bluuebunny/crossref_metadata_2025_split huggingface dataset:
Y‐chromosome identification in circulating cell‐free fetal DNA using surface plasmon resonance
Authors: Breveglieri Giulia , Bassi Elisabetta , Carlassara Silvia , Cosenza Lucia Carmela , Pellegatti Patrizia , Guerra Giovanni , Finotti Alessia , Gambari Roberto , Borgatti Monica 
Published: February 2016

AbstractObjectiveSince the discovery of cell‐free fetal DNA (cffDNA) in maternal plasma, diagnostic non‐invasive prenatal methods have been developed or optimized for fetal sex determination and identification of genetic diseases. As far as fetal sex determination, this might be important for therapeutic intervention on sex‐associated pathologies such as Duchenne muscular dystrophy, hemophilia and congenital adrenal hyperplasia. Surface plasmon resonance (SPR)‐based biosensors might be useful for these studies, because they allow to monitor the molecular interactions in real‐time providing qualitative and quantitative information, through kinetics, affinity and concentration analyses.MethodsThe BiacoreTM X100 has been applied to identify Y‐chromosome sequence in cffDNA obtained from plasma samples of 26 pregnant women at different gestational ages. We have performed SPR‐based analysis of SRY PCR products using SRY‐specific probes immobilized on the sensor chip.ResultsWe have demonstrated that there is a statistically significant difference between samples collected by pregnancies carrying male or female fetuses. Moreover, cffDNA obtained at early gestational ages and not detectable by conventional quantitative real‐time PCR can be discriminated with high accuracy and reliability using SPR‐based biosensors.ConclusionsThese data, in addition to their direct applicability in more extensive diagnostic trials, should be considered as the basis of future developments. © 2016 John Wiley & Sons, Ltd.
Model for the allosteric regulation of the Na+/Ca2+ exchanger NCX
Authors: Abiko Layara Akemi , Vitale Phelipe M. , Favaro Denize C. , Hauk Pricila , Li Da‐Wei , Yuan Jiaqi , Bruschweiler‐Li Lei , Salinas Roberto K. , Brüschweiler Rafael 
Published: February 2016

ABSTRACTThe Na+/Ca2+ exchanger provides a major Ca2+ extrusion pathway in excitable cells and plays a key role in the control of intracellular Ca2+ concentrations. In Canis familiaris, Na+/Ca2+ exchanger (NCX) activity is regulated by the binding of Ca2+ to two cytosolic Ca2+‐binding domains, CBD1 and CBD2, such that Ca2+‐binding activates the exchanger. Despite its physiological importance, little is known about the exchanger's global structure, and the mechanism of allosteric Ca2+‐regulation remains unclear. It was found previously that for NCX in the absence of Ca2+ the two domains CBD1 and CBD2 of the cytosolic loop are flexibly linked, while after Ca2+‐binding they adopt a rigid arrangement that is slightly tilted. A realistic model for the mechanism of the exchanger's allosteric regulation should not only address this property, but also it should explain the distinctive behavior of Drosophila melanogaster's sodium/calcium exchanger, CALX, for which Ca2+‐binding to CBD1 inhibits Ca2+ exchange. Here, NMR spin relaxation and residual dipolar couplings were used to show that Ca2+ modulates CBD1 and CBD2 interdomain flexibility of CALX in an analogous way as for NCX. A mechanistic model for the allosteric Ca2+ regulation of the Na+/Ca2+ exchanger is proposed. In this model, the intracellular loop acts as an entropic spring whose strength is modulated by Ca2+‐binding to CBD1 controlling ion transport across the plasma membrane. Proteins 2016; 84:580–590. © 2016 Wiley Periodicals, Inc.
Anti‐inflammatory effects of the novel inhaled phosphodiesterase type 4 inhibitor CHF6001 on virus‐inducible cytokines
Authors: Edwards Michael R. , Facchinetti Fabrizio , Civelli Maurizio , Villetti Gino , Johnston Sebastian L. 
Published: February 2016

AbstractRespiratory virus infections precipitate asthma and chronic obstructive pulmonary disease (COPD) exacerbations, with most exacerbations due to rhinovirus infection. Both asthma and COPD exacerbations are not well controlled by steroid therapies, and there is a much research interest in finding improved therapies or combinations of therapies for controlling exacerbations. CHF6001 is a new, inhaled highly potent and selective phosphodiesterase type 4 (PDE4) inhibitor. Using in vitro human bronchial epithelial cells (BEAS‐2B), we investigated the potential anti‐inflammatory effects of CHF6001 on rhinovirus (RV1B)‐induced cytokines. Cytokine mRNA was measured by real‐time PCR, while protein release was measured by ELISA. CHF6001 was used in a 7‐point dose–response curve (1000–0.001 nmol/L) as a 1.5‐h pretreatment prior to infection in comparison with roflumilast. Both roflumilast and CHF6001 reduced RV1B‐induced IL‐8, IL‐29, IP‐10, and RANTES mRNA and protein in a concentration‐dependent manner. Generally, CHF6001 was 13‐ to 16‐fold more potent (subnanomolar EC50 values) than roflumilast at reducing IL‐8, IL‐29, IP‐10, and RANTES mRNA and protein release, but had similar efficacies. In combination with the steroid fluticasone propionate (1 nmol/L), CHF6001 had additive effects, significantly reducing RV‐induced cytokines when compared with steroid or CHF6001 alone. Combined low‐dose steroid and low‐dose CHF6001 had a similar efficacy as high‐dose steroid or CHF6001 alone, indicating the combination had steroid and PDE4 inhibitor sparing effects. Overall results indicate that PDE4 inhibitors have anti‐inflammatory activity against virus‐induced inflammatory mediators and that CHF6001 is more potent than roflumilast.
Lack of cross‐resistance to FF‐10501, an inhibitor of inosine‐5′‐monophosphate dehydrogenase, in azacitidine‐resistant cell lines selected from SKM‐1 and MOLM‐13 leukemia cell lines
Authors: Murase Motohiko , Iwamura Hiroyuki , Komatsu Kensuke , Saito Motoki , Maekawa Toshihiko , Nakamura Takaaki , Yokokawa Takuya , Shimada Yasuhiro 
Published: February 2016

AbstractResistance to azacitidine is a major issue in the treatments of myelodysplastic syndrome and acute myeloid leukemia, and previous studies suggest that changes in drug metabolism are involved in the resistance. Therefore, drugs with mechanisms resistant or alternative to such metabolic changes have been desired for the treatment of resistant disease. We generated azacitidine‐resistant cells derived from SKM‐1 and MOLM‐13 leukemia cell lines in vitro, analyzed the mechanisms, and examined the impact on the efficacy of other antimetabolic drugs. It appeared that the cell growth‐inhibitory effect of azacitidine, expression levels of uridine–cytidine kinase 2, and the concentrations of azacitidine triphosphate were remarkably decreased in the resistant cells compared with those in parent cells. These results were consistent with previous observations that azacitidine resistance is derived from metabolic changes. Cross‐resistance of greater than 10‐fold (shift in IC50 value) was observed in azacitidine‐resistant cells for decitabine and for cytarabine, but not for gemcitabine or the inosine‐5′‐monophosphate dehydrogenase (IMPDH) inhibitors FF‐10501 and mycophenolate mofetil (cross‐resistance to 5‐fluorouracil was cell line dependent). The IMPDH inhibitors maintained their cell growth‐inhibitory activities in the azacitidine‐resistant cell lines, in which the levels of adenine phosphoribosyltransferase (which converts FF‐10501 to its active form, FF‐10501 ribosylmonophosphate [FF‐10501RMP]), FF‐10501RMP, and the target enzyme, IMPDH, were equivalent to those in the parent cell lines. These results suggest that an IMPDH inhibitor such as FF‐10501 could be an alternative therapeutic treatment for leukemia patients with acquired resistance to azacitidine.
Slow drug delivery decreased total body clearance and altered bioavailability of immediate‐ and controlled‐release oxycodone formulations
Authors: Li Yan , Sun Duxin , Palmisano Maria , Zhou Simon 
Published: February 2016

AbstractOxycodone is a commonly used analgesic with a large body of pharmacokinetic data from various immediate‐release or controlled‐release formulations, under different administration routes, and in diverse populations. Longer terminal half‐lives from extravascular administration as compared to IV administration have been attributed to flip‐flop pharmacokinetics with the rate constant of absorption slower than elimination. However, PK parameters from the extravascular studies showed faster absorption than elimination. Sustained release formulations guided by the flip‐flop concept produced mixed outcomes in formulation development and clinical studies. This research aims to develop a mechanistic knowledge of oxycodone ADME, and provide a consistent interpretation of diverging results and insight to guide further extended release development and optimize the clinical use of oxycodone. PK data of oxycodone in human studies were collected from literature and digitized. The PK data were analyzed using a new PK model with Weibull function to describe time‐varying drug releases/ oral absorption, and elimination dependent upon drug input to the portal vein. The new and traditional PK models were coded in NONMEM. Sensitivity analyses were conducted to address the relationship between rates of drug release/absorption and PK profiles plus terminal half‐lives. Traditional PK model could not be applied consistently to describe drug absorption and elimination of oxycodone. Errors were forced on absorption, elimination, or both parameters when IV and PO profiles were fitted separately. The new mechanistic PK model with Weibull function on absorption and slower total body clearance caused by slower absorption adequately describes the complex interplay between oxycodone absorption and elimination in vivo. Terminal phase of oxycodone PK profile was shown to reflect slower total body drug clearance due to slower drug release/absorption from oral formulations. Mechanistic PK models with Weibull absorption functions, and release rate‐dependent saturable total body clearance well described the diverging oxycodone absorption and elimination kinetics in the literature. It showed no actual drug absorption during the terminal phase, but slower drug clearance caused by slower release/absorption producing the appearance of flip‐flop and offered new insight for the development of modified release formulations and clinical use of oxycodone.
Role of endoplasmic reticulum stress in drug‐induced toxicity
Authors: Foufelle Fabienne , Fromenty Bernard 
Published: February 2016

AbstractDrug‐induced toxicity is a key issue for public health because some side effects can be severe and life‐threatening. These adverse effects can also be a major concern for the pharmaceutical companies since significant toxicity can lead to the interruption of clinical trials, or the withdrawal of the incriminated drugs from the market. Recent studies suggested that endoplasmic reticulum (ER) stress could be an important event involved in drug liability, in addition to other key mechanisms such as mitochondrial dysfunction and oxidative stress. Indeed, drug‐induced ER stress could lead to several deleterious effects within cells and tissues including accumulation of lipids, cell death, cytolysis, and inflammation. After recalling important information regarding drug‐induced adverse reactions and ER stress in diverse pathophysiological situations, this review summarizes the main data pertaining to drug‐induced ER stress and its potential involvement in different adverse effects. Drugs presented in this review are for instance acetaminophen (APAP), arsenic trioxide and other anticancer drugs, diclofenac, and different antiretroviral compounds. We also included data on tunicamycin (an antibiotic not used in human medicine because of its toxicity) and thapsigargin (a toxic compound of the Mediterranean plant Thapsia garganica) since both molecules are commonly used as prototypical toxins to induce ER stress in cellular and animal models.
Random points in halfspheres
Authors: Bárány Imre , Hug Daniel , Reitzner Matthias , Schneider Rolf 
Published: February 2016

AbstractA random spherical polytope Pn in a spherically convex set as considered here is the spherical convex hull of n independent, uniformly distributed random points in K. The behaviour of Pn for a spherically convex set K contained in an open halfsphere is quite similar to that of a similarly generated random convex polytope in a Euclidean space, but the case when K is a halfsphere is different. This is what we investigate here, establishing the asymptotic behaviour, as n tends to infinity, of the expectation of several characteristics of Pn, such as facet and vertex number, volume and surface area. For the Hausdorff distance from the halfsphere, we obtain also some almost sure asymptotic estimates. © 2016 Wiley Periodicals, Inc. Random Struct. Alg., 50, 3–22, 2017
Großbrücken in Deutschland
Authors: Reitz Dieter 
Published: February 2016

AbstractDie aktuelle Marktsituation steht im Bereich der Infrastruktur im Allgemeinen und vor allem im Brückenbau in einem besonderen Kontext. Waren die beiden großen Epochen des Stahlbrückenbaus, der Beginn der Massenmobilität in den 60er und Anfang der 70er Jahre des vergangenen Jahrhunderts sowie die Maßnahmen nach der Wiedervereinigung mit Ende der 90er und zu Beginn des 21. Jahrhunderts, geprägt durch Neubauten weitestgehend auf der grünen Wiese, so stehen wir heute womöglich am Anfang einer weiteren Phase mit wesentlich geänderten Rahmenbedingungen. Der Neubau von Fernstraßen wird sich auf einige Lückenschlüsse reduzieren. Dafür gilt es, das vorhandene Straßennetz fit für das nächste Jahrhundert zu machen. Nicht nur die in die Jahre gekommenen Bauwerke, auch die zu keiner Zeit erwartete Zunahme des Verkehrs und insbesondere des Schwerverkehrs führt zu Sanierungen und Erneuerungsmaßnahmen in nie gedachtem Umfang. Dabei stellt das Bauen unter der Aufrechterhaltung des Verkehrs eine besondere Herausforderung dar.Major bridges in Germany – Yesterday – today – tomorrow. The current market situation in the field of infrastructure in general and especially the superstructure are placed in a special context. Whereas the history of steel bridges started with the beginning of mass mobility in the 60s and early 70s of the last century as well as the reunification in the late 90s, our century is now battled with completely different conditions. A new area has started and is challenging us with all its pros and cons. The construction of new highways will be reduced to the closing of gaps. Therefore it is really important to focus on the existing road network and its future. Not only are old constructions a huge problem, especially the increase of traffic and particularly heavy load traffic are leading to redevelopment and renewal in never existing extent. All this leads to a very special challenge during the execution of all road and construction works.
Knockdown of DNMT1 and DNMT3a Promotes the Angiogenesis of Human Mesenchymal Stem Cells Leading to Arterial Specific Differentiation
Authors: Zhang Rui , Wang Nan , Zhang Li-Nan , Huang Na , Song Tie-Feng , Li Zheng-Zheng , Li Man , Luo Xue-Gang , Zhou Hao , He Hong-Peng , Zhang Xiao-Yu , Ma Wenjian , Zhang Tong-Cun 
Published: February 2016

Abstract
Human mesenchymal stem cells (hMSCs) possess the potential to differentiate into endothelial cells (EC). DNA methylation plays an important role in cell differentiation during development. However, the role of the DNA methyltransferases Dnmt1 and Dnmt3a in specific arterial differentiation of hMSCs is not clear. Here, we show that the CpG islands in the promoter regions of the EC specification and arterial marker genes were highly methylated in hMSCs based on bisulfite genomic sequencing. Treatment with the DNMT inhibitor 5-aza-dc induced the reactivation of EC specification and arterial marker genes by promoting demethylation of these genes as well as stimulating tube-like structure formation. The hMSCs with stable knockdown of Dnmt1/Dnmt3a were highly angiogenic and expressed several arterial specific transcription factors and marker genes. A Matrigel plug assay confirmed that Dnmt1/Dnmt3a stable knockdown hMSCs enhanced blood vessel formation compared with WT MSCs. We also identified that the transcription factor E2F1 could upregulate the transcription of arterial marker genes by binding to the promoters of arterial genes, suggesting its critical role for arterial specification. Moreover, miRNA gain/loss-of-function analyses revealed that miR152 and miR30a were involved in endothelial differentiation of hMSCs by targeting Dnmt1 and Dnmt3a, respectively. Taken together, these data suggest that Dnmt1 and Dnmt3a are critical regulators for epigenetic silencing of EC marker genes and that E2F1 plays an important role in promoting arterial cell determination.
Concise Review: Heteroplasmic Mitochondrial DNA Mutations and Mitochondrial Diseases: Toward iPSC-Based Disease Modeling, Drug Discovery, and Regenerative Therapeutics
Authors: Hatakeyama Hideyuki , Goto Yu-ichi 
Published: February 2016

Abstract
Mitochondria contain multiple copies of their own genome (mitochondrial DNA; mtDNA). Once mitochondria are damaged by mutant mtDNA, mitochondrial dysfunction is strongly induced, followed by symptomatic appearance of mitochondrial diseases. Major genetic causes of mitochondrial diseases are defects in mtDNA, and the others are defects of mitochondria-associating genes that are encoded in nuclear DNA (nDNA). Numerous pathogenic mutations responsible for various types of mitochondrial diseases have been identified in mtDNA; however, it remains uncertain why mitochondrial diseases present a wide variety of clinical spectrum even among patients carrying the same mtDNA mutations (e.g., variations in age of onset, in affected tissues and organs, or in disease progression and phenotypic severity). Disease-relevant induced pluripotent stem cells (iPSCs) derived from mitochondrial disease patients have therefore opened new avenues for understanding the definitive genotype-phenotype relationship of affected tissues and organs in various types of mitochondrial diseases triggered by mtDNA mutations. In this concise review, we briefly summarize several recent approaches using patient-derived iPSCs and their derivatives carrying various mtDNA mutations for applications in human mitochondrial disease modeling, drug discovery, and future regenerative therapeutics.
Jun-Mediated Changes in Cell Adhesion Contribute to Mouse Embryonic Stem Cell Exit from Ground State Pluripotency
Authors: Veluscek Giulia , Li Yaoyong , Yang Shen-Hsi , Sharrocks Andrew D. 
Published: February 2016

Abstract
Embryonic stem cells (ESC) are able to give rise to any somatic cell type. A lot is known about how ESC pluripotency is maintained, but comparatively less is known about how differentiation is promoted. Cell fate decisions are regulated by interactions between signaling and transcriptional networks. Recent studies have shown that the overexpression or downregulation of the transcription factor Jun can affect the ESC fate. Here we have focussed on the role of the Jun in the exit of mouse ESCs from ground state pluripotency and the onset of early differentiation. Transcriptomic analysis of differentiating ESCs reveals that Jun is required to upregulate a programme of genes associated with cell adhesion as ESCs exit the pluripotent ground state. Several of these Jun-regulated genes are shown to be required for efficient adhesion. Importantly this adhesion is required for the timely regulated exit of ESCs from ground state pluripotency and the onset of early differentiation events.
Cellular Ontogeny and Hierarchy Influence the Reprogramming Efficiency of Human B Cells into Induced Pluripotent Stem Cells
Authors: Muñoz-López Álvaro , van Roon Eddy. H.J. , Romero-Moya Damià , López-Millan Belén , Stam Ronald W. , Colomer Dolors , Nakanishi Mahito , Bueno Clara , Menendez Pablo 
Published: February 2016

Abstract
Although B cells have been shown to be refractory to reprogramming into pluripotency, induced pluripotent stem cells (iPSCs) have been very recently generated, at very low efficiency, from human cord blood (CB)- and peripheral blood (PB)-derived CD19+CD20 + B cells using nonintegrative tetracistronic OSKM-expressing Sendai Virus (SeV). Here, we addressed whether cell ontogeny and hierarchy influence the reprogramming efficiency of the B-cell compartment. We demonstrate that human fetal liver (FL)-derived CD19 + B cells are 110-fold easier to reprogram into iPSCs than those from CB/PB. Similarly, FL-derived CD34+CD19 + B progenitors are reprogrammed much easier than mature B cells (0.46% vs. 0.11%). All FL B-cell iPSCs carry complete VDJH rearrangements while 55% and 45% of the FL B-progenitor iPSCs carry incomplete and complete VDJH rearrangements, respectively, reflecting the reprogramming of developmentally different B progenitors (pro-B vs. pre-B) within a continuous differentiation process. Finally, our data suggest that successful B-cell reprogramming relies on active cell proliferation, and it is MYC-dependent as identical nonintegrative polycistronic SeV lacking MYC (OSKL or OSKLN) fail to reprogram B cells. The ability to efficiently reprogram human fetal primary B cells and B precursors offers an unprecedented opportunity for studying developmental B-lymphopoiesis and modeling B-cell malignances.
Pharmacological Regulation of In Situ Tissue Stem Cells Differentiation for Soft Tissue Calcification Treatment
Authors: Hu Jia-Jie , Yin Zi , Shen Wei-Liang , Xie Yu-Bin , Zhu Ting , Lu Ping , Cai You-Zhi , Kong Min-Jian , Heng Boon Chin , Zhou Yi-Ting , Chen Wei-Shan , Chen Xiao , Ouyang Hong-Wei 
Published: February 2016

AbstractCalcification of soft tissues, such as heart valves and tendons, is a common clinical problem with limited therapeutics. Tissue specific stem/progenitor cells proliferate to repopulate injured tissues. But some of them become divergent to the direction of ossification in the local pathological microenvironment, thereby representing a cellular target for pharmacological approach. We observed that HIF-2alpha (encoded by EPAS1 inclined form) signaling is markedly activated within stem/progenitor cells recruited at calcified sites of diseased human tendons and heart valves. Proinflammatory microenvironment, rather than hypoxia, is correlated with HIF-2alpha activation and promoted osteochondrogenic differentiation of tendon stem/progenitor cells (TSPCs). Abnormal upregulation of HIF-2alpha served as a key switch to direct TSPCs differentiation into osteochondral-lineage rather than teno-lineage. Notably, Scleraxis (Scx), an essential tendon specific transcription factor, was suppressed on constitutive activation of HIF-2alpha and mediated the effect of HIF-2alpha on TSPCs fate decision. Moreover, pharmacological inhibition of HIF-2alpha with digoxin, which is a widely utilized drug, can efficiently inhibit calcification and enhance tenogenesis in vitro and in the Achilles's tendinopathy model. Taken together, these findings reveal the significant role of the tissue stem/progenitor cells fate decision and suggest that pharmacological regulation of HIF-2alpha function is a promising approach for soft tissue calcification treatment.