen , Yang Ting , Liu Guangmang
Published: October 2013

MicroRNAs (miRNAs) are a class of endogenous non‐coding RNAs that are critically involved in roles in various aspects of skeletal myogenesis. microRNA miR‐27a promotes myoblast proliferation by targeting myostatin, a critical inhibitor of skeletal muscle development, but its mode of action in myoblast differentiation remains unclear. We have found that expression of miR‐27a and myostatin were upregulated and downregulated, respectively, during myoblast differentiation. Overexpression of miR‐27a increased the number of myosin heavy chain (MHC)‐positive cells and upregulated mRNA and protein of MyoD and myogenin. These findings indicate that miR‐27a plays a role in enhancing myoblast differentiation.
Comparing the effect of equiaxial cyclic mechanical stimulation on GATA4 expression in adipose‐derived and bone marrow‐derived mesenchymal stem cells
Authors: Amin Susan , Banijamali Seyede Elnaz , Tafazoli‐Shadpour Mohammad , Shokrgozar Mohammad Ali , Dehghan Mohammad Mehdi , Haghighipour Nooshin , Mahdian Reza , Bayati Vahid , Abbasnia Pegah
Published: October 2013

Myocardium is prone to mechanical stimuli among which pulsatile blood flow exerts both radial and longitudinal strains on the heart. Recent studies have shown that mechanical stimulation can notably influence regeneration of cardiac muscle cells. GATA4 is a cardiac‐specific transcription factor that plays an important role in late embryonic heart development. Our study aimed at investigating the effect of equiaxial cyclic strain on GATA4 expression in adipose‐derived (ASCs) and bone marrow‐derived (BMSCs) mesenchymal stem cells. For this reason, both ASCs and BMSCs were studied in four distinct groups of chemical, mechanical, mechano‐chemical and negative control. According to this categorisation, the cells were exposed to cyclic mechanical loading and/or 5‐azacytidine as the chemical factor. The level of GATA4 expression was then quantified using real‐time PCR method on the first, fourth and seventh days. The results show that: (1) equiaxial cyclic stimulation of mesenchymal stem cells could promote GATA4 expression from the early days of induction and as it went on, its combination with chemical factor elevated expression; (2) cyclic strain could accelerate GATA4 expression compared to the chemical factor; (3) in this regard, these results indicate a higher capacity of ASCs than BMSCs to express GATA4.
Incidence and implications of idiopathic thrombocytopenia following transcatheter aortic valve replacement with the Edwards Sapien© valves: A single center experience
Authors: McCabe James M. , Huang Pei‐Hsiu , Riedl Lauren A. , Devireddy Srikanth R. , Grondell Jennifer , Connors Annie C. , Davidson Michael J. , Eisenhauer Andrew C. , Welt Frederick G. P.
Published: October 2013

Objectives

To determine the incidence and etiology of thrombocytopenia following transcatheter Aortic valve replacement (TAVR).

Background

the use of TAVR in the United States has grown rapidly. Anecdotally, thrombocytopenia following TAVR with the Sapien valves has been observed, though little is known about this phenomenon.

Methods

All patients treated with TAVR using a Sapien valve or who underwent isolated balloon aortic valvuloplasty (BAV) at Brigham and Women's Hospital from October 2009 through November 2012 were analyzed. Post‐procedure thrombocytopenia severity was stratified as none (>150,000 cells/μL), mild (100–150,000), and moderate to severe (<100,000). Summary statistics and simple comparisons were evaluated. Linear regression models were used to identify patient or procedural factors associated with platelet count nadir.

Results

112 TAVR and 105 BAV patients were analyzed. Following TAVR the prevalence of thrombocytopenia was 69% and the incidence of new thrombocytopenia was 45% compared to 37% and 17% respectively following BAV (P < 0.01). Similar results were found across all strata of thrombocytopenia severity. Post‐TAVR platelet nadirs were greater by 1,840 cells/μL for each 1% increase in STS score (P = 0.03) and 670 cells/μL greater for each 1 mmHg increase in pre‐TAVR mean aortic stenosis gradient. Among TAVR patients, thrombocytopenia appears to spontaneously resolve an average of 8 days post‐procedure. No differences in clinical outcomes based on thrombocytopenia severity were observed except for an increased use of blood products (P = 0.05).

Conclusions

Thrombocytopenia following TAVR with the Edwards' Sapien valves is a frequent but generally self‐limited process. The etiology of this phenomenon is unknown. © 2013 Wiley Periodicals, Inc.
Working through complexities of radial and brachial vasculature during transradial approach
Authors: Patel Tejas , Shah Sanjay , Pancholy Samir , Radadiya Rajnikant , Deora Surender , Vyas Chirayu , Hamon Martial , Gilchrist Ian C.
Published: October 2013

Objectives

The aim of this review is to discuss different methods of working through radial artery (RA) and brachial artery (BA) vascular complexities to increase the success rate of transradial approach (TRA).

Background

Anatomical complexities of RA and BA vasculature are an important reason for failure of TRA. There are few current data describing methods to overcome these challenges and reduce TRA failure.

Methods

A series of the primary RA and BA anatomical complexities are identified and management techniques to overcome these variants are discussed.

Results

RA and BA vascular complexities can be divided into three subsets including (1) radial artery spasm (RAS); (2) variant anatomy including tortuosity, loops, and anomalous origin of RA; (3) acquired abnormalities including perforations, atherosclerotic lesions, and calcification of RA. In‐depth discussion with supportive examples for the identification and management of these challenges are provided. A classification of RAS and perforations is shown and simple algorithms that have been developed for management of RAS, perforations and loops are explained.

Conclusions

Despite lower rates of bleeding and vascular complications as compared to transfemoral approach (TFA), the adoption has been relatively slow particularly due to higher failure rates. Anatomical complexities of RA and BA vasculature play an important role for TRA failure cases. Using a simply framework to classify the anatomical or functional problem, and then approach these challenges in a logical sequence should facilitate management and increase success rate for TRA. © 2013 Wiley Periodicals, Inc.