Reaction of oligoglia to spinal cord injury in rats and transplantation of human olfactory ensheathing cells

In experiments on rats with lateral TVIIIhemisection of the spinal cord and transplantation of ensheating olfactory cells, we studied structural changes at the lesion site and adjacent rostral and dorsal regions of the spinal cord. The state of oligodendrocytes and myelin fibers and motor function in experimental animal were analyzed. Open field testing (BBB test) showed that motor functions steadily increased (by 13% on average) within the interval from day 21 to day 53 after transplantation. Histological examination showed that groups of transplanted cells carrying human nuclear marker (HNu + cells) were still present at the lesion site 30 days after surgery. Some of these cells migrated in the rostral and caudal directions from the injection site to a distance up to 6 mm. At the initial period after hemisection, the number of oligodendrocytes (O4+-cells) in the immediate vicinity to the lesion site decreased 2-fold, but no signifi cant changes in the number of neurons were found in the rostral and dorsal fragments of the spinal cord compared to the corresponding parameter in controls. Sixty days after transplantation, the cross-section area in the rostral part of the spinal cord at a distance of 3 mm from damage site increased by 15.3% compared to the control. The number of O4+- cells at the lesion site and in adjacent rostral and caudal parts of the spinal cord by 22.8% surpassed that in the control group. The number of remyelinated axons also increased. These findings suggest the absence of pronounced structural changes in the rostral and caudal parts of the spinal cord compared to lesion site at early stages after damage and cell transplantation. At the same time, pronounced activation of oligodendrocytes in this region suggests their involvement together with Schwann cells into remyelination of regenerating axons, which can serve as a factor of partial restoration of motor functions after spinal cord injury. © 2010 Springer Science+Business Media, Inc

Increased Expression of Clusterin in Kidney of a Fatal Nephropathia Epidemica Case

© 2016, Springer Science+Business Media New York.Nephropathia epidemica (NE), a mild form of hemorrhagic fever with renal syndrome (HFRS), is endemic in the European part of Russia. This disease is characterized by an acute onset followed by symptoms related to kidney impairment. Histologically, NE is defined as a tubulointerstitial nephritis with the prominent leukocyte infiltration and interstitial hemorrhages. Also, the presence of IgM, complement component C3, and fibrin deposits along the basal surface of the tubular epithelial cells has been reported. The pathogenesis of immunoglobulin and complement deposition in the kidney tubules remains unknown. We found clusterin deposits in the kidney tissue from fatal NE. Clusterin was co-localized with hantavirus antigens and C3 complement on the basal side of the kidney tubules. Additionally, kidney infiltrating leukocytes were positive for clusterin. It has previously been documented that the expression of clusterin is restricted to epithelial cells and is often found on tubular cells of the injured kidney tissue. Additionally, clusterin can bind to immunoglobulins and complement components. We propose that upregulation of clusterin expression in the NE kidney is associated with damage to the tissue and may cause deposition of immunoglobulins and complement

Considerations and strategies in L2 vocabulary acquisition among Japanese 1st year university students.

This article proposes a framework of strategies1） for L2 vocabulary acquisition among low-to-mid level L2 learners in their first year at Kansai University. The framework relies upon considerations posited by Kudo (1999), as well as, Hunt and Beglar’s (2005) model for developing EFL reading vocabulary, although the objective for our target learners is tofacilitate vocabulary acquisition in a learner-centered communicative context, where possible. This paper firstly underlines the critical role of vocabulary in second language acquisition, while raising awareness of the surrounding pedagogic climate in Japanese secondary education.研究ノー

Multiplex Analysis of the Activation of the Immune System after Transection of the Rat’s Sciatic Nerve

© 2016, Springer Science+Business Media New York.In this study, using the model of sciatic nerve transection in rats, the cytokine profile of peripheral blood serum and rat sciatic nerve fragments was measured 3, 7 and 14 days after the injury. Histological analysis of sciatic nerve longitudinal sections was carried out at the same periods of time. The cytokine profile of peripheral blood serum of an intact rat as well as the cytokine profile of an intact homogenized sciatic nerve of a rat was examined. Comparative study of changes in quantitative measures before and after the injury was carried out. It was shown that in peripheral blood serum after the sciatic nerve transection, there was a change in quantitative measures of IL­1a, Leptin, IL6, MCP1, and MIP­2. In the samples of the rat sciatic nerve, the changes were found in the levels of EGF, Fractalkine, G­CSF, IFNg, IL10, IL17A, IL18, IL2, IL6, IP10, Leptin, LIX, MCP1, MIP­1a, MIP­2, and RANTES

Assessment of glial scar, tissue sparing, behavioral recovery and axonal regeneration following acute transplantation of genetically modified human umbilical cord blood cells in a rat model of spinal cord contusion

©2016 Mukhamedshina et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.Objective and Methods This study investigated the potential for protective effects of human umbilical cord blood mononuclear cells (UCB-MCs) genetically modified with the VEGF and GNDF genes on contusion spinal cord injury (SCI) in rats. An adenoviral vector was constructed for targeted delivery of VEGF and GDNF to UCB-MCs. Using a rat contusion SCI model we examined the efficacy of the construct on tissue sparing, glial scar severity, the extent of axonal regeneration, recovery of motor function, and analyzed the expression of the recombinant genes VEGF and GNDF in vitro and in vivo. Results Transplantation of UCB-MCs transduced with adenoviral vectors expressing VEGF and GDNF at the site of SCI induced tissue sparing, behavioral recovery and axonal regeneration comparing to the other constructs tested. The adenovirus encoding VEGF and GDNF for transduction of UCB-MCs was shown to be an effective and stable vehicle for these cells in vivo following the transplantation into the contused spinal cord. Conclusion Our results show that a gene delivery using UCB-MCs-expressing VEGF and GNDF genes improved both structural and functional parameters after SCI. Further histological and behavioral studies, especially at later time points, in animals with SCI after transplantation of genetically modified UCB-MCs (overexpressing VEGF and GDNF genes) will provide additional insight into therapeutic potential of such cells

Surgical Procedure for Extracting Pig Teeth for Isolation and Cultivation of Mesenchymal Stem Cells from Dental Pulp for Regenerative Therapy Applications

© 2016, Springer Science+Business Media New York.Mesenchymal stem cells (MSCs) are considered the most versatile cells for cell therapy—particularly for repair of injuries to the central nervous system. Recently, the use of dental pulp MSCs (DP-MSCs) for spinal cord regeneration has became especially important. We describe a surgical procedure for extracting pig teeth to obtain DP-MSCs using protocols for direct and enzymatic isolation of DP-MSCs followed by cultivation. Our study shows that primary pulp cultures of DP-MSCs are established 5 days after enzymatic digestion and 7–10 days following attachment of the digested minced tissue to the bottom of a plate. Though in the first few days the rate of primary expansion for cultures generated by direct isolation was lower than the rate of enzymatic digestion, this difference leveled off on days 14–18 of culture. For DP-MSCs isolation, we recommend the use of deciduous succedaneous lateral incisors and canine teeth of pigs as well as deciduous premolars from the first dentition of young pigs (up to 3 months)

Use of Gene-Activated Demineralized Bone Allograft in the Therapy of Ulnar Pseudarthrosis. Case Report

© 2016, Springer Science+Business Media New York.This paper presents a clinical case of successful ulnar pseudarthrosis treatment using a gene-activated bone allograft containing VEGF (vascular endothelial growth factor) and BMP2 (bone morphogenetic protein 2) in the form of a multicystron plasmid. Demineralized bone matrix with applied recombinant plasmid DNA was grafted into the bone defect using the classical open surgical approach. Two months after the surgery, the patient noticed the disappearance of pain including pain during activity. On X-rays of ulna, signs of union in the form of callus formation were found

Immunomodulating and Revascularizing Activity of Kalanchoe pinnata Synergize with Fungicide Activity of Biogenic Peptide Cecropin P1

© 2017 N. S. Zakharchenko et al. Previously transgenic Kalanchoe pinnata plants producing an antimicrobial peptide cecropin P1 (CecP1) have been reported. Now we report biological testing K. pinnata extracts containing CecP1 as a candidate drug for treatment of wounds infected with Candida albicans. The drug constitutes the whole juice from K. pinnata leaves (not ethanol extract) sterilized with nanofiltration. A microbicide activity of CecP1 against an animal fungal pathogen in vivo was demonstrated for the first time. However, a favorable therapeutic effect of the transgenic K. pinnata extract was attributed to a synergism between the fungicide activity of CecP1 and wound healing (antiscar), revascularizing, and immunomodulating effect of natural biologically active components of K. pinnata. A commercial fungicide preparation clotrimazole eliminated C. albicans cells within infected wounds in rats with efficiency comparable to CecP1-enriched K. pinnata extract. But in contrast to K. pinnata extract, clotrimazole did not exhibit neither wound healing activity nor remodeling of the scar matrix. Taken together, our results allow assumption that CecP1-enriched K. pinnata extracts should be considered as a candidate drug for treatment of dermatomycoses, wounds infected with fungi, and bedsores

Bactericide, Immunomodulating, and Wound Healing Properties of Transgenic Kalanchoe pinnata Synergize with Antimicrobial Peptide Cecropin P1 In Vivo

Procedure of manufacturing K. pinnata water extracts containing cecropin P1 (CecP1) from the formerly described transgenic plants is established. It included incubation of leaves at +4°C for 7 days, mechanical homogenization of leaves using water as extraction solvent, and heating at +70°C for inactivating plant enzymes. Yield of CecP1 (after heating and sterilizing filtration) was 0.3% of total protein in the extract. The water extract of K. pinnata + CecP1 exhibits favorable effect on healing of wounds infected with S. aureus (equal to Cefazolin) and with a combination of S. aureus with P. aeruginosa (better than Cefazolin). Wild-type K. pinnata extract exhibited evident microbicide activity against S. aureus with P. aeruginosa but it was substantially strengthened in K. pinnata + CecP1 extract. K. pinnata extracts (both wild-type and transgenic) did not exhibit general toxicity and accelerated wound recovery. Due to immunomodulating activity, wild-type K. pinnata extract accelerated granulation of the wound bed and marginal epithelialization even better than K. pinnata + CecP1 extract. Immunomodulating and microbicide activity of K. pinnata synergizes with microbicide activity of CecP1 accelerating elimination of bacteria

Regeneration of rat sciatic nerve using tubulation and allogeneic transplantation of adipose-derived mesenchymal stromal cells

The effectiveness of application of biodegradable conduit (NeuraGen; INTEGRA, USA) in combination with adipose derived multipotent mesenchymal stem cells and fibrin sealant (Tissucol-Kit; Baxter AG, Austria) on posttraumatic peripheral nerve regeneration was investigated using a rat model of sciatic nerve injury. We demonstrate that our proposed cell therapy confers a neuroprotective effect under our experimental design