bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison of Three Methods for Estimating Land Surface Temperature from Landsat 8-TIRS Sensor Data
Authors: García-Santos Vicente , Cuxart Joan , Martínez-Villagrasa Daniel , Jiménez Maria , Simó Gemma
Published: September 2018

After Landsat 8 was launched in 2013, it was observed that for Thermal Infrared sensor (TIRS) bands, radiance from outside of an instrument’s field-of-view produced a non-uniform ghost signal across the focal plane that varied depending on the out-of-scene content (i.e., the stray light effect). A new stray light correction algorithm (SLCA) is currently operational and has been implemented into the United States Geological Survey (USGS) ground system since February 2017. The SLCA has also been applied to reprocess historical Landsat 8 scenes. After approximately two years of SLCA implementation, more land surface temperature (LST) validation studies are required to check the effect of correction in the estimation of LST from different retrieval algorithms. For this purpose, three different LST estimation method algorithms (i.e., the radiative transfer equation (RTE), single-channel algorithm (SCA), and split-window algorithm (SWA)) have been assessed. The study site is located on the campus of the University of Balearic Islands on the island of Mallorca (Spain) in the western Mediterranean Sea. The site is considered a heterogeneous area that is composed of different types of surfaces, such as buildings, asphalt roads, farming areas, sloped terrains, orange fields, almond trees, lawns, and some natural vegetation regions. Data from 21 scenes, which were acquired by the Landsat 8-TIRS sensor and extracted from a 100 × 100 m2 pixel, were used to retrieve the LST with different algorithms; then, they were compared with in situ LST measurements from a broadband thermal infrared radiometer located on the same Landsat 8 pixel. The results show good performances of the three methods, with the SWA showing the lowest observed RMSE (within 1.6–2 K), whereas the SCA applied to the TIRS band 10 (10 µm) was also appropriate, with a RMSE ranging within 2.0–2.3 K. The LST estimates using the RTE algorithm display the highest observed RMSE values (within 2.0–3.6 K) of all of the compared methods, but with an almost unbiased value of −0.1 K for the case of techniques applied to band 10 data. The SWAs are the preferred method to estimate the LST in our study area. However, further validation studies around the world are required.
Mapping Maize Evapotranspiration at Field Scale Using SEBAL: A Comparison with the FAO Method and Soil-Plant Model Simulations
Authors: Grosso Carla , Manoli Gabriele , Martello Marco , Chemin Yann H. , Pons Diego H. , Teatini Pietro , Piccoli Ilaria , Morari Francesco
Published: September 2018

The surface energy balance algorithm for land (SEBAL) has been successfully applied to estimate evapotranspiration (ET) and yield at different spatial scales. However, ET and yield patterns have never been investigated under highly heterogeneous conditions. We applied SEBAL in a salt-affected and water-stressed maize field located at the margin of the Venice Lagoon, Italy, using Landsat images. SEBAL results were compared with estimates of evapotranspiration by the Food and Agriculture Organization (FAO) method (ETc) and three-dimensional soil-plant simulations. The biomass production routine in SEBAL was then tested using spatially distributed crop yield measurements and the outcomes of a soil-plant numerical model. The results show good agreement between SEBAL evapotranspiration and ETc. Instantaneous ET simulated by SEBAL is also consistent with the soil-plant model results (R2 = 0.7047 for 2011 and R2 = 0.6689 for 2012). Conversely, yield predictions (6.4 t/ha in 2011 and 3.47 t/ha in 2012) are in good agreement with observations (8.64 t/ha and 3.86 t/ha, respectively) only in 2012 and the comparison with soil-plant simulations (8.69 t/ha and 5.49 t/ha) is poor. In general, SEBAL underestimates land productivity in contrast to the soil-plant model that overestimates yield in dry years. SEBAL provides accurate predictions under stress conditions due to the fact that it does not require knowledge of the soil/root characteristics.
Fringing Electric Field Sensors for Anti-Attack at System-Level Protection
Authors: Gao Xiang , Zhao Yiqiang , Ma Haocheng
Published: September 2018

Information system security has been in the spotlight of individuals and governments in recent years. Integrated Circuits (ICs) function as the basic element of communication and information spreading, therefore they have become an important target for attackers. From this perspective, system-level protection to keep chips from being attacked is of vital importance. This paper proposes a novel method based on a fringing electric field (FEF) sensor to detect whether chips are dismantled from a printed circuit board (PCB) as system-level protection. The proposed method overcomes the shortcomings of existing techniques that can be only used in specific fields. After detecting a chip being dismantled from PCB, some protective measures like deleting key data can be implemented to be against attacking. Fringing electric field sensors are analyzed through simulation. By optimizing sensor’s patterns, areas and geometrical parameters, the methods that maximize sensitivity of fringing electric field sensors are put forward and illustrated. The simulation is also reproduced by an experiment to ensure that the method is feasible and reliable. The results of experiments are inspiring in that they prove that the sensor can work well for protection of chips and has the advantage of universal applicability, low cost and high reliability.
Designing Efficient Low-Cost Paper-Based Sensing Plasmonic Nanoplatforms
Authors: Susu Laurentiu , Campu Andreea , Craciun Ana Maria , Vulpoi Adriana , Astilean Simion , Focsan Monica
Published: September 2018

Paper-based platforms can be a promising choice as portable sensors due to their low-cost and facile fabrication, ease of use, high sensitivity, specificity and flexibility. By combining the qualities of these 3D platforms with the optical properties of gold nanoparticles, it is possible to create efficient nanodevices with desired biosensing functionalities. In this work, we propose a new plasmonic paper-based dual localized surface plasmon resonance–surface-enhanced Raman scattering (LSPR-SERS) nanoplatform with improved detection abilities in terms of high sensitivity, uniformity and reproducibility. Specifically, colloidal gold nanorods (GNRs) with a well-controlled plasmonic response were firstly synthesized and validated as efficient dual LSPR-SERS nanosensors in solution using the p-aminothiophenol (p-ATP) analyte. GNRs were then efficiently immobilized onto the paper via the immersion approach, thus obtaining plasmonic nanoplatforms with a modulated LSPR response. The successful deposition of the nanoparticles onto the cellulose fibers was confirmed by LSPR measurements, which demonstrate the preserved plasmonic response after immobilization, as well as by dark-field microscopy and scanning electron microscopy investigations, which confirm their uniform distribution. Finally, a limit of detection for p-ATP as low as 10−12 M has been achieved by our developed SERS-based paper nanoplatform, proving that our optimized plasmonic paper-based biosensing design could be further considered as an excellent candidate for miniaturized biomedical applications.
Improvement of Left Ventricular Ejection Time Measurement in the Impedance Cardiography Combined with the Reflection Photoplethysmography
Authors: Liu Shing-Hong , Wang Jia-Jung , Su Chun-Hung , Cheng Da-Chuan
Published: September 2018

Cardiac stroke volume (SV) is an essential hemodynamic indicator that can be used to assess whether the pump function of the heart is normal. Non-invasive SV measurement is currently performed using the impedance cardiography (ICG). In this technology, left ventricular ejection time (LVET) is an important parameter which can be determined from the ICG signals. However, the ICG signals are inherently susceptible to artificial noise interference, which leads to an inaccurate LVET measurement and then yields an error in the calculation of SV. Therefore, the goal of the study was to measure LVETs using both the transmission and reflection photoplethysmography (PPG), and to assess whether the measured LVET was more accurate by the PPG signal than the ICG signal. The LVET measured by the phonocardiography (PCG) was used as the standard for comparing with those by the ICG and PPG. The study recruited ten subjects whose LVETs were simultaneously measured by the ICG using four electrodes, the reflection PPG using neck sensors (PPGneck) and the transmission PPG using finger sensors (PPGfinger). In each subject, ten LVETs were obtained from ten heartbeats selected properly from one-minute recording. The differences of the measured LVETs between the PCG and one of the ICG, PPGneck and PPGfinger were −68.2 ± 148.6 ms, 4.8 ± 86.5 ms and −7.0 ± 107.5 ms, respectively. As compared with the PCG, both the ICG and PPGfinger underestimated but the PPGneck overestimated the LVETs. Furthermore, the measured LVET by the PPGneck was the closest to that by the PCG. Therefore, the PPGneck may be employed to improve the LVET measurement in applying the ICG for continuous monitoring of SV in clinical settings.
CNN-Based Multimodal Human Recognition in Surveillance Environments
Authors: Koo Ja Hyung , Cho Se Woon , Baek Na Rae , Kim Min Cheol , Park Kang Ryoung
Published: September 2018

In the current field of human recognition, most of the research being performed currently is focused on re-identification of different body images taken by several cameras in an outdoor environment. On the other hand, there is almost no research being performed on indoor human recognition. Previous research on indoor recognition has mainly focused on face recognition because the camera is usually closer to a person in an indoor environment than an outdoor environment. However, due to the nature of indoor surveillance cameras, which are installed near the ceiling and capture images from above in a downward direction, people do not look directly at the cameras in most cases. Thus, it is often difficult to capture front face images, and when this is the case, facial recognition accuracy is greatly reduced. To overcome this problem, we can consider using the face and body for human recognition. However, when images are captured by indoor cameras rather than outdoor cameras, in many cases only part of the target body is included in the camera viewing angle and only part of the body is captured, which reduces the accuracy of human recognition. To address all of these problems, this paper proposes a multimodal human recognition method that uses both the face and body and is based on a deep convolutional neural network (CNN). Specifically, to solve the problem of not capturing part of the body, the results of recognizing the face and body through separate CNNs of VGG Face-16 and ResNet-50 are combined based on the score-level fusion by Weighted Sum rule to improve recognition performance. The results of experiments conducted using the custom-made Dongguk face and body database (DFB-DB1) and the open ChokePoint database demonstrate that the method proposed in this study achieves high recognition accuracy (the equal error rates of 1.52% and 0.58%, respectively) in comparison to face or body single modality-based recognition and other methods used in previous studies.
Pixel-Wise Crack Detection Using Deep Local Pattern Predictor for Robot Application
Authors: Li Yundong , Li Hongguang , Wang Hongren
Published: September 2018

Robotic vision-based crack detection in concrete bridges is an essential task to preserve these assets and their safety. The conventional human visual inspection method is time consuming and cost inefficient. In this paper, we propose a robust algorithm to detect cracks in a pixel-wise manner from real concrete surface images. In practice, crack detection remains challenging in the following aspects: (1) detection performance is disturbed by noises and clutters of environment; and (2) the requirement of high pixel-wise accuracy is difficult to obtain. To address these limitations, three steps are considered in the proposed scheme. First, a local pattern predictor (LPP) is constructed using convolutional neural networks (CNN), which can extract discriminative features of images. Second, each pixel is efficiently classified into crack categories or non-crack categories by LPP, using as context a patch centered on the pixel. Lastly, the output of CNN—i.e., confidence map—is post-processed to obtain the crack areas. We evaluate the proposed algorithm on samples captured from several concrete bridges. The experimental results demonstrate the good performance of the proposed method.
Understanding the Living Conditions of Chinese Urban Neighborhoods through Social Infrastructure Configurations: The Case Study of Tianjin
Authors: Sun Xuan , Wang Weikai , Sun Tao , Wang Ya Ping
Published: September 2018

Under the unprecedented wave of urbanization and pace of economic development, the living conditions of residents have been unevenly changed in Chinese cities. To understand the diversified living conditions in Chinese urban neighborhoods from the supply-side point of view, new spatial and residential data were gathered to investigate the social infrastructure configurations around the residential communities. Using Tianjin as a case study, the research focuses on six important categories of human needs: education, healthcare, leisure, culture, entertainment, and transportation. Based on the cumulative accessibility measurements of 25 types of facilities, the social infrastructure configurations within neighborhoods are statistically and spatially analyzed and compared. The study discovered that: (1) despite the great diversity of living conditions in the city, the neighborhoods that have better services and strong associations with entertainment and education are prominent; (2) the neighborhoods with advantageous living conditions in different aspects tend to cluster at different places; (3) the neighborhoods of different types of communities, belonging to different administrative districts, or in different urban zones, all have distinctive characteristics in living conditions.
Scientific Information System for Silk Road Education Study
Authors: Kim TaeYoung , An NamIl , Lim JongBeom
Published: September 2018

In this paper, we present a scientific information system for the Silk Road education study. The proposed information system includes martial arts, dance, and play of seven countries (Korea, Japan, China, Mongolia, Kazakhstan, Uzbekistan, and Iran) of the Silk Road. The purpose of the information system is to promote convergence education for university students by providing a fundamental framework of the information system and traditional cultures. The basic concept of the information system can help university students to develop information and communications technology skills and to develop their own applications by collaborating with each other as a team. In addition, while developing the information system of martial arts, dance, and play of seven countries of the Silk Road, university students will understand the connection between traditional cultures and modern cultures of the Silk Road.
A Survey of Techniques for Constructing Chinese Knowledge Graphs and Their Applications
Authors: Wu Tianxing , Qi Guilin , Li Cheng , Wang Meng
Published: September 2018

With the continuous development of intelligent technologies, knowledge graph, the backbone of artificial intelligence, has attracted much attention from both academic and industrial communities due to its powerful capability of knowledge representation and reasoning. In recent years, knowledge graph has been widely applied in different kinds of applications, such as semantic search, question answering, knowledge management and so on. Techniques for building Chinese knowledge graphs are also developing rapidly and different Chinese knowledge graphs have been constructed to support various applications. Under the background of the “One Belt One Road (OBOR)” initiative, cooperating with the countries along OBOR on studying knowledge graph techniques and applications will greatly promote the development of artificial intelligence. At the same time, the accumulated experience of China in developing knowledge graphs is also a good reference to develop non-English knowledge graphs. In this paper, we aim to introduce the techniques of constructing Chinese knowledge graphs and their applications, as well as analyse the impact of knowledge graph on OBOR. We first describe the background of OBOR, and then introduce the concept and development history of knowledge graph and typical Chinese knowledge graphs. Afterwards, we present the details of techniques for constructing Chinese knowledge graphs, and demonstrate several applications of Chinese knowledge graphs. Finally, we list some examples to explain the potential impacts of knowledge graph on OBOR.
Products and Mechanistic Investigations on the Reactions of Hydrazines with Ozone in Gas-Phase
Authors: Huang Dan , Liu Xiangxuan , Xie Zheng , Wang Xuanjun , Gao Xin , Yang Yuxue
Published: September 2018

The toxic transformation products of hydrazines are of great concern. These products’ properties combined with their formation mechanisms are needed to assess their potential environmental and human impacts. In this study, the gas-phase reaction of hydrazine (N2H4), monomethyldrazine (MMH) and unsymmetrical dimethyhydrazine (UDMH) with O3 have been studied at varying reactant ratios, both in the presence and absence of a radical trap. Gas chromatography-mass spectroscopy (GC-MS) has been implied to follow reactant consumption and product formation. Apart from the reported products detected by Fourier transform infrared spectroscopy (FT-IR), the newly found compounds (hydrazones, formamides, dimethylamine, 1,1,4,4-tetramethyl-1,2-tetrazene,dimethylamino-acetonitrile, N2, H2O, et al.) are identified by GC-MS. The relative yields of the organic products vary considerably at different O3/MMH or UDMH ratios. UDMH and MMH are confirmed as high potential precursors of N-nitrosodimethylamine (NDMA). The presence of hydroxyl radicals (HO·) hinders NDMA formation in MMH-O3 system. Meanwhile, it increases NDMA formation in UDMH-O3 system. The suggested reaction mechanisms which account for the observed products are discussed.
Adsorption of Aclonifen, Alachlor, Cd and Cu onto Natural River Suspended Matter in the Context of Multi-Pollutions: Influence of Contaminant Co-Presence and Order of Input into the Aqueous Solution
Authors: El Azzi Desiree , Laurent François , Roussiez Vincent , Chou Lei , Guiresse Maritxu , Probst Jean-Luc
Published: September 2018

In the environment, a pollutant is rarely present alone, and other contaminants can influence its fate. To investigate the influence of the presence of other pollutants on the sorption of pesticides and trace metals (TM), the adsorption of Aclonifen, alachlor, Cd and Cu onto suspended particulate matter (SPM) was studied. SPM was isolated during the flood event of May 2010 in the Save agricultural watershed of SW France. Adsorption equilibrium was reached after 2 h of contact with SPM for pesticides and 24 h for TM. To simulate natural conditions, the SPM load allowing a maximum adsorption of pollutants was set at 1 g L−1 and the concentration of pollutants at 10 μg L−1. These factors being established, the co-presence experiments showed that most contaminants were influenced by the presence of other pollutant(s) in the water, trace metals to a lesser extent than pesticides. The mutual influence can be either competition for the same adsorption sites or formation of new complexes between pollutants. These phenomena can modify the adsorption capacities of each pollutant. The order of introduction into the aqueous solution also influenced the amount of adsorption of pollutants onto SPM. These results open new perspectives on the fate of pollutants.
Characterization of an Important Enzymatic Component in Collagenase that is Essential for the Effective Digestion of the Human and Porcine Pancreas
Authors: Chen Ruo L. , James Roger F. L.
Published: September 2018

Recent clinical results from Edmonton have demonstrated the feasibility of achieving normoglycemia in type I diabetic patients by islet transplantation. One of the key issues in obtaining this success was transplanting sufficient numbers of islets by sequential transplants. Although the development of semipurified endotoxin-free Clostridium histolyticum-derived collagenase (Liberase) has improved islet yields from the human pancreas, batch-to-batch variation and loss of activity with time still hampers progress in obtaining consistent islet preparations. In order to define key components of crude collagenase, a panel of monoclonal antibodies (McAbs) was raised against crude collagenase. Monoclonal antibodies were generated by fusions between splenocytes of BALB/c mice immunized with Boheringer P collagenase and the myeloma cell line NS-0. These monoclonal antibodies were used as probes to study molecular differences between effective and ineffective collagenase batches using Western blotting. Two monoclonal antibodies (LDS71 and LDS81) were raised and characterized as recognizing separate epitopes on a 125-kDa component. Western blotting indicated that the 125-kDa band was rapidly broken down by storage or by dialysis in the presence of dithiothreitol. However, this breakdown could be prevented by the addition of leupeptin (a protease inhibitor) to the dialysis buffer. On testing fractions at 5-min intervals from the “Ricordi” digestion circuit during porcine and human pancreas digestion, the 125-kDa component was rapidly broken down in relatively ineffective collagenase batches but in effective batches was present throughout the digestion process. The correlation between the presence of the 125-kDa band and effectiveness of pancreas digestion suggests that this may be a key component in the formulation of C. histolyticum collagenase.
Semiautomatic Macroencapsulation of Fresh or Cryopreserved Porcine Hepatocytes Maintain Their Ability for Treatment of Acute Liver Failure
Authors: Sarkis R. , Honiger J. , Chafai N. , Baudrimont M. , Sarkis K. , Delelo R. , Becquemont L. , Benoist S. , Balladur P. , Capeau J. , Nordlinger B.
Published: September 2018

We have previously demonstrated that fresh or cryopreserved xenogeneic hepatocytes manually macroencapsulated in AN69 polymer and transplanted intraperitoneally in rats were able to improve the survival rate after 95% hepatectomy without immunosuppression. In addition, we developed a semiautomatic device where porcine hepatocytes were coextruded with AN69 hydrogel in order to macroencapsulate large amounts of cells. The purpose of the present study was to 1) test whether transplanted porcine hepatocytes macroencapsulated in this device remained functional as evaluated by their ability to prevent death from acute liver failure, and 2) compare the efficiency of cryopreserved or freshly isolated hepatocytes. Fresh or cryopreserved porcine hepatocytes were macroencapsulated in the semiautomatic device by coextrusion in AN69 polymer in 2-m minitubes containing 6 × 107 cells. Acute liver failure was induced in rats by two-step 95% hepatectomy. At the time of completion of liver resection, rats were either not transplanted with minitubes (control group I, n = 13), or were implanted with two minitubes containing culture medium (control group II, n = 11), hepatocytes killed by heat treatment (control group III, n = 10), coextruded fresh hepatocytes (group IV, n = 11), or coextruded cryopreserved hepatocytes (group V, n = 11), without immunosuppression. The survival rate at day 7 was between 0% and 31% in the three control groups. By contrast, coextruded fresh hepatocytes significantly improved the survival rate (group IV, 82%) as did cryopreserved cells (group V, 91% survival). In surviving rats, minitubes were explanted after 20 days: either fresh or cryopreserved hepatocytes appeared morphologically viable and their ultrastructure was preserved. Their detoxification capacities evaluated by the activity of the cyt P450 CYP3A4 were partly maintained. In conclusion, porcine hepatocytes macroencapsulated by coextrusion using a semiautomatic device and transplanted without immunosuppression were able to prevent death from acute liver failure in rats. Cryopreserved cells were as efficient as fresh hepatocytes.
Effect of Flow on the Detoxification Function of Rat Hepatocytes in a Bioartificial Liver Reactor
Authors: Roy Partha , Washizu Junji , Tilles Arno W. , Yarmush Martin L. , Toner Mehmet
Published: September 2018

Ethoxyresorufin-o-deethylation (EROD) can be used as a sensitive measure of hepatic detoxification function. In this study, we employed a fluorescence assay based on EROD to study the effect of varying Peclet number (or flow) on hepatic function in a microchannel flat-plate bioartificial liver (BAL) reactor containing a coculture of hepatocytes and fibroblasts. Static culture and reactor flow experiments established that: 1) a pseudo-steady-state detoxification rate could be attained at each Peclet number, 2) the steady-state detoxification rate increased nonlinearly with Peclet number (ranging from 167 to 2500), 3) the uptake rate of substrate was a linear function of cell surface substrate concentration (<1 μM), and 4) a shear stress of 10 dyne/cm2 did not adversely affect hepatic function for at least 12 h. A convection–diffusion–reaction model supports the conclusion that increased convective mass transfer of substrate to the cell surface is the primary cause of the observed increase in EROD rate with Peclet number. Our results suggest that detoxification rates can be enhanced by an order of magnitude by choosing an appropriate Peclet number. For our bioreactor configuration, this optimum corresponds to a Peclet number range of 1000–2000 at a Damkohler number of 0.55. The usefulness of the mathematical model is discussed in the context of scale-up to a clinical BAL reactor for human application.
Absence of CSF-1-Dependent Macrophages Does Not Improve Function of Transplanted Islets of Langerhans
Authors: Berney Thierry , Molano R. Damaris , Pileggi Antonello , Cattan Pierre , Vizzardelli Caterina , Ricordi Camillo , Inverardi Luca
Published: September 2018

A role of macrophage-mediated inflammatory events in early islet graft loss is increasingly acknowledged. Osteopetrotic mice (op/op) have a complete absence of CSF-1, and thus of most tissue macrophages. We have investigated whether the absence of CSF-1-dependent macrophages in the graft itself or at the transplant site could decrease the delay to function of a syngeneic marginal islet mass. Islets transplanted into op/op or control recipients reversed diabetes in 59 days vs. 10 days (p = 0.28, NS). Islets isolated from op/op or control mice reversed diabetes in 11 days vs. 10 days. IL-1 and TNF-α release by cultured islets was markedly decreased for op/op islets compared with control islets (IL-1: 0 vs. 4.2 pg/ml, p = 0.07; TNF-α: 67 vs. 311 pg/ml, p = 0.002). In contrast, IL-6 release by op/op islets was significantly increased (11.1 vs. 4.3 ng/ml, p = 0.006). CSF-1-dependent tissue macrophages may not be critical in the inflammatory insult to islet transplants. Alternate patterns of intraislet release of deleterious proinflammatory cytokines may exist.
Inhibition of Cytokine Production and Cytokine-Stimulated T-Cell Activation by FK506 (Tacrolimus)1
Authors: Almawi Wassim Y. , Assi Joumana W. , Chudzik Dagmara M. , Jaoude Maroun M. Abou , Rieder Michael J.
Published: September 2018

Insofar as it exerted its immunosuppressive effect by inhibiting cytokine expression, we assessed the effect of FK506 (Tacrolimus) on cytokine-stimulated T-cell activation. Human T cells, treated with FK506, or controls were stimulated with the mitogens PHA + PMA, Con A, and the “CD3-bypass” stimulation regimen, PMA + ionomycin. T-cell proliferation was quantitated by measuring the uptake of tritiated thymidine, and mRNA expression was assessed by RT-PCR. FK506, in a concentration-dependent fashion, inhibited T-cell proliferation and steady-state mRNA expression of IL-2 and IL-7; half-maximal suppression was obtained at 10-7 to 5 × 10-8 M. We tested whether FK506 antiproliferative effect could be overcome with exogenously reconstituted rIL-2 and/or rIL-7. Neither rIL-2 nor rIL-7, individually in conjunction with suboptimal concentrations of PHA or Con A, or in combination without any costimulus, was capable of abrogating FK506 antiproliferative effect, indicating that FK506 also acted by inhibiting cytokine-stimulated T-cell activation. To confirm this, T cells were treated with FK506 and stimulated by rIL-2 and rIL-7, individually in conjunction with suboptimal concentration of PHA and Con A. In addition, T cells were stimulated with rIL-2 and rIL-7 without any costimuli. FK506 inhibited T-cell activation stimulated by rIL-2 and by rIL-7, individually and in combination. This confirms that, in exerting its antiproliferative effect, FK506 acts at two levels, by inhibiting cytokine availability and by suppressing cytokine effect on target cells, and explains the beneficial effect of FK506 in attenuating ongoing immune responses.
Prolongation of Islet Allograft Survival in Mice by Combined Treatment with Pravastatin and Low-Dose Cyclosporine
Authors: Arita Seiji , Kasraie Ali , Une Satoshi , Ohtsuka Satoki , Smith Craig V. , Mullen Yoko
Published: September 2018

Pravastatin, a 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitor, is known to have suppressive effects on immune and inflammatory cells. We have previously shown in mice and dogs that this agent prevents primary nonfunction of islet iso- and autografts by reducing inflammation at the graft site. The present study was designed to further investigate whether pravastatin has a synergistic effect with cyclosporine (Cs) to prolong islet allograft survival in mice. Unpurified 3000 BALB/c newborn islets were transplanted under the renal capsule of a streptozotocin-diabetic C57BL/6 mouse. Pravastatin and Cs were administered for 10 days starting on the day of grafting (day 0). Five groups were set up based on the treatment protocol: group 1, treatment with 40 mg/kg pravastatin; group 2, 30 mg/kg Cs; group 3, 50 mg/ kg Cs; group 4, 40 mg/kg pravastatin and 30 mg/kg Cs; group 5, vehicle alone. Graft survival was indicated by blood glucose levels sustained at <200 mg/dl, and graft rejection by >250 mg/dl for 2 consecutive days. Hyperglycemia persisted in six of the eight (75%) mice and grafts were rejected in 3.6 ± 0.5 days (mean ± SD) in group 5. In group 1, grafts were also rejected in 3.8 ± 0.8 days, but blood glucose was transiently <200 mg/dl in three of the five mice. Despite Cs, grafts were rejected between 7 and 15 days (10.3 ± 2.4 days) in group 2. Among six mice in group 3, one maintained euglycemia for >60 days, the other rejected the graft on day 15, and the remaining four died with functioning grafts between 9 and 13 days due to Cs toxicity. A combination of a low dose of Cs and pravastatin (group 4) prolonged graft survival for >19 days in five of the eight mice, and for 7–13 days in the remaining three mice. Histological examination of the grafts in this group showed significantly reduced local inflammation. Results indicate a synergistic effect of pravastatin and Cs on prevention of islet allograft rejection.
A Molecular Basis of Cryopreservation Failure and its Modulation to Improve Cell Survival
Authors: Baust John M. , Vogel Martin J. , Van Buskirk Robert , Baust John G.
Published: September 2018

The requirement for more effective cryopreservation (CP) methodologies in support of the emerging fields of cell bioprocessing and cell therapy is now critical. Current CP strategies appropriately focus on minimizing the damaging actions of physicochemical stressors and membrane disruption associated with extra- and intracellular ice formation that occurs during the freeze–thaw process. CP protocols derived from this conceptual paradigm, however, yield suboptimal survival rates. We now provide the first report on the identification of delayed-onset cell death following CP and the significance of modulating molecular biological aspects of the cellular responses (apoptosis) to low temperature as an essential component to improve postthaw outcome. In this study we quantitatively examined the molecular basis of cell death associated with CP failure in a canine renal cell model. In addition, we report on the significant improvement in CP outcome through the modulation of these molecular mechanisms by the utilization of an organ preservation solution, HypoThermosol®. Further, the utilization of HypoThermosol® as the preservation medium and the modulation of molecular-based cell death have led to a paradigm shift in biologic preservation methodologies. The recognition of molecular mechanisms associated with CP-induced cell death offers the promise of improved CP of more complex and/or fragile biological systems such as stem cells, engineered tissues, and human organs.
In Vivo Estimation of Bioartificial Liver with Recombinant HepG2 Cells Using Pigs with Ischemic Liver Failure
Authors: Enosawa Shin , Miyashita Tomoyuki , Fujita Yuji , Suzuki Seiichi , Amemiya Hiroshi , Omasa Takeshi , Hiramatsu Shinya , Suga Kenichi , Matsumura Toshiharu
Published: September 2018

Biological efficacy of a recombinant human hepatic cell line, glutamine synthetase transfected HepG2 (GS-HepG2), was examined with large-scale culture in a circulatory flow bioreactor and in pigs with ischemic liver failure. GS-HepG2 cells were cultured in a circulatory flow bioreactor from 5 × 107 to 4 × 109 cells for 109 days. The cells showed ammonia removal activity even under substrate (glutamic acid)-free medium, suggesting that the GS catalyzed the activity using intracellular glutamic acid that had been pooled during conventional culture. When GS-HepG2 bioartificial liver (BAL) was applied to pigs with ischemic liver failure, survival time was prolonged to 18.8 ± 6.1 h (mean ± SD, n = 4) from 13.8 ± 5.4 h (n = 6) and 10.7 ± 4.1 h (n = 6) (groups treated with cell-free BAL and treated with plasma exchange and continuous hemodia-filtration, respectively). Laboratory data indicated the tendency for improvement in increase of blood ammonia level and decline of blood coagulation indices in the GS-HepG2 BAL-treated group. The advantages and potential for the cell line as a bioreactor in BAL is also discussed, comparing to those of isolated porcine hepatocytes.
The Influence of the Anticomplement Synthetic Sulfonic Polymers on the Function of Pancreatic Islets: An In Vitro Study
Authors: Xu Baoyou , Gu Yuanjun , Miyamoto Masaaki , Balamurugan A. N. , Cui Wanxing , Imamura Masayuki , Iwata Hiroo , Inoue Kazutomo
Published: September 2018

In a previous experiment, we demonstrated the anticomplementary efficacy of poly(stryrene sulfonic acid) (PSSa) and poly(2-acrylamido-2-methyl propane sulfonic acid) (PAMPS). The aim of this study was to examine their influence on the function of pancreatic islets in vitro. In this study, after culturing the rat islets with RPMI-1640 culture medium containing different concentrations of soluble PSSa or PAMPS for 24 h at 37°C, we performed morphological and functional examination of the rat islets. We found that the islets maintained their normal morphology regardless of whether they were in the PSSa or PAMPS groups when the concentrations of soluble PSSa or PAMPS in the media were below 1 g/dl. In the static incubation study, the islets cultured in the PAMPS groups showed significantly high insulin secretory response to glucose challenge but those in the PSSa groups lost the response when the concentrations of soluble PSSa or PAMPS in the media were below 1 g/dl. The PAMPS not only had strong anticomplementry effect, but also maintained the good insulin secretory capacity of the islets. These results indicated that PAMPS is a promising bioartificial material for future clinical application of biohybrid artificial pancreas preparation. It is well suitable for xenotransplantation experiments.
Effect of Basic Fibroblast Growth Factor on Insulin Secretion from Microencapsulated Pancreatic Islets: An In Vitro Study
Authors: Wang Wenjing , Gu Yuanjun , Miyamoto Masaaki , Hori Hiroshi , Nagata Natsuki , Balamurugan A. N. , Touma Maki , Sakurai Tomonori , Inoue Kazutomo
Published: September 2018

Microencapsulation of pancreatic islets represents a potentially effective method to prevent graft rejection in allotransplantation or xenotransplantation without the need of immunosuppression. Adequate insulin secretion and glucose responsiveness of microencapsulated pancreatic islets has been regarded as a prerequisite for successful transplantation. The microencapsulated pancreatic islets were respectively cultured in bFGF+ RPMI-1640 medium (bFGF+) or bFGF- RPMI-1640 medium (bFGF-) for 21 days. The functional activities of microencapsulated pancreatic islets were assessed by measuring basal insulin secretion and stimulated insulin release at different time points. The results revealed that microencapsulated pancreatic islets in the presence of bFGF demonstrated an increase in basal insulin secretion. Furthermore, microencapsulated pancreatic islets in the presence of bFGF demonstrated a marked stimulated insulin release and relative stability of stimulation indices (SI). The results in the perifusion study showed that microencapsulated pancreatic islets in the presence of bFGF maintained good glucose responsiveness over the course of culture period as well. These results indicate that bFGF has a beneficial effect on insulin secretion from microencapsulated pancreatic islets during in vitro culture. New strategies for preserving and improving function of microencapsulated pancreatic islets prior to transplantation may be developed by application of growth factors or other factors.
Evaluation of Insulin Secretion of Isolated Rat Islets Cultured in Extracellular Matrix
Authors: Nagata Natsuki , Gu Yuanjun , Hori Hiroshi , Balamurugan A. N. , Touma Maki , Kawakami Yoshiyuki , Wang Wenjing , Baba Tomomi T. , Satake Akira , Nozawa Masumi , Tabata Yasuhiko , Inoue Kazutomo
Published: September 2018

Islet isolation involves enzymatic digestion of the interstitial matrix and mechanical disruption of the tissue. It is possible that a fundamental change of islet biology resulting from the loss of critical factors required for islet function or survival will occur. Extracellular matrix (ECM) is one of the most important components of the islet microenvironment. Reconstruction of the cell–matrix relationship seems to be effective for improving the loss of differentiated islet structure and function. The purpose of this study was to characterize and compare the effects of collagen gel mixture or Matrigel on β-cell function and islet cell survival. After isolation by the collagenase digestion technique, rat islets were divided and cultured with various types of collagen gel mixture. They were assessed for their glucose-stimulated insulin secretion and cell viability. Glucose-induced insulin secretion of islets cultured with collagen type I gel or a mixture of collagen type I and IV was improved after 11 days in culture. In conclusion, a type of gel composed of collagen type I and/ or type IV as an islet microenvironment is sufficient to maintain glucose responsiveness and may be useful for islet transplantation.
Successful Retroviral Gene Transfer of Simian Virus 40 T Antigen and Herpes Simplex Virus-Thymidine Kinase into Human Hepatocytes
Authors: Kobayashi Naoya , Noguchi Hirofumi , Westerman Karen A. , Watanabe Takamasa , Matsumura Toshihisa , Totsugawa Toshinori , Fujiwara Toshiyoshi , Leboulch Philippe , Tanaka Noriaki
Published: September 2018

Current clinical reports have indicated that hepatocyte transplantation (HTX) could be used in patients with liver failure and in children with liver-based metabolic diseases. One of the major limiting factors of HTX is a serious shortage of donor livers for hepatocyte isolation. To address this issue, we immortalized adult human hepatocytes with a retroviral vector SSR#69 expressing the genes of simian virus 40 large T antigen and herpes simplex virus-thymidine kinase simultaneously. One of the resulting clones, NKNT-3, grew steadily in chemically defined serum-free medium without any obvious crisis and showed the gene expression of differentiated liver functions. Under the administration of 5 μM ganciclovir, NKNT-3 cells stopped proliferation and died in in vitro experiments. We have established a tightly regulated immortal human hepatocyte cell line. The cells could allow the need for immediate availability of consistent and functionally uniform cells in sufficient quantity and adequate quality.
Cytological Examination of Rat Amniotic Epithelial Cells and Cell Transplantation to the Liver
Authors: Nakajima Tatsuya , Enosawa Shin , Mitani Tasuku , Li Xiao-Kang , Suzuki Seiichi , Amemiya Hiroshi , Koiwai Osamu , Sakuragawa Norio
Published: September 2018

It is hoped that amniotic epithelial cells can be useful in cell-mediated gene therapy. We report here an experimental cell transplantation model of amniotic cells in rats. There is an anatomical difference between human and rodent embryos. We established a method to isolate amniotic cells that are equivalent to human amniotic epithelial cells. An amniotic membrane distinct from the yolk sac was carefully collected and teased in saline containing deoxyribonuclease and hyaluronidase, followed by collagenase digestion. The cell yield was approximately 106 cells per pregnant female (105 cells per fetus), roughly in proportion to the age of fetus used, and 60% of the isolated cells were attached to the dish under culture conditions. Telomerase activity was higher in the cells isolated from fetuses in the middle stage (day 13.5 to 15.5) than in the late stage (day 17.5 to 21.5). Adherent cells exhibited two to three times more cell division, resulting in a ninefold increase in the number of cells. Immunohistochemical analysis revealed that approximately half of the adherent cells were albumin positive and formed clusters. The senescent cells survived for 2 months without apparent morphological changes. The adherent cells were able to be stored in liquid nitrogen and had a viability of 70% when thawed. Gene transduction with adenovirus vector was highly effective for rat amniotic cells. Transplantation of lacZ transfected amniotic cells into syngeneic rat liver resulted in the integration of the transplanted cells in the liver structure and the cells survived for at least 30 days.
Widespread Distribution of Adenovirus-Transduced Monkey Amniotic Epithelial Cells after Local Intracerebral Injection: Implication for Cell-Mediated Therapy for Lysosome Storage Disorders
Authors: Kosuga Motomichi , Takahashi Satoru , Tanabe Akiko , Fujino Masayuki , Li Xiao-Kang , Suzuki Seiichi , Yamada Masao , Kakishita Kohji , Ono Fumiko , Sakuragawa Norio , Okuyama Torayuki
Published: September 2018

Cell-mediated therapy for mucopolysaccharidosis type VII (MPSVII) was studied using monkey amniotic epithelial cells (mAEC). The cells were transduced with a recombinant adenovirus expressing human β-glucuronidase (GUSB), and cells overexpressing GUSB were generated. The cells expressed 2000-fold higher activities than the endogenous GUSB activities of nontransduced mAEC, demonstrating that mAEC were successfully transduced with adenoviral vectors. These cells also secreted high levels of GUSB. To clarify the cross-correction of GUSB secreted from mAEC, the conditioned medium containing high levels of GUSB was added into the medium for culturing human or murine fibroblasts established from an MPSVII patient or a mouse model of the disease. Dramatic increases in GUSB activities were observed in both fibroblasts. We then transplanted the cells transduced with an adenovirus expressing LacZ into the caudate-putamen of monkey brain. Survival and distribution of the transplanted cells 1 month after the treatment were evaluated. Histochemical analysis showed that LacZ-positive cells were widely distributed in the brain, suggesting that the transplanted cells had migrated and were distributed even at regions far from the implantation site. These findings suggest that local intracerebral engraftment of genetically engineered amniotic epithelial cells is favorable for the treatment of lysosome storage disorders, whose pathological abnormalities are not restricted to specific regions of the brain.
Chitosan-Polyvinyl Pyrrolidone Hydrogel does Not Activate Macrophages: Potentials for Transplantation Applications
Authors: Risbud Makarand , Bhonde Mandar , Bhonde Ramesh
Published: September 2018

We have shown previously that chitosan-polyvinyl pyrrolidone (PVP) hydrogels are blood compatible, islet compatible, and noncytotoxic to various cell types. Because of these potential applications of chitosan-PVP hydrogel, the present study was designed to investigate its effect on macrophage activation. Macrophages did not adhere to hydrogel in culture but maintained their viability and did not undergo apoptosis as confirmed by trypan blue staining and absence of DNA ladder. Hydrogel leach-out products did not exhibit cytotoxic effects on macrophage functionality at mitochondrial and lysosomal level as confirmed by tetrazolium reduction (MTT) and neutral red uptake (NRU) assay. On exposure to hydrogels, macrophages showed comparable expression of activation markers such as CD11b/CD18 (Mac-1), CD45, and CD14 to those cultured in the presence of PTFE, a known biocompatible control, indicating its nonactivating nature. Macrophage activation was also assessed by checking the level of messenger RNA of inflammatory cytokines such as IL-6 and TNF-α by reverse transcriptase polymerase chain reaction (RT-PCR), which did not show stastistically significant difference (p > 0.05) in the expression of these transcripts in both control and hydrogel-exposed macrophages. The nonimmunogenic nature of the hydrogel was further confirmed by the lack of induced proliferation of mouse splenic lymphocytes after exposure to hydrogel leach-outs. All these results point out that chitosan-PVP hydrogel did not activate macrophages and thus is immunocompatible. Our results indicate that this hydrogel could be a potential candidate for transplantation studies by virtue of its biocompatibility and imunocompatibility.