bluuebunny/crossref_metadata_2025 huggingface dataset:
BcCluster: A Bladder Cancer Database at the Molecular Level
Authors: Bhat Akshay , Mokou Marika , Zoidakis Jerome , Jankowski Vera , Vlahou Antonia , Mischak Harald
Published: February 2016

Background:

Bladder Cancer (BC) has two clearly distinct phenotypes. Non-muscle invasive BC has good prognosis and is treated with tumor resection and intravesical therapy whereas muscle invasive BC has poor prognosis and requires usually systemic cisplatin based chemotherapy either prior to or after radical cystectomy. Neoadjuvant chemotherapy is not often used for patients undergoing cystectomy. High-throughput analytical omics techniques are now available that allow the identification of individual molecular signatures to characterize the invasive phenotype. However, a large amount of data produced by omics experiments is not easily accessible since it is often scattered over many publications or stored in supplementary files .

Objective:

To develop a novel open-source database, BcCluster ( http://www.bccluster.org/ ), dedicated to the comprehensive molecular characterization of muscle invasive bladder carcinoma.

Materials:

A database was created containing all reported molecular features significant in invasive BC. The query interface was developed in Ruby programming language (version 1.9.3) using the web-framework Rails (version 4.1.5) ( http://rubyonrails.org/ ).

Results:

BcCluster contains the data from 112 published references, providing 1,559 statistically significant features relative to BC invasion. The database also holds 435 protein-protein interaction data and 92 molecular pathways significant in BC invasion. The database can be used to retrieve binding partners and pathways for any protein of interest. We illustrate this possibility using survivin, a known BC biomarker.

Conclusions:

BcCluster is an online database for retrieving molecular signatures relative to BC invasion. This application offers a comprehensive view of BC invasiveness at the molecular level and allows formulation of research hypotheses relevant to this phenotype.
Inhibiting Immune Checkpoints for the Treatment of Bladder Cancer
Authors: Bidnur S. , Savdie R. , Black P.C.
Published: February 2016

Background:

Increasing evidence supporting the role of immune checkpoint blockade in cancer management has been bolstered by recent reports demonstrating significant and durable clinical responses across multiple tumour types, including metastatic urothelial carcinoma (mUC). The majority of these results are achieved via blockade of the programmed death (PD) axis, which like CTLA-4 blockade permits T-cell activation and immune-mediated anti-tumour activity- essentially harnessing the patient’s own immune system to mount an anti-neoplastic response. However, while clinical responses can be striking, our understanding of the biology of immune checkpoint blockade is only beginning to shed light on how to maximize and even improve patient outcomes with immune checkpoint blockade, especially in UC.

Methods:

We performed a literature review for immune checkpoint blockade with a focus on rationale for checkpoint therapy and outcomes in UC. We also highlight the advances made in other tumour types, with a focus on the recent 2015 meeting of the American Society for Clinical Oncology.

Results:

In heavily pre-treated UC, trials are suggesting objective response rates above 30% . These impressive results are seen across multiple different tumour types, especially those with high burden of DNA level mutations. Identification of prognostic biomarkers is currently under investigation, in order to improve patient selection. Interestingly, response to PD-1 directed therapy is seen even in patients with no evidence of PD-1 positivity on immunohistochemistry. This has led to the development of enhanced biomarkers including assessing DNA mutation rates and immune gene signatures, to improve patient selection.

Conclusions:

Immune checkpoint blockade is an exciting cancer treatment modality which is demonstrating impressive clinical results across multiple tumour types. For UC, anti-PD directed therapy represents a much needed treatment in the metastatic, post chemotherapy context. Potential for these agents to have clinical utility in non-metastatic UC is still to be assessed.
Comparison of Guidelines on Non-Muscle Invasive Bladder Cancer (EAU, CUA, AUA, NCCN, NICE)
Authors: Power Nicholas E. , Izawa Jonathan
Published: February 2016

Background:

Non-muscle invasive bladder cancer (NMIBC) represents a considerably diverse patient group and the management of this complex disease is debatable. A number of panels from Europe and North America have convened on the topic and recently released guideline documents.

Objective:

The purpose was to compare and contrast the NMIBC guideline recommendations from the EAU (Europe), CUA (Canada), NCCN (United States), AUA (United States), and NICE (United Kingdom).

Methods:

All unabridged guideline documents were reviewed by the authors and comparisons were completed according to major topics in NMIBC.

Results:

Despite a paucity of high level evidence regarding the majority of management topics in NMIBC, there was general agreement among the various guideline panels. Differences mainly centered on the categories of evidence synthesized and grades of recommendations. Each document offers a unique presentation of the available literature and guideline recommendation.

Conclusions:

The guidelines for NMIBC from the EAU, CUA, AUA, NCCN, and NICE provide considerable consensus regarding the management of this often difficult disease. Clinicians are encouraged to familiarize themselves with all of the guidelines in order to determine which style of presentation would be most useful to their current practice.
Concordance in Biomarker Status Between Bladder Tumors at Time of Transurethral Resection and Subsequent Radical Cystectomy: Results of a 5-year Prospective Study
Authors: Passoni Niccolò M. , Shariat Shahrokh F. , Bagrodia Aditya , Francis Franto , Rachakonda Varun , Xylinas Evanguelos , Kapur Payal , Sagalowsky Arthur I. , Lotan Yair
Published: February 2016

Purpose:

To assess the concordance rate in alterations of molecular markers at the time of transurethral resection (TUR) and subsequent radical cystectomy (RC) among patients with high-grade urothelial carcinoma of the bladder (UCB).

Methods:

We prospectively performed immunohistochemical staining p53, p21, p27, Ki-67 and cyclin E1 on TUR and on RC specimens from 102 patients treated with RC and bilateral lymphadenectomy for high-grade UCB. We analyzed the concordance rate of individual markers and of the number of altered markers. Concordant and discordant findings were reported in the overall population and according to clinical stage.

Results:

Median patient age was 74 years (IQR 67–79) and mostly male (86%). Median time from TUR to RC was 1.5 months (IQR 1.0–2.4). Clinical stage at time of RC was cTa/Tis/T1 in 50% , cT2 in 47% , and cT4 in 1% of patients Nine (9%) patients received neoadjuvant chemotherapy. The concordance of biomarkers between TUR and RC specimens was 92.2% , 77.5% , 80.4% , 77.5% , and 83.3% for cyclin E1, p21, p27, p53 and Ki-67, respectively. The concordance between number of altered biomarkers was 51.0%.

Conclusions:

The rate of individual marker alterations at time of TUR closely approximates that found at RC specimens. However, the correlation of number of altered markers is lower. Molecular marker status at TUR could help predict the marker status at RC and may help guide multimodal therapeutic planning.
Subjective visual and haptic vertical in young and elderly
Authors: Čakrt Ondřej , Slabý Kryštof , Kmet' Jan , Kolář Pavel , Jeřábek Jaroslav
Published: February 2016

Perception of verticality can be assessed in two different ways: measuring of subjective visual vertical (SVV) and subjective haptic vertical (SHV). The evidence on aging of SVV and SVH is not conclusive and there is just little knowledge focused on this issue. The aim of this study was to compare accuracy of perception of SVV and SHV between groups of young and elderly healthy subjects. SVV examination was performed using the bucket test. An experimental tactile device was used to assess perceived SHV. Measurements of SVV and SHV were made in 27 young healthy and 30 elderly healthy subjects, both groups were right-handed due to self-report. SHV estimated position was significantly different (p< 0.01) in young and elderly (counterclockwise shift of 0.72 ± 3.70° and 3.51 ± 3.99°, respectively) and the SHV range (4.17 ± 5.40° and 9.64 ± 7.42°, respectively) was also different (p< 0.01). The differences were caused by significant difference in the supination maneuver (clockwise rod rotation, p< 0.001) which resulted in counterclockwise shift of 2.80 ± 4.90° and 8.33 ± 4.62° in young and elderly respectively. Pronation part of the SHV task (counterclockwise rod rotation) did not significantly differ between groups. SVV estimated position and range were not statistically different between young and elderly. These results provide evidence that the ability to detect SVV is not impaired, SHV seems also not to be affected by aging but there may be methodologic issues in SHV testing in elderly which should be controled for in future studies. Results of both pronation and supination maneuvers should be provided along with SHV position.
The activities-specific balance confidence scale and berg balance scale: Reliability and validity in Arabic-speaking vestibular patients
Authors: Alghwiri Alia A. , Alghadir Ahmad H. , Al-momani Murad O. , Whitney Susan L.
Published: February 2016

Persons with vestibular disorders are susceptible to imbalance. The Activities-specific Balance Confidence (ABC) scale and Berg Balance Scale (BBS) have been validated in persons with vestibular disorders. The purpose of this study was to examine the reliability and validity of the Arabic versions of ABC and BBS among Arabic-speaking persons with vestibular disorders in Jordan and Saudi Arabia. Therefore, the A-ABC and A-BBS were administered to a convenience sample of 82 persons with vestibular disorders (age = 43 ± 14), (56% female). The test-retest reliability of the A-ABC item and total score as well as the inter-rater and intra-rater reliability of the A-BBS total score reflected high agreement. Significant and large correlations were found between the A-ABC and the A-BBS (r= 0.54, P< 0.05), the A-ABC and the Arabic Dizziness Handicap Inventory (A-DHI) (r= -0.76, P< 0.05), and the A-BBS and the A-DHI (r= -0.56, P< 0.05). The A-ABC and the A-BBS demonstrated good reliability and validity and can be utilized with Arabic-speaking persons with vestibular disorders.
Examining work-education intersections: the production of learning reals in and through practice
Authors: Harman Kerry
Published: February 2016

Working within an assemblage analytic, this paper examines work – education intersections using the notion of learning reals. The learning real examined is learning as mastery and skills development. The concepts of embodiment and performativity guide the exploration. The paper draws on interview and observational data collected during a three year research project exploring the everyday learning (of employees) in a post-secondary education institution in Australia. The project was an industryuniversity collaboration between a group of professional developers from the organisation and a group of workplace learning academics. The assemblages making up learning as mastery are traced through examining the enactment of this real by a group of trade teachers, one of the workgroups participating in the project. I propose that this learning real was produced and made durable in and through the practices of the trade teachers. Furthermore, the ongoing performing of mastery produced particular effects, including the separation of theory and practice in the trade school. The notion of learning reals enables an exploration of the way particular ways of conceiving learning are made durable in particular workplaces as well as opening up the space to examine the partial connections between workplaces and educational institutions.
Vehicle Speed Estimation and Forecasting Methods Based on Cellular Floating Vehicle Data
Authors: Lai Wei-Kuang , Kuo Ting-Huan , Chen Chi-Hua
Published: February 2016

Traffic information estimation and forecasting methods based on cellular floating vehicle data (CFVD) are proposed to analyze the signals (e.g., handovers (HOs), call arrivals (CAs), normal location updates (NLUs) and periodic location updates (PLUs)) from cellular networks. For traffic information estimation, analytic models are proposed to estimate the traffic flow in accordance with the amounts of HOs and NLUs and to estimate the traffic density in accordance with the amounts of CAs and PLUs. Then, the vehicle speeds can be estimated in accordance with the estimated traffic flows and estimated traffic densities. For vehicle speed forecasting, a back-propagation neural network algorithm is considered to predict the future vehicle speed in accordance with the current traffic information (i.e., the estimated vehicle speeds from CFVD). In the experimental environment, this study adopted the practical traffic information (i.e., traffic flow and vehicle speed) from Taiwan Area National Freeway Bureau as the input characteristics of the traffic simulation program and referred to the mobile station (MS) communication behaviors from Chunghwa Telecom to simulate the traffic information and communication records. The experimental results illustrated that the average accuracy of the vehicle speed forecasting method is 95.72%. Therefore, the proposed methods based on CFVD are suitable for an intelligent transportation system.
Wear Behaviour of A356/TiAl3 in Situ Composites Produced by Mechanical Alloying
Authors: Çam Seda , Demir Vedat , Özyürek Dursun
Published: February 2016

In this study, the effects of in situ TiAl3 particles on dry sliding wear behavior of A356 aluminum alloy (added Ti) composites were investigated. The wear samples were prepared by adding different amounts of Ti (4%, 6%, and 8%) into A356 powder alloy by mechanical alloying. The mechanically alloyed powders were cold pressed at 600 MPa and sintered 530 °C for 1 h in argon atmosphere and cooled in the furnace. After the sintering process, the samples were characterized. The results show that AlTi and TiAl3 intermetallic phases were formed and their amount increased depending on the amount of Ti added into A356 powder alloy. Out of the samples sintered with different titanium amounts (1 h at 530 °C), the highest hardness value and, accordingly, the lowest wear amount, were observed in the alloy containing 8% Ti.
Facile Functionalization of Electrospun Poly(ethylene-co-vinyl alcohol) Nanofibers via the Benzoxaborole-Diol Interaction
Authors: Kotsuchibashi Yohei , Ebara Mitsuhiro
Published: February 2016

A facile functionalization method of poly(ethylene-co-vinyl alcohol) (EVOH) nanofiber meshes was demonstrated by utilizing the benzoxaborole-diol interaction between EVOH and benzoxaborole-based copolymers (BOP). EVOH and BOP were firstly mixed to prepare the quasi-gel-state solution with enough viscosity for electro-spinning. The fiber morphology was controlled via changing the mixing ratio of EVOH and BOP. The prepared EVOH/BOP nanofiber mesh showed good stability in aqueous solution. Over 97% of the nanofibers remained after the immersion test for 24 h in acid or alkali aqueous solutions without changing their morphology. Temperature and pH-responsive moieties were copolymerized with BOP, and cationic dye was easily immobilized into the nanofiber mesh via an electrostatic interaction. Therefore, the proposed functionalization technique is possible to perform on multi-functionalized molecule-incorporated nanofibers that enable the fibers to show the environmental stimuli-responsive property for the further applications of the EVOH materials.
The Role of Rail Transit Systems in Reducing Energy and Carbon Dioxide Emissions: The Case of The City of Rio de Janeiro
Authors: Andrade Carlos , D’Agosto Márcio
Published: February 2016

The city of Rio de Janeiro, Brazil, has established, by municipal law, a goal of reducing greenhouse gas emissions of the transport system by 20% until 2020 compared to 2005. In order to reach this goal, the city’s public transport has been restructured with an emphasis on rail transit systems. The city will host the 2016 Olympic Games and this has encouraged the transformation of public transportation. One of the new projects is the construction of a new metro line, Line 4, to connect the downtown area to the city’s fastest growing part, the western region, which will also be a venue for many events during the Olympic Games. This article presents and applies a procedure for calculating energy use and emissions avoided by Line 4 by attracting users from other transport modes in the period from 2016 to 2040. The procedure uses a detailed demand forecast for this period and considers the local transport profile and the different fuels used. The net amount of carbon dioxide avoided was 55,449 tonnes per year and 44.53 grams per passenger kilometer. The avoided energy reached 0.76 MJ per passenger kilometer.
Teratogenicity of Ochratoxin A and the Degradation Product, Ochratoxin α, in the Zebrafish (Danio rerio) Embryo Model of Vertebrate Development
Authors: Haq Mehreen , Gonzalez Nelson , Mintz Keenan , Jaja-Chimedza Asha , De Jesus Christopher , Lydon Christina , Welch Aaron , Berry John
Published: February 2016

Ochratoxins, and particularly ochratoxin A (OTA), are toxic fungal-derived contaminants of food and other agricultural products. Growing evidence supports the degradation of OTA by chemical, enzymatic and/or microbial means as a potential approach to remove this mycotoxin from food products. In particular, hydrolysis of OTA to ochratoxin α (OTα) and phenylalanine is the presumptive product of degradation in most cases. In the current study, we employed the zebrafish (Danio rerio) embryo, as a model of vertebrate development to evaluate, the teratogenicity of OTA and OTα. These studies show that OTA is potently active in the zebrafish embryo toxicity assay (ZETA), and that toxicity is both concentration- and time-dependent with discernible and quantifiable developmental toxicity observed at nanomolar concentrations. On the other hand, OTα had no significant effect on embryo development at all concentrations tested supporting a decreased toxicity of this degradation product. Taken together, these results suggest that ZETA is a useful, and highly sensitive, tool for evaluating OTA toxicity, as well as its degradation products, toward development of effective detoxification strategies. Specifically, the results obtained with ZETA, in the present study, further demonstrate the toxicity of OTA, and support its degradation via hydrolysis to OTα as an effective means of detoxification.
A KDEL Retrieval System for ER-Golgi Transport of Japanese Encephalitis Viral Particles
Authors: Wang Robert , Wu Yu-Jen , Chen Han-Shan , Chen Chih-Jung
Published: February 2016

Evidence has emerged that RNA viruses utilize the host secretory pathway for processing and trafficking mature viral particles and for exiting the infected cells. Upon completing the complex assembly process, the viral particles take advantage of the cellular secretory trafficking machinery for their intracellular trafficking toward the Golgi organelle and budding or export of virions. In this study, we showed that Japanese encephalitis virus (JEV)-induced extracellular GRP78 contains no KDEL motif using an anti-KDEL-specific antibody. Overexpression of the KDEL-truncated GRP78 in the GPR78 knocked down cells significantly reduced JEV infectivity, suggesting that the KDEL motif is required for GRP78 function in the release of JE viral particles. In addition, we demonstrated the KDELR protein, an ER-Golgi retrieval system component, is associated with viral envelope proteins and is engaged in the subcellular localization of viral particles in Golgi. More importantly, accumulation of intracellular virions was observed in the KDELR knocked down cells, indicating that the KDELR protein mediated the intracellular trafficking of JE viral particles. Altogether, we demonstrated that intracellular trafficking of JE assembled viral particles was mediated by the host ER-Golgi retrieval system prior to exit by the secretory pathway.
Big Data Offers Novel Insights for Oncolytic Virus Immunotherapy
Authors: Swift Stephanie , Stojdl David
Published: February 2016

Large-scale assays, such as microarrays, next-generation sequencing and various “omics” technologies, have explored multiple aspects of the immune response following virus infection, often from a public health perspective. Yet a lack of similar data exists for monitoring immune engagement during oncolytic virus immunotherapy (OVIT) in the cancer setting. Tracking immune signatures at the tumour site can create a snapshot or longitudinally analyse immune cell activation, infiltration and functionality within global populations or individual cells. Mapping immune changes over the course of oncolytic biotherapy—from initial infection to tumour stabilisation/regression through to long-term cure or escape/relapse—has the potential to generate important therapeutic insights around virus-host interactions. Further, correlating such immune signatures with specific tumour outcomes has significant value for guiding the development of novel oncolytic virus immunotherapy strategies. Here, we provide insights for OVIT from large-scale analyses of immune populations in the infection, vaccination and immunotherapy setting. We analyse several approaches to manipulating immune engagement during OVIT. We further explore immunocentric changes in the tumour tissue following immunotherapy, and compile several immune signatures of therapeutic success. Ultimately, we highlight clinically relevant large-scale approaches with the potential to strengthen future oncolytic strategies to optimally engage the immune system.
Kidney versus Islet Allograft Survival after Induction of Mixed Chimerism with Combined Donor Bone Marrow Transplantation
Authors: Oura Tetsu , Ko Dicken S. C. , Boskovic Svjetlan , O'neil John J. , Chipashvili Vaja , Koulmanda Maria , Hotta Kiyohiko , Kawai Kento , Nadazdin Ognjenka , Smith R. Neal , Cosimi A. B. , Kawai Tatsuo
Published: February 2016

We have previously reported successful induction of transient mixed chimerism and long-term acceptance of renal allografts in MHC mismatched nonhuman primates. In this study, we attempted to extend this tolerance induction approach to islet allografts. A total of eight recipients underwent MHC mismatched combined islet and bone marrow (BM) transplantation after induction of diabetes by streptozotocin. Three recipients were treated after a nonmyeloablative conditioning regimen that included low-dose total body and thymic irradiation, horse Atgam (ATG), six doses of anti-CD154 monoclonal antibody (mAb), and a 1-month course of cyclosporine (CyA) (Islet A). In Islet B, anti-CD8 mAb was administered in place of CyA. In Islet C, two recipients were treated with Islet B, but without ATG. The results were compared with previously reported results of eight cynomolgus monkeys that received combined kidney and BM transplantation (Kidney A) following the same conditioning regimen used in Islet A. The majority of kidney/BM recipients achieved long-term renal allograft survival after induction of transient chimerism. However, prolonged islet survival was not achieved in similarly conditioned islet/BM recipients (Islet A), despite induction of comparable levels of chimerism. In order to rule out islet allograft loss due to CyA toxicity, three recipients were treated with anti-CD8 mAb in place of CyA. Although these recipients developed significantly superior mixed chimerism and more prolonged islet allograft survival (61, 103, and 113 days), islet function was lost soon after the disappearance of chimerism. In Islet C recipients, neither prolonged chimerism nor islet survival was observed (30 and 40 days). Significant improvement of mixed chimerism induction and islet allograft survival were achieved with a CyA-free regimen that included anti-CD8 mAb. However, unlike the kidney allograft, islet allograft tolerance was not induced with transient chimerism. Induction of more durable mixed chimerism may be necessary for induction of islet allograft tolerance.
Dimethyl Sulfoxide (DMSO) Increases Percentage of CXCR4+ Hematopoietic Stem/Progenitor Cells, Their Responsiveness to an SDF-1 Gradient, Homing Capacities, and Survival
Authors: Jarocha Danuta , Zuba-Surma Ewa , Majka Marcin
Published: February 2016

Cryopreservation of bone marrow (BM), mobilized peripheral blood (mPB), and cord blood (CB) hematopoietic stem/progenitor cells (HSPCs) is a routine procedure before transplantation. The most commonly used cryoprotectant for HSPCs is dimethyl sulfoxide (DMSO). The objective of this study was to evaluate the influence of DMSO on surface receptor expression and chemotactic activities of HSPCs. We found that 10 min of incubation of human mononuclear cells (MNCs) with 10% DMSO significantly increases the percentage of CXCR4+, CD38+, and CD34+ cells, resulting in an increase of CD34+, CD34+CXCR4+, and CD34+CXCR4+CD38– subpopulations. Furthermore, DMSO significantly increased chemotactic responsiveness of MNCs and CXCR4+ human hematopoietic Jurkat cell line to a stromal cell-derived factor-1 (SDF-1) gradient. Furthermore, we demonstrated enhanced chemotaxis of human clonogenic progenitor cells to an SDF-1 gradient, which suggests that DMSO directly enhances the chemotactic responsiveness of early human progenitors. DMSO preincubation also caused lower internalization of the CXCR4 receptor. In parallel experiments, we found that approximately 30% more of DMSO-preincubated human CD45+ and CD45+CD34+ cells homed to the mouse BM 24 h after transplantation in comparison to control cells. Finally, we demonstrated considerably higher (25 days) survival of mice transplanted with DMSO-exposed MNCs than those transplanted with the control cells. We show in this study an unexpected beneficial influence of DMSO on HSPC homing and suggest that a short priming with DMSO before transplantation could be considered a new strategy to enhance cell homing and engraftment.
Optimization and Scale-up Isolation and Culture of Neonatal Porcine Islets: Potential for Clinical Application
Authors: Ellis Cara , Lyon James G. , Korbutt Gregory S.
Published: February 2016

One challenge that must be overcome to allow transplantation of neonatal porcine islets (NPIs) to become a clinical reality is defining a reproducible and scalable protocol for the efficient preparation of therapeutic quantities of clinical grade NPIs. In our standard protocol, we routinely isolate NPIs from a maximum of four pancreases, requiring tissue culture in 16 Petri dishes (four per pancreas) in Ham's F10 and bovine serum albumin (BSA). We have now developed a scalable and technically simpler protocol that allows us to isolate NPIs from a minimum of 12 pancreases at a time by employing automated tissue chopping, collagenase digestion in a single vessel, and tissue culture/media changes in 75% fewer Petri dishes. For culture, BSA is replaced with human serum albumin and supplemented with Z-VAD-FMK general caspase inhibitor and a protease inhibitor cocktail. The caspase inhibitor was added to the media for only the first 90 min of culture. NPIs isolated using the scalable protocol had significantly more cellular insulin recovered (56.9 ± 1.4 μg) when compared to the standard protocol (15.0 ± 0.5 μg; p < 0.05). Compared to our standard protocol, recovery of β-cells (6.0 × 106 ± 0.2 vs. 10.0 × 106 ± 0.4; p < 0.05) and islet equivalents (35,135 ± 186 vs. 41,810 ± 226; p < 0.05) was significantly higher using the scalable protocol. During a static glucose stimulation assay, the SI of islets isolated by the standard protocol were significantly lower than the scale-up protocol (4.3 ± 0.2 vs. 5.5 ± 0.1; p < 0.05). Mice transplanted with NPIs using the scalable protocol had significantly lower blood glucose levels than the mice that receiving NPIs from the standard protocol ( p < 0.01) and responded significantly better to a glucose tolerance test. Based on the above findings, this improved simpler scalable protocol is a significantly more efficient means for preparing therapeutic quantities of clinical grade NPIs.
Cell-Based Therapy in TBI: Magnetic Retention of Neural Stem Cells in Vivo
Authors: Shen Wei-Bin , Plachez Céline , Tsymbalyuk Orest , Tsymbalyuk Natalya , Xu Su , Smith Aaron M. , Michel Sarah L. J. , Yarnell Deborah , Mullins Roger , Gullapalli Rao P. , Puche Adam , Simard J. Marc , Fishman Paul S. , Yarowsky Paul
Published: February 2016

Stem cell therapy is under active investigation for traumatic brain injury (TBI). Noninvasive stem cell delivery is the preferred method, but retention of stem cells at the site of injury in TBI has proven challenging and impacts effectiveness. To investigate the effects of applying a magnetic field on cell homing and retention, we delivered human neuroprogenitor cells (hNPCs) labeled with a superparamagnetic nanoparticle into post-TBI animals in the presence of a static magnetic field. We have previously devised a method of loading hNPCs with ultrasmall superparamagnetic iron oxide (USPIO) nanoparticles Molday ION Rhodamine B (MIRB™). Labeling of hNPCs (MIRB-hNPCs) does not affect hNPC viability, proliferation, or differentiation. The 0.6 tesla (T) permanent magnet was placed ~4 mm above the injured parietal cortex prior to intracarotid injection of 4 × 104MIRB-hNPCs. Fluorescence imaging, Perls' Prussian blue histochemistry, immunocytochemistry with SC121, a human-specific antibody, and T2-weighted magnetic resonance imaging ex vivo revealed there was increased homing and retention of MIRB-hNPCs in the injured cortex as compared to the control group in which MIRB-hNPCs were injected in the absence of a static magnetic field. Fluoro-Jade C staining and immunolabeling with specific markers confirmed the viability status of MIRB-hNPCs posttransplantation. These results show that increased homing and retention of MIRB-hNPCs post-TBI by applying a static magnetic field is a promising technique to deliver cells into the CNS for treatment of neurological injuries and neurodegenerative diseases.
Clinical Trials with Mesenchymal Stem Cells: An Update
Authors: Squillaro Tiziana , Peluso Gianfranco , Galderisi Umberto
Published: February 2016

In the last year, the promising features of mesenchymal stem cells (MSCs), including their regenerative properties and ability to differentiate into diverse cell lineages, have generated great interest among researchers whose work has offered intriguing perspectives on cell-based therapies for various diseases. Currently the most commonly used adult stem cells in regenerative medicine, MSCs, can be isolated from several tissues, exhibit a strong capacity for replication in vitro, and can differentiate into osteoblasts, chondrocytes, and adipocytes. However, heterogeneous procedures for isolating and cultivating MSCs among laboratories have prompted the International Society for Cellular Therapy (ISCT) to issue criteria for identifying unique populations of these cells. Consequently, the isolation of MSCs according to ISCT criteria has produced heterogeneous, nonclonal cultures of stromal cells containing stem cells with different multipotent properties, committed progenitors, and differentiated cells. Though the nature and functions of MSCs remain unclear, nonclonal stromal cultures obtained from bone marrow and other tissues currently serve as sources of putative MSCs for therapeutic purposes, and several findings underscore their effectiveness in treating different diseases. To date, 493 MSC-based clinical trials, either complete or ongoing, appear in the database of the US National Institutes of Health. In the present article, we provide a comprehensive review of MSC-based clinical trials conducted worldwide that scrutinizes biological properties of MSCs, elucidates recent clinical findings and clinical trial phases of investigation, highlights therapeutic effects of MSCs, and identifies principal criticisms of the use of these cells. In particular, we analyze clinical trials using MSCs for representative diseases, including hematological disease, graft-versus-host disease, organ transplantation, diabetes, inflammatory diseases, and diseases in the liver, kidney, and lung, as well as cardiovascular, bone and cartilage, neurological, and autoimmune diseases.
Optimal Timing of Mesenchymal Stem Cell Therapy for Neonatal Intraventricular Hemorrhage
Authors: Park Won Soon , Sung Se In , Ahn So Yoon , Sung Dong Kyung , Im Geun Ho , Yoo Hye Soo , Choi Soo Jin , Chang Yun Sil
Published: February 2016

We recently showed that intraventricular transplantation of human umbilical cord blood (UCB)-derived mesenchymal stem cells (MSCs) significantly attenuated posthemorrhagic hydrocephalus (PHH) and brain injury after severe intraventricular hemorrhage (IVH) in newborn rat pups. The purpose of this study was to optimize the timing of MSC transplantation for severe IVH. Severe IVH was induced by injecting 100 μl of blood into each ventricle of Sprague–Dawley rats on postnatal day 4 (P4). Human UCB-derived MSCs (1 × 105cells in 10 μl of normal saline) were transplanted intraventricularly under stereotaxic guidance either early at P6 or late at P11. Serial brain MRIs and behavioral function tests, such as negative geotaxis and rotarod tests, were performed. At P32, brain tissue samples were obtained for histological and biochemical analyses. Intracerebroventricular transplantation of MSCs significantly attenuated the development of PHH, behavioral impairment, increased apoptosis and astrogliosis, reduced corpus callosum thickness and brain myelination, and upregulated inflammatory cytokines including interleukin (IL)-1α, IL-1β, IL-6, and tumor necrosis factor-α (TNF-α) at P6 but not at P11 after induction of severe IVH. Intracerebroventricular transplantation of human UCB-derived MSCs attenuated PHH and brain injury after severe IVH in newborn rats in a time-dependent manner. Significant neuroprotection was only demonstrated when administered early at 2 days after induction but not late at 7 days after induction of severe IVH.
The Effect of Fetal Bovine Serum (FBS) on Efficacy of Cellular Reprogramming for Induced Pluripotent Stem Cell (iPSC) Generation
Authors: Kwon Daekee , Kim Jin-Su , Cha Byung-Hyun , Park Kwang-Sook , Han Inbo , Park Kyung-Soon , Bae Hojae , Han Myung-Kwan , Kim Kwang-Soo , Lee Soo-Hong
Published: February 2016

Induced pluripotent stem cells (iPSCs) are pivotal to the advancement of regenerative medicine. However, the low efficacy of iPSC generation and insufficient knowledge about the reprogramming mechanisms involved in somatic cell/adult stem cell reversion to a pluripotent phenotype remain critical hurdles to the therapeutic application of iPSCs. The present study investigated whether the concentration of fetal bovine serum (FBS), a widely employed cell culture additive, can influence the cellular reprogramming efficacy (RE) of human adipose-derived stem cells (hADSCs) to generate iPSCs. Compared with the typically employed concentration of FBS (10%), high concentrations (20% and 30%) increased the RE of hADSCs by approximately twofold, whereas a low concentration (5%) decreased the RE by the same extent. Furthermore, cell counting kit-8 (CCK-8), bromodeoxyuridine (BrdU) incorporation, and fluorescence-activated cell sorting (FACS) assays showed that hADSC proliferation during reprogramming was significantly enhanced by FBS at 20% and 30%, whereas quantitative polymerase chain reaction (qPCR) and Western blotting assays revealed a concomitant decrease in p53, p51, and p21 expression. In addition, the efficacy of retrovirus-mediated transduction into hADSCs was increased by approximately 10% at high concentrations of FBS. It was confirmed that platelet-derived growth factor in the FBS enhanced proliferation and reprogramming efficacy. Finally, the generated iPSCs showed a normal karyotype, the same fingerprinting pattern as parental hADSCs, a genome-wide transcriptome pattern similar to that of human embryonic stem cells (hESCs), and in vivo pluripotency. In conclusion, the current investigation demonstrated that high concentrations of FBS can modulate molecular and cellular mechanisms underlying the reprogramming process in hADSCs, thereby augmenting the cellular RE for iPSC generation.
The Orphan Receptor Tyrosine Kinase ROR2 Facilitates MSCs to Repair Lung Injury in ARDS Animal Model
Authors: Cai Shi-Xia , Liu Ai-Ran , Chen Song , He Hong-Li , Chen Qi-Hong , Xu Jing-Yuan , Pan Chun , Yang Yi , Guo Feng-Mei , Huang Ying-Zi , Liu Ling , Qiu Hai-Bo
Published: February 2016

There are some limitations to the therapeutic effects of mesenchymal stem cells (MSCs) on acute respiratory distress syndrome (ARDS) due to their low engraftment and differentiation rates in lungs. We found previously that noncanonical Wnt5a signaling promoted the differentiation of mouse MSCs (mMSCs) into type II alveolar epithelial cells (AT II cells), conferred resistance to oxidative stress, and promoted migration of MSCs in vitro. As receptor tyrosine kinase-like orphan receptor 2 (ROR2) is an essential receptor for Wnt5a, it was reasonable to deduce that ROR2 might be one of the key molecules for the therapeutic effect of MSCs in ARDS. The mMSCs that stably overexpressed ROR2 or the green fluorescent protein (GFP) control were transplanted intratracheally into the ARDS mice [induced by intratracheal injection of lipopolysaccharide (LPS)]. The results showed that ROR2-overexpressing mMSCs led to more significant effects than the GFP controls, including the retention of the mMSCs in the lung, differentiation into AT II cells, improvement of alveolar epithelial permeability, improvement of acute LPS-induced pulmonary inflammation, and, finally, reduction of the pathological impairment of the lung tissue. In conclusion, MSCs that overexpress ROR2 could further improve MSC-mediated protection against epithelial impairment in ARDS.
Transferring Xenogenic Mitochondria Provides Neural Protection against Ischemic Stress in Ischemic Rat Brains
Authors: Huang Po-Jui , Kuo Chi-Chung , Lee Hsiu-Chin , Shen Ching-I , Cheng Fu-Chou , Wu Shih-Fang , Chang Jui-Chih , Pan Hung-Chuan , Lin Shinn-Zong , Liu Chin-San , Su Hong-Lin
Published: February 2016

Transferring exogenous mitochondria has therapeutic effects on damaged heart, liver, and lung tissues. Whether this protective effect requires the symbiosis of exogenous mitochondria in host cells remains unknown. Here xenogenic mitochondria derived from a hamster cell line were applied to ischemic rat brains and rat primary cortical neurons. Isolated hamster mitochondria, either through local intracerebral or systemic intra-arterial injection, significantly restored the motor performance of brain-ischemic rats. The brain infarct area and neuronal cell death were both attenuated by the exogenous mitochondria. Although internalized mitochondria could be observed in neurons and astrocytes, the low efficacy of mitochondrial internalization could not completely account for the high rate of rescue of the treated neural cells. We further illustrated that disrupting electron transport or ATPase synthase in mitochondria significantly attenuated the protective effect, suggesting that intact respiratory activity is essential for the mitochondrial potency on neural protection. These results emphasize that nonsymbiotic extracellular mitochondria can provide an effective cell defense against acute injurious ischemic stress in the central nervous system.
Induction of Endothelial Phenotype from Wharton's Jelly-Derived MSCs and Comparison of Their Vasoprotective and Neuroprotective Potential with Primary WJ-MSCs in CA1 Hippocampal Region Ex Vivo
Authors: Obtulowicz Patrycja , Lech Wioletta , Strojek Lukasz , Sarnowska Anna , Domanska-Janik Krystyna
Published: February 2016

Ischemic stroke results in violent impairment of tissue homeostasis leading to severe perturbation within the neurovascular unit (NVU) during the recovery period. The aim of this study was to assess the potential of mesenchymal stem cells (MSCs) originating from Wharton's jelly (WJ) to differentiate into functionally competent cells of endothelial lineage (WJ-EPCs). The protective effect(s) of either primary WJ-MSCs or induced WJ-EPCs was investigated and compared after oxygen–glucose deprivation (OGD) of hippocampal organotypic slices (OHC) in the indirect coculture model. WJ-MSCs, primed in EGM-2 (Lonza commercial medium) under 5% O2, acquired cobblestone endothelial-like morphology, formed capillary-like structures and actively took up DiI-Ac-LDL. Both cell types (WJ-MSCs and WJ-EPCs) were positive for CD73, CD90, CD105, VEGFR-2, and VEGF, but only endothelial-like culture expressed vWF and PECAM-1 markers at significant levels. In the presence of either WJ-MSCs or WJ-EPCs in the compartment below OGD-injured slices, cell death and vascular atrophy in the hypoxia-sensitive CA1 region were substantially decreased. This suggests that a paracrine mechanism may mediate WJ-MSC- and WJ-EPC-dependent protection. Thus, finally, we estimated secretion of the neuro/angio/immunomodulatory molecules IL-6, TGF-β1, and VEGF by these cell cultures. We have found that release of TGF-β1 and IL-6 was TLR ligand [LPS and Poly(I:C)] concentration dependent and stronger in WJ-EPC than WJ-MSC cultures. Simultaneously, the uneven pattern of TLR receptors and modulatory cytokine gene expression was confirmed also on qRT-PCR level, but no significant differences were noticed between WJ-EPC and primary WJ-MSC cultures.
The Cytoprotective Effects of Human Endothelial Progenitor Cell-Conditioned Medium against an Ischemic Insult Are Not Dependent on VEGF and IL-8
Authors: Di Santo Stefano , Fuchs Anna-Lena , Periasamy Ramesh , Seiler Stefanie , Widmer Hans Rudolf
Published: February 2016

Endothelial progenitor cells (EPCs) promote revascularization and tissue repair mainly by paracrine actions. In the present study, we investigated whether EPC-secreted factors in the form of conditioned medium (EPC-CM) can protect cultured brain microvascular endothelial cells against an ischemic insult. Furthermore, we addressed the type of factors that are involved in the EPC-CM-mediated functions. For that purpose, rat brain-derived endothelial cells (rBCEC4 cell line) were exposed to EPC-CM pretreated with proteolytic digestion, heat inactivation, and lipid extraction. Moreover, the involvement of VEGF and IL-8, as canonical angiogenic factors, was investigated by means of neutralizing antibodies. We demonstrated that EPC-CM significantly protected the rBCEC4 cells against an ischemic insult mimicked by induced oxygen–glucose deprivation followed by reoxygenation. The cytoprotective effect was displayed by higher viable cell numbers and reduced caspase 3/7 activity. Heat inactivation, proteolytic digestion, and lipid extraction resulted in a significantly reduced EPC-CM-dependent increase in rBCEC4 viability, tube formation, and survival following the ischemic challenge. Notably, VEGF and IL-8 neutralization did not affect the actions of EPC-CM on rBCEC4 under both standard and ischemic conditions. In summary, our findings show that paracrine factors released by EPCs activate an angiogenic and cytoprotective response on brain microvascular cells and that the activity of EPC-CM relies on the concerted action of nonproteinaceous and proteinaceous factors but do not directly involve VEGF and IL-8.
Induced Pluripotent Stem Cells with Six Reprogramming Factors from Prairie Vole, Which is an Animal Model for Social Behaviors
Authors: Katayama Masafumi , Hirayama Takashi , Horie Kengo , Kiyono Tohru , Donai Kenichiro , Takeda Satoru , Nishimori Katsuhiko , Fukuda Tomokazu
Published: February 2016

Prairie voles show strong pair bonding with their mating partners, and they demonstrate parental behavior toward their infants, indicating that the prairie vole is a unique animal model for analysis of molecular mechanisms of social behavior. Until a recent study, the signaling pathway of oxytocin was thought to be critical for the social behavior of prairie voles, but neuron-specific functional research may be necessary to identify the molecular mechanisms of social behavior. Prairie vole pluripotent stem cells of high quality are essential to elucidate the molecular mechanisms of social behaviors. Generation of high-quality induced pluripotent stem cells (iPSCs) would help to establish a genetically modified prairie vole, including knockout and knock-in models, based on the pluripotency of iPSCs. Thus, we attempted to establish high-quality prairie vole-derived iPSCs (pv-iPSCs) in this study. We constructed a polycistronic reprogramming vector, which included six reprograming factors (Oct3/4, Sox2, Klf4, c-myc, Lin28, and Nanog). Furthermore, we evaluated the effect of six reprogramming factors, which included Oct3/4 with the transactivation domain (TAD) of MyoD. Implantation of the pv-iPSCs into immunodeficient mice caused a teratoma with three germ layers. Furthermore, the established pv-iPSCs tested positive for stem cell markers, including alkaline phosphatase activity (ALP), stage-specific embryonic antigen (SSEA)-1, and dependence on leukemia inhibitory factor (LIF). Our data indicate that our newly established pv-iPSCs may be a useful tool for genetic analysis of social behavior.
Induction of Neurorestoration from Endogenous Stem Cells
Authors: Yu Ji Hea , Seo Jung-Hwa , Lee Ji Yong , Lee Min-Young , Cho Sung-Rae
Published: February 2016

Neural stem cells (NSCs) persist in the subventricular zone lining the ventricles of the adult brain. The resident stem/progenitor cells can be stimulated in vivo by neurotrophic factors, hematopoietic growth factors, magnetic stimulation, and/or physical exercise. In both animals and humans, the differentiation and survival of neurons arising from the subventricular zone may also be regulated by the trophic factors. Since stem/progenitor cells present in the adult brain and the production of new neurons occurs at specific sites, there is a possibility for the treatment of incurable neurological diseases. It might be feasible to induce neurogenesis, which would be particularly efficacious in the treatment of striatal neurodegenerative conditions such as Huntington's disease, as well as cerebrovascular diseases such as ischemic stroke and cerebral palsy, conditions that are widely seen in the clinics. Understanding of the molecular control of endogenous NSC activation and progenitor cell mobilization will likely provide many new opportunities as therapeutic strategies. In this review, we focus on endogenous stem/progenitor cell activation that occurs in response to exogenous factors including neurotrophic factors, hematopoietic growth factors, magnetic stimulation, and an enriched environment. Taken together, these findings suggest the possibility that functional brain repair through induced neurorestoration from endogenous stem cells may soon be a clinical reality.
Interactions of cyclodextrins and their derivatives with toxic organophosphorus compounds
Authors: Letort Sophie , Balieu Sébastien , Erb William , Gouhier Géraldine , Estour François
Published: February 2016

The aim of this review is to provide an update on the current use of cyclodextrins against organophosphorus compound intoxications. Organophosphorus pesticides and nerve agents play a determinant role in the inhibition of cholinesterases. The cyclic structure of cyclodextrins and their toroidal shape are perfectly suitable to design new chemical scavengers able to trap and hydrolyze the organophosphorus compounds before they reach their biological target.