bluuebunny/crossref_metadata_2025 huggingface dataset:
P2X7R‐mediated Ca2+‐independent d‐serine release via pannexin‐1 of the P2X7R‐pannexin‐1 complex in astrocytes
Authors: Pan Han‐Chi , Chou Yun‐Chia , Sun Synthia H.
Published: January 2015

d‐serine is a coagonist of N‐methyl‐d‐aspartate (NMDA) subtype of glutamate receptor and plays a role in regulating activity‐dependent synaptic plasticity. In this study, we examined the mechanism by which extracellular ATP triggers the release of d‐serine from astrocytes and discovered a novel Ca2+‐independent release mechanism mediated by P2X7 receptors (P2X7R). Using [3H] d‐serine, which was loaded into astrocytes via the neutral amino acid transporter 2 (ASCT2), we observed that ATP and a potent P2X7R agonist, 2′(3′)‐O‐(4‐benzoylbenzoyl)adenosine‐5′‐triphosphate (BzATP), stimulated [3H]D‐serine release and that were abolished by P2X7R selective antagonists and by shRNAs, whereas enhanced by removal of intracellular or extracellular Ca2+. The P2X7R‐mediated d‐serine release was inhibited by pannexin‐1 antagonists, such as carbenoxolone (CBX), probenecid (PBN), and 10Panx‐1 peptide, and shRNAs, and stimulation of P2X7R induced P2X7R‐pannexin‐1 complex formation. Simply incubating astrocytes in Ca2+/Mg2+‐free buffer also induced the complex formation, and that enhanced basal d‐serine release through pannexin‐1. The P2X7R‐mediated d‐serine release assayed in Ca2+/Mg2+‐free buffer was enhanced as well, and that was inhibited by CBX. Treating astrocytes with general protein kinase C (PKC) inhibitors, such as chelerythrine, GF109203X, and staurosporine, but not Ca2+‐dependent PKC inhibitor, Gö6976, inhibited the P2X7R‐mediated d‐serine release. Thus, we conclude that in astrocytes, P2X7R‐pannexin‐1 complex formation is crucial for P2X7R‐mediated d‐serine release through pannexin‐1 hemichannel. The release is Ca2+‐independent and regulates by a Ca2+‐independent PKC. The activated P2X7R per se is also functioned as a permeation channel to release d‐serine in part. This P2X7R‐mediated d‐serine release represents an important mechanism for activity‐dependent neuron‐glia interaction. GLIA 2015;63:877–893
Orientation Training and Job Satisfaction: A Sector and Gender Analysis
Authors: Tabvuma Vurain , Georgellis Yannis , Lange Thomas
Published: January 2015

Using data from the British Household Panel Survey (BHPS), we investigate how various types of job training impact upon employees’ job satisfaction and its domains. We find that orientation training exerts a significant positive effect on newcomer male employees’ job satisfaction in both the private and public sectors, but it increases the job satisfaction of newcomer female employees only in the public sector. Other types of job training have only a weak effect on job satisfaction. We attribute the predominance of orientation training as a strong predictor of job satisfaction to its important function of facilitating the workplace socialization of new employees by reducing the uncertainty about aspects of the job that are not always easily contractible. © 2015 Wiley Periodicals, Inc.
Parameter identification and uncertainty analysis for simulating streamflow in a river basin of Eastern India
Authors: Uniyal Bhumika , Jha Madan K. , Verma Arbind K.
Published: January 2015

Uncertainty is inherent in modelling studies. However, the quantification of uncertainties associated with a model is a challenging task, and hence, such studies are somewhat limited. As distributed or semi‐distributed hydrological models are being increasingly used these days to simulate hydrological processes, it is vital that these models should be equipped with robust calibration and uncertainty analysis techniques. The goal of the present study was to calibrate and validate the Soil and Water Assessment Tool (SWAT) model for simulating streamflow in a river basin of Eastern India, and to evaluate the performance of salient optimization techniques in quantifying uncertainties. The SWAT model for the study basin was developed and calibrated using Parameter Solution (ParaSol), Sequential Uncertainty Fitting Algorithm (SUFI‐2) and Generalized Likelihood Uncertainty Estimation (GLUE) optimization techniques. The daily observed streamflow data from 1998 to 2003 were used for model calibration, and those for 2004–2005 were used for model validation. Modelling results indicated that all the three techniques invariably yield better results for the monthly time step than for the daily time step during both calibration and validation. The model performances for the daily streamflow simulation using ParaSol and SUFI‐2 during calibration are reasonably good with a Nash–Sutcliffe efficiency and mean absolute error (MAE) of 0.88 and 9.70 m3/s for ParaSol, and 0.86 and 10.07 m3/s for SUFI‐2, respectively. The simulation results of GLUE revealed that the model simulates daily streamflow during calibration with the highest accuracy in the case of GLUE (R2 = 0.88, MAE = 9.56 m3/s and root mean square error = 19.70 m3/s). The results of uncertainty analyses by SUFI‐2 and GLUE were compared in terms of parameter uncertainty. It was found that SUFI‐2 is capable of estimating uncertainties in complex hydrological models like SWAT, but it warrants sound knowledge of the parameters and their effects on the model output. On the other hand, GLUE predicts more reliable uncertainty ranges (R‐factor = 0.52 for daily calibration and 0.48 for validation) compared to SUFI‐2 (R‐factor = 0.59 for daily calibration and 0.55 for validation), though it is computationally demanding. Although both SUFI‐2 and GLUE appear to be promising techniques for the uncertainty analysis of modelling results, more and more studies in this direction are required under varying agro‐climatic conditions for assessing their generic capability. Copyright © 2015 John Wiley & Sons, Ltd.
Nitrate fluctuations at the water table: implications for recharge processes and solute transport in the Chalk aquifer
Authors: Sorensen James P. R. , Butcher Andrew S. , Stuart Marianne E. , Townsend Barry R.
Published: January 2015

This study investigates fluctuations in nitrate concentration at the water table to improve understanding of unsaturated zone processes in the Chalk aquifer. Sampling was conducted using a novel multi‐level sampler during periods of water table rise over 5 years at a vertical resolution of 0.05 m. Nitrate concentration increased as the water table seasonally recovered, with similar inter‐annual trends with depth. The rising water table activated horizontal fractures facilitating the delivery of water elevated by up to 10 mg/l of nitrate with respect to the adjacent groundwater below. These fractures are considered to activate via piston displacement of water from the adjoining matrix. Hydrograph analysis identified 16 events which significantly perturbed the water table within 24–48 h of rainfall. Consistent nitrate concentrations indicate recharge through persistent fracture flow from the surface was not generally the primary driver of the rapid water table response during these events. Instead, the response was attributed to the piston displacement of porewater immediately above the water table. However, a single event in November 2012 delivered relatively dilute recharge indicating rapid persistent fracture flow following rainfall was possible to a depth of 14–15 m. Decreases in porewater nitrate concentration around fracture horizons and the dilution of many groundwater samples with respect to porewaters indicate a fresher source of water at depth. This was considered most likely to be a result of near surface water bypassing the matrix because of widespread mineralization on fracture surfaces, which retard water and solute exchange. Therefore, persistent fracture flow maybe considered a frequent process, operating independently of the matrix, and is not necessarily event driven. Copyright © 2015 John Wiley & Sons, Ltd.
A review of low‐cost space‐borne data for flood modelling: topography, flood extent and water level
Authors: Yan Kun , Di Baldassarre Giuliano , Solomatine Dimitri P. , Schumann Guy J.‐P.
Published: January 2015

During the last two decades, remote sensing data have led to tremendous progress in advancing flood inundation modelling. In particular, low‐cost space‐borne data can be invaluable for large‐scale flood studies in data‐scarce areas. Various satellite products yield valuable information such as land surface elevation, flood extent and water level, which could potentially contribute to various flood studies. An increasing number of research studies have been dedicated to exploring those low‐cost data towards building, calibration and evaluation, and remote‐sensed information assimilation into hydraulic models. This paper aims at reviewing these recent scientific efforts on the integration of low‐cost space‐borne remote sensing data with flood modelling. Potentials and limitations of those data in flood modelling are discussed. This paper also introduces the future satellite missions and anticipates their likely impacts in flood modelling. Copyright © 2015 John Wiley & Sons, Ltd.
Gray level entropy matrix is a superior predictor than multiplex ELISA in the detection of reactive stroma and metastatic potential of high‐grade prostatic adenocarcinoma
Authors: Jia Xiaopeng , Sun Yanan , Wang Baozhi
Published: January 2015

Recent reports have indicated that not only the primary glandular tissue but also the surrounding stromal tissue plays an active role in the progression of carcinoma. Such is true for cancer tissues arising in the prostate. However, the precise role of stromal tissue in benign prostatic hyperplasia (BPH) and prostate adenocarcinoma is not well described. We undertook this current investigation to examine the changes in orientation of the extracellular matrix and correlate with prostatic cancer progression. We used a novel form of image analysis called gray level entropy matrix (GLEM) texture analysis to evaluate morphometric changes in stromal tissues. We used normal prostatic tissue obtained from cadaveric specimen and compared with BPH, prostatic intraepithelium neoplastic, hormone responsive prostatic adenocarcinoma and castration‐resistant prostatic adenocarcinoma tissues. GLEM showed higher entropy in disease‐resistant prostatic tissues, compared with benign forms of all spectra of pathologically diagnosed prostatic tissues (P < 0.05, ANOVA, between groups). Higher entropy is reflective of the disorganized morphological organization of the stroma, possibly reflecting the reactive matrix. In contrast, ELISA revealed that although individually correlated with the progressive stages of benign and carcinomatous prostatic tissues and trend correlation between groups, intergroup comparisons failed to arrive at statistical significance of comparisons between markers of neovasculogenesis, vascular endothelial growth factor, epithelial–mesenchymal transition (beta1‐integrin, E‐cadherin, MMP3) and osteogenic metastasis (RANKL and osteoprotegerin). The results of our study demonstrate the potential of GLEM entropy of gray level pixel in providing quasiquantitative estimate of a reactive stroma in advance stages of prostatic adenocarcinoma and thus can be routinely used in clinical decision making. © 2015 IUBMB Life, 66(12):847–853, 2014
Inflammation in arterial diseases
Authors: Carbone Federico , Montecucco Fabrizio
Published: January 2015

The pathophysiology of some inflammatory arterial diseases (such as vasculitis, atherogenesis and aneurysms) has been widely investigated in the last decades. Among different soluble molecules, proinflammatory cytokines (such as TNF‐α, IL‐1 and IL‐6) were shown to trigger critical pathways regulating these arterial diseases. Together with these cytokines, chemokines were also associated with endothelial dysfunction and intima injury in arterial diseases. Recently, autoantibodies have been also described to pathophysiologically influence not only autoimmune vasculitis but also atherogenesis and more in general vascular inflammation. These soluble mediators actively trigger inflammatory functions of leukocytes and vascular cells. For instance, B and T lymphocytes, macrophages and neutrophils were shown to actively participate in inflammatory processes within the arterial wall in vasculitis, atherogenesis and aneurysms. The aim of this narrative review is to provide an overview of pathophysiology and treatments targeting arterial inflammation in these diseases. © 2015 IUBMB Life, 67(1):18–28, 2015
Aquaporin‐5 is expressed in adipocytes with implications in adipose differentiation
Authors: Madeira Ana , Mósca Andreia F. , Moura Teresa F. , Soveral Graça
Published: January 2015

Aquaporins (AQPs) are membrane channels widely distributed in nature. Typically, multiple isoforms are expressed in a single tissue. The adipose tissue is no exception where several AQP members have been identified. The importance of overlapped AQPs expression is unclear, yet interisoforms interactions might be required for key cellular functions. Recently, AQP5 was described as a regulator of other AQP isoforms. Therefore, we hypothesized for a role of AQP5 in adipocyte biology. Gene expression analysis revealed the presence of AQP5 in both 3T3‐L1 fibroblasts and adipocytes, being more abundant in the later. AQP5 depletion impaired adipocyte differentiation, which was confirmed by decreased expression of specific differentiation markers. By overexpressing the human AQP5 in mature adipocytes it was possible to ascertain its role as a water channel in a gain‐of‐function scenario. To our knowledge, this is the first time that AQP5 is reported on adipose tissue. Our data revealed AQP5 as a new player in adipose tissue biology. © 2015 IUBMB Life, 67(1):54–60, 2015
DONATIONS TO THE NEEDY: CORRELATES OF FINANCIAL CONTRIBUTIONS TO THE DESTITUTE
Authors: Bryan James H. , Davenport Michael
Published: January 2015

The New York Times' 100 Neediest Cases were analyzed for a variety of factors that might be expected to affect the number of contributions to the needy. Indices of psychological disturbance and of moral transgression in such cases were negatively associated with the number of contributors. The presence of physical illness, however, was positively associated with number of contributions.
AN EXPLORATORY STUDY OF WORD ASSOCIATIONS OF NEGRO COLLEGE STUDENTS1,2
Authors: Belcher Leon H. , Campbell Joel T.
Published: January 2015

Two word association lists of 50 words were each administered to 50 Negro college students. Forty‐one words were taken from the Kent‐Rosanoff list, 29 from the Palermo‐Jenkins list, and 30 were words used in analogy items of the Scholastic Aptitude Test. Comparisons with previous normative studies showed generally similar results. The present study did result in slightly smaller proportions of matching from class primary responses to noun, pronoun, and adverb stimulus words, and of opposite responses to “opposite‐evoking stimuli.” A number of the responses indicated reading difficulty or misunderstanding of the word.
TRANSFORMATIONS WHICH RENDER CURVES PARALLEL1
Authors: Levine Michael V.
Published: January 2015

Eight examples are given to indicate the desirability of having a general theory treating sets of curves which can be transformed into parallel curves. With this theory it is shown that a set of curves can be rendered parallel if and only if every pair of functions in the completion of a particular group associated with the curves is uncrossed. Under general conditions any two transformations rendering a set of curves parallel are related by a linear transformation. Methods for calculating transformations are proposed. Several structural properties of sets of curves which can be rendered parallel are proven.
ACTIONS SPEAK LOUDER THAN WORDS: MODEL INCONSISTENCY AND ITS EFFECT ON SELF‐SACRIFICE1
Authors: Bryan James H.
Published: January 2015

This experiment was designed to assess the effects of model inconsistency upon children's altruistic behavior. Thirty‐eight boys and 53 girls were exposed to a model (M) who, when provided the opportunity to donate gift certificates to the needy, either did so (practiced charity) or refused to do so (modeled greed), In addition, within each of these two treatments, subjects (Ss) heard M either exhort charity, preach greed, or maintain a “neutral” conversation. Ss were then left alone and were provided the opportunity to donate gift certificates won while playing a bowling game. While the amount donated was not significantly related to the treatments, the number of donors were. Children imitated the M's actions but were not affected by the exhortations. Inconsistency in words and deeds did not affect subsequent emission of the altruistic behavior.
CRITERION SCALING OF QUESTIONNAIRE ITEMS FOR REGRESSION ANALYSIS1,2
Authors: Beaton Albert E.
Published: January 2015

Criterion scaling is a technique for scaling questionnaire item responses so as to maximize their correlation with an external criterion. The correlation between the criterion and the scaled item is equal to the correlation ratio or η coefficient. The results of rescaling the ninth grade questionnaire of the “Equal Educational Opportunity Survey” is presented and compared to the subjective scaling used in the Report. Criterion scaling increased all correlations but the change is not substantial enough to change the interpretation of the Report.
DEGREES AND DIMENSIONS OF SOCIAL INTERACTION IN TRIBAL GROUPINGS: A SOCIOMETRIC STUDY
Authors: Irvine S. H.
Published: January 2015

This study reports the results of administering an inventory of 15 sociometric questions to 11 groups of students, 274 in all, in primary, secondary, teacher‐training and mine youth schools on the Copperbelt of Zambia. Choices made were analysed with tribal identification as a classification within class groupings and over all respondents. Two conclusions emerge. First, tribal identity represents a significant category for social interaction within institutionalised groups when social distance is pronounced. Second, the inventory's dimensions, when subjected to factor analysis based on the tribal groupings of the respondents, lay on three axes. They were hypothesised as leadership within an institutionalised system, social acceptance by peers, and ethnic identity. The results, when discussed with reference to other work in social distance, suggest modifications of current theory.
ON THE INTERPRETATION OF STUDENT PERCEPTIONS OF THEIR COLLEGE ENVIRONMENT
Authors: Centra John A. , Linn Robert L.
Published: January 2015

The purpose of this study was to explore further the relationships between student perceived college environment as measured by Pace's College and University Environment Scales (CUES) and objective institutional characteristics. CUES scores for 75 colleges (administered in 1965) were related to college mean scores of 1964 entering freshmen on SAT‐V and SAT‐M, sex composition of students in attendance, religious affiliation, and size of entering class.

Although multiple correlations were all relatively high (particularly the .80 for the Propriety scale), there remained substantial variance on the five CUES scales that was not predictable from the set of five institutional characteristics. Comparing colleges with CUES scores that were much larger than predicted to colleges with CUES scores that were much smaller than predicted in stepwise regression analyses revealed systematic differences on two CUES scales: Scholarship and Practicality. For the Scholarship scale the systematic differences were geographical, with Northeastern colleges scoring lower than predicted. A “frame of reference” hypothesis is advanced as one possible explanation for systematic differences found.
STYLISTIC FACTORS IN LEARNING AND RETENTION
Authors: McKenna Virgil V.
Published: January 2015

The purposes of this investigation were, first, to determine the extent of individual consistency in the tendencies to err either by omission or commission in tasks requiring learning and retention, and, secondly, to determine the relationship between any such consistent tendencies and relevant cognitive, personality, and ability variables.

A battery of 26 tests from which 86 variables were generated was administered to 147 college students, 70 men and 77 women, by individual and group testing. Information concerning scholastic ability and achievement, as well as control measures of socioeconomic status and seating position in the testing room, were included in the analysis. The 86 variables were intercorrelated, and 44 were factor analyzed by the method of principal components. Thirteen obtained factors were rotated to oblique simple structure by the Promax method plus three visual rotations. Loadings for the remaining 42 variables were determined by extension methods. Thirteen first order factors and four second order were then cast into a Schmid‐Leiman hierarchical solution.

Four first order factors were interpreted as representing error styles in learning and retention. The clearest stylistic factor obtained at this level, Factor V, was characterized by omission tendencies in span memory for primarily numerical stimuli. Other stylistic factors were interpreted as commission tendencies in connected discourse, Factor IV; omission tendencies
The relationship between Cd‐induced autophagy and lysosomal activation in WRL‐68 cells
Authors: Meng Su‐Fang , Mao Wei‐Ping , Wang Fang , Liu Xiao‐Qian , Shao Luan‐Luan
Published: January 2015

This study shows that Cd induces autophagy in the human's embryonic normal liver cell line (WRL‐68). The expression of LC3B‐II and the mature cathepsin L were analyzed by Western blotting. The autophagosomes and lysosomes were directly visualized by electron microscopy and confocal microscopy analysis in Cd‐exposed WRL‐68 cells. In this study, we first found that autophagy induced the activation of lysosomal function in WRL‐68 cells. The lysosomal activation was markedly decreased when the cells were co‐treated with 3‐MA (an inhibitor of autophagy). Secondly, we provided the evidence that the activation of lysosomal function depended on autophagosome–lysosome fusion. The colocalization of lysosome‐associated membrane protein‐2 (LAMP2) and GFP‐LC3 was significantly reduced, when they were treated with thapsigargin (an inhibitor of autophagosome–lysosome fusion). We demonstrated that deletion or blockage of the autophagosome–lysosome fusion process effectively diminished lysosomal activation, which suggests that lysosomal activation occurring in the course of autophagy is dependent on autophagosome–lysosome fusion. Thirdly, we provided evidence that the activation of lysosomal function was associated with lysosomal acid. We investigated the relationship between autophagosome–lysosome fusion and pH in acidic compartments by visualizing fusion process in WRL‐68 cells. This suggests that increasing pH in acidic compartments in WRL‐68 cells inhibits the autophagosome–lysosome fusion. Finally, we found that the activation of lysosomal function was associated with Ca2+ stores and the intracellular Ca2+ channels or pumps were possibly pH‐dependent. Copyright © 2015 John Wiley & Sons, Ltd.
Effects of cylindrospermopsin on the phagocytic cells of the common carp (Cyprinus carpio L.)
Authors: Sieroslawska Anna , Rymuszka Anna , Adaszek Łukasz
Published: January 2015

Cylindrospermopsin is a cyanotoxin with cytotoxic activity. It is released into water during and after cyanobacterial water blooms and thus poses a threat to the health of fish. There is very little information available concerning the effects of the toxin on fish immune cells. In this study, we assessed the potential impact of cylindrospermopsin on the basic functions of phagocytic cells from common carp (Cyprinus carpio L.), including phagocytosis, reactive oxygen and nitrogen species production, and the structure of microfilaments and selected cytokine expression. Phagocytic cells, isolated from fish head kidneys, were exposed to the toxin at concentrations of 0.05, 0.1, 0.5 or 1 µg ml−1, for up to 24 h. Cytotoxicity, detected by lactate dehydrogenase release, was observed at the highest studied concentration. A decrease in phagocytic activity and changes in actin cytoskeletal structures were observed after the cell exposure to the toxin at 0.5 and 1 µg ml−1. Moreover, at all tested concentrations, cylindrospermopsin increased the production of reactive oxygen and nitrogen species. It also evidently influenced the expression of genes of proinflammatory cytokines interleukin‐1β and tumour necrosis factor‐α and, to a minor extent, anti‐inflammatory transforming growth factor‐β, but had no effects on interleukin‐10. The results indicated that the cyanotoxin cylindrospermopsin is able to modify basic features of carp phagocytic cells, which might result in adverse consequences for fish health. Copyright © 2015 John Wiley & Sons, Ltd.
An automated multidimensional thin film stretching device for the generation of anisotropic polymeric micro‐ and nanoparticles
Authors: Meyer Randall A. , Meyer Randall S. , Green Jordan J.
Published: January 2015

Anisotropic polymeric particles are of growing interest for biomaterials applications due to their unique properties. These include the ability for these particles to evade nonspecific cellular uptake and to have enhanced targeted cellular uptake and interaction. One of the most widely used methods for generating anisotropic polymeric particles is the thin film stretching procedure. Despite its theoretical simplicity, this procedure, as it has been implemented to date, can be difficult due to the inconsistent nature of the manual operation of machinery used to stretch the film. We have constructed an automated thin film stretcher for control over biomaterials via thin film stretching in 1D and 2D and as a result, have enabled precise generation of anisotropic polymeric particles. We demonstrate that this device can be utilized to produce anisotropic biodegradable particles of different size, shape, and material consistency. Furthermore, we show that this machine has enabled the scaled up and rapid production of anisotropic polymeric particles, including polymeric microparticles that mimic the shape of red blood cells. Further application of this automated thin film stretching device could allow for significant impact to diverse biomaterial and biomedical applications such as biomimetic particles for immunoengineering and long‐circulating particles for controlled release of drugs. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 103A: 2747–2757, 2015.
In vitro neurite guidance effects induced by polylysine pinstripe micropatterns with polylysine background
Authors: Joo Sunghoon , Kang Kyungtae , Nam Yoonkey
Published: January 2015

Engineered culture substrates with chemical neurite guidance cues have been used for studying the mechanism of axon pathfinding at cellular level. In this study, we designed a novel poly‐l‐lysine (PLL) micropattern (“pinstripe micropattern”) to investigate how the same biomolecules with slightly different surface concentration can affect in vitro neuronal growth. The pinstripe micropattern was fabricated by stamping PLL on a PLL‐coated glass coverslip, which resulted in denser PLL lines and a less‐dense PLL background. There were two effects of the substrate on cultured primary hippocampal neuron: neurite initiation and growth cone turning. Although the whole surface was permissive for neurite outgrowth, we observed that the growth direction of neurites had a strong tendency to follow the stamped PLL line patterns with PLL background. However, the micropattern did not affect the spreading of cell body on the substrate. According to these investigations, we concluded that the PLL pinstripe pattern with PLL background, which had the step difference of polylysine concentrations, would be very useful for designing novel cell assays for the investigation of neurite guidance mechanisms, and suggested it as a new design method for controlling the direction of neurite growth on in vitro neural network. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 103A: 2731–2739, 2015.
Influence of ceramic disk material, surface hemispheres, and SBF volume on in vitro mineralization
Authors: Urquia Edreira Eva R. , Wolke Joop G. C. , Jansen John A. , van den Beucken Jeroen J. J. P.
Published: January 2015

Calcium phosphate ceramics are the main mineral constituents of bone and teeth and have therefore been extensively investigated for bone regenerative applications. In the current study, the effect of disk material, surface geometry, and SBF volume on mineralization capacity was investigated. Hemispherical concavities were created on the surfaces of disks made of different materials (i.e., hydroxyapatite (HA), β‐tricalcium phosphate (β‐TCP), biphasic calcium phosphate (BCP) and titanium (Ti)) which were sintered at 1200 °C. Mineralization of CaP was assessed on disk surfaces after immersion of the samples in different volumes of simulated body fluid (SBF) up to 14 days by means of calcium assay and scanning electron microscopy (SEM). This study showed that different SBF volumes have different effects on mineralization, with an optimum material/liquid ratio of 5 mL of SBF per cm2. Additionally, at this volume, apparent differences based on disk material became obvious. Evidently, surface hemispherical concavities acted as initiator areas for nucleation and crystal growth. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 103A: 2740–2746, 2015.
An electrically conductive 3D scaffold based on a nonwoven web of poly(l‐lactic acid) and conductive poly(3,4‐ethylenedioxythiophene)
Authors: Niu Xufeng , Rouabhia Mahmoud , Chiffot Nicolas , King Martin W. , Zhang Ze
Published: January 2015

This study was to demonstrate that an extremely thin coating of poly(3,4‐ethylenedioxythiophene) (PEDOT) on nonwoven microfibrous poly(l‐lactic acid) (PLLA) web is of sufficient electrical conductivity and stability in aqueous environment to sustain electrical stimulation (ES) to cultured human skin fibroblasts. The PEDOT imparted the web a surface resistivity of approximately 0.1 KΩ/square without altering the web morphology. X‐ray photoelectron spectroscopy demonstrated that the surface chemistry of the PLLA/PEDOT is characteristic of both PLLA and PEDOT. The PEDOT‐coated web also showed higher hydrophilicity, lower glass transition temperature and unchanged fiber crystallinity and thermal stability compared with the PLLA web. The addition of PEDOT to the web marginally increased the web's tensile strength and lowered the elongation. An electrical stability test showed that the PLLA/PEDOT structure was more stable than a polypyrrole treated PLLA fabric, showing only a slow deterioration in conductivity when exposed to culture medium. The cytotoxicity test showed that the PLLA/PEDOT scaffold was not cytotoxic and supported human dermal fibroblast adhesion, migration, and proliferation. Preliminary ES experiments have demonstrated that this conductive web mediated effective ES to fibroblasts. Therefore, this new conductive biodegradable scaffold may be used to electrically modulate cellular activity and tissue regeneration. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 103A: 2635–2644, 2015.
A ferulic acid (FA)‐eluting system for biodegradable magnesium stent: Cells response of HUVECs
Authors: Zhang Erlin , Shen Feng
Published: January 2015

A new drug‐eluting system was designed for biodegradable magnesium stents, in which ferulic acid (FA) was used as drug due to its promotion function to endothelial cells and PHBHHx as drug delivery due to its biodegradability and biocompatibility. A 5 and 10% FA were added in PHBHHx to prepare FA containing PHBHHX films. The cell adhesion, spreading and proliferation on these films was investigated in order to assess cell response of HUVECs. It was also found that FA enhanced the adhesion, spreading and proliferation of HUVECs in a dose‐dependent manner. Cell viability after H2O2 injury and NO production of HUVECs were also studied. The results indicated that FA effectively inhibited H2O2‐induced injury and promotes NO production. It was also shown that alkali treatment improved the cell adhesion, spreading and proliferation while the treatment reduces the FA release and in turn reduces the inhibition on H2O2‐induced injury and NO production. However, alkali treatment itself had no influence on the H2O2 induced injure and NO production. The tensile shear strength between the FA containing coating and Mg substrate was also tested. All results demonstrated that FA containing PHBHHx films exhibited strong promotion to the endothelialization and could be a choice for surface modification of magnesium stent. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 103A: 2758–2769, 2015.
Human periosteum cell osteogenic differentiation enhanced by ionic silicon release from porous amorphous silica fibrous scaffolds
Authors: Odatsu Tetsurou , Azimaie Taha , Velten Megan F. , Vu Michael , Lyles Mark B. , Kim Harry K. , Aswath Pranesh B. , Varanasi Venu G.
Published: January 2015

Current synthetic grafts for bone defect filling in the sinus can support new bone formation but lack the ability to stimulate or enhance osteogenic healing. To promote such healing, osteoblast progenitors such as human periosteum cells must undergo osteogenic differentiation. In this study, we tested the hypothesis that degradation of porous amorphous silica fibrous (PASF) scaffolds can enhance human periosteum cell osteogenic differentiation. Two types of PASF were prepared and evaluated according to their densities (PASF99, PASF98) with 99 and 98% porosity, respectively. Silicon (Si) ions were observed to rapidly release from both scaffolds within 24 hin vitro. PASF99 Si ion release rate was estimated to be nearly double that of PASF98 scaffolds. Mechanical tests revealed a lower compressive strength in PASF99 as compared with PASF98. Osteogenic expression analysis showed that PASF99 scaffolds enhanced the expression of activating transcription factor 4, alkaline phosphatase, and collagen (Col(I)α1, Col(I)α2). Scanning electron microscopy showed cellular and extracellular matrix (ECM) ingress into both scaffolds within 16 days and the formation of Ca‐P precipitates within 85 days. In conclusion, this study demonstrated that PASF scaffolds enhance human periosteum cell osteogenic differentiation by releasing ionic Si, and structurally supporting cellular and ECM ingress. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 103A: 2797–2806, 2015.
Spontaneous gene transfection of human bone cells using 3D mineralized alginate–chitosan macrocapsules
Authors: Green David W. , Kim Eun‐Jung , Jung Han‐Sung
Published: January 2015

The effectiveness of nonviral gene therapy remains uncertain because of low transfection efficiencies and high toxicities compared with viral‐based strategies. We describe a simple system for transient transfection of continuous human cell lines, with low toxicity, using mineral‐coated chitosan and alginate capsules. As proof‐of‐concept, we demonstrate transfection of Saos‐2 and MG63 human osteosarcoma continuous cell lines with gfp, LacZ reporter genes, and a Sox‐9 carrying plasmid, to illustrate expression of a functional gene with therapeutic relevance. We show that continuous cell lines transfect with significant efficiency of up to 65% possibly through the interplay between chitosan and DNA complexation and calcium/phosphate‐induced translocation into cells entrapped within the 3D polysaccharide based environment, as evidenced by an absence of transfection in unmineralized and chitosan‐free capsules. We demonstrated that our transfection system was equally effective at transfection of primary human bone marrow stromal cells. To illustrate, the Sox‐9, DNA plasmid was spontaneously expressed in primary human bone marrow stromal cells at 7 days with up to 90% efficiency in two repeats. Mineralized polysaccharide macrocapsules are gene delivery vehicles with a number of biological and practical advantages. They are highly efficient at self‐transfecting primary bone cells, with programmable spatial and temporal delivery prospects, premineralized bone‐like environments, and have no cytotoxic effects, as compared with many other nonviral systems. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 103A: 2855–2863, 2015.
Dynamically Regulated CFTR Expression and Its Functional Role in Cutaneous Wound Healing
Authors: Dong Jianda , Jiang Xiaohua , Zhang Xiaohu , Liu Kai Sheng , Zhang Jieting , Chen Jing , Yu Mei Kuen , Tsang Lai Ling , Chung Yiu Wa , Wang Yanrong , Zhou Wen‐liang , Chan Hsiao Chang
Published: January 2015

The physiological role of cystic fibrosis transmembrane conductance regulator (CFTR) in keratinocytes and skin wound healing is completely unknown. The present study shows that CFTR is expressed in the multiple layers of keratinocytes in mouse epidermis and exhibits a dynamic expression pattern in a dorsal skin wound healing model, with diminishing levels observed from day 3 to day 5 and re‐appearing from day 7 to day 10 after wounding. Knockdown of CFTR in cultured human keratinocytes promotes cell migration but inhibits differentiation, while overexpression of CFTR suppresses migration but enhances differentiation, indicating an important role of CFTR in regulating keratinocyte behavior. In addition, we have demonstrated a direct association of CFTR with epithelial junction formation as knockdown of CFTR downregulates the expression of adhesion molecules, such as E‐cadherin, ZO‐1 and β‐catenin, and disrupts the formation of cell junction, while overexpression of CFTR enhances cell junction formation. More importantly, we have shown that ΔF508cftr−/− mice with defective CFTR exhibit delayed wound healing as compared to wild type mice, indicating that normal function of CFTR is critical for wound repair. Taken together, the present study has revealed a previously undefined role of CFTR in regulating skin wound healing processes, which may have implications in injury repair of other epithelial tissues. J. Cell. Physiol. 230: 2049–2058, 2015. © 2015 Wiley Periodicals, Inc.
Identification of Genes Selectively Regulated in Human Hepatoma Cells by Treatment With Dyslipidemic Sera and PUFAs
Authors: De Rosa Maria Caterina , Caputo Mariella , Zirpoli Hylde , Rescigno Tania , Tarallo Roberta , Giurato Giorgio , Weisz Alessandro , Torino Gaetano , Tecce Mario Felice
Published: January 2015

Serum composition is linked to metabolic diseases not only to understand their pathogenesis but also for diagnostic purposes. Quality and quantity of nutritional intake can affect disease risk and serum composition. It is then possible that diet derived serum components directly affect pathogenetic mechanisms. To identify involved factors, we evaluated the effect on gene expression of direct addition of dyslipidemic human serum samples to cultured human hepatoma cells (HepG2). Sera were selected on the basis of cholesterol level, considering this parameter as mostly linked to dietary intake. Cells were treated with 32 sera from hypercholesterolemic and normocholesterolemic subjects to identify differentially regulated mRNAs using DNA microarray analysis. We identified several mRNAs with the highest modulations in cells treated with dyslipidemic sera versus cells treated with normal sera. Since the two serum groups had variable polyunsaturated fatty acids (PUFAs) contents, selected mRNAs were further assessed for their regulation by docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA) and arachidonic acid (AA). Four genes resulted both affected by serum composition and PUFAs: 3‐hydroxy‐3‐methylglutaryl‐CoenzymeA synthase 2 (HMGCS2), glutathione S‐transferase alpha 1 (GSTA1), liver expressed antimicrobial peptide 2 (LEAP2) and apolipoprotein M (ApoM). HMGCS2 expression appears the most relevant and was also found modulated via transcription factors peroxysome proliferator activated receptor α (PPARα) and forkhead box O1 (FoxO1). Our data indicate that expression levels of the selected mRNAs, primarily of HMGCS2, could represent a reference of nutritional intake, PUFAs effects and dyslipidemic diseases pathogenesis. J. Cell. Physiol. 230: 2059–2066, 2015. © 2015 Wiley Periodicals, Inc.
Y‐box Binding Protein‐1 Is Part of a Complex Molecular Network Linking ΔNp63α to the PI3K/akt Pathway in Cutaneous Squamous Cell Carcinoma
Authors: Troiano Annaelena , Lomoriello Irene Schiano , di Martino Orsola , Fusco Sabato , Pollice Alessandra , Vivo Maria , La Mantia Girolama , Calabrò Viola
Published: January 2015

Cutaneous squamous cell carcinomas (SCCs) typically lack somatic oncogene‐activating mutations and most of them contain p53 mutations. However, the presence of p53 mutations in skin premalignant lesions suggests that these represent early events during tumor progression and additional alterations may be required for SCC development. SCC cells frequently express high levels of ΔNp63α and Y‐box binding 1 (YB‐1 or YBX1) oncoproteins. Here, we show that knockdown of YB‐1 in spontaneously immortalized HaCaT and non‐metastatic SCC011 cells led to a dramatic decrease of ΔNp63α, cell detachment and death. In highly metastatic SCC022 cells, instead, YB‐1 silencing induces PI3K/AKT signaling hyperactivation which counteracts the effect of YB‐1 depletion and promotes cell survival. In summary, our results unveil a functional cross‐talk between YB‐1, ΔNp63α and the PI3K/AKT pathway critically governing survival of squamous carcinoma cells. J. Cell. Physiol. 230: 2067–2074, 2015. © 2015 Wiley Periodicals, Inc.
β2‐Adrenoreceptor‐Mediated Proliferation Inhibition of Embryonic Pluripotent Stem Cells
Authors: Sun Fan , Yang Xin‐Jie , Lv Hao‐Yu , Tang Ya‐Bin , An Shi‐Min , Ding Xu‐Ping , Li Wen‐Bin , Teng Lin , Shen Ying , Chen Hong‐Zhuan , Zhu Liang
Published: January 2015

Adrenoreceptors (ARs) are widely expressed and play essential roles throughout the body. Different subtype adrenoceptors elicit distinct effects on cell proliferation, but knowledge remains scarce about the subtype‐specific effects of β2‐ARs on the proliferation of embryonic pluripotent stem (PS) cells that represent different characteristics of proliferation and cell cycle regulation with the somatic cells. Herein, we identified a β2‐AR/AC/cAMP/PKA signaling pathway in embryonic PS cells and found that the pathway stimulation inhibited proliferation and cell cycle progression involving modulating the stem cell growth and cycle regulatory machinery. Embryonic stem (ES) cells and embryonal carcinoma stem (ECS) cells expressed functional β‐ARs coupled to AC/cAMP/PKA signaling. Agonistic activation of β‐ARs led to embryonic PS cell cycle arrest and proliferation inhibition. Pharmacological and genetic analyzes using receptor subtype blocking and RNA interference approaches revealed that this effect selectively depended on β2‐AR signaling involving the regulation of AKT, ERK, Rb, and Cyclin E molecules. Better understanding of the effects of β2‐ARs on embryonic PS cell proliferation and cycle progression may provide new insights into stem cell biology and afford the opportunity for exploiting more selective ligands targeting the receptor subtype for the modulation of stem cells. J. Cell. Physiol. 9999: 2640–2646, 2015. © 2015 Wiley Periodicals, Inc.
Effects of maturation and aging on the pressure‐bearing region of the plantaris longus tendon of the bullfrog (Lithobates catesbeianus)
Authors: Zorel Valdenilson José , Morais Jymenez De , de Aro Andrea Aparecida , Gomes Laurecir , Esquisatto Marcelo Augusto Marretto
Published: January 2015

The plantaris longus tendon (PLT) in bullfrog develops a fibrocartilage‐like tissue in the area that is functionally subject to compressive forces. The aim of this study was to analyze the modifications of the pressure‐bearing region in bullfrog PLT with different ages (7, 180, and 1,080 days after the end of metamorphosis) using histomorphometric, ultrastructural and biochemical methods. Weak basophilia and cells with a fibroblastic phenotype were observed in the compression region at 7 days of age. On the other hand, a large area of intense tissue basophilia associated with a chondroblast‐like cell population was noted at the other ages. Collagen fibers exhibited a three‐dimensional network‐like arrangement at all ages. The number of connective tissue cells increased between 7 and 180 days of age and was reduced in older animals. The 180‐day‐old animals presented a well‐developed pericellular matrix rich in proteoglycans. The mean diameter of collagen fibrils increased from 7 to 180 days and was the same at 1,080 days. Glycosaminoglycan content was higher in 7‐day‐old animals. A higher amount of hydroxyproline was observed at 180 and 1,080 days. The swelling test showed a significant increase of wet weight in 7‐day‐old animals. In conclusion, the alterations that occur in the pressure‐bearing of bullfrog PLT are the result of physiological alterations of the animal with the maturation and aging. Microsc. Res. Tech. 77:797–805, 2014. © 2014 Wiley Periodicals, Inc.
Prediction of protein–protein interaction sites from weakly homologous template structures using meta‐threading and machine learning
Authors: Maheshwari Surabhi , Brylinski Michal
Published: January 2015

The identification of protein–protein interactions is vital for understanding protein function, elucidating interaction mechanisms, and for practical applications in drug discovery. With the exponentially growing protein sequence data, fully automated computational methods that predict interactions between proteins are becoming essential components of system‐level function inference. A thorough analysis of protein complex structures demonstrated that binding site locations as well as the interfacial geometry are highly conserved across evolutionarily related proteins. Because the conformational space of protein–protein interactions is highly covered by experimental structures, sensitive protein threading techniques can be used to identify suitable templates for the accurate prediction of interfacial residues. Toward this goal, we developed eFindSitePPI, an algorithm that uses the three‐dimensional structure of a target protein, evolutionarily remotely related templates and machine learning techniques to predict binding residues. Using crystal structures, the average sensitivity (specificity) of eFindSitePPI in interfacial residue prediction is 0.46 (0.92). For weakly homologous protein models, these values only slightly decrease to 0.40–0.43 (0.91–0.92) demonstrating that eFindSitePPI performs well not only using experimental data but also tolerates structural imperfections in computer‐generated structures. In addition, eFindSitePPI detects specific molecular interactions at the interface; for instance, it correctly predicts approximately one half of hydrogen bonds and aromatic interactions, as well as one third of salt bridges and hydrophobic contacts. Comparative benchmarks against several dimer datasets show that eFindSitePPI outperforms other methods for protein‐binding residue prediction. It also features a carefully tuned confidence estimation system, which is particularly useful in large‐scale applications using raw genomic data. eFindSitePPI is freely available to the academic community at http://www.brylinski.org/efindsiteppi. Copyright © 2014 John Wiley & Sons, Ltd.