bluuebunny/crossref_metadata_2025 huggingface dataset:
Investigations on the Effects of Seasonal Temperature Changes on the Electrical Resistance of Living Trees
Authors: Yue Xiaoquan , Wang Lihai , Shi Xiaolong , Xu Mingxian , Zhu Zhiming
Published: September 2018

In order to use the electrical resistance method to accurately and timely detect and evaluate the internal decay defects of living trees, the effects of the seasonal temperature and moisture content on the electrical resistance of standing trees were investigated. At the Northeast Forestry University Experimental Forest Farm, Harbin, Heilongjiang Province of China, Populus simonii Populus simonii Carr. and Larix gmelinii (Rupr.) Rupr. were selected as the objects and the electrical resistance of standing trees was tested through different seasons from December 2016 to December 2017. Meanwhile, the effects of changes in the seasonal temperatures (−20 to −10 °C, −10 to −5 °C, −5 to 0 °C, 0 to 5 °C, 5 to 10 °C, 10 to 15 °C, 15 to 25 °C) as well as changes in the moisture content (MC) (Populus simonii, MC ≥ 103%; Larix gmelinii, MC ≥ 77.5%) on the electrical resistance in the cross-sections of living trees were studied. The influence of temperature at different moisture contents, the moisture content at different temperatures, and their combined effects on electrical resistance were analyzed, following which a regression model was also established. The obtained results indicated that ambient temperature had a significant effect on the average value of electrical resistance in the cross-section of living trees when temperatures were below the freezing point. There was a sudden discontinuity near the freezing point, and logR (logarithm value of electrical resistance) in the cross-sections of sound trees and decayed trees changed in a similar trend with variations in the temperature. While the effect of moisture content on logR in the cross-sections of threes was insignificant at different temperatures because of the moisture content above FSP (fiber saturation point). It indicated that the temperature and moisture content had interactive effects on logR in the cross-sections. The binary linear regression model between moisture content, temperature, and logR was highly fitted with a correlation coefficient (R2) higher than 0.8. The outcome of this investigation indicates that when non-destructive testing is performed on living trees using electrical resistance at different seasonal temperatures, the measured results need to consider both the temperature and moisture content. For practical work, it is not recommended to consider testing living trees near the freezing point temperature using the electrical resistive tomography. Below the freezing point, the electrical resistance changes with temperature greatly relative to the normal temperature. Therefore, when performing the detection of electrical resistance, it is necessary to calibrate the effects of temperature
70 Years of Land Use/Land Cover Changes in the Apennines (Italy): A Meta-Analysis
Authors: Malandra Francesco , Vitali Alessandro , Urbinati Carlo , Garbarino Matteo
Published: September 2018

Land use science usually adopts a case study approach to investigate landscape change processes, so we considered a meta-analysis an appropriate tool for summarizing general patterns and heterogeneous findings across multiple case studies over a large geographic area. Mountain landscapes in the Apennines (Italy) have undergone significant variations in the last century due to regional and national socio-economic changes. In this work, we reviewed 51 manuscripts from different databases and examined 57 case studies. We explored heterogeneous data sets, adopting a stepwise approach to select the case studies: Step 1, a general overview of the main studies; Step 2, an analysis of the features of the study sites and of land-use/cover transitions; Step 3, a landscape pattern analysis. We standardized the processing methods to obtain a new set of homogeneous data suitable for comparative analysis. After some pre-processing of the selected paper due to the broad heterogeneity of the data, we calculated common landscape metrics ex novo. We obtained digital images used to perform automatic segmentation with eCognition Developer 64 software. Our review indicated that most case studies were in Central and Southern Italy, 83% were examined at local scale, 77% carried out change detection, but only 38% included both change detection and landscape spatial pattern analysis. The results revealed a clear trend of forest expansion (+78%) and the reduction of croplands (−49%) and grasslands (−19%). We did not find significant changes in the landscape spatial patterns.
Quantitative Assessment of Surface Runoff and Base Flow Response to Multiple Factors in Pengchongjian Small Watershed
Authors: Ouyang Lei , Liu Shiyu , Ye Jingping , Liu Zheng , Sheng Fei , Wang Rong , Lu Zhihong
Published: September 2018

Quantifying the impacts of multiple factors on surface runoff and base flow is essential for understanding the mechanism of hydrological response and local water resources management as well as preventing floods and droughts. Despite previous studies on quantitative impacts of multiple factors on runoff, there is still a need for assessment of the influence of these factors on both surface runoff and base flow in different temporal scales at the watershed level. The main objective of this paper was to quantify the influence of precipitation variation, evapotranspiration (ET) and vegetation restoration on surface runoff and base flow using empirical statistics and slope change ratio of cumulative quantities (SCRCQ) methods in Pengchongjian small watershed (116°25′48″–116°27′7″ E, 29°31′44″–29°32′56″ N, 2.9 km2), China. The results indicated that, the contribution rates of precipitation variation, ET and vegetation restoration to surface runoff were 42.1%, 28.5%, 29.4% in spring; 45.0%, 37.1%, 17.9% in summer; 30.1%, 29.4%, 40.5% in autumn; 16.7%, 35.1%, 48.2% in winter; and 35.0%, 38.7%, 26.3% in annual scale, respectively. For base flow they were 33.1%, 41.9%, 25.0% in spring; 39.3%, 51.9%, 8.8% in summer; 40.2%, 38.2%, 21.6% in autumn; 24.3%, 39.4%, 36.3% in winter; and 24.4%, 47.9%, 27.7% in annual scale, respectively. Overall, climatic factors, including precipitation and ET change, affect surface runoff generation the most, while ET affects the dynamic change of annual base flowthe most. This study highlights the importance of optimizing forest management to protect the water resource.
Paleo-Drainage Network, Morphotectonics, and Fluvial Terraces: Clues from the Verde Stream in the Middle Sangro River (Central Italy)
Authors: Miccadei Enrico , Carabella Cristiano , Paglia Giorgio , Piacentini Tommaso
Published: September 2018

This work analyzes the role of paleo-drainage network, morphotectonics, and surface processes in landscape evolution in a sector of the transition zone between the chain and the piedmont area of Central Apennines. Particularly, it focuses on the Verde Stream, a tributary of the middle Sangro River valley, which flows in the southeastern Abruzzo area at the boundary with the Molise region. The Verde Stream was investigated through a drainage basin scale geomorphological analysis incorporating the morphometry of the orography and hydrography, structural geomorphological field mapping, and the investigation of morphological field evidence of tectonics with their statistical azimuthal distributions. The local data obtained were compared with the analysis of the middle Sangro River valley and the tectonic features of the Abruzzo–Molise area. This approach led us to also provide relevant clues about the definition of the role of karst features and paleo-landscapes in the general setting of the study area and to identify the impact of active tectonics, confirmed by recent and active seismicity. In conclusion, the paper contributes to defining the main stages of the geomorphological evolution of this area, driven by uplift and local tectonics and due to a combination of fluvial, karst, and landslide processes.
δ13C and δ18O Stable Isotope Analysis Applied to Detect Technological Variations and Weathering Processes of Ancient Lime and Hydraulic Mortars
Authors: Dotsika Elissavet , Kyropoulou Dafni , Christaras Vassilios , Diamantopoulos Georgios
Published: September 2018

Samples of mortars were collected from lime and hydraulic mortars affected by environmental degradation. A total of 63 samples were obtained from Hellenistic, Late Roman and Byzantine historic constructions located at Kavala, Drama and Makrygialos in North Greece. Samples were collected in sections from the surface up to 6 cm deep using a drill-core material. The first sample was collected from the external layer, while the internal samples were collected each 1cm beeper from the previous, in order to monitor the moisture ingress. Isotopic data will make it possible to create an ideal Hellenistic and Byzantine mortar layer and to provide weathering gradients. The isotopic values comprise a range of δ13C and δ18O values from −17.1‰ to 1.2‰ and −25.9‰ to −2‰, respectively. The weathering process of Hellenistic and Byzantine are expressed, by the regression lines δ18Ocalcite matrix = 0.6 × δ13Ccalcite matrix − 1.9 and δ18Ocalcite matrix = 0.6 × δ13Ccalcite matrix − 2.0 for hydraulic and Lime mortars respectively. Pronounced isotopic shift to heavy or light δ13C and δ18O in the carbonate matrix was attributed to the primary source of CO2 (atmospheric versus biogenic) and H2O (evaporation of local primary water), in residual limestone and in secondary processes such as recrystallization of calcite with pore water and salts attack. Exogenic processes related to biological growth are responsible for further alterations of δ18O and δ13C in lime mortars. This study indicated that stable isotope analysis is an excellent tool to fingerprint the origin of carbonate, the environmental setting conditions of mortar, origin of CO2 and water during calcite formation and to determine the weathering depth and the potential secondary degradation mechanisms.
Have Studies that Measure Lumbar Kinematics and Muscle Activity Concurrently during Sagittal Bending Improved Understanding of Spinal Stability and Sub-System Interactions? A Systematic Review
Authors: Du Rose Alister
Published: September 2018

In order to improve understanding of the complex interactions between spinal sub-systems (i.e., the passive (ligaments, discs, fascia and bones), the active (muscles and tendons) and the neural control systems), it is necessary to take a dynamic approach that incorporates the measurement of multiple systems concurrently. There are currently no reviews of studies that have investigated dynamic sagittal bending movements using a combination of electromyography (EMG) and lumbar kinematic measurements. As such it is not clear how understanding of spinal stability concepts has advanced with regards to this functional movement of the spine. The primary aim of this review was therefore to evaluate how such studies have contributed to improved understanding of lumbar spinal stability mechanisms. PubMed and Cochrane databases were searched using combinations of the keywords related to spinal stability and sagittal bending tasks, using strict inclusion and exclusion criteria and adhering to PRISMA guidelines. Whilst examples of the interactions between the passive and active sub-systems were shown, typically small sample sizes meant that results were not generalizable. The majority of studies used regional kinematic measurements, and whilst this was appropriate in terms of individual study aims, the studies could not provide insight into sub-system interaction at the level of the spinal motion segment. In addition, the heterogeneity in methodologies made comparison between studies difficult. The review suggests that since Panjabi’s seminal spinal control papers, only limited advancement in the understanding of these theories has been provided by the studies under review, particularly at an inter-segmental level. This lack of progression indicates a requirement for new research approaches that incorporate multiple system measurements at a motion segment level.
Characterization of Children’s Exposure to Extremely Low Frequency Magnetic Fields by Stochastic Modeling
Authors: Bonato Marta , Parazzini Marta , Chiaramello Emma , Fiocchi Serena , Le Brusquet Laurent , Magne Isabelle , Souques Martine , Röösli Martin , Ravazzani Paolo
Published: September 2018

In this study, children’s exposure to extremely low frequency magnetic fields (ELF-MF, 40–800 Hz) is investigated. The interest in this thematic has grown due to a possible correlation between the increased risk of childhood leukemia and a daily average exposure above 0.4 µT, although the causal relationship is still uncertain. The aim of this paper was to present a new method of characterizing the children’s exposure to ELF-MF starting from personal measurements using a stochastic approach based on segmentation (and to apply it to the personal measurements themselves) of two previous projects: the ARIMMORA project and the EXPERS project. The stochastic model consisted in (i) splitting the 24 h recordings into stationary events and (ii) characterizing each event with four parameters that are easily interpretable: the duration of the event, the mean value, the dispersion of the magnetic field over the event, and a final parameter characterizing the variation speed. Afterward, the data from the two databases were divided in subgroups based on a characteristic (i.e., children’s age, number of inhabitants in the area, etc.). For every subgroup, the kernel density estimation (KDE) of each parameter was calculated and the p-value histogram of the parameters together was obtained, in order to compare the subgroups and to extract information about the children’s exposure. In conclusion, this new stochastic approach allows for the identification of the parameters that most affect the level of children’s exposure.
Health Impacts of Exposure to Gaseous Pollutants and Particulate Matter in Beijing—A Non-Linear Analysis Based on the New Evidence
Authors: Cheng Yunfei , Ermolieva Tatiana , Cao Gui-Ying , Zheng Xiaoying
Published: September 2018

This paper aimed to estimate health risks focusing on respiratory diseases from exposure to gaseous multi-pollutants based on new data and revealed new evidence after the most stringent air pollution control plan in Beijing which was carried out in 2013. It used daily respiratory diseases outpatient data from a hospital located in Beijing with daily meteorological data and monitor data of air pollutants from local authorities. All data were collected from 2014 to 2016. Distributed lag non-linear model was employed. Results indicated that NO2 and CO had positive association with outpatients number on the day of the exposure (1.045 (95% confidence interval (CI): 1.003, 1.089) for CO and 1.022 (95% CI: 1.008, 1.036) for NO2) (and on the day after the exposure (1.026 (95% CI: 1.005, 1.048) for CO and 1.013 (95% CI: 1.005, 1.021) for NO2). Relative risk (RR) generally declines with the number of lags; ozone produces significant effects on the first day (RR = 0.993 (95% CI: 0.989, 0.998)) as well as second day (RR = 0.995 (95% CI: 0.991, 0.999)) after the exposure, while particulate pollutants did not produce significant effects. Effects from the short-term exposure to gaseous pollutants were robust after controlling for particulate matters. Our results contribute to a comprehensive understanding of the dependencies between the change of air pollutants concentration and their health effects in Beijing after the implementation of promising air regulations in 2013. Results of the study can be used to develop relevant measures minimizing the adverse health consequences of air pollutants and supporting sustainable development of Beijing as well as other rapidly growing Asian cities.
Understanding the Role of Mobile Internet-Based Health Services on Patient Satisfaction and Word-of-Mouth
Authors: Gu Dongxiao , Yang Xuejie , Li Xingguo , Jain Hemant K. , Liang Changyong
Published: September 2018

With the rapid advancement of Web 2.0 technologies, Internet medicine, and mobile healthcare, the influence of the use of patient-oriented Mobile Internet-based Health Services (MIHS) on patient satisfaction and the electronic word-of-mouth (WOM) of health service agencies is becoming the focus of the academic research community. Many large hospitals, including some Internet hospitals, have provided various online healthcare service platforms that enable patients to expediently consult with physicians and obtain healthcare services in an online to offline format. The purpose of this study is to analyze the main mechanisms of how the features and users’ experiences of MIHS influenced patient satisfaction and continuous use behaviors of the system to generate additional WOM dissemination behaviors. Based on post-adoption behavior and Expectation Confirmation Model of Information Technology Continuance (ECM-IT), this study conducted an empirical study through data collection from users (patients) from a large hospital providing online healthcare services. A total of 494 pieces of data were collected and analyzed using SmartPLS2.0(SmartPLS GmbH, Hamburg, Gernmany). The results show that: (1) patient satisfaction with MIHS and their intentions to continue use of MIHS have significantly positive influences on WOM; (2) patient satisfaction with MIHS is positively influenced by perceived usefulness and confirmation of MIHS performance expectations; (3) and patient intentions to continue use of MIHS are also affected by some technology factors, such as facilitating conditions and perceived risk, as well as some subjective feelings, such as perceived usefulness and perceived interactivity. The results of this study provide important implications for both research and practice of public health.
An Improved Shapley Value Method for a Green Supply Chain Income Distribution Mechanism
Authors: Xu Zhongwen , Peng Zixuan , Yang Ling , Chen Xudong
Published: September 2018

Low-carbon development and environmental remediation are key factors for green resource-based supply chains in China. With this aim in mind, by applying game theory under uncertain market demand, this paper incorporates low-carbon development and environmental remediation into a resource-based supply chain coordination model for decentralized and centralized markets. The results show that a centralized market can lead to improvement in total profit. Furthermore, based on an improved Shapley value method, a theoretical model for the centralized market income distribution mechanism is developed that incorporates three corrective risk factors, ecological investment, and technological level. Finally, a numerical analysis is conducted using a MATLAB simulation to obtain intuitive results, which, in turn, show the validity of incentive income distribution mechanisms for green supply chain development in China.
Identification of Novel Quantitative Trait Loci Linked to Crown Rot Resistance in Spring Wheat
Authors: Erginbas-Orakci Gul , Sehgal Deepmala , Sohail Quahir , Ogbonnaya Francis , Dreisigacker Susanne , Pariyar Shree R. , Dababat Abdelfattah A.
Published: September 2018

Crown rot (CR), caused by various Fusarium species, is a major disease in many cereal-growing regions worldwide. Fusarium culmorum is one of the most important species, which can cause significant yield losses in wheat. A set of 126 advanced International Maize and Wheat Improvement Center (CIMMYT) spring bread wheat lines were phenotyped against CR for field crown, greenhouse crown and stem, and growth room crown resistance scores. Of these, 107 lines were genotyped using Diversity Array Technology (DArT) markers to identify quantitative trait loci linked to CR resistance by genome-wide association study. Results of the population structure analysis grouped the accessions into three sub-groups. Genome wide linkage disequilibrium was large and declined on average within 20 cM (centi-Morgan) in the panel. General linear model (GLM), mixed linear model (MLM), and naïve models were tested for each CR score and the best model was selected based on quarantine-quarantine plots. Three marker-trait associations (MTAs) were identified linked to CR resistance; two of these on chromosome 3B were associated with field crown scores, each explaining 11.4% of the phenotypic variation and the third MTA on chromosome 2D was associated with greenhouse stem score and explained 11.6% of the phenotypic variation. Together, these newly identified loci provide opportunity for wheat breeders to exploit in enhancing CR resistance via marker-assisted selection or deployment in genomic selection in wheat breeding programs.
A One-Step Extraction and Luminescence Assay for Quantifying Glucose and ATP Levels in Cultured HepG2 Cells
Authors: Csepregi Rita , Temesfői Viktória , Sali Nikolett , Poór Miklós , W. Needs Paul , A. Kroon Paul , Kőszegi Tamás
Published: September 2018

A fluorescence-based enzymatic microplate intracellular glucose assay was designed and fully validated. The method was tested in a hepatocellular cancer cell line (HepG2). Our novel one-step extraction reagent gave stable cell lysates for glucose, adenosine triphosphate (ATP), and total protein determination from the same sample. Limit of detection for glucose was 0.13 µM (26 pmol/well), which is superior to commercially available glucose assays. Both intra- and interday assay imprecision in HepG2 cultures were less than 12% coefficient of variance (CV). In cell lysates spiked with glucose, recovery at two levels varied between 83.70% and 91.81%, and both linearity and stability were acceptable. HepG2 cells treated with agents affecting glucose uptake/metabolism (phloretin, quercetin, quercetin-3′-sulfate, NaF, 3-bromopyruvate, NaN3, oligomycin A, ochratoxin A, cytochalasin B, and anti-GLUT1 antibody) showed dose-dependent changes in glucose and ATP levels without total protein (cell) loss. Finally, we performed flow cytometric glucose uptake measurement in the treated cells using 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxyglucose fluorescent glucose analog. Glucose uptake did not always mirror the intracellular glucose levels, which most likely reflects the differences between the two methodologies. However, interpreting data obtained by both methods and taking ATP/protein levels at the same time, one can get information on the mode of action of the compounds.
Biocatalysis of Platycoside E and Platycodin D3 Using Fungal Extracellular β-Glucosidase Responsible for Rapid Platycodin D Production
Authors: Ahn Hyung , You Hyun , Park Myung , Johnston Tony , Ku Seockmo , Ji Geun
Published: September 2018

Platycodi radix (i.e., Platycodon grandiflorum root) products (e.g., tea, cosmetics, and herbal supplements) are popular in East Asian nutraceutical markets due to their reported health benefits and positive consumer perceptions. Platycosides are the key drivers of Platycodi radixes’ biofunctional effects; their nutraceutical and pharmaceutical activities are primarily related to the number and varieties of sugar side-chains. Among the various platycosides, platycodin D is a major saponin that demonstrates various nutraceutical activities. Therefore, the development of a novel technology to increase the total platycodin D content in Platycodi radix extract is important, not only for consumers’ health benefits but also producers’ commercial applications and manufacturing cost reduction. It has been reported that hydrolysis of platycoside sugar moieties significantly modifies the compound’s biofunctionality. Platycodi radix extract naturally contains two major platycodin D precursors (platycoside E and platycodin D3) which can be enzymatically converted to platycodin D via β-d-glucosidase hydrolysis. Despite evidence that platycodin D precursors can be changed to platycodin D in the Platycodi radix plant, there is little research on increasing platycodin D concentrations during processing. In this work, platycodin D levels in Platycodi radix extracts were significantly increased via extracellular Aspergillus usamii β-d-glucosidase (n = 3, p < 0.001). To increase the extracellular β-d-glucosidase activity, A. usamii was cultivated in a culture media containing cellobiose as its major carbon source. The optimal pH and temperature of the fungal β-d-glucosidase were 6.0 and 40.0 °C, respectively. Extracellular A. usamii β-d-glucosidase successfully converted more than 99.9% (w/v, n = 3, p < 0.001) of platycoside E and platycodin D3 into platycodin D within 2 h under optimal conditions. The maximum level of platycodin D was 0.4 mM. Following the biotransformation process, the platycodin D was recovered using preparatory High Performance Liquid Chromatography (HPLC) and applied to in vitro assays to evaluate its quality. Platycodin D separated from the Platycodi radix immediately following the bioconversion process showed significant anti-inflammatory effects from the Lipopolysaccharide (LPS)-induced macrophage inflammatory responses with decreased nitrite and IL-6 production (n = 3, p < 0.001). Taken together, these results provide evidence that biocatalysis of Platycodi radix extracts with A. usamii may be used as an efficient method of platycodin D-enriched extract production and novel Platycodi radix products may thereby be created.
Multiple Aspects of PIP2 Involvement in C. elegans Gametogenesis
Authors: Ulicna Livia , Rohozkova Jana , Hozak Pavel
Published: September 2018

One of the most studied phosphoinositides is phosphatidylinositol 4,5-bisphosphate (PIP2), which localizes to the plasma membrane, nuclear speckles, small foci in the nucleoplasm, and to the nucleolus in mammalian cells. Here, we show that PIP2 also localizes to the nucleus in prophase I, during the gametogenesis of C. elegans hermaphrodite. The depletion of PIP2 by type I PIP kinase (PPK-1) kinase RNA interference results in an altered chromosome structure and leads to various defects during meiotic progression. We observed a decreased brood size and aneuploidy in progeny, defects in synapsis, and crossover formation. The altered chromosome structure is reflected in the increased transcription activity of a tightly regulated process in prophase I. To elucidate the involvement of PIP2 in the processes during the C. elegans development, we identified the PIP2-binding partners, leucine-rich repeat (LRR-1) protein and proteasome subunit beta 4 (PBS-4), pointing to its involvement in the ubiquitin–proteasome pathway.
Inhibition of Wnt3a/FOXM1/β-Catenin Axis and Activation of GSK3β and Caspases are Critically Involved in Apoptotic Effect of Moracin D in Breast Cancers
Authors: Hwang Sung Min , Lee Hyo-Jung , Jung Ji Hoon , Sim Deok Yong , Hwang Jisung , Park Ji Eon , Shim Bum Sang , Kim Sung-Hoon
Published: September 2018

Although Moracin D derived from Morus alba was known to have anti-inflammatory and antioxidant activities, the underlying antitumor mechanism of Moracin D has not been unveiled thus far. Thus, in the recent study, the apoptotic mechanism of Moracin D was elucidated in breast cancer cells. Herein, Moracin D exerted significant cytotoxicity in MDA-MB-231 and MCF-7 cells. Furthermore, Moracin D increased sub G1 population; cleaved poly (Adenosine diphosphate (ADP-ribose)) polymerase (PARP); activated cysteine aspartyl-specific protease 3 (caspase 3); and attenuated the expression of c-Myc, cyclin D1, B-cell lymphoma 2 (Bcl-2), and X-linked inhibitor of apoptosis protein (XIAP) in MDA-MB231 cells. Of note, Moracin D reduced expression of Forkhead box M1 (FOXM1), β-catenin, Wnt3a, and upregulated glycogen synthase kinase 3 beta (GSK3β) on Tyr216 along with disturbed binding of FOXM1 with β-catenin in MDA-MB-231 cells. Conversely, GSK3β inhibitor SB216763 reversed the apoptotic ability of Moracin D to reduce expression of FOXM1, β-catenin, pro-caspase3, and pro-PARP in MDA-MB-231 cells. Overall, these findings provide novel insight that Moracin D inhibits proliferation and induces apoptosis via suppression of Wnt3a/FOXM1/β-catenin signaling and activation of caspases and GSK3β.
The Protective Effect of a Long-Acting and Multi-Target HM-3-Fc Fusion Protein in Rheumatoid Arthritis
Authors: Huang Ruijing , Li Jian , Wang Yibo , Zhang Lihua , Ma Xiaohui , Wang Hongyu , Li Wenlei , Cao Xiaodan , Xu Hanmei , Hu Jialiang
Published: September 2018

Current treatment of rheumatoid arthritis (RA) is limited by relative shortage of treatment targets. HM-3 is a novel anti-RA polypeptide consisting of 18 amino acids with integrin αVβ3 and α5β1 as targets. Previous studies confirmed that HM-3 effectively inhibited the synovial angiogenesis and the inflammatory response. However, due to its short half-life, the anti-RA activity was achieved by frequent administration. To extend the half-life of HM-3, we designed a fusion protein with name HM-3-Fc, by combination of modified Fc segment of immunoglobulin 4 (IgG4) with HM-3 polypeptide. In vitro cell experiments demonstrated that HM-3-Fc inhibited the proliferation of splenic lymphocytes and reduced the release of TNF-α from macrophages. The pharmacodynamics studies on mice paw in Collagen-Induced Arthritis (CIA) model demonstrated that HM-3-Fc administered once in 5 days in the 50 and 25 mg/kg groups, or once in 7 days in the 25 mg/kg group showed a better protective effect within two weeks than the positive control adalimumab and HM-3 group. Preliminary pharmacokinetic studies in cynomolgus confirmed that the in vivo half-life of HM-3-Fc was 15.24 h in comparison with 1.32 min that of HM-3, which demonstrated that an Fc fusion can effectively increase the half-life of HM-3 and make it possible for further reduction of subcutaneous injection frequency. Fc-HM-3 is a long-acting active molecule for RA treatment.
Uremic Toxins Activates Na/K-ATPase Oxidant Amplification Loop Causing Phenotypic Changes in Adipocytes in In Vitro Models
Authors: Bartlett David E. , Miller Richard B. , Thiesfeldt Scott , Lakhani Hari Vishal , Khanal Tilak , D. Pratt Rebecca , Cottrill Cameron L. , Klug Rebecca L. , Adkins Nathaniel Seth , Bown Paul C. , Nease D. Blaine , Shapiro Joseph I. , Sodhi Komal
Published: September 2018

Background: Oxidant stress plays a key role in the development of chronic kidney disease (CKD). Experimental CKD leads to accumulation of uremic toxins (UT) in the circulation resulting in increased ROS production, which in turn, is known to activate the Na/K-ATPase/ROS amplification loop. Studies in a murine model of obesity have shown that increased oxidative stress in plasma is due to increased ROS and cytokine production from dysfunctional adipocytes. Therefore, we hypothesized that adipocytes exposed to UTs will activate the Na/K-ATPase oxidant amplification loop causing redox imbalance and phenotypic alterations in adipocytes. We also aimed to demonstrate that the Na/K-ATPase signaling antagonist, pNaKtide, attenuates these pathophysiological consequences. Methods: In the first set of experiments, 3T3-L1 murine pre-adipocytes were treated with varying concentrations of UTs, indoxyl sulfate (IS) (50, 100 and 250 µM) and p-cresol (50, 100 and 200 µM), with or without pNaKtide (0.7 µM) for five days in adipogenic media, followed by Oil Red O staining to study adipogenesis. RT-PCR analysis was performed to study expression of adipogenic, apoptotic and inflammatory markers, while DHE staining evaluated the superoxide levels in UT treated cells. In a second set of experiments, visceral fat was obtained from the West Virginian population. MSCs were isolated and cultured in adipogenic media for 14 days, which was treated with indoxyl sulfate (0, 25, 50 and 100 µM) with or without pNaKtide (1 µM). MSC-derived adipocytes were evaluated for morphological and molecular analysis of the above markers. Results: Our results demonstrated that 3T3-L1 cells and MSCs-derived adipocytes, treated with UTs, exhibited a significant decrease in adipogenesis and apoptosis through activation of the Na/K-ATPase/ROS amplification loop. The treatment with pNaKtide in 3T3-L1 cells and MSC-derived adipocytes negated the effects of UTs and restored cellular redox in adipocytes. We noted a varying effect of pNaKtide, in adipocytes treated with UTs, on inflammatory markers, adipogenic marker and superoxide levels in 3T3-L1 cells and MSC-derived adipocytes. Conclusions: This study demonstrates for the first time that the Na/K-ATPase/ROS amplification loop activated by elevated levels of UTs has varying effect on phenotypic alterations in adipocytes in various in vitro models. Thus, we propose that, if proven in humans, inhibition of Na/K-ATPase amplification of oxidant stress in CKD patients may ultimately be a novel way to combat adipocyte dysfunction and metabolic imbalance in these patients.
The Role of Sugarcane Catalase Gene ScCAT2 in the Defense Response to Pathogen Challenge and Adversity Stress
Authors: Sun Tingting , Liu Feng , Wang Wenju , Wang Ling , Wang Zhuqing , Li Jing , Que Youxiong , Xu Liping , Su Yachun
Published: September 2018

Catalases, which consist of multiple structural isoforms, catalyze the decomposition of hydrogen peroxide in cells to prevent membrane lipid peroxidation. In this study, a group II catalase gene ScCAT2 (GenBank Accession No. KF528830) was isolated from sugarcane genotype Yacheng05-179. ScCAT2 encoded a predicted protein of 493 amino acid residues, including a catalase active site signature (FARERIPERVVHARGAS) and a heme-ligand signature (RVFAYADTQ). Subcellular localization experiments showed that the ScCAT2 protein was distributed in the cytoplasm, plasma membrane, and nucleus of Nicotiana benthamiana epidermal cells. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis indicated that the ScCAT2 gene was ubiquitously expressed in sugarcane tissues, with expression levels from high to low in stem skin, stem pith, roots, buds, and leaves. ScCAT2 mRNA expression was upregulated after treatment with abscisic acid (ABA), sodium chloride (NaCl), polyethylene glycol (PEG), and 4 °C low temperature, but downregulated by salicylic acid (SA), methyl jasmonate (MeJA), and copper chloride (CuCl2). Moreover, tolerance of Escherichia coli Rosetta cells carrying pET-32a-ScCAT2 was enhanced by NaCl stress, but not by CuCl2 stress. Sporisorium scitamineum infection of 10 different sugarcane genotypes showed that except for YZ03-258, FN40, and FN39, ScCAT2 transcript abundance in four smut-resistant cultivars (Yacheng05-179, YZ01-1413, YT96-86, and LC05-136) significantly increased at the early stage (1 day post-inoculation), and was decreased or did not change in the two smut-medium-susceptibility cultivars (ROC22 and GT02-467), and one smut-susceptible cultivar (YZ03-103) from 0 to 3 dpi. Meanwhile, the N. benthamiana leaves that transiently overexpressed ScCAT2 exhibited less severe disease symptoms, more intense 3,3′-diaminobenzidine (DAB) staining, and higher expression levels of tobacco immune-related marker genes than the control after inoculation with tobacco pathogen Ralstonia solanacearum or Fusarium solani var. coeruleum. These results indicate that ScCAT2 plays a positive role in immune responses during plant–pathogen interactions, as well as in salt, drought, and cold stresses.
Melatonin Inhibits the Progression of Hepatocellular Carcinoma through MicroRNA Let7i-3p Mediated RAF1 Reduction
Authors: Wang Tong-Hong , Hsueh Chuen , Chen Chin-Chuan , Li Wan-Syuan , Yeh Chau-Ting , Lian Jang-Hau , Chang Junn-Liang , Chen Chi-Yuan
Published: September 2018

Melatonin is the main pineal hormone that relays light/dark-cycle information to the circadian system. Recent studies have examined the intrinsic antitumor activity of melatonin in various cancers, including hepatocellular carcinoma (HCC), the primary life-threatening malignancy in both sexes in Taiwan. However, the detailed regulatory mechanisms underlying melatonin’s anti-HCC activity remain incompletely understood. Here, we investigated the mechanisms by which the anti-HCC activity of melatonin is regulated. Human hepatoma cell lines were treated with 1 and 2 mM melatonin, and functional assays were used to dissect melatonin’s antitumor effect in HCC; small-RNA sequencing was performed to identify the microRNAs (miRNAs) involved in the anti-HCC activity of melatonin; and quantitative RT-PCR and Western blotting were used to elucidate how miRNAs regulate melatonin-mediated HCC suppression. Melatonin treatment at both doses strongly inhibited the proliferation, migration and invasion capacities of Huh7 and HepG2 cell lines, and melatonin treatment markedly induced the expression of the miRNA let7i-3p in cells. Notably, transfection of cells with a let7i-3p mimic drastically reduced RAF1 expression and activation of mitogen-activated protein kinase signaling downstream from RAF1, and rescue-assay results demonstrated that melatonin inhibited HCC progression by modulating let7i-3p-mediated RAF1 suppression. Our findings support the view that melatonin treatment holds considerable promise as a therapy for HCC.
Role of Cannabinoids in Obesity
Authors: Rossi Francesca , Punzo Francesca , Umano Giuseppina Rosaria , Argenziano Maura , Miraglia Del Giudice Emanuele
Published: September 2018

Obesity is an increasing health problem worldwide. Its related comorbidities imply a high cost for the National Health System and diminish a patient’s life quality. Adipose tissue is composed of three types of cells. White adipocytes are involved in fat storage and secretion of hormones. Brown adipocytes are involved in thermogenesis and caloric expenditure. Beige adipocytes are transitional adipocytes that in response to various stimuli can turn from white to brown and could be protective against the obesity, enhancing energy expenditure. The conversion of white in beige adipose tissue is a potential new therapeutic target for obesity. Cannabinoid receptors (CB) regulate thermogenesis, food intake and inflammation. CB1 ablation or inhibition helps reducing body weight and food intake. Stimulation of CB2 limits inflammation and promotes anti-obesity effects by reducing food intake and weight gain. Its genetic ablation results in adiposity development. CB receptors are also responsible for transforming white adipose tissue towards beige or brown adipocytes, therefore their modulation can be considered potential anti-obesity target. CB1 principal localization in central nervous system represents an important limit. Stimulation of CB2, principally localized on peripheral cells instead, should facilitate the anti-obesity effects without exerting remarkable psychotropic activity.
The “Frail” Brain Blood Barrier in Neurodegenerative Diseases: Role of Early Disruption of Endothelial Cell-to-Cell Connections
Authors: Maiuolo Jessica , Gliozzi Micaela , Musolino Vincenzo , Scicchitano Miriam , Carresi Cristina , Scarano Federica , Bosco Francesca , Nucera Saverio , Ruga Stefano , Zito Maria Caterina , Mollace Rocco , Palma Ernesto , Fini Massimo , Muscoli Carolina , Mollace Vincenzo
Published: September 2018

The main neurovascular unit of the Blood Brain Barrier (BBB) consists of a cellular component, which includes endothelial cells, astrocytes, pericytes, microglia, neurons, and oligodendrocytes as well as a non-cellular component resulting from the extracellular matrix. The endothelial cells are the major vital components of the BBB able to preserve the brain homeostasis. These cells are situated along the demarcation line between the bloodstream and the brain. Therefore, an alteration or the progressive disruption of the endothelial layer may clearly impair the brain homeostasis. The proper functioning of the brain endothelial cells is generally ensured by two elements: (1) the presence of junction proteins and (2) the preservation of a specific polarity involving an apical-luminal and a basolateral-abluminal membrane. This review intends to identify the molecular mechanisms underlying BBB function and their changes occurring in early stages of neurodegenerative processes in order to develop novel therapeutic strategies aimed to counteract neurodegenerative disorders.
Targeting CD46 Enhances Anti-Tumoral Activity of Adenovirus Type 5 for Bladder Cancer
Authors: Do Manh-Hung , To Phuong Kim , Cho Young-Suk , Kwon Se-Young , Hwang Eu Chang , Choi Chan , Cho Sang-Hee , Lee Sang-Jin , Hemmi Silvio , Jung Chaeyong
Published: September 2018

CD46 is generally overexpressed in many human cancers, representing a prime target for CD46-binding adenoviruses (Ads). This could help to overcome low anti-tumoral activity by coxsackie-adenoviral receptor (CAR)-targeting cancer gene therapy viruses. However, because of scarce side-by-side information about CAR and CD46 expression levels in cancer cells, mixed observations of cancer therapeutic efficacy have been observed. This study evaluated Ad-mediated therapeutic efficacy using either CAR-targeting Ad5 or CD46-targeting Ad5/35 fiber chimera in bladder cancer cell lines. Compared with normal urothelia, bladder cancer tissue generally overexpressed both CAR and CD46. While CAR expression was not correlated with disease progression, CD46 expression was inversely correlated with tumor grade, stage, and risk grade. In bladder cancer cell lines, expression levels of CD46 and CAR were highly correlated with Ad5/35- and Ad5-mediated gene transduction and cytotoxicity, respectively. In a human EJ bladder cancer xenograft mouse model, with either overexpressed or suppressed CD46 expression levels, Ad5/35-tk followed by ganciclovir (GCV) treatment significantly affected tumor growth, whereas Ad5-tk/GCV had only minimal effects. Overall, our findings suggest that bladder cancer cells overexpress both CAR and CD46, and that adenoviral cancer gene therapy targeting CD46 represents a more suitable therapy option than a CAR-targeting therapy, especially in patients with low risk bladder cancers.
Can Intradetrusor OnabotulinumtoxinA Injections Alter Heart Function in Patients with Cardiac Arrhythmia?
Authors: Miotla Pawel , Olejniczak Pawel , Futyma Konrad , Wrobel Andrzej , Tomaszewski Michal , Bogusiewicz Michal , Wawrysiuk Sara , Markut-Miotla Ewa , Rechberger Tomasz
Published: September 2018

The prevalence of overactive bladder (OAB) increases with age and can be associated with other co-morbidities, such as cardiac arrhythmia. Unfortunately, commonly used anticholinergic drugs for OAB can affect the cardiovascular system, leading to tachycardia. However, there are no data, which consider the influence of intradetrusor onabotulinumtoxinA injections on heart function in idiopathic OAB patients. The aim of the present study was to evaluate the influence of intradetrusor onabotulinumtoxinA injections on electrocardiogram (ECG) parameters. Additionally, changes in ECG were analyzed in OAB patients without cardiac arrhythmia. Thirty-one patients with cardiac arrhythmia and 31 participants without irregular heart rate (HR) completed the study. ECG measurements were performed in supine positions 2 h before onabotulinumtoxinA injections, 1 h after treatment, and at 2 weeks of follow-up. At week 6, a phone-call survey was conducted to collect data about adverse events. OnabotulinumtoxinA injections were performed with rigid cystoscopy under local anesthesia. We did not observe any clinically significant changes in the analyzed ECG parameters between consecutive measurements. While a slight increase of HR was observed in patients without cardiac arrhythmia, it remained within normal range. Intravesical onabotulinumtoxinA injections are, hence, safe for female patients with cardiac arrhythmia and do not significantly influence changes in ECG.
Translational Medicine in Pulmonary-Renal Crosstalk: Therapeutic Targeting of p-Cresyl Sulfate Triggered Nonspecific ROS and Chemoattractants in Dyspneic Patients with Uremic Lung Injury
Authors: Chang Jia-Feng , Liang Shih-Shin , Thanasekaran Pounraj , Chang Hsueh-Wei , Wen Li-Li , Chen Chung-Hua , Liou Jian-Chiun , Yeh Jih-Chen , Liu Shih-Hao , Dai Huei-Min , Lin Wei-Ning
Published: September 2018

Molecular mechanisms and pathological features of p-Cresyl sulfate (PCS)-induced uremic lung injury (ULI) in chronic kidney disease (CKD) remain unclear. We analyzed pleural effusions (PE) from CKD and non-CKD patients for uremic toxins, reactive oxygen species (ROS), and chemotactic cytokines. Correlations between PE biomarkers and serum creatinine were also studied. Cell viability and inflammatory signaling pathways were investigated in PCS-treated human alveolar cell model. To mimic human diseases, CKD-ULI mouse model was developed with quantitative comparison of immunostaining and morphometric approach. PE from CKD patients enhance expressions of uremic toxins, hydroxyl radicals, and IL-5/IL-6/IL-8/IL-10/IL-13/ENA-78/GRO α/MDC/thrombopoietin/VEGF. PE concentrations of ENA-78/VEGF/IL-8/MDC/PCS/indoxyl sulphate correlate with serum creatinine concentrations. In vitro, PCS promotes alveolar cell death, cPLA2/COX-2/aquaporin-4 expression, and NADPH oxidase/mitochondria activation-related ROS. Intracellular ROS is abrogated by non-specific ROS scavenger N-acetyl cysteine (NAC), inhibitors of NADPH oxidase and mitochondria-targeted superoxide scavenger. However, only NAC protects against PCS-induced cell death. In vivo, expressions of cPLA2/COX2/8-OHdG, resident alveolar macrophages, recruited leukocytes, alveolar space, interstitial edema and capillary leakage increase in lung tissues of CKD-ULI mice, and NAC pretreatment ameliorates alveolar–capillary injury. PCS causes alveolar–capillary injury through triggering intracellular ROS, downstream prostaglandin pathways, cell death, and activating leukocytes to release multiplex chemoattractants and extracellular ROS. Thus PCS and nonspecific ROS serve as potential therapeutic targets.
An Experimental Investigation of FRCC Shear Walls Reinforced with Steel and GFRP Bars
Authors: Tolou Kian Mohammad J. , Ghazizadeh Sina , Cruz Noguez Carlos A.
Published: September 2018

Contemporary structures can resist earthquakes as they deform and dissipate energy. However, during strong ground motions, these structures can sustain significant concrete damage and overall permanent deformations. Therefore, it is of great benefit if earthquake-resisting structures can deform and dissipate energy, and yet sustain mitigated damage. This paper illustrates the findings of an experimental study focused on the mitigation of damage and reduction of residual displacements in reinforced concrete (RC) shear walls. In this study, the cyclic properties of two innovative shear walls—a slender and a squat wall—which were cast with fiber-reinforced cementitious composites and reinforced with steel and glass fiber reinforced polymer bars are investigated. Then, the improvements of the innovative specimens with respect to two conventional RC shear walls are discussed in terms of damage propagation, self-centering, stiffness retention and energy dissipation. As the experiments showed, the innovative walls sustained mitigated concrete damage and less residual drift ratios while illustrating significant stiffness and energy dissipation capacities.
Clock Topologies for Molecular Quantum-Dot Cellular Automata
Authors: Blair Enrique , Lent Craig
Published: September 2018

Quantum-dot cellular automata (QCA) is a low-power, non-von-Neumann, general-purpose paradigm for classical computing using transistor-free logic. Here, classical bits are encoded on the charge configuration of individual computing primitives known as “cells.” A cell is a system of quantum dots with a few mobile charges. Device switching occurs through quantum mechanical inter-dot charge tunneling, and devices are interconnected via the electrostatic field. QCA devices are implemented using arrays of QCA cells. A molecular implementation of QCA may support THz-scale clocking or better at room temperature. Molecular QCA may be clocked using an applied electric field, known as a clocking field. A time-varying clocking field may be established using an array of conductors. The clocking field determines the flow of data and calculations. Various arrangements of clocking conductors are laid out, and the resulting electric field is simulated. It is shown that that control of molecular QCA can enable feedback loops, memories, planar circuit crossings, and versatile circuit grids that support feedback and memory, as well as data flow in any of the ordinal grid directions. Logic, interconnect and memory now become indistinguishable, and the von Neumann bottleneck is avoided.