Non-Thermal Atmospheric-Pressure Plasma Possible Application in Wound Healing

Non-thermal atmospheric-pressure plasma, also named cold plasma, is defined as a partly ionized gas. Therefore, it cannot be equated with plasma from blood; it is not biological in nature. Non-thermal atmospheric-pressure plasma is a new innovative approach in medicine not only for the treatment of wounds, but with a wide-range of other applications, as e.g. topical treatment of other skin diseases with microbial involvement or treatment of cancer diseases. This review emphasizes plasma effects on wound healing. Non-thermal atmospheric-pressure plasma can support wound healing by its antiseptic effects, by stimulation of proliferation and migration of wound relating skin cells, by activation or inhibition of integrin receptors on the cell surface or by its pro-angiogenic effect. We summarize the effects of plasma on eukaryotic cells, especially on keratinocytes in terms of viability, proliferation, DNA, adhesion molecules and angiogenesis together with the role of reactive oxygen species and other components of plasma. The outcome of first clinical trials regarding wound healing is pointed out.

CTLA-4-Tg/CD-28-KO Mice Exhibit Reduced T Cell Proliferation in vivo Compared to CD-28-KO Mice in a Graft-versus-host Disease Model

Activated T cells express inhibitory receptors such as CTLA-4 that can downregulate immune responses. Blockade of or genetic deficiency in CTLA-4 can result in autoimmunity. Therefore, strategies to increase the inhibitory function of CTLA-4 may be attractive in settings of undesirable T cell responses such as autoimmunity or transplant rejection. We have tested the hypothesis that transgenic constitutive expression of CTLA-4 can further attenuate immune responses when compared with normal inducible expression. Our results indicate that transgenic expression of CTLA-4 in mouse T cells (CTLA-4-Tg T cells) results in reduced cell cycle progression and increased apoptosis of TCR-stimulated T cells. CTLA-4-Tg T cells display reduced T cell proliferation in an in vivo model of graft versus host disease (GVHD). These results further our understanding of how CTLA-4 can be manipulated to inhibit immune responses and may help development of new therapeutic strategies for clinical settings of autoimmunity and transplantation.

Method for analysis of surface molecule alteration upon phagocytosis by flow cytometry.

In this study, we introduce an application of flow cytometry for the concurrent detection of phagocytotic cells and surface molecules involved in the phagocytic process. E. coli expressing green fluorescent protein (GFP) were applied as the phagocytosable particles. Blood samples were incubated with E. coli expressing GFP, followed by indirect immunofluorescence using four candidate monoclonal antibodies (mAbs). Granulocytes that had phagocytosed E. coli exhibited high levels of GFP intensity, in contrast to the non-phagocytosed cells. By comparing the level of expression of molecules expressed on phagocytosed granulocytes with that of non-phagocytosed cells by flow cytometry, it enabled the determination of the expression and alteration of the cell surface molecules upon phogocytosis. Of the four mAbs used in this study, upon phagocytosis, molecules recognized by mAbs WK13, COSA5A and COSA33NL were up-regulated. However, CD15 recognized by mAb VIMD5 was down-regulated. The proposed method will benefit the study of phagocytic mechanisms in the future.

The Role Of Cells, Neurotrophins, Extracellular Matrix And Cell Surface Molecules In Peripheral Nerve Regeneration

Wallerian degeneration is a complicated process whereby axons and myelin sheaths undergo degeneration, and eventually are phagocytosed by macrophages and Schwann cells following nerve damage. Schwann cells proliferate and the endoneural tubes persist. In addition, neurotrophins, neural cell adhesion molecules, cytokines and other soluble factors are upregulated to facilitate regeneration. The important role of cellular components, neurotrophins, and extracellular matrix components, including cell surface molecules involved in this regenerative process, is highlighted and discussed in this review.

Evaluation of ginger (Zingiber officinale Roscoe) bioactive compounds in increasing the ratio of T-cell surface molecules of CD3+CD4+:CD3+CD8+ in-vitro

The potential ability of ginger bioactive compounds in increasing the ratio of T-cell surface molecules of CD3+CD4+:CD3+CD8+ was investigated using dual tagging FITC and PE of monoclonal antibody anti-human with its fluorescence measured by flow cytometer. Oleoresin was extracted using sinkhole distillation technique. Its components namely, gingerol in fraction-1, shogaol in fraction 2 and zingeron in fraction-3 were separated by column vacuum chromatography method. The doses of oleoresin, gingerol, shogaol, and zingeron tested were 50, 100,150, 200, and 250 μg/ml. Lymphocytes (2x106 cell/ml) from human peripheral blood were isolated using ficoll density gradient technique, and cultured in the presence of the compounds in RPMI-1640 medium and phytohemaglutinin (PHA) mitogen for 96 h under normal conditions. Percentages of T-cell surface molecules (CD4+ and CD8+) were determined using dual-tagging FITC and PE fluorescents labeled on monoclonal antibody anti human. The fluorescence-labeled bands on the T-cell surface molecules were counted using flow cytometer. The experiment revealed that oleoresin and its three fractions increased the percentage of CD3+CD4+. The compound in fraction 3 of oleoresin at 200 μg/ml increased by the highest percentage of CD3+CD4+ of 9%, but slightly decreased the percentage of CD3+CD8+. These ginger bioactive compounds increased the ratio of CD3+CD4:CD3+CD8+ T-cells with the highest increment of 30% from effects of 200 μg/ml fraction 3 of oleoresin. This in vitro finding revealed that ginger bioactive compounds potentially increased cellular and humoral immune response. Further clinical studies are needed to confirm the benefits of these ginger bioactive compounds as a potential functional food for testing on HIV infected patients.

Histopaghologic Finding and Modulation of Expression of The Cell Surface Molecules in The Primary Syphilitic Sedions of a Rabbit Infected with Treponema pallidum / 대한피부과학회지

BACKGROUND: When T. pallidum invades through skin and mucous membrane, there may be some changes in the expression of cell surface molecules and cell adhesion molecules on the keratinocytes and vascular endothelial cells. OBJECTIVES: The purpose of this study was to investigate histologic changes and changes in the expression of cell surface molecules and cell adhesion molecules on the keratinocytes and vascular endothelial cells according to the time course of primary syphilic lesion. METHODS: We obtained primary syphilitic lesions by inoculation of T. pallidum into the back skin of the rabbit. Biopsies of the syphilitic lesions were performed according to the stages. and H&E and immunohistochemical stains for cell surface molecules were done. RESULTS: 1. Out of the 39 injected sites(103 T. pallidum were inoculated into the back skin of the rabbit), 24(61.5%) primary syphilitic lesion could be found. The duration for the developement of papules, ulcers, and softenings is an average 15 days, 27 days, and 47 days respectively. 2. H & E findings :Acanthosis, spongiosis, and exocytosis in the epidermis were observed in the papule of the primary syphilitic lesion. Infiltration of inflammatory cells, including many lymphocytes, a few histiocytes and plasma cells was also observed. Some cases showed endothelial cell swelling of vessels. Compared to papules, the number of lymphocytes in the ulcer reduced but the number of histiocytes increased. Softened lesion showed infiltrating cells, consisted of lymphocytes and histiocytes, and fibrosis. 3. Immunohistochemical findings :Keratinocytes of the lower epidermis, upper portion of hair follicles, vessel, and infiltrating inflammatory cells in papules and ulcers showed expression of the MHC class II molecule. Most of the infiltrating cells in all cases of papules showed CD5 expression. Keratinocytes, inflammatory cells, and vascular endothelial cells showed positive reaction to ICAM-1 stain in papules and ulcers. VCAM-1 showed the positive reaction to the vascular endothelial cells in the papules and ulcers. In softened lesions, the intensity of the positive reaction to MHC class II, ICAM-1 and VCAM-1 was weakened. CONCLUSION: The skin of the rabbit which was invaded by T. pallidum increased the expression of the cell surface molecules and cell adhesion molecules of MHC class I, MHC class II, ICAM-1, and VCAM-1. We believe that these expressions of cell surface molecules and cell adhesion molecules by T. pallidum, inflammatory cells, activated keratinocytes, and vascular endothelial cells play important roles in the host defence mechanism and the T. pallidum infection.