bluuebunny/crossref_metadata_2025 huggingface dataset:
Four-Year Follow-Up of Children in the LEAP Randomized Trial: Some Planned and Accidental Findings
Authors: Strain Phillip S.
Published: June 2017

This article reports on a 4-year follow-up study from the Learning Experiences and Alternative Program for Preschoolers and Their Parents (LEAP) randomized trial of early intervention for young children with autism. Overall, participants from LEAP classes were marginally superior to comparison class children on elementary school outcomes specific to communication, adaptive behavior, social, academic, and cognitive skills. Statistically significant group differences were noted in cognitive development and social skills. However, when placement was treated as an independent variable, very large effects were seen across all outcome measures, including autism symptoms, for children who were enrolled in inclusive settings. Data from adult family members confirmed important changes in perceived quality of life.
Dermatomyositis with renal infarction: a case report and literature review
Authors: Zhang Ti , Liu Xin , Xu Huji
Published: June 2017

Renal infarction is a rare clinical entity that is not easily detected by low-sensitivity ultrasound. We herein report a case of dermatomyositis with renal infarction detected during corticosteroid therapy. The patient was followed up for 18 months. A woman who was clinically diagnosed with dermatomyositis complained of severe pain in the right flank of the low back and abdomen, accompanied by nausea and vomiting during corticosteroid therapy. Based on the findings of routine blood tests, abdominal X-ray radiography, and abdominal ultrasound, the patient was diagnosed with acute gastroenteritis and treated with levofloxacin. However, her symptoms were not relieved. Abdominal contrast-enhanced computed tomography revealed renal infarction. Clinicians should be alert to the occurrence of thrombosis, especially when it manifests as vasculitis in patients with rheumatic disease who complain of severe abdominal pain, because it may suggest the presence of renal infarction.
Microarray-Based Gene Expression Analysis for Veterinary Pathologists: A Review
Authors: Raddatz Barbara B. , Spitzbarth Ingo , Matheis Katja A. , Kalkuhl Arno , Deschl Ulrich , Baumgärtner Wolfgang , Ulrich Reiner
Published: June 2017

High-throughput, genome-wide transcriptome analysis is now commonly used in all fields of life science research and is on the cusp of medical and veterinary diagnostic application. Transcriptomic methods such as microarrays and next-generation sequencing generate enormous amounts of data. The pathogenetic expertise acquired from understanding of general pathology provides veterinary pathologists with a profound background, which is essential in translating transcriptomic data into meaningful biological knowledge, thereby leading to a better understanding of underlying disease mechanisms. The scientific literature concerning high-throughput data-mining techniques usually addresses mathematicians or computer scientists as the target audience. In contrast, the present review provides the reader with a clear and systematic basis from a veterinary pathologist’s perspective. Therefore, the aims are (1) to introduce the reader to the necessary methodological background; (2) to introduce the sequential steps commonly performed in a microarray analysis including quality control, annotation, normalization, selection of differentially expressed genes, clustering, gene ontology and pathway analysis, analysis of manually selected genes, and biomarker discovery; and (3) to provide references to publically available and user-friendly software suites. In summary, the data analysis methods presented within this review will enable veterinary pathologists to analyze high-throughput transcriptome data obtained from their own experiments, supplemental data that accompany scientific publications, or public repositories in order to obtain a more in-depth insight into underlying disease mechanisms.
A Controversial Warplane
Authors: Coticchia Fabrizio
Published: June 2017

The literature on strategic narratives has started to pay growing attention to the concept of “narrative dominance,” stressing the role played by counternarratives in hindering a wider acceptance of a specific message. However, limited consideration has been devoted to counternarratives, which have seldom been assessed in a systematic way. The aim of this article is to fill these gaps by examining the underrated case of Italy. The article investigates the main content of narratives and counternarratives developed by parties and peace movements regarding the decision to acquire the F-35. The article, which is based on primary and secondary sources, adopts a multidisciplinary approach, combining security studies and social movement studies.
STS as science or politics?
Authors: Collins Harry , Evans Robert , Weinel Martin
Published: June 2017

In a recent editorial for this journal, Sergio Sismondo makes two claims. First, he states that STS bears no responsibility for the emergence of post-truth politics. Second, he claims that debates about the nature of expertise that take place within STS are irrelevant in this context. In contrast, we argue that, whether or not STS had a causal influence on the emergence of post-truth politics, there is a clear resonance between the two positions and that the current political climate makes the empirically informed and scientific analysis of expertise and the form of life of science more important than ever. We argue that treating the contribution of STS to these matters as essentially political rather than scientific surrenders any special role we have as experts on the organization and values of science and leaves STS as just one political actor among others.
Myths of Sexual Economics Theory
Authors: Rudman Laurie A.
Published: June 2017

The authors of sexual economics theory (Baumeister & Twenge, 2002; Baumeister & Vohs, 2004) argue that sex is a female commodity that women exchange for men’s resources; therefore, women (not men) are responsible for the cultural suppression of sexuality, ostensibly to preserve the value of sex. In this article, I describe the central tenets of sexual economics theory and summarize a growing body of research contradicting them. I also explain the negative implications of the claims of sexual economics theory for gender equality and heterosexual relationships. Researchers, clinicians, and educators engaged in understanding human sexuality may use the arguments provided in this article to counteract gender myths. This article also serves as a case study of how feminist scholars can employ empirical evidence to weaken a popularized, patriarchal theory.
A Practitioner Implementation of a Tier 2 First-Grade Mathematics Intervention
Authors: Strand Cary Mari G. , Clarke Ben , Doabler Christian T. , Smolkowski Keith , Fien Hank , Baker Scott K.
Published: June 2017

We report on a practitioner implementation of Fusion, a first-grade mathematics intervention. Studies such as this evaluation of a loose implementation under realistic conditions are important to curriculum developers’ understanding of how evidence-based programs and tools work under a variety of implementation scenarios. In this quasi-experimental study, first-grade students ( n = 253) in 10 schools were assigned to treatment ( n = 154) or control ( n = 99). Rather than randomly assigning students, schools assigned those students most at risk to treatment and, typically, those less at risk to control. School staff administered pre- and postassessments and led Fusion sessions approximately 30 min per day, 3 days per week. The intervention resulted in a significant positive effect on a researcher-developed first-grade math measure. The implementation of Fusion and feedback from school staff provided insights to guide the curriculum development process.
Examination of the Psychometric Properties of the Geriatric Fear of Falling Measure Among Community-Dwelling Older Adults in the United States
Authors: Chen Tuo-Yu , Edwards Jerri D. , Janke Megan
Published: June 2017

This current study investigated the construct validity and reliability of the Geriatric Fear of Falling Measure (GFFM) among community-dwelling older adults in the United States. Eighty-eight participants were assessed on the GFFM together with demographics, falls, and fear of falling and falls-efficacy measures at baseline and an 8-week follow-up visit. Cronbach’s alpha, regression analyses, and correlation analyses were used to examine the psychometric properties of the GFFM. The results showed that the GFFM demonstrated good construct validity and reliability among community-dwelling older adults in the United States. Our findings provide evidence for the validity and reliability of the GFFM. Further study with a larger and diverse sample is needed to determine whether the GFFM has potential as a quick screening tool of fear of falling in clinical settings.
Meta-Analysis of Social Cognition in Mild Cognitive Impairment
Authors: Bora Emre , Yener Görsev G.
Published: June 2017

Social cognitive abilities are impaired in Alzheimer disease and other dementias. Recent studies suggested that social cognitive abilities might be also impaired in mild cognitive impairment (MCI). Current meta-analysis aimed to summarize available evidence for deficits in theory of mind (ToM) and emotion recognition in MCI. In this meta-analysis of 17 studies, facial emotion recognition and ToM performances of 513 individuals with MCI and 693 healthy controls were compared. Mild cognitive impairment was associated with significant impairments falling in the medium effect sizes range in ToM ( d = 0.63) and facial emotion recognition ( d = 0.58). Among individual emotions, recognition of fear and sadness were particularly impaired. There were no significant between-group differences in recognition of disgust, happiness, and surprise. Social cognitive deficits were more severe in multidomain MCI. There is a need for longitudinal studies investigating the potential role of social cognitive impairment in predicting conversion to dementia.
Adverse Childhood Circumstances and Functional Status Throughout Adult Life
Authors: Laditka James N. , Laditka Sarah B.
Published: June 2017

Objective:We studied the association of childhood adversity with adult functional status. Method: With data from the Panel Study of Income Dynamics and the 2014 Childhood Retrospective Circumstances Study (1992-2013; N = 6,705; 62,885 person-years), we estimated functional status transition probabilities associated with childhood adversity, with multinomial logistic Markov models adjusted for age, sex, race/ethnicity, and education. Microsimulation then estimated functional status outcomes throughout adulthood for African American, Hispanic, and non-Hispanic White women and men. Results: Adversity was significantly associated with functional status. Of White women without adversities, 2.3% had difficulty doing activities of daily living at age 30, compared with 8.2% with high adversity; comparable results were 3.7% and 8.7% for African Americans, 0.9% and 11.5% for Hispanics (all p < .01). Patterns were similar at other ages, for men, and when adjusted for midlife health conditions and health behavior. Discussion: Childhood adversity may substantially increase functional impairment throughout adult life.
Religious Participation and Biological Functioning in Mexico
Authors: Hill Terrence D. , Rote Sunshine M. , Ellison Christopher G.
Published: June 2017

Objective:Although several studies suggest that religious involvement tends to favor healthy biological functioning, most of this work has been conducted in the United States. This study explores the association between religious participation and biological functioning in Mexico. Method: The data are drawn from two waves of the Mexican Health and Aging Study (2003-2012) to assess continuous and categorical biomarker specifications. Results: Across specifications, religious participation in 2003 is associated with lower levels of waist-to-hip ratio, total cholesterol, pulse rate, and overall allostatic load in 2012. Respondents who increased their participation over the study period also exhibit a concurrent reduction in pulse rate. Depending on the specification, participation is also associated with lower levels of diastolic blood pressure and C-reactive protein. Participation is generally unrelated to body mass index, glycosylated hemoglobin, and systolic blood pressure. Discussion: Our results confirm that religious participation is associated with healthier biological functioning in Mexico.
Suspected Lateral Periodontal Cyst Presenting Concurrently with Canine Acanthomatous Ameloblastoma in a 2-Year-Old Standard Poodle
Authors: Tjepkema Jennifer , Soukup Jason W. , Bell Cynthia
Published: June 2017

Lateral periodontal cysts (LPCs) are odontogenic epithelial cysts composed of nonkeratinized epithelial cells that are in the category of developmental cysts, rather than inflammatory cysts. Lateral periodontal cysts are rare both in people and domestic animals; they are associated with vital teeth and located lateral to a tooth root. Lateral periodontal cysts are typically asymptomatic lesions that are characterized radiographically as a unilocular lucency with well-defined corticated borders. Canine acanthomatous ameloblastoma (CAA) is the most common odontogenic neoplasm in dogs and rarely presents as a cystic lesion. This case report describes the diagnosis and treatment of a cyst that occurred as a swelling apical to a gingival mass that was diagnosed histologically as CAA. Surgical management by conservative gingivectomy, cyst enucleation, and bone grafting was an effective treatment in this patient.
A new method for rapid shock loss evaluation and reduction for the optimization design of a supersonic compressor cascade
Authors: Liu Baojie , Shi Hengtao , Yu Xianjun
Published: June 2017

A one-dimensional analytical shock loss prediction method was proposed to tailor the shock system, i.e. the strength of the first and second passage shock, and reduce the shock loss in a supersonic cascade. To develop the one-dimensional analytical model, the shock system in a supersonic cascade was divided into four processes which can be seen in most supersonic compressor cascade, i.e. the flow upstream the extending-external shock, the flow between the extending-external shock and the first passage shock, the accelerating flow from the first passage shock to the second passage shock, and the flow downstream of the second passage shock. Based on some flow assumptions and experimental empirical correlations, the complex flows, containing the shock system, in the blade passage of a supersonic compressor cascade could be described with one-dimensional relationships, which can be used to predict shock losses along the flow passage rapidly and determine the shock system improving direction for achieving lower shock loss while keeping the same cascade static pressure ratio. In order to validate the one-dimensional analytical method, the shock system of two supersonic cascades ARL-SL19 and DLR-PAV-1.5 are modified based on the analysis of the model. The modified cascades achieved about 29% and 25% reduction of shock loss at redesign point compared with baseline cascades, respectively.
A dual-system cooperative co-evolutionary algorithm for satellite equipment layout optimization
Authors: Cui Feng-Zhe , Xu Zhi-Zheng , Wang Xiu-Kun , Zhong Chong-Quan , Teng Hong-Fei
Published: June 2017

This paper develops a new dual-system cooperative co-evolutionary algorithm for multi-modules (or multi-bearing plate) satellite equipment layout optimization problem, based upon the Potter’s cooperative co-evolutionary framework. Firstly, a new dual-system framework based on the Potter’s cooperative co-evolutionary is constructed and then, corresponding system decomposition rule, matrix analysis method and coordination mechanism are presented. Finally, the way of matching algorithms (e.g. evolutionary algorithms and swarm intelligence algorithms) with systems A and B in the dual system is presented. The purpose is to enhance the computational accuracy and robustness of the developed algorithm for satellite equipment layout optimization problem. The experimental results show that the developed algorithm has better computational accuracy and robustness (computational success ratio and standard deviation) as compared with four dual-system algorithms and two single-system algorithms based upon Potter’s cooperative co-evolutionary.
High-resolution simulation and parametric research on helicopter rotor vortex flowfield with TAMI control in hover
Authors: Ye Zhou , Xu Guohua , Shi Yongjie
Published: June 2017

In order to investigate the effect of active jet control on the rotor vortex flowfield in hover, a high-accuracy numerical method is presented in this paper, and a novel computer code is developed to calculate rotor flowfield with tip air mass injection. In the present computational fluid dynamics method, the Navier–Stokes equation is selected as the governing equation. For the sake of reducing numerical diffusion in calculation, the upwind Roe scheme with a fifth-order WENO scheme is employed to spatial discretization. A dual-time method is utilized in time marching and the high-efficiency implicit LU-SGS scheme is applied to every pseudo time step. The Baldwin–Lomax turbulence model is employed for eddy viscosity modeling. A surface boundary condition, which may effectively simulate the effect of tip air mass injection, is introduced into the current method. The overset grid method, which can veritably simulate the rotor blade motion, is adopted to exchange the flowfield information between the rotor blade and background grids. By the established method, the rotor flowfield with jet control of different jet angles is simulated. Furthermore, parametric analyses of jet velocity are carried out at different tip March numbers and collective pitches. It is demonstrated that tip air mass injection can make the tip vortex rolled up and generated in advance compared with the clean rotor, which can change the position and strength of the vortex core greatly. It is also shown that the rotor aerodynamic characteristics, such as blade surface pressure and radial lift distributions, are more sensitive to tip air mass injection control at the condition with a lower Mach number or a lower collective pitch.
Selective laser melting–enabled electrospinning: Introducing complexity within electrospun membranes
Authors: Paterson Thomas E , Beal Selina N , Santocildes-Romero Martin E , Sidambe Alfred T , Hatton Paul V , Asencio Ilida Ortega
Published: June 2017

Additive manufacturing technologies enable the creation of very precise and well-defined structures that can mimic hierarchical features of natural tissues. In this article, we describe the development of a manufacturing technology platform to produce innovative biodegradable membranes that are enhanced with controlled microenvironments produced via a combination of selective laser melting techniques and conventional electrospinning. This work underpins the manufacture of a new generation of biomaterial devices that have significant potential for use as both basic research tools and components of therapeutic implants. The membranes were successfully manufactured and a total of three microenvironment designs (niches) were chosen for thorough characterisation. Scanning electron microscopy analysis demonstrated differences in fibre diameters within different areas of the niche structures as well as differences in fibre density. We also showed the potential of using the microfabricated membranes for supporting mesenchymal stromal cell culture and proliferation. We demonstrated that mesenchymal stromal cells grow and populate the membranes penetrating within the niche-like structures. These findings demonstrate the creation of a very versatile tool that can be used in a variety of tissue regeneration applications including bone healing.
Analysis of variability in additive manufactured open cell porous structures
Authors: Evans Sam , Jones Eric , Fox Pete , Sutcliffe Chris
Published: June 2017

In this article, a novel method of analysing build consistency of additively manufactured open cell porous structures is presented. Conventionally, methods such as micro computed tomography or scanning electron microscopy imaging have been applied to the measurement of geometric properties of porous material; however, high costs and low speeds make them unsuitable for analysing high volumes of components. Recent advances in the image-based analysis of open cell structures have opened up the possibility of qualifying variation in manufacturing of porous material. Here, a photogrammetric method of measurement, employing image analysis to extract values for geometric properties, is used to investigate the variation between identically designed porous samples measuring changes in material thickness and pore size, both intra- and inter-build. Following the measurement of 125 samples, intra-build material thickness showed variation of ±12%, and pore size ±4% of the mean measured values across five builds. Inter-build material thickness and pore size showed mean ranges higher than those of intra-build, ±16% and ±6% of the mean material thickness and pore size, respectively. Acquired measurements created baseline variation values and demonstrated techniques suitable for tracking build deviation and inspecting additively manufactured porous structures to indicate unwanted process fluctuations.
Late Neolithic to Middle Bronze Age (around 4900–3100 cal. BP) lake-level fluctuations at Lake Neuchâtel (Switzerland) as reflected by the sediment sequence of the site of Colombier/Les Plantées de Rive: Palaeoclimatic and archaeological implications
Authors: Deák Judit , Magny Michel , Wüthrich Sonia
Published: June 2017

Recent archaeological investigations at the site of Colombier/Les Plantées de Rive (COL-PR) on the northwestern shore of Lake Neuchâtel (Switzerland) revealed an exceptional concentration of anthropic structures from the Late Neolithic to the Late Bronze Age interbedded within an alternation of terrestrial and lacustrine deposits. The chronology is based on radiocarbon dates from terrestrial material and on artefacts found in archaeological structures. Using various proxies, the sedimentological analysis of the COL-PR sediment sequence allows the reconstruction of lake-level fluctuations for the period 4900–3300 cal. BP. Three major lake-level highstands occurring at ca. 4900 (Late Neolithic), 3650 (Early Bronze Age) and 3400 cal. BP (Middle Bronze Age) have been related to the possible influences of decreases in the solar activity sometimes accentuated by increases in volcanic activity. The COL-PR site provides robust data to document the climatic deterioration assumed to be responsible for the abandonment of the lake-dwellings north of the Alps during the Middle and the early Late Bronze Age. In addition, the interbedding of successive monuments within terrestrial as well as lacustrine deposits suggests a significant role played over hundred years by the collective memory in the perpetuation of the ceremonial and funeral vocation of the site and its spatial organisation at the interface between lacustrine and terrestrial domains, despite interruptions provoked by major lake-level rises associated with worsening of climatic conditions.
The Risks of Total Pancreatectomy and Splenic Islet Autotransplantation
Authors: White Steven A. , London Nicholas J. M. , Johnson Paul R. V. , Davies Joanne E. , Pollard Cris , Contractor Harold H. , Hughes David P. , Robertson Gavin S. M. , Musto Patrick P. , Dennison Ashley R.
Published: June 2017

The intraportal site is the most common site for islet transplantation. Many other sites have been tried experimentally, including the spleen, which has successfully lead to insulin independence in a number of animal models. Nevertheless, there are no detailed reports of total pancreatectomy and splenic islet autotransplantation in humans. Five patients underwent total pancreatectomy and splenic islet autotransplantation for chronic pancreatitis. Four patients had a pylorus-preserving total pancreatectomy and one patient a duodenal-preserving pancreatectomy. In three cases islets were embolized into both the portal vein and spleen. Two patients received splenic islet transplants alone. Islets were transplanted by retrograde venous infusion via the short gastric veins (n = 3), splenic vein stump (n = 1), and the left gastroepiploic vein (n = 1). The total volumes of transplanted pancreatic digest in those receiving combined intraportal and splenic autografts (n = 3) were 15.8, 13.0, and 13.5 ml. The volumes in those receiving a splenic-alone autograft (n = 2) were 12.0 and 5 ml. The mean rise in portal pressure was 18 cm of water. Complications related to the splenic autograft included a wedge splenic infarct, an emergency splenectomy, and a portal vein thrombosis in one patient having a combined intraportal and splenic autograft. Two patients developed insulin independence, two patients were still insulin independent at 1-year follow-up, and all had normal HbA1c levels (mean 5.6, range 5.2–6.3). Splenic islet autotransplantation, after total pancreatectomy, does lead to insulin independence. However, in our experience the combined procedure has a high morbidity because of splenic infarction and venous thrombosis.
Neural Stem Cells: From Cell Biology to Cell Replacement
Authors: Armstrong Richard J. E. , Svendsen Clive N.
Published: June 2017

A large number of crippling neurological conditions result from the loss of certain cell populations from the nervous system through disease or injury, and these cells are not intrinsically replaced. Mounting evidence now suggests that replacement of depleted cell populations by transplantation may be of functional benefit in many such diseases. A diverse range of cell populations is vulnerable, and the loss of specific populations results in circumscribed deficits in different conditions. This diversity presents a considerable challenge if cell replacement therapy is to become widely applicable in the clinical domain, because each condition has specific requirements for the phenotype, developmental stage, and number of cells required. An ideal cell for universal application in cell replacement therapy would possess several key properties: it would be highly proliferative, allowing the ex vivo production of large numbers of cells from minimal donor material; it would also remain immature and phenotypically plastic such that it could differentiate into appropriate neural and glial cell types on, or prior to, transplantation. Critically, both proliferation and differentiation would be controllable. This review considers some of the evidence that stem cells exist in the central nervous system and that they may possess characteristics that make them ideal for broad application in cell replacement therapy.
Towards Neurotransplantation in Multiple System Atrophy: Clinical Rationale, Pathophysiological Basis, and Preliminary Experimental Evidence
Authors: Wenning Gregor Karl , Tison Francois , Scherfler Christoph , Puschban Zoe , Waldner Regina , Granata Roberta , Ghorayeb Imad , Poewe Werner
Published: June 2017

Multiple system atrophy (MSA) is a neurodegenerative disorder that occurs sporadically and causes parkinsonism, cerebellar, autonomic, urinary, and pyramidal dysfunction in many combinations. Progressive L-dopa-unresponsive parkinsonism due to underlying striatonigral degeneration dominates the clinical syndrome in the majority of cases (MSA-P subtype). MSA-P is characterized pathologically by degenerative changes in somatotopically related areas of the substantia nigra pars compacta and of the putamen. Furthermore, oligodendroglial cytoplasmic inclusions (GCIs) are observed throughout the cortico-striato-pallido-cortical loops and may contribute to the basal ganglia dysfunction. Neurotransplantation strategies are of potential interest in this disease, which causes marked and early disability and dramatically reduces life expectancy. A number of experimental MSA-P models have been employed to evaluate neurotransplantation approaches. Sequential nigral and striatal lesions using 6-hydroxydopamine and quinolinic acid (double toxin–double lesion approach) indicate that apomorphine-induced contralateral rotation is abolished by a secondary striatal lesion. Intrastriatal injection of mitochondrial respiratory chain toxins produces secondary excitotoxic striatal lesions combined with retrograde nigral degeneration and therefore provides an alternative single toxin–double lesion approach. Neurotransplantation in MSA-P animal models has been used to improve functional deficits by replacing lost nigral and/or striatal circuitry (neuroregenerative approach). The available data indicate that embryonic mesencephalic grafts alone or combined with striatal grafts partially reverse drug-induced rotation asymmetries without improving deficits of complex motor function. The potential neuroprotective efficacy of embryonic striatal grafts against striatal excitotoxicity is presently under investigation in the double toxin–double lesion MSA-P rat model. Anecdotal clinical evidence in one MSA-P patient misdiagnosed as Parkinson's disease indicates that embryonic mesencephalic grafts produce incomplete clinical benefit. Striatal co-grafts may increase functional improvement. Further experimental studies are required prior to the clinical application of embryonic neurotransplantation in MSA-P. Future research strategies should explore the effect of neurotransplantation in partial MSA-P rat models with less severe nigral and striatal degeneration, the feasibility of a primate model closely mimicking the human disease, and the replication of oligodendroglial dysfunction.
Exposure to Tissue Culture Conditions Can Adversely Affect Myoblast Behavior in Vivo in Whole Muscle Grafts: Implications for Myoblast Transfer Therapy
Authors: Smythe Gayle M. , Grounds Miranda D.
Published: June 2017

The effects of tissue culture conditions on the viability of myoblasts in whole muscles transplanted in vivo were investigated. Whole male (SJL/J) donor muscles were exposed to various tissue culture reagents and proteolytic enzymes, and allografted into female (SJL/J) host mice. Desmin immunohistochemistry was used to assess the numbers of myogenic cells (as an index of myoblast viability and the extent of regeneration) in tissue sections of whole-muscle grafts sampled on days 7 and 14. DNA quantitation with a Y-chromosome-specific probe was used to determine the total Y-1 sequence DNA (as an index of myoblast survival and proliferation) in whole-muscle grafts sampled on days 1, 3, and 7. In grafts exposed to serum-free medium, there was a delay in myoblast fusion at 7 days that was recovered by 14 days, but exposure to serum (10% or 20%) had a prolonged adverse effect on myotube formation at 14 days. DNA quantitation demonstrated that either serum-free culture medium or 10% serum enhanced the number of male cells within whole-muscle grafts at 7 days. Proteolytic digestion (even for 5 min) of whole muscles prior to grafting was extremely detrimental to myoblast survival and viability at 7 and 14 days. The unexpected finding of adverse effects of tissue culture conditions on the regeneration of whole-muscle grafts in vivo appears to parallel the major problem of the rapid death of isolated cultured donor myoblasts after injection in myoblast transfer therapy. The use of whole-muscle grafts provides an alternative and sensitive model to analyze the crucial effects of various tissue culture components on the subsequent survival and proliferation of myogenic cells in vivo.
Evaluation of an Intrathecal Immune Response in Amyotrophic Lateral Sclerosis Patients Implanted with Encapsulated Genetically Engineered Xenogeneic Cells
Authors: Zurn A. D. , Henry H. , Schluep M. , Aubert V. , Winkel L. , Eilers B. , Bachmann C. , Aebischer P.
Published: June 2017

A phase I/II clinical trial has been performed in 12 amyotrophic lateral sclerosis (ALS) patients to evaluate the safety and tolerability of intrathecal implants of encapsulated genetically engineered baby hamster kidney (BHK) cells releasing human ciliary neurotrophic factor (CNTF). These patients have been assessed for a possible intrathecal or systemic immune response against the implanted xenogeneic cells. Hundreds of pg CNTF/ml could be detected for several weeks in the cerebrospinal fluid (CSF) of 9 out of 12 patients, in 2 patients up to 20 weeks after capsule implantation. Slightly elevated leukocyte counts were observed in 6 patients. Clear evidence for a delayed humoral immune response was found in the CSF of only 3 patients out of 12 (patients #4, #6, and #10). Characterization of the antigen(s) recognized by the antibodies present in these CSF samples allowed to identify bovine fetuin as the main antigenic component. The defined medium used for maintaining the capsules in vitro before implantation contains bovine fetuin. Fetuin may therefore still be adsorbed to the surface of the cells and/or the polymer membrane, or be present in the medium surrounding the encapsulated cells at the time of implantation. Because of the insufficient availability of CSF samples, as well as the relatively poor sensitivity of the assays used, a weak humoral immune response against components of the implanted cells themselves cannot be excluded. However, the present study demonstrates that encapsulated xenogeneic cells implanted intrathecally can survive for up to 20 weeks in the absence of immunosuppression and that neither CNTF nor the presence of antibodies against bovine fetuin elicit any adverse side effects in the implanted patients.
Why Do Cultured Transplanted Myoblasts Die in Vivo? DNA Quantification Shows Enhanced Survival of Donor Male Myoblasts in Host Mice Depleted of CD4+ and CD8+ Cells or NK1.1+ Cells
Authors: Hodgetts Stuart I. , Beilharz Manfred W. , Scalzo Anthony A. , Grounds Miranda D.
Published: June 2017

Overcoming the massive and rapid death of injected donor myoblasts is the primary hurdle for successful myoblast transfer therapy (MTT), designed as a treatment for the lethal childhood myopathy Duchenne muscular dystrophy. The injection of male myoblasts into female host mice and quantification of surviving male DNA using the Y-chromosome-specific (Y1) probe allows the speed and extent of death of donor myoblasts to be determined. Cultured normal C57BL/10Sn male donor myoblasts were injected into untreated normal C57BL/10Sn and dystrophic mdx female host mice and analyzed by slot blots using a 32P-labeled Y1 probe. The amount of male DNA from donor myoblasts showed a remarkable decrease within minutes and by 1 h represented only about 10–18% of the 2.5 × 105 cells originally injected (designated 100%). This declined further over 1 week to approximately 1–4%. The host environment (normal or dystrophic) as well as the extent of passaging in tissue culture (early “P3” or late “P15–20” passage) made no difference to this result. Modulation of the host response by CD4+/CD8+-depleting antibodies administered prior to injection of the cultured myoblasts dramatically enhanced donor myoblast survival in dystrophic mdx hosts (15-fold relative to untreated hosts after 1 week). NK1.1 depletion also dramatically enhanced donor myoblast survival in dystrophic mdx hosts (21-fold after 1 week) compared to untreated hosts. These results provide a strategic approach to enhance donor myoblast survival in clinical trials of MTT.
Grafting of Nigral Tissue Hibernated with Tirilazad Mesylate and Glial Cell Line-Derived Neurotrophic Factor
Authors: Petersén Åsa , Hansson Oskar , Emgård Mia , Brundin Patrik
Published: June 2017

Transplantation of embryonic ventral mesencephalon is a potential therapy for patients with Parkinson's disease. As only around 5–10% of embryonic dopaminergic neurons survive grafting into the adult striatum, it is considered necessary to use multiple donor embryos. To increase the survival of the grafted dopaminergic neurons, the clinical transplantation program in Lund currently employs the lipid peroxidation inhibitor, tirilazad mesylate, in all solutions used during tissue storage, preparation, and transplantation. However, the difficulty in obtaining a sufficient number of donor embryos still remains an important limiting factor for the clinical application of neural transplantation. In many clinical transplantation programs, it would be a great advantage if human nigral donor tissue could be stored for at least 1 week. This study was performed in order to investigate whether storage of embryonic tissue at 4°C for 8 days can be applied clinically without creating a need to increase the number of donors. We compared the survival of freshly grafted rat nigral tissue, prepared according to the clinical protocol, with tissue transplanted after hibernation. Thus, in all groups tirilazad mesylate was omnipresent. One group of rats was implanted with fresh tissue and three groups with hibernated tissue with or without addition of glial cell line-derived neurotrophic factor (GDNF) in the hibernation medium and/or the final cell suspension. Earlier studies have suggested that GDNF improves the survival of hibernated nigral transplants. We found no statistically significant difference between the groups regarding graft survival after 3 weeks. However, there was a nonsignificant trend for fewer surviving dopaminergic neurons in grafts from hibernated tissue compared to fresh controls. Furthermore, we show that the addition of GDNF to the hibernation medium and/or to the final cell suspension does not significantly increase the survival of the dopaminergic neurons.
Strong, Long-Term Transgene Expression in Rat Liver Using Chicken β-Actin Promoter Associated with Cytomegalovirus Immediate-Early Enhancer (CAG Promoter)
Authors: Kosuga Motomichi , Enosawa Shin , Li Xiao-Kang , Suzuki Seiichi , Matsuo Nobutake , Yamada Masao , Roy-Chowdhury Jayanta , Koiwai Osamu , Okuyama Torayuki
Published: June 2017

For successful gene therapy in hepatic enzyme deficiencies, it is essential to use promoters that can maintain strong transcriptional activity for the long term in the liver. Using Gunn rats, a model animal for Crigler-Najjar syndrome type I, the long-term transcriptional function of the CAG promoter (a combination of chicken β-actin promoter and cytomegalovirus immediate-early enhancer) was evaluated in the rat liver. We constructed a plasmid pCAGGHUGT, containing expression cassettes of human bilirubin UDP-glucurono-syltransferase (BUGT) and hygromycin phosphotransferase, under the control of the CAG promoter and murine phosphoglycerate kinase promoter, respectively. Conditionally immortalized Gunn rat hepatocytes (IGRH), which had been established using mutant SV40 large T antigen (TST), were transfected with pCAGGHUGT. A stably transfected clone IGRHUGT, expressing a high level of BUGT, was obtained after selection with hygromycin. At 33°C, the cells doubled in number in approximately 72 h; however, at 37°C, cell proliferation stopped, indicating that the characteristic of temperature-dependent proliferation was retained in this clone. Ten million cells were injected into the spleen of syngeneic Gunn rats five times at 10-day intervals. Serum bilirubin levels were reduced by 45–50% at 70 days after the first transplantation and remained so throughout the duration of the study (120 days). These results suggested that the CAG promoter was able to maintain strong transcriptional activity in rat liver for at least 120 days.
Clonogenic Colony-Forming Ability of Flow Cytometrically Isolated Hepatic Progenitor Cells in the Murine Fetal Liver
Authors: Taniguchi Hideki , Kondo Reika , Suzuki Atsushi , Zheng Yun-Wen , Takada Yasutsugu , Fukunaga Kiyoshi , Seino Ken-Ichiro , Yuzawa Kenji , Otsuka Masaaki , Fukao Katashi , Nakauchi Hiromitsu
Published: June 2017

Stem cells are defined as cells having multilineage differentiation potential and self-renewal capability. Hepatic stem cells have aroused considerable interest not only because of their developmental importance but also for their therapeutic potential. However, their presence in the liver has not yet been demonstrated. With the use of a fluorescence-activated cell sorter (FACS) and monoclonal antibodies, we attempted to ascertain whether hepatic stem cells are present in the murine fetal liver. For this purpose, we optimized a cell isolation technique for FACS sorting of fetal liver cells. When isolated CD45–TER119– cells (the non-blood cell fraction in the fetal liver) were tested for their clonogenic colony-forming ability, mechanical dissociation (pipetting) was the most suitable cell isolation technique for FACS sorting. We confirmed that these colonies contained not only cells expressing hepatocyte markers but also cells expressing cholangiocyte markers. To identify hepatic stem cells, studies must focus on CD45–TER119– cells in the murine fetal liver.
Inducing Proliferation of Human Amniotic Epithelial (HAE) Cells for Cell Therapy
Authors: Terada Satoshi , Matsuura Keiko , Enosawa Shin , Miki Masao , Hoshika Akinori , Suzuki Seiichi , Sakuragawa Norio
Published: June 2017

Probably because amnion is derived from the fetus and is exposed to the maternal immune system, human amniotic epithelial (HAE) cells do not express the HLA-A, -B, -C, or -DR antigens on their surfaces, suggesting that HAE cells do not induce rejection (immune reaction) after allotransplantation. And the amnion, like the placenta, is useless to the mother and child after birth. Therefore, HAE cells or tissues were expected to be suitable for allotransplantation. Because HAE cells produce large amounts of enzymes, amnion transplantation has been carried out in order to correct inborn errors of metabolism by supplementing lysosomal enzyme deficiencies. However, several problems remain before amnion allotransplantation can be accepted as effective. The HAE cell population is limited, because the maximum number of HAE cells obtainable from one donor is about 2 × 108 cells, and HAE cells proliferate poorly in in vitro culture. In this study, we aimed at increasing the HAE cell population in vitro. First, we investigated the effect of several cytokines on HAE cell proliferation and found that hepatocyte growth factor (HGF), epidermal growth factor (EGF), and transforming growth factor-β stimulated it, whereas IL-6 and LIF inhibited it. Second, we investigated the effects of amniotic fluid on HAE cell proliferation and observed that IL-6 in amniotic fluid inhibits it. Then, to inhibit the dying of cells, we attempted to inhibit apoptosis (one mode of cell death). Treatment with caspase III inhibitor increased the cell viability of HAE cells by 20%.
Modified Subcutaneous Tissue with Neovascularization is Useful as the Site for Pancreatic Islet Transplantation
Authors: Kawakami Yoshiyuki , Iwata Hiroo , Gu Yuanjung , Miyamoto Masaaki , Murakami Yoshinobu , Yamasaki Toru , Cui Wanxing , Ikada Yoshito , Imamura Masayuki , Inoue Kazutomo
Published: June 2017

The success rate of subcutaneous transplantation of pancreatic islets has been extremely low. Insufficient oxygen supply to the grafted islets is one possible major obstacle to the preservation of graft function. This study attempted to use basic fibroblast growth factor (bFGF) in subcutaneous transplantation to induce neovascularization and a sufficient blood flow around the space formed for grafted islets in the subcutaneous tissues. A bFGF-releasing device was designed enclosing bFGF in a polyethylene terephthalate mesh bag coated with polyvinylalcohol hydrogel. In the vascularized group (n = 5), two bFGF-releasing devices were implanted bilaterally into the subcutaneous tissue of the back of streptozotocin-induced diabetic Lewis rats. One week after implantation, isolated rat islets (5000) were syngeneically transplanted subcutaneously after the removal of the devices. In the control group (n = 5), no devices were implanted and the same number of rat islets was transplanted directly. One week after the implantation of the devices into the test animals, a thick, well-vascularized capsule was observed in the subcutaneous site. All vascularized recipient rats showed significant decreases in nonfasting blood glucose and maintained normoglycemia for more than 1 month after islet transplantation. However, in the control group, all rats failed to achieve normoglycemia after transplantation. This study provides evidence that the subcutaneous tissue is a promising site for pancreatic islet transplantation, offering convincing advantages in acceptability for diabetic recipients. Establishment of this subcutaneous islet transplantation technique will afford some new perspectives on successful clinical islet transplantation.
Hibernated Human Fetal Striatal Tissue: Successful Transplantation in a Rat Model of Huntington's Disease
Authors: Hurelbrink Carrie B. , Armstrong Richard J. E. , Barker Roger A. , Dunnett Stephen B. , Rosser Anne E.
Published: June 2017

The use of fresh human fetal tissue in neural transplantation presents considerable logistical difficulties and limits the clinical applicability of this promising therapy. This study compared the survival of human fetal striatal tissue that had been stored for 24 h in a defined hibernation medium with that of fresh human fetal striatal tissue following xenotransplantation in a rat model of Huntington's disease (HD). Six to 7 weeks postgrafting, there was no significant difference between fresh and hibernated grafts in volume or in various striatal phenotypic markers, although there was a trend towards decreased graft volume. We conclude that short-term hibernation of this tissue is without significant adverse effects on the survival of grafted human fetal striatal tissue. This has important implications for the practical implementation of clinical neural transplant programs in HD.
No Evidence of Infection with Porcine Endogenous Retrovirus in Recipients of Encapsulated Porcine Islet Xenografts
Authors: Elliott R. B. , Escobar L. , Garkavenko O. , Croxson M. C. , Schroeder B. A. , Mcgregor M. , Ferguson G. , Beckman N. , Ferguson S.
Published: June 2017

Transplantation of pig tissues into humans has the potential for cotransferring pig infections. Knowledge of the epidemiology of pig infections transmissible to humans allows the development of risk limitation strategies at the source herd level, but potentially infectious pig endogenous retrovirus (PERV) is ubiquitous in all domestic pigs and therefore is not avoidable. Using a specific and sensitive RT-PCR and nested PCR for PERV nucleic acids with primers, the screening of pigs from New Zealand herds for the presence and expression of the PERV was conducted. The presence of PERV proviral DNA (pol and env region) and viral RNA was demonstrated in all tested pig tissues including pancreas, liver, spleen, brain, heart, and PBMC. Using the same assays it was established that different tissues (liver, spleen, and heart) of nude and nonobese diabetic (NOD) mice previously transplanted with nonencapsulated pig islets were PERV DNA and RNA negative. Alginate polylysine capsules prepared with encapsulated pig islets were tested for possible leakage of viral particles or viral nucleic acids. RNA was extracted from the supernatant of viable encapsulated pig islet cells grown in culture for 2 months. No evidence of PERV RNA or of cellular nucleic acids could be found. Two adult type I diabetic subjects were transplanted with 1 × 106 neonatal pig islets encased in alginate capsules into the peritoneal cavity. One patient was immunosuppressed. Both showed evidence of graft function (up to 34% reduction in insulin dose, corresponding increase in serum pig C-peptide) for up to 2 years. DNA and RNA were extracted from PBMC and blood plasma of both patients at 19 months posttransplant. No evidence of PERV proviral DNA or RNA could be detected. Piglet islets contain PERV DNA and RNA, but this does not traverse the capsules used or produce any evidence of infection in nude and nonobese diabetic (NOD) mice or humans.