bluuebunny/crossref_metadata_2025 huggingface dataset:
Using new non-invasive quick method to detect Borrelia Burgdorferi (B.B.) infection from specific parts of the heart in “seemingly normal” ECGs, and from the ECGs of Atrial Fibrillation (AF), a majority of AF ECGs are found to have: 1) Significant B.B. infection,
2) Markedly increased ANP, 3) Increased Cardiac Troponin I & 4) Markedly reduced Taurine. These 4 factors were mainly localized at infected areas of the SA node area, R-&L-Atria & pulmonary veins at the L-atrium
Authors: Omura Yoshiaki , Lu Dominic , Jones Marilyn K. , Nihrane Abdallah , Duvvi Harsha , Yapor Dario , Shimotsuura Yasuhiro , Ohki Motomu
Published: February 2016

Lyme disease is found in a majority of people we tested. Once Borrelia Burgdorferi (B.B.) spirochete enters human body, it not only causes pain by infecting joints, but it also often enters the brain and the heart. Infection of brain can be quickly detected from the pupil and
infection of the heart by ECGs non-invasively. By evaluating recorded ECGs of atrial fibrillation (AF), using U.S. patented non-invasive highly sensitive electromagnetic field (EMF) resonance phenomenon between 2 identical molecules or between a molecule and its antibody, we examined 25 different
AF patients' ECGs and found the majority of them suffer from various degrees of B.B. spirochete infection in SA node areas, also in the right & left atria, and pulmonary vein near and around its junction at left atrium & lesser degrees of infection at the AV node & His Bundle.
When B.B. infection reaches over 224∼600ng or higher at these areas, AF often appears in the majority of all AF analyzed. In order to develop AF, the 4 abnormal factors must be present simultaneously: 1) B.B. infection must be increased to 224∼600ng or higher, 2) Atrial Natriuretic
Peptide (ANP) must be markedly reduced from normal value of less than 4ng to over 100∼400ng, 3) A significant increase of Cardiac Troponin I from normal value of less than 3ng to over 12ng and 4) Taurine must also be markedly reduced from normal value of 4∼6ng to 0.25ng. These 4 changes
were mainly found only at infected sites of the SA node area, both atria and between the end of the T wave & the beginning of the SA node area, which corresponds to U waves at recorded ECG. Origin of the U wave is mainly due to abnormal electrical potential of pulmonary vein at L-atrium.
If all 4 factors do not occur at the infection site, no AF will develop. In seemingly normal ECGs, if using this method, one can detect invisible B.B. infection in early stages. Long before AF appears, AF can be prevented by improved treatment with Amoxicillin 500ng 3 times/day + Taurine 175mg
x3 times/day, with or without EPA 180mg & DHA 120mg, to avoid serious current limitations in the use of Doxycycline 100mg 2 times/day, for 4 weeks.
Electro-Acupuncture at GV.4 Improves Functional Recovery in paralyzed rats after a Traumatic Spinal Cord Injury
Authors: JuarezBecerril Oswaldo , SalgadoCeballos Hermelinda , AnguianoSolis Candy , AlvaradoSanchez Belen , Hernandez Monica Elba Lopez , DiazRuiz Araceli , TorresCastillo Sergio
Published: February 2016

In the present study, the effect of electro-acupuncture (EA) on the oxidative stress, the spinal cord tissue preservation and the recovery of motor function was evaluated after a traumatic spinal cord injury (TSCI). Long Evans rats were randomized into five groups: 1. Sham; 2. TSCI
without treatment; 3. TSCI + EA (acupoint GV.4); 4. TSCI + EA (acupoint GV.26) and 5. TSCI + EA (GV.4 + GV.26). The EA was performed with an Electro-Acupunctoscope, AWQ-104L Digital, wave dense-dispersed, current intensity 2.5mA and frequency 2-100Hz for 30 minutes. The biochemical results
showed a significant increase in the hydroxyl radical concentration in group 2 (3.1±1.4nmol) compared with groups 1 (1.8±0.5nmol) and 4 (2.4±1.1nmol) (p<0.05), whereas in group 4 (4.8±1.8nmol), there was a significant increase in lipid peroxidation when compared
with group 1 (1.7±0.5nmol) (p<0.05). The BBB motor function score in the paralyzed hind limbs (normal BBB=21points) was greater in groups 3 (15.2 points) and 5 (13.5 points) in comparison with groups 2 (11.4 points) and 4 (9.3 points) (p<0.05). The quantity of preserved spinal
cord tissue was greater in group 3 (6582.7±20μm2) than in groups 2 (5262.4±20μm2), 4 (3995.6±26μm2) and 5 (4266.7±22μm2). Although EA in GV.26 decreases hydroxyl radical concentration (50%), it significantly
increases lipid peroxidation (45%), while stimulation of GV.4 decreases oxidative stress (15%), preserves spinal cord tissue (25%) and improves recovery of motor function in the hind limbs of rats with paralysis (18.1%) compared with untreated group. These findings suggest that EA in GV.4
may be a therapeutic alternative on TSCI.
Characterization of Innate Immunity in an Extended Whole Blood Model of Human Islet Allotransplantation
Authors: Hårdstedt Maria , Lindblom Susanne , Karlsson-Parra Alex , Nilsson Bo , Korsgren Olle
Published: February 2016

The instant blood-mediated inflammatory reaction (IBMIR) has been studied in whole blood models of human allo-islet transplantation for short periods (< 6 h). Beyond this time frame the innate response to intraportally transplanted islets is less well described. A novel whole blood model was applied to study blood–islet–graft interactions up to 48 h. Heparinized polyvinyl chloride tubing was sealed into small bags containing venous blood together with allogeneic human islets and exocrine tissue, respectively. The bags were attached to a rotating wheel (37°C). Concentrated glucose and sodium hydrogen carbonate were added every 12 h to maintain physiological limits for sustained immune cell functions. Plasma was collected at repeated time points for analyses of coagulation/complement activation and chemokine/cytokine production. Immune cell infiltration was analyzed using immunohistochemistry. Coagulation and platelet activation markers, thrombin–antithrombin complex (TAT) and soluble CD40 ligand (sCD40L) showed early high concentrations (at 6–12 h). sC5b-9 steadily increased over 48 h. At 6 h neutrophils and monocytes surrounded the clotted cellular grafts with a following massive infiltration of neutrophils. High and increasing concentrations of CXCR1/2 ligands [IL-8 and growth-regulated oncogene α/β/γ (Gro-α/β/γ)] and IL-6 were produced in response to human islets and exocrine tissue. The CCR2 ligand monocyte chemoattractant protein 1 (MCP-1) exhibited increasing concentrations in response to exocrine tissue. The CXCR3 ligand interferon-inducible T cell a chemoattractant (I-TAC) was produced in response to both human islets and exocrine tissue from 6 h. Monokine induced by g interferon (Mig) and interferon γ-induced protein 10 (IP-10) showed a later response, preferentially to exocrine tissue and with larger variations among preparations. An extended blood model of clinical islet transplantation allowed characterization of early immune activation in response to human islets and exocrine tissue. Increased production of chemokines targeting CXCR1/2, CCR2, and CXCR3 was observed, accompanied by massive intraislet neutrophil infiltration over 48 h. The model proved to be useful in exploring early blood-mediated reactions to cellular transplants and has relevance for evaluation of pharmacological interventions to prevent graft loss.
Human Fetal Progenitor Tenocytes for Regenerative Medicine
Authors: Grognuz A. , Scaletta C. , Farron A. , Raffoul W. , Applegate L. A.
Published: February 2016

Tendon injuries are very frequent and affect a wide and heterogeneous population. Unfortunately, the healing process is long with outcomes that are not often satisfactory due to fibrotic tissue appearance, which leads to scar and adhesion development. Tissue engineering and cell therapies emerge as interesting alternatives to classical treatments. In this study, we evaluated human fetal progenitor tenocytes (hFPTs) as a potential cell source for treatment of tendon afflictions, as fetal cells are known to promote healing in a scarless regenerative process. hFPTs presented a rapid and stable growth up to passage 9, allowing to create a large cell bank for off-the-shelf availability. hFPTs showed a strong tenogenic phenotype with an excellent stability, even when placed in conditions normally inducing cells to differentiate. The karyotype also indicated a good stability up to passage 12, which is far beyond that necessary for clinical application (passage 6). When placed in coculture, hFPTs had the capacity to stimulate human adult tenocytes (hATs), which are responsible for the deposition of a new extracellular matrix during tendon healing. Finally, it was possible to distribute cells in porous or gel scaffolds with an excellent survival, thus permitting a large variety of applications (from simple injections to grafts acting as filling material). All of these results are encouraging in the development of an off-the-shelf cell source capable of stimulating tendon regeneration for the treatment of tendon injuries.
Plasma Derived from Human Umbilical Cord Blood Modulates Mitogen-Induced Proliferation of Mononuclear Cells Isolated from the Peripheral Blood of ALS Patients
Authors: Eve David J. , Ehrhart Jared , Zesiewicz Theresa , Jahan Israt , Kuzmin-Nichols Nicole , Sanberg Cyndy Davis , Gooch Clifton , Sanberg Paul R. , Garbuzova-Davis Svitlana
Published: February 2016

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease characterized by degeneration of motor neurons in the spinal cord and brain. This disease clinically manifests as gradual muscular weakness and atrophy leading to paralysis and death by respiratory failure. While multiple interdependent factors may contribute to the pathogenesis of ALS, increasing evidence shows the possible presence of autoimmune mechanisms that promote disease progression. The potential use of plasma derived from human umbilical cord blood (hUCB) as a therapeutic tool is currently in its infancy. The hUCB plasma is rich in cytokines and growth factors that are required for growth and survival of cells during hematopoiesis. In this study, we investigated the effects of hUCB plasma on the mitogen-induced proliferation of mononuclear cells (MNCs) isolated from the peripheral blood of ALS patients and apoptotic activity by detection of caspase 3/7 expression of the isolated MNCs in vitro. Three distinct responses to phytohemagglutinin (PHA)-induced proliferation of MNCs were observed, which were independent of age, disease duration, and the ALS rating scale: Group I responded normally to PHA, Group II showed no response to PHA, while Group III showed a hyperactive response to PHA. hUCB plasma attenuated the hyperactive response (Group III) and potentiated the normal response in Group I ALS patients, but did not alter that of the nonresponders to PHA (Group II). The elevated activity of caspase 3/7 observed in the MNCs from ALS patients was significantly reduced by hUCB plasma treatment. Thus, study results showing different cell responses to mitogen suggest alteration in lymphocyte functionality in ALS patients that may be a sign of immune deficiency in the nonresponders and autoimmunity alterations in the hyperactive responders. The ability of hUCB plasma to modulate the mitogen cell response and reduce caspase activity suggests that the use of hUCB plasma alone, or with stem cells, may prove useful as a therapeutic in ALS patients.
In Vivo Tracking of Human Neural Progenitor Cells in the Rat Brain Using Magnetic Resonance Imaging is Not Enhanced by Ferritin Expression
Authors: Bernau Ksenija , Lewis Christina M. , Petelinsek Anna M. , Reagan Matthew S. , Niles David J. , Mattis Virginia B. , Meyerand M. Elizabeth , Suzuki Masatoshi , Svendsen Clive N.
Published: February 2016

Rapid growth in the field of stem cell research has generated a lot of interest in their therapeutic use, especially in the treatment of neurodegenerative diseases. Specifically, human neural progenitor cells (hNPCs), unique in their capability to differentiate into cells of the neural lineage, have been widely investigated due to their ability to survive, thrive, and migrate toward injured tissues. Still, one of the major roadblocks for clinical applicability arises from the inability to monitor these cells following transplantation. Molecular imaging techniques, such as magnetic resonance imaging (MRI), have been explored to assess hNPC transplant location, migration, and survival. Here we investigated whether inducing hNPCs to overexpress ferritin (hNPCsFer), an iron storage protein, is sufficient to track these cells long term in the rat striatum using MRI. We found that increased hypointensity on MRI images could establish hNPCFer location. Unexpectedly, however, wild-type hNPC transplants were detected in a similar manner, which is likely due to increased iron accumulation following transplantation-induced damage. Hence, we labeled hNPCs with superparamagnetic iron oxide (SPIO) nanoparticles to further increase iron content in an attempt to enhance cell contrast in MRI. SPIO-labeling of hNPCs (hNPCs-SPIO) achieved increased hypointensity, with significantly greater area of decreased T2* compared to hNPCFer ( p < 0.0001) and all other controls used. However, none of the techniques could be used to determine graft rejection in vivo, which is imperative for understanding cell behavior following transplantation. We conclude that in order for cell survival to be monitored in preclinical and clinical settings, another molecular imaging technique must be employed, including perhaps multimodal imaging, which would utilize MRI along with another imaging modality.
Pluripotent Stem Cells to Rebuild a Kidney: The Lymph Node as a Possible Developmental Niche
Authors: Francipane Maria Giovanna , Lagasse Eric
Published: February 2016

Kidney disease poses a global challenge. Stem cell therapy may offer an alternative therapeutic approach to kidney transplantation, which is often hampered by the limited supply of donor organs. While specific surface antigen markers have yet to be identified for the analysis and purification of kidney stem/progenitor cells for research or clinical use, the reprogramming of somatic cells to pluripotent cells and their differentiation into the various kidney lineages might represent a valuable strategy to create a renewable cell source for regenerative purposes. In this review, we first provide an overview of kidney development and explore current knowledge about the role of extra- and intrarenal cells in kidney repair and organogenesis. We then discuss recent advances in the 1) differentiation of rodent and human embryonic stem cells (ESCs) into renal lineages; 2) generation of induced pluripotent stem cells (iPSCs) from renal or nonrenal (kidney patient-derived) adult cells; 3) differentiation of iPSCs into renal lineages; and 4) direct transcriptional reprogramming of adult renal cells into kidney progenitor cells. Finally, we describe the lymph node as a potential three-dimensional (3D) in vivo environment for kidney organogenesis from pluripotent stem cells.
Injection of Dental Pulp Stem Cells Promotes Healing of Damaged Bladder Tissue in a Rat Model of Chemically Induced Cystitis
Authors: Hirose Yujiro , Yamamoto Tokunori , Nakashima Misako , Funahashi Yasuhito , Matsukawa Yoshihisa , Yamaguchi Masaya , Kawabata Shigetada , Gotoh Momokazu
Published: February 2016

Dental pulp stem cells (DPSCs) are reported as sources of mesenchymal stem cells (MSCs). MSCs are used as cell therapy options for various diseases. The present study examined the healing effects of DPSC injection on damaged bladder tissue in a chemically induced cystitis rat model. Cystitis was induced by hydrochloride injection into the bladder of female F344/NSlc rats. On the following day, DPSCs suspended in phosphate-buffered saline (PBS) were injected into the bladder and maintained for 1 h (DPSC injection group), while PBS alone was injected as the standard for comparison (PBS injection group). After 2 days following injection, considerable submucosal edema, vascular structure destruction, hemorrhage, and inflammatory cell invasion were observed both in the DPSC and PBS injection groups, with no difference in their degree of submucosal edema and hemorrhage. Six days after injection, vascular structure regeneration was observed in both groups; however, unlike the DPSC injection group, the PBS injection group showed traces of submucosal edema and hemorrhage. These results correlated with tissue concentrations of myeloperoxidase (MPO) and the inflammatory cytokines IL-1β, IL-6, and TNF-α. Furthermore, the intercontraction interval was prolonged, and the frequency of nociceptive behaviors was reduced in the DPSC injection group compared with the PBS injection group. DPSCs were found on the bladder epithelium until day 3 after injection. In the DPSC-conditioned media (CM), the trophic factors FGF-2, VEGF, and the C-C and C-X-C families of chemokines were detected. The results of DPSC injection into the cystitis rat model suggested that the injected cells promote the healing of the damaged bladder tissue by exerting various trophic effects while localizing on the bladder epithelium and that MSC injection is a potential novel therapy for interstitial cystitis/painful bladder syndrome.
High Seeding Density Induces Local Hypoxia and Triggers a Proinflammatory Response in Isolated Human Islets
Authors: Brandhorst Daniel , Brandhorst Heide , Mullooly Niamh , Acreman Samuel , Johnson Paul R. V.
Published: February 2016

Hypoxia is the main threat to morphological and functional integrity of isolated pancreatic islets. Lack of oxygen seems to be of particular importance for functionality of encapsulated islets. The present study was initiated as an experimental model for the environment experienced by human islets in a confined space present during culture, shipment, and in an implanted macrodevice. Quadruplicate aliquots of isolated human islets ( n = 12) were cultured for 24 h at 37°C under normoxic conditions using 24-well plates equipped with 8-μm pore size filter inserts and filled with islet aliquots adjusted to obtain a seeding density of 75, 150, 300, or 600 IEQ/cm2. After culture viability, glucose-stimulated insulin release, DNA content as well as Bax and Bcl-2 gene expression were measured. Culture supernatants were collected to determine production of VEGF and MCP-1. Viability correlated inversely with IEQ seeding density ( r = −0.71, p < 0.001), while the correlation of VEGF and MCP-1 secretion with seeding density was positive ( r = 0.78, p < 0.001; r = 0.54, p < 0.001). Decreased viability corresponded with a significant increase in the Bax/Bcl-2 mRNA ratio at 300 and 600 IEQ/cm2and with a sigificantly reduced glucose-stimulated insulin secretion and insulin content compared to 75 or 150 IEQ/cm2( p < 0.01). The present study demonstrates that the seeding density is inversely correlated with islet viability and in vitro function. This is associated with a significant increase in VEGF and MCP-1 release suggesting a hypoxic and proinflammatory islet microenvironment.
Digital Image Analysis to Assess Quantity and Morphological Quality of Isolated Pancreatic Islets
Authors: Wang Ling-Jia , Kaufman Dixon B.
Published: February 2016

Quantity and quality assessment of human pancreatic islets are essential processes to define a safe and potent quality product used for clinical transplantation. The conventional method of manual assessment has been used in the field for longer than two decades. The high degree of variability in product quantity and lack of archival imaging records of the product for verification are two major disadvantages of using the manual method for quantity and quality assessment of human pancreatic islets. Investigators have developed promising new methods for technical improvement. In this study, we briefly review the published methods and highlight the advantages of digital imaging analysis (DIA) when compared to the manual method. The application of DIA reduces measurement variability and increases the precision of islet equivalent (IEQ) determination for batch analysis. It produces images that can be archived for retrospective analysis and validation, and the data can be transmitted electronically for off-site analysis. These features are important for quality pancreatic islet assessment and are consistent with FDA requirements of current good manufacturing practice for clinical islet transplantation.
Mesenchymal Stem Cells and Their Clinical Applications in Osteoarthritis
Authors: Chang Yu-Hsun , Liu Hwan-Wun , Wu Kun-Chi , Ding Dah-Ching
Published: February 2016

Osteoarthritis is a chronic degenerative joint disorder characterized by articular cartilage destruction and osteophyte formation. Chondrocytes in the matrix have a relatively slow turnover rate, and the tissue itself lacks a blood supply to support repair and remodeling. Researchers have evaluated the effectiveness of stem cell therapy and tissue engineering for treating osteoarthritis. All sources of stem cells, including embryonic, induced pluripotent, fetal, and adult stem cells, have potential use in stem cell therapy, which provides a permanent biological solution. Mesenchymal stem cells (MSCs) isolated from bone marrow, adipose tissue, and umbilical cord show considerable promise for use in cartilage repair. MSCs can be sourced from any or all joint tissues and can modulate the immune response. Additionally, MSCs can directly differentiate into chondrocytes under appropriate signal transduction. They also have immunosuppressive and anti-inflammatory paracrine effects. This article reviews the current clinical applications of MSCs and future directions of research in osteoarthritis.
Direct Comparison of Rat- and Human-Derived Ganglionic Eminence Tissue Grafts on Motor Function
Authors: Lelos Mariah J. , Roberton Victoria H. , Vinh Ngoc-Nga , Harrison Carl , Eriksen Peter , Torres Eduardo M. , Clinch Susanne P. , Rosser Anne E. , Dunnett Stephen B.
Published: February 2016

Huntington's disease (HD) is a debilitating, genetically inherited neurodegenerative disorder that results in early loss of medium spiny neurons from the striatum and subsequent degeneration of cortical and other subcortical brain regions. Behavioral changes manifest as a range of motor, cognitive, and neuropsychiatric impairments. It has been established that replacement of the degenerated medium spiny neurons with rat-derived fetal whole ganglionic eminence (rWGE) tissue can alleviate motor and cognitive deficits in preclinical rodent models of HD. However, clinical application of this cell replacement therapy requires the use of human-derived (hWGE), not rWGE, tissue. Despite this, little is currently known about the functional efficacy of hWGE. The aim of this study was to directly compare the ability of the gold standard rWGE grafts, against the clinically relevant hWGE grafts, on a range of behavioral tests of motor function. Lister hooded rats either remained as unoperated controls or received unilateral excitotoxic lesions of the lateral neostriatum. Subsets of lesioned rats then received transplants of either rWGE or hWGE primary fetal tissue into the lateral striatum. All rats were tested postlesion and postgraft on the following tests of motor function: staircase test, apomorphine-induced rotation, cylinder test, adjusting steps test, and vibrissae-evoked touch test. At 21 weeks postgraft, brain tissue was taken for histological analysis. The results revealed comparable improvements in apomorphine-induced rotational bias and the vibrissae test, despite larger graft volumes in the hWGE cohort. hWGE grafts, but not rWGE grafts, stabilized behavioral performance on the adjusting steps test. These results have implications for clinical application of cell replacement therapies, as well as providing a foundation for the development of stem cell-derived cell therapy products.
Intracranial Transplantation of Hypoxia-Preconditioned iPSC-Derived Neural Progenitor Cells Alleviates Neuropsychiatric Defects after Traumatic Brain Injury in Juvenile Rats
Authors: Wei Zheng Zachory , Lee Jin Hwan , Zhang Yongbo , Zhu Yan Bing , Deveau Todd C. , Gu Xiaohuan , Winter Megan M. , Li Jimei , Wei Ling , Yu Shan Ping
Published: February 2016

Traumatic brain injury (TBI) is a common cause of mortality and long-term morbidity in children and adolescents. Posttraumatic stress disorder (PTSD) frequently develops in these patients, leading to a variety of neuropsychiatric syndromes. Currently, few therapeutic strategies are available to treat juveniles with PTSD and other developmental neuropsychiatric disorders. In the present investigation, postnatal day 14 (P14) Wistar rats were subjected to TBI induced by a controlled cortical impact (CCI) (velocity = 3 m/s, depth = 2.0 mm, contact time = 150 ms). This TBI injury resulted in not only cortical damages, but also posttrauma social behavior deficits. Three days after TBI, rats were treated with intracranial transplantation of either mouse iPSC-derived neural progenitor cells under normal culture conditions (N-iPSC-NPCs) or mouse iPSC-derived neural progenitor cells pretreated with hypoxic preconditioning (HP-iPSC-NPCs). Compared to TBI animals that received N-iPSC-NPCs or vehicle treatment, HP-iPSC-NPC-transplanted animals showed a unique benefit of improved performance in social interaction, social novelty, and social transmission of food preference tests. Western blotting showed that HP-iPSC-NPCs expressed significantly higher levels of the social behavior-related genes oxytocin and the oxytocin receptor. Overall, HP-iPSC-NPC transplantation exhibits a great potential as a regenerative therapy to improve neuropsychiatric outcomes after juvenile TBI.
Assessment of Platelet Activation and Immature Platelet Fraction as Predictors of Platelet Engraftment after Hematopoietic Stem Cell Transplantation
Authors: Jaing Tang-Her , Chen Shih-Hsiang , Wen Yu-Chuan , Chang Tsung-Yen , Huang Jing-Long , Tsay Pei-Kwei
Published: February 2016

Delayed platelet engraftment is a well-known complication of umbilical cord blood transplantation (CBT). Megakaryocytes derived from cord blood (CB) in vitro are smaller than megakaryocytes derived from bone marrow (BM) in adults. A small megakaryocyte size might contribute to delayed megakaryocytic maturation. This study included 37 patients undergoing hematopoietic stem cell transplantation (HSCT) at Chang Gung Children's Hospital between July 2011 and June 2013. Blood samples were obtained at different times: preconditioning and post-HSCT days 56 and 97. To test whether platelet activation persists posttransplantation, two commonly used platelet activation marker antibodies, CD62P (P-selectin) and CD42b, were evaluated using whole blood flow cytometry, combining thiazole orange and anti-CD41a staining, to assess reticulated platelets. Serial peripheral blood (PB) samples were obtained posttransplantation from patients undergoing CBT (CBT group; n = 15) and mobilized peripheral blood transplantation (PBT group; n = 22). Platelet activation in the postengraftment samples was considerably higher in the PBT group than the CBT group. Moreover, immature platelet fractions (IPF) were higher in the CBT group. Our results emphasize the role of IPF for dynamic prediction of platelet engraftment in CBT.
Stroke Serum Priming Modulates Characteristics of Mesenchymal Stromal Cells by Controlling the Expression miRNA-20a
Authors: Kim Eun Hee , Kim Dong Hee , Kim Hye Ree , Kim Soo Yoon , Kim Hyeon Ho , Bang Oh Young
Published: February 2016

Transplantation of mesenchymal stem cells (MSCs) expanded with fetal bovine serum (FBS) has some limitations, including the requirement of a long culture period to obtain a sufficient amount of stem cells. Priming of MSCs with serum from patients with ischemic stroke (stroke serum) increased the proliferation rate and the neurorestorative capacity of MSCs. We hypothesized that this novel priming method increases the proliferation rate of MSCs via the regulation of microRNAs (miRs). Thus, we investigated miR profiling in stroke serum-primed MSCs and tested whether the regulation of certain miRs may affect the proliferation rate of rat MSCs. The proliferation rate of MSCs cultured with stroke serum was higher than that of MSCs cultured with normal serum or FBS. Using miR microarray analysis, we compared the miR expression profiles between MSCs cultured in FBS and in stroke serum. Among miRs associated with cell proliferation, miR-20a was most significantly increased. Similarly, miR-20a was increased in MSCs obtained from the bone marrow of stroke rats compared with MSCs from normal rats. Furthermore, the deregulation of miR-20a by the transfection of MSCs with pre-miR-20a or anti-miR-20a was significantly correlated with the increased proliferation rate of MSCs. The overexpression of miR-20a in MSCs cultured in FBS improved the proliferation rate, while the knockdown of endogenous miR-20a decreased the proliferation rate. In addition, miR-20a promoted proliferation by suppressing the expression of p21 cyclin-dependent kinase inhibitor 1 (CDKN1A). A dual-luciferase reporter assay showed that CDKN1A is a target of miR-20a. Our findings indicate that stroke serum priming upregulated the expression of miR-20a, which promoted MSC proliferation by regulating the cell cycle inhibitor p21 CDKN1A, and suggest the possible roles of priming methods in modulating the characteristics of MSCs by controlling the expression of miR in MSCs.
Analgesic Effect of Recombinant GABAergic Cells in a Model of Peripheral Neuropathic Pain
Authors: Jergova Stanislava , Gajavelli Shyam , Varghese Mathew S. , Shekane Paul , Sagen Jacqueline
Published: February 2016

Chronic neuropathic pain represents a clinically challenging state with a poor response to current treatment options. Long-term management of chronic pain is often associated with the development of tolerance, addiction, and other side effects, reducing the therapeutic value of treatment. Alternative strategies based on cell therapy and gene manipulation, balancing the inhibitory and excitatory events in the spinal cord, may provide sustained pain relief in the long term. Transplantation of GABAergic cells has been successfully used to enhance inhibition and to restore physiological spinal pain processing. However, since the underlying mechanism of chronic pain development involves changes in several pain-signaling pathways, it is essential to develop an approach that targets several components of pain signaling. Recombinant cell therapy offers the possibility to deliver additional analgesic substances to the restricted area in the nervous system. The current study explores the analgesic potential of genetically modified rat embryonic GABAergic cells releasing a peptidergic NMDA receptor antagonist, Serine1-histogranin (SHG). Overactivation of glutamate NMDA receptors contributes to the hyperexcitability of spinal neurons observed in chronic pain models. Our approach allows us to simultaneously target spinal hyperexcitability and reduced inhibitory processes. Transplantable cells were transduced by viral vectors encoding either one or six copies of SHG cDNAs. The analgesic potential of recombinant cells after their intraspinal transplantation was evaluated in a model of peripheral nerve injury. Enhanced reduction of hypersensitivity to thermal and mechanical stimuli was observed in animals treated by recombinant cells compared to the nonrecombinant group. The recombinant peptide was detected in the spinal tissue, suggesting its successful production by transplanted cells. Our results demonstrate the feasibility of using recombinant cells releasing adjunct analgesic peptides in the therapy of neuropathic pain.
Experimental Support for the Ecoimmunity Theory: Distinct Phenotypes of Nonlymphocytic Cells in SCID and Wild-Type Mice
Authors: Ochayon David E. , Baranovski Boris M. , Malkin Peter , Schuster Ronen , Kalay Noa , Ben-Hamo Rotem , Sloma Ido , Levinson Justin , Brazg Jared , Efroni Sol , Lewis Eli C. , Nevo Uri
Published: February 2016

Immune tolerance toward “self” is critical in multiple immune disorders. While there are several mechanisms to describe the involvement of immune cells in the process, the role of peripheral tissue cells in that context is not yet clear. The theory of ecoimmunity postulates that interactions between immune and tissue cells represent a predator–prey relationship. A lifelong interaction, shaped mainly during early ontogeny, leads to selection of nonimmune cell phenotypes. Normally, therefore, nonimmune cells that evolve alongside an intact immune system would be phenotypically capable of evading immune responses, and cells whose phenotype falls short of satisfying this steady state would expire under hostile immune responses. This view was supported until recently by experimental evidence showing an inferior endurance of severe combined immunodeficiency (SCID)-derived pancreatic islets when engrafted into syngeneic immune-intact wild-type (WT) mice, relative to islets from WT. Here we extend the experimental exploration of ecoimmunity by searching for the presence of the phenotypic changes suggested by the theory. Immune-related phenotypes of islets, spleen, and bone marrow immune cells were determined, as well as SCID and WT nonlymphocytic cells. Islet submass grafting was performed to depict syngeneic graft functionality. Islet cultures were examined under both resting and inflamed conditions for expression of CD40 and major histocompatibility complex (MHC) class I/II and release of interleukin-1α (IL-1α), IL-1β, IL-6, tumor necrosis factor-α (TNF-α), IL-10, and insulin. Results depict multiple pathways that appear to be related to the sculpting of nonimmune cells by immune cells; 59 SCID islet genes displayed relative expression changes compared with WT islets. SCID cells expressed lower tolerability to inflammation and higher levels of immune-related molecules, including MHC class I. Accordingly, islets exhibited a marked increase in insulin release upon immunocyte depletion, in effect resuming endocrine function that was otherwise suppressed by resident immunocytes. This work provides further support of the ecoimmunity theory and encourages subsequent studies to identify its role in the emergence and treatment of autoimmune pathologies, transplant rejection, and cancer.
Mesenchymal Stem Cells as Treatment for Behavioral Deficits and Neuropathology in the 5xFAD Mouse Model of Alzheimer's Disease
Authors: Matchynski-Franks Jessica J. , Pappas Colleen , Rossignol Julien , Reinke Tiffany , Fink Kyle , Crane Andrew , Twite Alison , Lowrance Steven A. , Song Cheng , Dunbar Gary L.
Published: February 2016

Alzheimer's disease (AD) is characterized by a progressive loss of memory and other cognitive disturbances. The neuropathology of AD includes the major hallmarks of toxic amyloid-β oligomer accumulation and neurofibrillary tangles, as well as increased oxidative stress, cholinergic dysfunction, synapse loss, changes in endogenous neurotrophic factors, and overall degeneration of the brain. Adult mesenchymal stem cells (MSCs) offer the potential for a readily available treatment that would be long lasting, have low likelihood of rejection, and could target a variety of pathological deficits. MSCs have been shown to be effective in alleviating symptoms in some transgenic models of AD, but the optimal location for transplanting MSCs has yet to be determined. In the present study, the behavioral effects of transplantation of MSCs into the lateral ventricles, the hippocampus, or both of these regions were compared in the 5xFAD mouse model of AD. The results indicate that MSC transplants effectively reduce learning deficits in the 5xFAD mouse model and demonstrate a clear impact of MSCs on the levels of Aβ42 in the brains of 5xFAD mice. Overall, these findings support the hypothesis that MSCs may be a viable treatment for AD, especially when injected into the lateral ventricles.
A Chemically Defined Medium-Based Strategy to Efficiently Generate Clinically Relevant Cord Blood Mesenchymal Stromal Colonies
Authors: Barilani Mario , Lavazza Cristiana , Boldrin Valentina , Ragni Enrico , Parazzi Valentina , Crosti Mariacristina , Montelatici Elisa , Giordano Rosaria , Lazzari Lorenza
Published: February 2016

During the last decade it has been demonstrated that mesenchymal progenitors are present and can be isolated also from cord blood (CB). Recently, we managed to set up a standard protocol allowing the isolation of mesenchymal stromal cells (MSCs) with high proliferative potential and multiple differentiation capabilities, whereas the generation rate of MSC-initiating colonies could still be further improved. Herein, we strikingly succeeded in defining some simple and basic culture conditions based on the use of a chemically defined medium that increased the colony isolation efficiency up to almost 80% of processed CB units. Importantly, this result was achieved irrespective of CB unit white blood cell content and time elapsed from delivery, two limiting parameters involved with processing CB units. Thus, this high efficiency is guaranteed without strict selection of the starting material. In addition, since we are profoundly concerned about how different culture conditions can influence cell behavior, we devoted part of this study to in-depth characterization of the established CB-MSC populations to confirm their stemness features in this novel isolation and culture system. Therefore, an extended study of their immunophenotype, including classical pericytic markers, and a detailed molecular analysis addressing telomere length and also stemness-related microRNA contribution were performed. In summary, we propose a straightforward, extremely efficient, and reliable approach to isolate and expand thoroughly characterized CB-MSCs, even when poor-quality CB units are the only available source, or there is no space for an isolation to fail.
Human Fibroblast Sheet Promotes Human Pancreatic Islet Survival and Function in Vitro
Authors: Matsushima Hajime , Kuroki Tamotsu , Adachi Tomohiko , Kitasato Amane , Ono Shinichiro , Tanaka Takayuki , Hirabaru Masataka , Kuroshima Naoki , Hirayama Takanori , Sakai Yusuke , Soyama Akihiko , Hidaka Masaaki , Takatsuki Mitsuhisa , Kin Tatsuya , Shapiro James , Eguchi Susumu
Published: February 2016

In previous work, we engineered functional cell sheets using bone marrow-derived mesenchymal stem cells (BM-MSCs) to promote islet graft survival. In the present study, we hypothesized that a cell sheet using dermal fibroblasts could be an alternative to MSCs, and then we aimed to evaluate the effects of this cell sheet on the functional viability of human islets. Fibroblast sheets were fabricated using temperature-responsive culture dishes. Human islets were seeded onto fibroblast sheets. The efficacy of the fibroblast sheets was evaluated by dividing islets into three groups: the islets-alone group, the coculture with fibroblasts group, and the islet culture on fibroblast sheet group. The ultrastructure of the islets cultured on each fibroblast sheet was examined by electron microscopy. The fibroblast sheet expression of fibronectin (as a component of the extracellular matrix) was quantified by Western blotting. After 3 days of culture, islet viabilities were 70.2 ± 9.8%, 87.4 ± 5.8%, and 88.6 ± 4.5%, and survival rates were 60.3 ± 6.8%, 65.3 ± 3.0%, and 75.8 ± 5.6%, respectively. Insulin secretions in response to high-glucose stimulation were 5.1 ± 1.6, 9.4 ± 3.8, and 23.5 ± 12.4 μIU/islet, and interleukin-6 (IL-6) secretions were 3.0 ± 0.7, 5.1 ± 1.2, and 7.3 ± 1.0 ng/day, respectively. Islets were found to incorporate into the fibroblast sheets while maintaining a three-dimensional structure and well-preserved extracellular matrix. The fibroblast sheets exhibited a higher expression of fibronectin compared to fibroblasts alone. In conclusion, human dermal fibroblast sheets fabricated by tissue-engineering techniques could provide an optimal substrate for human islets, as a source of cytokines and extracellular matrix.
RESEARCH OF RAILWAY AGGREGATE SUB-BALLAST MIXTURE PROPERTIES / GELEŽINKELIO POBALASTINIO SLUOKSNIO IŠ UŽPILDŲ MIŠINIO SAVYBIŲ TYRIMAS
Authors: Navikas Deividas , Sivilevičius Henrikas
Published: February 2016

Railway sub-ballast layer is a connecting layer between the ballast and other track layers. This layer must ensure even load distribution into the subgrade, drainage surface water, prevent materials of layers mixing. Ensuring these properties allows to extend the explotation of all railway road construction and geometrical parameter of road maintenance. This article analyzes investigated railway sub-ballast mixture properties from large sample sizes (n = 49): gradation, water permeability and density statistical parameters. The regression equation showing the relationship between the fully percent passing through sieves averages and standard deviations is presented. Her coefficient of determination (R2 = 0.909). The minimum number of representative samples of these three properties research is determined. Irregularity in most cases of water permeability (ν = 88,3%), depending on the laboratory tests methodology errors, is indicated.
Geležinkelio pobalastinis sluoksnis yra jungiamasis sluoksnis tarp balasto ir grunto sluoksnių. Šis sluoksnis turi užtikrinti tolygų apkrovos perdavimą į sankasą, nudrenuoti paviršinį vandenį, neleisti maišytis sluoksnių medžiagoms. Šių savybių užtikrinimas leidžia prailginti visos kelio konstrukcijos eksploataciją ir geometrinių kelio parametrų išlaikymą. Šiame straipsnyje yra analizuojamos užpildų mišinio, naudojamo geležinkelio pobalastiniame sluoksnyje, iš didelės imties (n = 49) gautos savybės: granuliometrinės sudėties, pralaidumo vandeniui ir tankio statistiniai parametrai. Pateikta granuliometrinės sudėties regresijos lygtis, rodanti sąsają tarp pilnutinių išbirų per sietus vidurkių ir standartinių nuokrypių. Jos determinacijos koeficientas R2 = 0.909. Apskaičiuoti reprezentatyvūs minimalūs bandinių skaičiai šioms trims savybėms tirti. Gauta, kad pralaidumo vandeniui savybė yra labiausiai nevienoda dėl laboratorinių tyrimų metodikos paklaidų, variacijos koeficientas (88,3 %).
VEHICLES' SAMPLE GENERATION AND REALIZATION IN CAR-FOLLOWING MATHEMATICAL MODELS / AUTOMOBILIŲ IMTIES GENERAVIMAS BEI REALIZAVIMAS SEKIMO PASKUI LYDERĮ MATEMATINIUOSE MODELIUOSE
Authors: Danilevičius Algimantas , Junevičius Raimundas
Published: February 2016

The object of the article is the adjustment of car-following mathematical models according to collected traffic data. Here the problem of ineffectively burdened road section is solved by adjusting the speed of vehicles in order to reduce the distance between the cars to a safe distance. The paper analyzes the car-following models to measure the interaction between vehicles in the same lane. Experimental data processed in Matlab and traffic distribution histograms are created using the most appropriate distribution curve. Distribution curve is used to compile congestion scenario of road section. Applicable model uses fundamental diagrams, which are created from the kind of traffic flow measurements. The mathematical model allows to choose the optimal vehicle speed while maintaining safe distance between vehicles, and to make recommendations to improve the traffic as the process.
Straipsnio tyrimo objektas yra sukauptų automobilių srauto duomenų pritaikymas sekimo paskui lyderį matematiniuose modeliuose. Čia sprendžiama neefektyviai apkrautos kelio atkarpos problema. Siekiama sumažinti atstumą tarp automobilių iki saugaus atstumo, koreguojant automobilių greitį. Straipsnyje nagrinėjami sekimo paskui lyderį modeliai, pagal kuriuos įvertinama sąveika tarp toje pačioje eismo juostoje esančių transporto priemonių. Eksperimentiniai duomenys apdorojami taikant Matlab, sudaromos transporto srauto pasiskirstymo histogramos bei parenkama tinkamiausia skirstinio kreivė. Eksperimentinė skirstinio kreivė naudojama sudarant kelio atkarpos apkrovimo scenarijų – nustatoma modeliuojamos kelio atkarpos intensyvumo ir atstumų tarp transporto priemonių priklausomybė nuo laiko. Taikomame modelyje naudojamos fundamentalios diagramos, sudaromos pagal natūrinius eismo srauto matavimus. Matematinis modelis leidžia parinkti optimalų transporto priemonės greitį išlaikant saugų atstumą tarp transporto priemonių, taip pat juo remiantis galima teikti rekomendacijas, kaip gerinti automobilių eismą.
OVERVIEW OF NUMERICAL METHODS FOR SIMULATING RANQUE-HILSCH EFFECT WITHIN VORTEX TUBES / LIETUVOS KARJERŲ UŽPILDŲ POVEIKIO BETONO ŠARMINEI KOROZIJAI TYRIMAI
Authors: Nikitin Vadim , Bogdevičius Paulius , Bogdevičius Marijonas
Published: February 2016

The main goal of this paper is to present a summarized overview of the methods used for Ranque–Hilsch vortex tube numerical analysis using computational fluid dynamics (CFD). The overview is mainly focused on the most recently conducted studies that are both backed by experimental data and is evaluated by the authors as being in good agreement with experimental results. Generalized tendencies in computational vortex tube analysis are presented while focusing on researchers’ approach towards the geometric model used for the study and the governing equations. A brief introduction to the subject matter is presented followed by a short retrospective of the previous studies and related challenges. The conclusions are formed based on the most recent, as well as previously analysed, published research results.
Užpildų šarminė korozija betone vyksta reaguojant cemente esantiems natrio ir kalio hidroksidams (šarmams) su aktyviu SiO2, esančiu kai kuriuose užpilduose. Vykstant šiai reakcijai betone susidaro didelių vidinių įtempių, kurie sukelia betono deformacijas, pleišėjimą ir suirimą. Reakcija vyksta lėtai, betono irimo požymių atsiranda tik po kelių mėnesių ar metų. Tyrimams buvo naudojami dviejų skirtingų karjerų užpildai. Atlikus tyrimus nustatyta, kad Lietuvos žvyro karjerai užteršti reaktyviomis dalelėmis, turinčiomis amorfinio silicio dioksido, reaguojančio su cemente esančiais natrio ir kalio šarmais, ir sukeliančiomis betono šarminę koroziją. Nustatyta, kad pagal AAR 2 stambieji užpildai priskiriami II grupei – galimai reaktyviems užpildams, nes jų plėtra po 14 parų viršija 0,1 %.