bluuebunny/crossref_metadata_2025 huggingface dataset:
Improved Low-Temperature Activity of V2O5-WO3/TiO2 for Denitration Using Different Vanadium Precursors
Authors: Gan Lina , Guo Feng , Yu Jian , Xu Guangwen
Published: February 2016

This work tested two V2O5-WO3/TiO2 catalysts with different vanadium precursors for selective catalytic reduction (SCR) of flue gas NO using NH3 at 150–450 °C. While catalyst A was prepared using ammonium metavanadate (NH4VO3) through incipient impregnation, catalyst B was made according to the solvothermal method using vanadyl acetylacetonate (VO(acac)2) as the vanadium precursor. The catalytic evaluation for denitration was in a laboratory fixed bed reactor using simulated flue gas under conditions of a gas hourly space velocity (GHSV) of 40,000 h−1 and an NH3/NO molar ratio of 0.8. Without SO2 and water vapor in the flue gas at 200 °C, the realized NO conversion was 56% for catalyst A but 80% for B. The presence of 350 ppm SO2 and 10 vol. % water vapor in the flue gas slightly reduced the NO conversion over catalyst B, and its activity was stable in a 108-h continuous test at temperatures varying from 450 °C to 220 °C. Via fourier transformation infrared spectroscopy (FT-IR) and thermogravimetric (TG) analysis, it was shown that over catalyst B a dynamic balance between the formation and decomposition of ammonium sulfite or sulfate is built possibly at temperatures as low as 220 °C. For this catalyst there was a higher surface atomic concentration of vanadium and a higher ratio of V4+/(V4+ + V5+), while the NH3 adsorption test revealed more acidic sites on catalyst B. The study discloses a potentially new approach to prepare a V2O5-WO3/TiO2 catalyst with good performance for SCR of flue gas NO at 220–300 °C.
5-Azido-4-dimethylamino-1-methyl-1,2,4-triazolium Hexafluoridophosphate and Derivatives
Authors: Laus Gerhard , Kahlenberg Volker , Schottenberger Herwig
Published: February 2016

5-Azido-4-(dimethylamino)-1-methyl-1,2,4-triazolium hexafluoridophosphate was synthesized from the corresponding 5-bromo compound with NaN3. Reaction with bicyclo[2.2.1]hept-2-ene yielded a tricyclic aziridine, addition of an N-heterocyclic carbene resulted in a triazatrimethine cyanine, and reduction with triphenylphosphane gave the 5-amino derivative. The crystal structures of three nitrogen-rich salts were determined. Thermoanalysis of the cationic azide and triazene showed exothermal decomposition. The triazene exhibited negative solvatochromism in polar solvents involving the dipolarity π* and hydrogen-bond donor acidity α of the solvent.
Above- and Belowground Biomass Models for Trees in the Miombo Woodlands of Malawi
Authors: Kachamba Daud , Eid Tron , Gobakken Terje
Published: February 2016

In this study we present general (multiple tree species from several sites) above- and belowground biomass models for trees in the miombo woodlands of Malawi. Such models are currently lacking in the country. The modelling was based on 74 trees comprising 33 different species with diameters at breast height (dbh) and total tree height (ht) ranging from 5.3 to 2 cm and from 3.0 to 25.0 m, respectively. Trees were collected from four silvicultural zones covering a wide range of conditions. We tested different models including dbh, ht and wood specific gravity ( ρ ) as independent variables. We evaluated model performance using pseudo-R2, root mean square error (RMSE), a covariance matrix for the parameter estimates, mean prediction error (MPE) and relative mean prediction error (MPE%). Computation of MPE% was based on leave-one-out cross-validation. Values of pseudo-R2 and MPE% ranged 0.82–0.97 and 0.9%–2.8%, respectively. Model performance indicated that the models can be used over a wide range of geographical and ecological conditions in Malawi.
Serotonin 2A Receptor SNP rs7330461 Association with Treatment Response to Pomaglumetad Methionil in Patients with Schizophrenia
Authors: Nisenbaum Laura , Downing AnnCatherine , Zhao Fangyi , Millen Brian , Munsie Leanne , Kinon Bruce , Adams David , Gomez Juan , Penny Michelle
Published: February 2016

This study aims to confirm the initial pharmacogenetic finding observed within the clinical proof-of-concept trial of an enhanced response to treatment with pomaglumetad methionil (LY2140023 monohydrate) in Caucasian schizophrenia patients homozygous for T/T at single nucleotide polymorphism rs7330461 in the serotonin (5-hydroxytryptamine) 2A receptor gene compared to A/A homozygous patients. The effect of the rs7330461 genotype on the response to pomaglumetad methionil treatment was assessed in three additional clinical trials and in an integrated analysis. Overall, this study includes data from 1115 Caucasian patients for whom genotyping information for rs7330461 was available, consisting of 513 A/A homozygous, 466 A/T heterozygous and 136 T/T homozygous patients. Caucasian T/T homozygous patients showed significantly (p ≤ 0.05) greater improvement in Positive and Negative Syndrome Scale (PANSS) total scores during treatment with pomaglumetad methionil 40 mg twice daily compared to A/A homozygous patients. Additionally, T/T homozygous patients receiving pomaglumetad methionil had significantly (p ≤ 0.05) greater improvements in PANSS total scores compared to placebo and similar improvements as T/T homozygous patients receiving standard-of-care (SOC) treatment. The findings reported here in conjunction with prior reports show that in Caucasian patients with schizophrenia, the T/T genotype at rs7330461 is consistently associated with an increased treatment response to pomaglumetad methionil compared to the A/A genotype.
Cellular Uptake and Photo-Cytotoxicity of a Gadolinium(III)-DOTA-Naphthalimide Complex “Clicked” to a Lipidated Tat Peptide
Authors: O’Malley William , Rubbiani Riccardo , Aulsebrook Margaret , Grace Michael , Spiccia Leone , Tuck Kellie , Gasser Gilles , Graham Bim
Published: February 2016

A new bifunctional macrocyclic chelator featuring a conjugatable alkynyl-naphthalimide fluorophore pendant group has been prepared and its Gd(III) complex coupled to a cell-penetrating lipidated azido-Tat peptide derivative using Cu(I)-catalysed “click” chemistry. The resulting fluorescent conjugate is able to enter CAL-33 tongue squamous carcinoma cells, as revealed by confocal microscopy, producing a very modest anti-proliferative effect (IC50 = 93 µM). Due to the photo-reactivity of the naphthalimide moiety, however, the conjugate’s cytotoxicity is significantly enhanced (IC50 = 16 µM) upon brief low-power UV-A irradiation.
Effect of NaCl on the Lifetime of Micro- and Nanobubbles
Authors: Uchida Tsutomu , Liu Shu , Enari Masatoshi , Oshita Seiichi , Yamazaki Kenji , Gohara Kazutoshi
Published: February 2016

Micro- and nanobubbles (MNBs) are potentially useful for industrial applications such as the purification of wastewater and the promotion of physiological activities of living organisms. To develop such applications, we should understand their properties and behavior, such as their lifetime and their number density in solution. In the present study, we observed oxygen MNBs distributed in an electrolyte (NaCl) solution using a transmission electron microscope to analyze samples made with the freeze-fracture replica method. We found that MNBs in a 100 mM NaCl solution remain for at least 1 week, but at higher concentrations decay more quickly. To better understand their lifetimes, we compared measurements of the solution's dissolved oxygen concentration and the ζ-potential of the MNBs. Our detailed observations of transmission electron microscopy (TEM) images allows us to conclude that low concentrations of NaCl stabilize MNBs due to the ion shielding effect. However, higher concentrations accelerate their disappearance by reducing the repulsive force between MNBs.
Light Emission Properties of a Cross-Conjugated Fluorene Polymer: Demonstration of Its Use in Electro-Luminescence and Lasing Devices
Authors: Romero-Servin Sergio , Lozano-Hernández Luis-Abraham , Maldonado José-Luis , Carriles Ramón , Ramos-Ortíz Gabriel , Pérez-Gutiérrez Enrique , Scherf Ullrich , Zolotukhin Mikhail
Published: February 2016

Light emission properties of a fluorene cross-conjugated polymer (PF–1) based on the monomer 4,7-bis[2-(9,9-dimethyl)fluorenyl] benzo[1,2,5]thiadiazole are reported. This polymer exhibits solubility at high concentrations, good processability into thin solid films of good quality and a broad emission band with a fluorescence quantum yield of approximately 1. Based on these features, in this paper we implemented the use of PF–1 as an active layer in polymer light-emitting diodes (PLEDs) and as a laser gain medium in solution. To get insight on the conducting properties of PF–1, two different electron injectors, poly [(9,9-bis(3′-(N,N-dimethylamino) propyl)-2,7-fluorene)-alt-2,7-(9,9–dioctylfluorene)] (PFN) and lithium fluoride (LiF), were used in a simple PLED architecture. PLEDs with the PFN film were found to exhibit better performance with a maximum luminous efficiency of 40 cd/A, a turn-on voltage (Von) of approximately 4.5 V and a luminance maximum of 878 cd/m2 at 5.5 V, with a current density of 20 A/m2. For the lasing properties of PF–1, we found a lasing threshold of around 75 μJ and a tunability of 20 nm. These values are comparable with those of rhodamine 6G, a well-known laser dye.
Religious Groups as Interest Groups: The United States Catholic Bishops in the Welfare Reform Debate of 1995–1996 and the Health Care Reform Debate of 2009–20101
Authors: Cammisa Anne , Manuel Paul
Published: February 2016

The United States has a long history of religious influence on public policy: the anti-slavery movement, progressivism, prohibition, civil rights, abortion, school vouchers, school prayer and nuclear disarmament are all issues that have involved religion and religious groups in policymaking. In recent decades, the number of religious interest groups (as well as interest groups in general) has greatly expanded, but the role that the religious organizations play as interest groups in the policy arena has received relatively little attention. How are they similar to and different from other interest groups? What tactics do they use? How successful are they? Under what conditions is success or failure more likely? This article examines Roman Catholic religious groups as interest groups in the congressional policymaking process. First, it places Catholic interest groups in the context of the interest group literature, and second, it examines Catholic interest groups’ activity in the passage of welfare reform in 1996 and in the passage of health care reform in 2010. In both cases, they played a greater role in context-setting than in actually changing provisions.
PSInSAR Analysis in the Pisa Urban Area (Italy): A Case Study of Subsidence Related to Stratigraphical Factors and Urbanization
Authors: Solari Lorenzo , Ciampalini Andrea , Raspini Federico , Bianchini Silvia , Moretti Sandro
Published: February 2016

Permanent Scatterer Interferometry (PSI) has been used to detect and characterize the subsidence of the Pisa urban area, which extends for 33 km2 within the Arno coastal plain (Tuscany, Italy). Two SAR (Synthetic Aperture Radar) datasets, covering the time period from 1992 to 2010, were used to quantify the ground subsidence and its temporal evolution. A geotechnical borehole database was also used to make a correspondence with the detected displacements. Finally, the results of the SAR data analysis were contrasted with the urban development of the eastern part of the city in the time period from 1978 to 2013. ERS 1/2 (European Remote-Sensing Satellite) and Envisat SAR data, processed with the PSInSAR (Permanent Scatterer InSAR) algorithm, show that the investigated area is divided in two main sectors: the southwestern part, with null or very small subsidence rates (<2 mm/year), and the eastern portion which shows a general lowering with maximum deformation rates of 5 mm/year. This second area includes deformation rates higher than 15 mm/year, corresponding to small groups of buildings. The case studies in the eastern sector of the urban area have demonstrated the direct correlation between the age of construction of buildings and the registered subsidence rates, showing the importance of urbanization as an accelerating factor for the ground consolidation process.
Mechanical Modulation of Phonon-Assisted Field Emission in a Silicon Nanomembrane Detector for Time-of-Flight Mass Spectrometry
Authors: Park Jonghoo , Blick Robert
Published: February 2016

We demonstrate mechanical modulation of phonon-assisted field emission in a free-standing silicon nanomembrane detector for time-of-flight mass spectrometry of proteins. The impacts of ion bombardment on the silicon nanomembrane have been explored in both mechanical and electrical points of view. Locally elevated lattice temperature in the silicon nanomembrane, resulting from the transduction of ion kinetic energy into thermal energy through the ion bombardment, induces not only phonon-assisted field emission but also a mechanical vibration in the silicon nanomembrane. The coupling of these mechanical and electrical phenomenon leads to mechanical modulation of phonon-assisted field emission. The thermal energy relaxation through mechanical vibration in addition to the lateral heat conduction and field emission in the silicon nanomembrane offers effective cooling of the nanomembrane, thereby allowing high resolution mass analysis.
Geographical Detector Model for Influencing Factors of Industrial Sector Carbon Dioxide Emissions in Inner Mongolia, China
Authors: Wu Rina , Zhang Jiquan , Bao Yuhai , Zhang Feng
Published: February 2016

Studying the influencing factors of carbon dioxide emissions is not only practically but also theoretically crucial for establishing regional carbon-reduction policies, developing low-carbon economy and solving the climate problems. Therefore, we used a geographical detector model which is consists of four parts, i.e., risk detector, factor detector, ecological detector and interaction detector to analyze the effect of these social economic factors, i.e., GDP, industrial structure, urbanization rate, economic growth rate, population and road density on the increase of energy consumption carbon dioxide emissions in industrial sector in Inner Mongolia northeast of China. Thus, combining with the result of four detectors, we found that GDP and population more influence than economic growth rate, industrial structure, urbanization rate and road density. The interactive effect of any two influencing factors enhances the increase of the carbon dioxide emissions. The findings of this research have significant policy implications for regions like Inner Mongolia.
A Two-Step Approach for Analytical Optimal Hedging with Two Triggers
Authors: Hu Tiesong , Zhang Xu-Zhao , Zeng Xiang , Wang Jing
Published: February 2016

Hedging is widely used to mitigate severe water shortages in the operation of reservoirs during droughts. Rationing is usually instituted with one hedging policy, which is based only on one trigger, i.e., initial storage level or current water availability. It may perform poorly in balancing the benefits of a release during the current period versus those of carryover storage during future droughts. This study proposes a novel hedging rule to improve the efficiency of a reservoir operated to supply water, in which, based on two triggers, hedging is initiated with three different hedging sub-rules through a two-step approach. In the first step, the sub-rule is triggered based on the relationship between the initial reservoir storage level and the level of the target rule curve or the firm rule curve at the end of the current period. This step is mainly concerned with increasing the water level or not in the current period. Hedging is then triggered under the sub-rule based on current water availability in the second step, in which the trigger implicitly considers both initial and ending reservoir storage levels in the current period. Moreover, the amount of hedging is analytically derived based on the Karush–Kuhn–Tucker (KKT) conditions. In addition, the hedging parameters are optimized using the improved particle swarm optimization (IPSO) algorithm coupled with a rule-based simulation. A single water-supply reservoir located in Hubei Province in central China is selected as a case study. The operation results show that the proposed rule is reasonable and significantly improves the reservoir operation performance for both long-term and critical periods relative to other operation policies, such as the standard operating policy (SOP) and the most commonly used hedging rules.
Models and Data Sources Used in Systems Medicine
Authors: Löpprich M. , Karmen C. , Ganzinger M. , Gietzelt M.
Published: February 2016

Background: Systems medicine is a new approach for the development and selection of treatment strategies for patients with complex diseases. It is often referred to as the application of systems biology methods for decision making in patient care. For systems medicine computer applications, many different data sources have to be integrated and included into models. This is a challenging task for Medical Informatics since the approach exceeds traditional systems like Electronic Health Records. To prioritize research activities for systems medicine applications, it is necessary to get an overview over modelling methods and data sources already used in this field.

Objectives: We performed a systematic literature review with the objective to capture current use of 1) modelling methods and 2) data sources in systems medicine related research projects.

Methods: We queried the MEDLINE and ScienceDirect databases for papers associated with the search term systems medicine and related terms. Papers were screened and assessed in full text in a two-step process according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement guidelines.

Results: The queries returned 698 articles of which 34 papers were finally included into the study. A multitude of modelling approaches such as machine learning and network analysis was identified and classified. Since these approaches are also used in other domains, no methods specific for systems medicine could be identified. Omics data are the most widely used data types followed by clinical data. Most studies only include a rather limited number of data sources.

Conclusions: Currently, many different modelling approaches are used in systems medicine. Thus, highly flexible modular solutions are necessary for systems medicine clinical applications. However, the number of data sources included into the models is limited and most projects currently focus on prognosis. To leverage the potential of systems medicine further, it will be necessary to focus on treatment strategies for patients and consider a broader range of data.
Islet Volume and Indexes of β-Cell Function in Humans
Authors: Nano Rita , Melzi Raffaella , Mercalli Alessia , Balzano Gianpaolo , Scavini Marina , Bonadonna Riccardo , Piemonti Lorenzo
Published: February 2016

Islet volume and endocrine pancreas architecture (islet size distribution) may be independent determinants of β-cell function. Furthermore, the accuracy of homeostatic model assessment (HOMA) indexes in predicting β-cell mass has never been assessed. Here we investigated the relationships between islet volume, islet density, and islet size distribution, estimated after pancreatic tissue digestion, with established indexes of β-cell function in humans. We included in this study 42 patients who were candidates for islet autotransplantation and had well-characterized glucose metabolism. Indexes of insulin secretion were calculated and compared with the islet volume, as a surrogate of β-cell mass, obtained after digestion of pancreas. Islet counting analysis showed considerable interindividual variation in islet density and size. Islet volume, but not density nor size, was the only independent determinant of β-cell function assessed by insulin HOMA β-cell. Islet volume was significantly reduced in the patients with overt hyperglycemia, but not in patients with impaired fasting glucose. Insulin HOMA β-cell predicted islet volume better than other measures of fasting insulin secretion. In conclusion, the present study documented a close direct relationship between indexes of β-cell function and islet volume in humans. The insulin HOMA β-cell provides a more reliable estimate of pancreatic islet volume than fasting glucose before islet isolation.
Pancreatic L-Glutamine Administration Protects Pig Islets from Cold Ischemic Injury and Increases Resistance toward Inflammatory Mediators
Authors: Brandhorst Heide , Theisinger Bastian , Guenther Bernhard , Johnson Paul R. , Brandhorst Daniel
Published: February 2016

The isolation and transplantation of porcine islets represent a future option for the treatment of type 1 diabetic patients. Stringent product release criteria and limited availability of transgenic and specific pathogen-free pigs will essentially require processing of explanted pig pancreata in specialized, possibly remote isolation facilities, whereby pancreata are exposed to cold ischemia due to prolonged tissue transit time. In the present study we investigated whether pancreas oxygenation can be efficiently combined with an antioxidant strategy utilizing intraductal L-glutamine administration. Pig pancreata were intraductally perfused after retrieval and after cold storage in oxygen-precharged perfluorohexyloctane utilizing University of Wisconsin solution supplemented with ( n = 16) or without ( n = 14) 5 mmol/L L-glutamine. After isolation purified islets were subjected to extensive quality assessment. Islet recovery postpurification was significantly higher in glutamine-treated pancreata (77.0 ± 3.3% vs. 60.3 ± 6.0%, p < 0.05). Glutamine administration increased intraislet content of reduced glutathione (117.8 ± 16.5 vs. 15.9 ± 2.8 ng/ng protein, p < 0.001) associated with increased islet recovery after culture (65.8 ± 12.1% vs. 40.3 ± 11.7%, p < 0.05), enhanced glucose stimulation index (1.82 ± 0.16 vs. 1.38 ± 0.10, p < 0.05), and improved posttransplant function in diabetic nude mice ( p < 0.05). Furthermore, intraductally administered glutamine increased pig islet resistance toward reactive oxygen species, nitric oxide, and high-dose proinflammatory cytokines. The present study demonstrates that quality and function of pig islets exposed to warm and cold ischemia can significantly be improved using intraductal L-glutamine administration. As the efficiency of the intraductal route may be inferior compared to intravascular administration further studies should aim on assessment of L-glutamine as supplement for pancreas perfusion during organ procurement.
Efficacy of Multilayered Hepatocyte Sheet Transplantation for Radiation-Induced Liver Damage and Partial Hepatectomy in a Rat Model
Authors: Baimakhanov Zhassulan , Yamanouchi Kosho , Sakai Yusuke , Koike Makiko , Soyama Akihiko , Hidaka Masaaki , Takatsuki Mitsuhisa , Fujita Fumihiko , Kanetaka Kengo , Kuroki Tamotsu , Eguchi Susumu
Published: February 2016

Although cell sheet technology has recently been developed for use in both animal experiments and in the clinical setting, it remains unclear whether transplanted hepatocyte sheets improve the liver function in vivo. Radiation-induced liver damage (RILD) combined with partial hepatectomy (PH) has been reported to suppress the proliferation of host hepatocytes and induce critical liver failure. The aim of this study was to improve the liver function in the above-mentioned diseased rat model (RILD + PH) using multilayered hepatocyte sheet transplantation. In this study, we used Fischer rats as a donor for primary hepatocytes and dermal fibroblast isolation. Cocultured multilayered hepatocyte sheets were generated by disseminating hepatocytes onto fibroblasts cultured beforehand on temperature-responsive culture dishes. Four cell sheets were transplanted into the recipient rats subcutaneously. Prior to transplantation, RILD (50 Gy) with 2/3PH was induced in the recipients. The same model was applied in the control group without transplantation. The serum was collected each week. The rats in both groups were sacrificed at 2 months after transplantation for the histological analysis. Consequently, the serum albumin concentrations were significantly higher in the transplant group than in the control group (54.3 ± 9.6 vs. 32.7 ± 5.7 mg/ml; p < 0.01) after 2 months and comparable to the serum albumin levels in the normal rats (58.1 ± 6.4 mg/ml). In addition, treatment with the transplanted sheets significantly improved the survival rate (57% vs. 22%, p < 0.05), and the hepatocyte sheets showed the storage of albumin, glycogen, and bile canaliculus structures. Some hepatocytes and fibroblasts were positive for Ki-67, and vascularization was observed around the cell sheets. Transplanted multilayered hepatocyte sheets can survive with additional proliferative activity, thereby maintaining the liver function in vivo for at least 2 months, providing metabolic support for rats with RILD.
Extensive Loss of Islet Mass beyond the First Day after Intraportal Human Islet Transplantation in a Mouse Model
Authors: Liljebäck Hanna , Grapensparr Liza , Olerud Johan , Carlsson Per-Ola
Published: February 2016

Clinical islet transplantation is characterized by a progressive deterioration of islet graft function, which renders many patients once again dependent on exogenous insulin administration within a couple of years. In this study, we aimed to investigate possible engraftment factors limiting the survival and viability of experimentally transplanted human islets beyond the first day after their transplantation to the liver. Human islets were transplanted into the liver of nude mice and characterized 1 or 30 days after transplantation by immunohistochemistry. The factors assessed were endocrine mass, cellular death, hypoxia, vascular density and amyloid formation in the transplanted islets. One day posttransplantation, necrotic cells, as well as apoptotic cells, were commonly observed. In contrast to necrotic death, apoptosis rates remained high 1 month posttransplantation, and the total islet mass was reduced by more than 50% between 1 and 30 days posttransplantation. Islet mass at 30 days posttransplantation correlated negatively to apoptotic death. Vascular density within the transplanted islets remained less than 30% of that in native human islets up to 30 days posttransplantation and was associated with prevailing hypoxia. Amyloid formation was rarely observed in the 1-day-old transplants, but was commonly observed in the 30-day-old islet transplants. We conclude that substantial islet cell death occurs beyond the immediate posttransplantation phase, particularly through apoptotic events. Concomitant low vascularization with prevailing hypoxia and progressive amyloid development was observed in the human islet grafts. Strategies to improve engraftment at the intraportal site or change of implantation site in the clinical setting are needed.
Pancreatic Tissue Transplanted in TheraCyte™ Encapsulation Devices Is Protected and Prevents Hyperglycemia in a Mouse Model of Immune-Mediated Diabetes
Authors: Boettler Tobias , Schneider Darius , Cheng Yang , Kadoya Kuniko , Brandon Eugene P. , Martinson Laura , Von Herrath Matthias
Published: February 2016

Type 1 diabetes (T1D) is characterized by destruction of glucose-responsive insulin-producing pancreatic β-cells and exhibits immune infiltration of pancreatic islets, where CD8 lymphocytes are most prominent. Curative transplantation of pancreatic islets is seriously hampered by the persistence of autoreactive immune cells that require high doses of immunosuppressive drugs. An elegant approach to confer graft protection while obviating the need for immunosuppression is the use of encapsulation devices that allow for the transfer of oxygen and nutrients, yet prevent immune cells from making direct contact with the islet grafts. Here we demonstrate that macroencapsulation devices (TheraCyte™) loaded with neonatal pancreatic tissue and transplanted into RIP-LCMV.GP mice prevented disease onset in a model of virus-induced diabetes mellitus. Histological analyses revealed that insulin-producing cells survived within the device in animal models of diabetes. Our results demonstrate that these encapsulation devices can protect from an immune-mediated attack and can contain a sufficient amount of insulin-producing cells to prevent overt hyperglycemia.
Transcriptomic Analysis of Cultured Corneal Endothelial Cells as a Validation for Their Use in Cell Replacement Therapy
Authors: Frausto Ricardo F. , Le Derek J. , Aldave Anthony J.
Published: February 2016

The corneal endothelium plays a primary role in maintaining corneal homeostasis and clarity and must be surgically replaced with allogenic donor corneal endothelium in the event of visually significant dysfunction. However, a worldwide shortage of donor corneal tissue has led to a search for alternative sources of transplantable tissue. Cultured human corneal endothelial cells (HCEnC) have been shown to restore corneal clarity in experimental models of corneal endothelial dysfunction in animal models, but characterization of cultured HCEnC remains incomplete. To this end, we utilized next-generation RNA sequencing technology to compare the transcriptomic profile of ex vivo human corneal endothelial cells (evHCEnC) with that of primary HCEnC (pHCEnC) and HCEnC lines and to determine the utility of cultured and immortalized corneal endothelial cells as models of in vivo corneal endothelium. Multidimensional analyses of the transcriptome data sets demonstrated that primary HCEnC have a closer relationship to evHCEnC than do immortalized HCEnC. Subsequent analyses showed that the majority of the genes specifically expressed in HCEnC (not expressed in ex vivo corneal epithelium or fibroblasts) demonstrated a marked variability of expression in cultured cells compared with evHCEnC. In addition, genes associated with either corneal endothelial cell function or corneal endothelial dystrophies were investigated. Significant differences in gene expression and protein levels were observed in the cultured cells compared with evHCEnC for each of the genes tested except for AGBL1 and LOXHD1, which were not detected by RNA-seq or qPCR. Our transcriptomic analysis suggests that at a molecular level pHCEnC most closely resemble evHCEnC and thus represent the most viable cell culture-based therapeutic option for managing corneal endothelial cell dysfunction. Our findings also suggest that investigators should perform an assessment of the entire transcriptome of cultured HCEnC prior to determination of their potential clinical utility for the management of corneal endothelial cell failure.
Hepatocyte-Like Cells Derived from Mouse Induced Pluripotent Stem Cells Produce Functional Coagulation Factor IX in a Hemophilia B Mouse Model
Authors: Wu Yao-Ming , Huang Yu-Jen , Chen Poda , Hsu Yu-Chen , Lin Shu-Wha , Lai Hong-Shiee , Lee Hsuan-Shu
Published: February 2016

Hemophilia B (HB) is an inherited deficiency in coagulation factor IX (FIX) that leads to prolonged bleeding after injury. Although hepatocyte transplantation has been demonstrated to be an effective therapeutic strategy for HB, the quality and sources of hepatocytes still limit their application. Recently, stem cells were proposed as an alternative source of donor cells for cell-based therapy. Much research has been devoted to the properties of stem cells that can be differentiated into functional hepatocytes, thereby providing a new cell source for cell-based therapy. Induced pluripotent stem cells (iPSCs) represent a renewable source of hepatocytes for cell-based therapy; these cells exhibit pluripotency and differentiation ability and can be derived from somatic cells. These iPSCs are highly similar to embryonic stem cells (ESCs). We hypothesized that hepatocyte-like cells derived from iPSCs would have therapeutic efficiency in a mouse model of HB. To test this hypothesis, we differentiated iPSCs toward hepatocytes by stepwise protocol and then transplanted these cells into HB mice. We found that these cells shared many characteristics with hepatocytes, such as albumin synthesis, metabolic capacity, glycogen storage, and ureagenesis. Moreover, iPSC-derived hepatocyte transplantation led to increased coagulation factor IX activity, improved thrombus generation, and better hemostasis parameters, and the transferred cells were localized in the liver in recipient HB mice. In conclusion, our results clearly demonstrate that hepatocyte-like cells derived from iPSCs represent a potential cell source for cell-based therapy in the treatment of HB.
Melatonin as an Antioxidant for Stroke Neuroprotection
Authors: Watson Nate , Diamandis Theo , Gonzales-Portillo Chiara , Reyes Stephanny , Borlongan Cesar V.
Published: February 2016

Melatonin ( N-acetyl-5-methoxytryptamine) is a hormone derived from the pineal gland that has a wide range of clinical applications. While melatonin was originally assessed as a hormone specializing in regulation of the normal circadian rhythm in mammals, it now has been shown to be an effective free radical scavenger and antioxidant. Current research has focused on central nervous system (CNS) disorders, stroke in particular, for potential melatonin-based therapeutics. As of now, the realm of potential therapy regimens is focused on three main treatments: exogenously delivered melatonin, pineal gland grafting, and melatonin-mediated stem cell therapy. All therapies contain both costs and benefits, and current research is still focused on finding the best treatment plan. While comprehensive research has been conducted, more research regarding the safety of such therapies is needed in order to transition into the clinical level of testing. Antioxidants such as traditional Chinese medicine, (–)-epigallocatechin-3-gallate (EGCG), and lavender oil, which have been used for thousands of years as treatment, are now gaining recognition as effective melatonin treatment alternatives. This review will further discuss relevant studies assessing melatonin-based therapeutics and provide evidence of other natural melatonin treatment alternatives for the treatment of stroke.
Extracardiac-Lodged Mesenchymal Stromal Cells Propel an Inflammatory Response against Myocardial Infarction via Paracrine Effects
Authors: Peng Yi , Pan Wei , Ou Yali , Xu Weifang , Kaelber Sussannah , Borlongan Cesario V. , Sun Meiqin , Yu Guolong
Published: February 2016

Transplantation of stem cells, including mesenchymal stromal cells (MSCs), improves the recovery of cardiac function after myocardial infarction (MI) in experimental studies using animal models and in patients. However, the improvement of cardiac function following MSC transplantation remains suboptimal in both preclinical and clinical studies. Understanding the mechanism of cell therapy may improve its therapeutic outcomes, but the mode of action mediating stem cell promotion of cardiac repair is complex and not fully understood. Recent studies suggest that the immunomodulatory effects of MSCs on the macrophage M1/M2 subtype transition allow the transplanted stem cells to inhibit inflammation-induced injury and promote cardiac repair in acute MI. However, equally compelling evidence shows that there is poor survival and minimal graft persistence of transplanted MSCs within the infarcted heart tissues, negating the view that graft survival per se is required for the observed high rate and long duration of the transition from proinflammatory M1 to reparative M2 macrophages in the infarcted myocardium. Therefore, we raised a novel hypothesis that the therapeutic effects of MSC transplantation for acute MI depends not primarily on the grafted cells in infarct myocardium, but that MSCs migrating to and being lodged in the extracardiac organs, demonstrating good graft survival and persistence, may render the therapeutic effects in MI. More specifically, MSC transplantation promotes the transition from M1 to M2 in extracardiac organs, such as spleen and bone marrow, and therapeutic effects are conferred to the infarcted myocardium via paracrine effects. In MSC transplantation, the conversion from proinflammatory M1 to anti-inflammatory M2 monocytes may occur remotely from the heart and may serve as one of the major pathways in regulating the dual effects of inflammation. This hypothesis, if proven valid, may represent an important new mechanism of action to be considered for the future of MSC transplantation in the treatment of MI.
Identification of Bone Marrow Cell Subpopulations Associated with Improved Functional Outcomes in Patients with Chronic Left Ventricular Dysfunction: An Embedded Cohort Evaluation of the FOCUS-CCTRN Trial
Authors: Taylor Doris A. , Perin Emerson C. , Willerson James T. , Zierold Claudia , Resende Micheline , Carlson Marjorie , Nestor Belinda , Wise Elizabeth , Orozco Aaron , Pepine Carl J. , Henry Timothy D. , Ellis Stephen G. , Zhao David X. M. , Traverse Jay H. , Cooke John P. , Schutt Robert C. , Bhatnagar Aruni , Grant Maria B. , Lai Dejian , Johnstone Brian H. , Sayre Shelly L. , Moyé Lem , Ebert Ray F. , Bolli Roberto , Simari Robert D. ,   for the Cardiovascular Cell Therapy Research Network (CCTRN), Cogle Christopher R.
Published: February 2016

In the current study, we sought to identify bone marrow-derived mononuclear cell (BM-MNC) subpopulations associated with a combined improvement in left ventricular ejection fraction (LVEF), left ventricular end-systolic volume (LVESV), and maximal oxygen consumption (VO2 max) in patients with chronic ischemic cardiomyopathy 6 months after receiving transendocardial injections of autologous BM-MNCs or placebo. For this prospectively planned analysis, we conducted an embedded cohort study comprising 78 patients from the FOCUS-Cardiovascular Cell Therapy Research Network (CCTRN) trial. Baseline BM-MNC immunophenotypes and progenitor cell activity were determined by flow cytometry and colony-forming assays, respectively. Previously stable patients who demonstrated improvement in LVEF, LVESV, and VO2 max during the 6-month course of the FOCUS-CCTRN study (group 1, n = 17) were compared to those who showed no change or worsened in one to three of these endpoints (group 2, n = 61) and to a subset of patients from group 2 who declined in all three functional endpoints (group 2A, n = 11). Group 1 had higher frequencies of B-cell and CXCR4+ BM-MNC subpopulations at study baseline than group 2 or 2A. Furthermore, patients in group 1 had fewer endothelial colony-forming cells and monocytes/macrophages in their bone marrow than those in group 2A. To our knowledge, this is the first study to show that in patients with ischemic cardiomyopathy, certain bone marrow-derived cell subsets are associated with improvement in LVEF, LVESV, and VO2 max at 6 months. These results suggest that the presence of both progenitor and immune cell populations in the bone marrow may influence the natural history of chronic ischemic cardiomyopathy—even in stable patients. Thus, it may be important to consider the bone marrow composition and associated regenerative capacity of patients when assigning them to treatment groups and evaluating the results of cell therapy trials.
Tolerogenic Dendritic Cells Generated by in Vitro Treatment with SAHA are not Stable in Vivo
Authors: Thewissen Kristof , Broux Bieke , Hendriks Jerome J. A. , Vanhees Mandy , Stinissen Piet , Slaets Helena , Hellings Niels
Published: February 2016

The aim of this study is to examine whether the histone deacetylase inhibitor, suberoylanilide hydroxamic acid (SAHA), can generate dendritic cells (DCs) with a stable tolerogenic phenotype to counteract autoimmune responses in an animal model of multiple sclerosis. We investigated if the tolerogenic potency of DCs could be increased by continuous treatment during in vitro differentiation toward DCs compared to standard 24-h in vitro treatment of already terminally differentiated DCs. We show that in vitro treatment with SAHA reduces the generation of new CD11c+ DCs out of mouse bone marrow. SAHA-generated DCs show reduced antigen-presenting function as evidenced by a reduction in myelin endocytosis, a decreased MHC II expression, and a failure to upregulate costimulatory molecules upon LPS challenge. In addition, SAHA-generated DCs display a reduction in proinflammatory cytokines and molecules involved in apoptosis induction, inflammatory migration, and TLR signaling, and they are less immunostimulatory compared to untreated DCs. We demonstrated that the underlying mechanism involves a diminished STAT1 phosphorylation and was independent of STAT6 activation. Although in vitro results were promising, SAHA-generated DCs were not able to alleviate the development of experimental autoimmune encephalomyelitis in mice. In vitro washout experiments demonstrated that the tolerogenic phenotype of SAHA-treated DCs is reversible. Taken together, while SAHA potently boosts tolerogenic properties in DCs during the differentiation process in vitro, SAHA-generated DCs were unable to reduce autoimmunity in vivo. Our results imply that caution needs to be taken when developing DC-based therapies to induce tolerance in the context of autoimmune disease.
Relationship of Grafted FGF-2-Overexpressing Neural Stem/Progenitor Cells with the Vasculature in the Cerebral Cortex
Authors: Tsupykov Oleg , Kanemitsu Michiko , Smozhanik Ekaterina , Skibo Galina , Dayer Alexandre G. , Kiss Jozsef Z.
Published: February 2016

Neural progenitor cells (NPCs) overexpressing fibroblast growth factor 2 (FGF-2) have the distinct tendency to associate with the vasculature and establish multiple proliferative clusters in the perivascular environment after transplantation into the cerebral cortex. Strikingly, the vascular clusters of progenitor cells give rise to immature neurons after ischemic injury, raising prospects for the formation of ectopic neurogenic niches for repair. We investigated the spatial relationship of perivascular clusters with the host vascular structures. FGF-2-GFP-transduced NPCs were transplanted into the intact somatosensory rat cortex. Confocal microscopic analysis revealed that grafted cells preferentially contacted venules at sites with aquaporin-4-positive astrocytic endfeet and avoided contacts with desmin-positive pericytes. Electron microscopic analysis confirmed that grafted cells preferentially made contact with astroglial endfeet, and only a minority of them reached the endothelial basal lamina. These results provide new insights into the fine structural and anatomical relationship between grafted FGF-2-transduced NPCs and the host vasculature.
Cell-Permeable Peptide Blocks TLR4 Signaling and Improves Islet Allograft Survival
Authors: Dong Huansheng , Zhang Yong , Song Lili , Kim Do-Sung , Wu Hongju , Yang Lijun , Li Shiwu , Morgan Katherine A. , Adams David B. , Wang Hongjun
Published: February 2016

Toll-like receptor 4 (TLR4) activation in pancreatic β cells activates aberrant islet graft cellular pathways and contributes to immune rejection in allogeneic islet transplantation. As an approach to overcoming this problem, we determined the capacity of a 33-amino acid peptide consisting of a protein transduction domain (PTD) from the Hph-1 virus and a fragment of the intracellular domain of TLR4 from the C3H mice (PTD-dnTLR4) to block TLR4 signaling and improve allogeneic islet survival in vitro and after transplantation. The efficacy of PTD-dnTLR4 in blocking TLR4 signaling was assessed in the Raw264.7 macrophage line, in the islets, and the βTC3 cell line. In Raw264.7 cells, preculture with the peptide reduced LPS-induced NF-κB activation and production of proinflammatory cytokines (IL-1β, TNF-α, iNOS, and IL-6). In islets and β cells, preincubation with PTD-dnTLR4 suppressed LPS-induced TNF-α expression via inhibition of NF-κB activation and protected them from stress-induced cell death. In vivo, preincubation of BALB/c (H-2d) islets with PTD-dnTLR4 resulted in significantly longer survival than control islets in a streptozotocin-induced diabetes model (two of seven grafts survived long term >100 days). PTD-dnTLR4-treated grafts exhibited reduced expression of TNF-α and iNOS and reduced macrophage infiltration posttransplant. The data indicate that PTD-dnTLR4 blocked TLR4 signaling in both macrophages and β cells, and prolonged allograft survival at least in part by suppressing inflammation and macrophage infiltration. This strategy for blocking TLR4 activity has potential utilization in the treatment of diseases where excessive TLR4 activation contributes to the pathologic cellular pathways such as islet transplantation.
Melanoma Immunotherapy in Mice Using Genetically Engineered Pluripotent Stem Cells
Authors: Haque Mohammad , Song Jianyong , Fino Kristin , Sandhu Praneet , Wang Youfei , Ni Bing , Fang Deyu , Song Jianxun
Published: February 2016

Adoptive cell transfer (ACT) of antigen (Ag)-specific CD8+ cytotoxic T lymphocytes (CTLs) is a highly promising treatment for a variety of diseases. Naive or central memory T-cell-derived effector CTLs are optimal populations for ACT-based immunotherapy because these cells have a high proliferative potential, are less prone to apoptosis than terminally differentiated cells, and have the higher ability to respond to homeostatic cytokines. However, such ACT with T-cell persistence is often not feasible due to difficulties in obtaining sufficient cells from patients. Here we present that in vitro differentiated HSCs of engineered PSCs can develop in vivo into tumor Ag-specific naive CTLs, which efficiently suppress melanoma growth. Mouse-induced PSCs (iPSCs) were retrovirally transduced with a construct encoding chicken ovalbumin (OVA)-specific T-cell receptors (TCRs) and survival-related proteins (i.e., BCL-xL and survivin). The gene-transduced iPSCs were cultured on the delta-like ligand 1-expressing OP9 (OP9-DL1) murine stromal cells in the presence of murine recombinant cytokines (rFlt3L and rIL-7) for a week. These iPSC-derived cells were then intravenously adoptively transferred into recipient mice, followed by intraperitoneal injection with an agonist α-Notch 2 antibody and cytokines (rFlt3L and rIL-7). Two weeks later, naive OVA-specific CD8+ T cells were observed in the mouse peripheral lymphatic system, which were responsive to OVA-specific stimulation. Moreover, the mice were resistant to the challenge of B16-OVA melanoma induction. These results indicate that genetically modified stem cells may be used for ACT-based immunotherapy or serve as potential vaccines.
miR-sc3, a Novel MicroRNA, Promotes Schwann Cell Proliferation and Migration by Targeting Astn1
Authors: Yi Sheng , Wang Shanshan , Zhao Qing , Yao Chun , Gu Yun , Liu Jie , Gu Xiaosong , Li Shiying
Published: February 2016

MicroRNAs (miRNAs, miRs) are small noncoding RNAs that regulate gene expression at the posttranscriptional level. We have previously identified a group of novel miRNAs in proximal sciatic nerve after sciatic nerve transection by Solexa sequencing, and miR-sc3 is a member of the group. In this study, we aimed to investigate the effects of miR-sc3 on phenotypic modulation of Schwann cells (SCs). miR-sc3 was highly expressed in the injured nerve after sciatic nerve transection. An increased and decreased expression of miR-sc3 promoted and reduced the proliferation and migration of primary SCs, respectively. miR-sc3 directly targeted astrotactin 1 ( Astn1) and led to translational suppression of Astn1. There was an inverse association between the time-dependent expressions of miR-sc3 and Astn1 in proximal sciatic nerve after sciatic nerve transection. Overall, miR-sc3 affected SC proliferation and migration by targeting Astn1, thus playing the regulatory role in peripheral nerve regeneration.