Secretome of Peripheral Blood Mononuclear Cells Enhances Wound Healing

Non-healing skin ulcers are often resistant to most common therapies. Treatment with growth factors has been demonstrated to improve closure of chronic wounds. Here we investigate whether lyophilized culture supernatant of freshly isolated peripheral blood mononuclear cells (PBMC) is able to enhance wound healing. PBMC from healthy human individuals were prepared and cultured for 24 hours. Supernatants were collected, dialyzed and lyophilized (SEC^PBMC). Six mm punch biopsy wounds were set on the backs of C57BL/6J-mice and SEC^PBMC containing emulsion or controls were applied daily for three days. Morphology and neo-angiogenesis were analyzed by H&E-staining and CD31 immuno-staining, respectively. In vitro effects on diverse skin cells were investigated by migration assays, cell cycle analysis, and tube formation assay. Signaling pathways were analyzed by Western blot analysis. Application of SEC^PBMC on 6 mm punch biopsy wounds significantly enhanced wound closure. H&E staining of the wounds after 6 days revealed that wound healing was more advanced after application of SEC^PBMC containing emulsion. Furthermore, there was a massive increase in CD31 positive cells, indicating enhanced neo-angiogenesis. In primary human fibroblasts (FB) and keratinocytes (KC) migration but not proliferation was induced. In endothelial cells (EC) SEC^PBMC induced proliferation and tube-formation in a matrigel-assay. In addition, SEC^PBMC treatment of skin cells led to the induction of multiple signaling pathways involved in cell migration, proliferation and survival. In summary, we could show that emulsions containing the secretome of PBMC derived from healthy individuals accelerates wound healing in a mouse model and induce wound healing associated mechanisms in human primary skin cells. The formulation and use of such emulsions might therefore represent a possible novel option for the treatment of non-healing skin ulcers.     

Enzymatic utilization of cotton oil soap stock

Enzymatic hydrolysis of neutral fat of cotton oil soap stock with a nonspecific lipase produced byOospora lactis F-500 was designed. The culture liquid and a preparation of enzyme obtained by precipitation with isopropanol from a filtrate of the culture liquid were used. Utilization of cotton oil soap stock as the only source of carbon during cultivation of the fungus was studied. The rate of hydrolysis of soap stock fat strongly depended on the way of biological conversion of cotton oil soap stock. The most effective utilization was observed during cultivation of the fungus in the medium containing soap stock.     

Phenothiazine derivatives as enhancers of peroxidase-dependent chemiluminescence

Some N-alkyl phenothiazines with different ionic groups were studied as enhancers of chemiluminescence catalyzed by soybean peroxidase. It was shown that under experimental conditions, the compounds with positively charged groups do not exhibit enhancing ability, while the addition of phenothiazines with negatively charged groups to a substrate mixture significantly increased the chemiluminescence intensity. The relationship between the enhancing activity of phenothiazines and their capacity for enzymatic oxidation by hydrogen peroxide was found. The enhancers discovered new opportunities for increasing the sensitivity of determination of analytes by chemiluminescent enzyme immunoassay.     

The level of leukotrienes B4 and C4 in the blood serum of rats subjected to thermal burns, x-irradiation and combined radiation-thermal injuries

In the present study the content of leukotrienes B4 and C4 (LTB and LTC) in rat serum was investigated. The changes in the leukotriene content after thermal burn was different from that after X-irradiation and combined effect of radiation and heat.     

Phenotypic characteristics of heat-resistant murine neuroblastoma cells

One-step selection of murine neuroblastoma (N18TG2 cell line) for the resistance to injuring action of higher temperature resulted in obtaining cell line NTR1 stably capable of proliferating at 40 degrees C. The thermoresistance is shown to be coupled with the changes in some phenotypic features of NTR1 cells: the multiplication rate, saturation density of cells, morphological features, inducibility of long neurite-like processes and adhesion. A possible significance of plasma membrane changes in genetically stable thermoresistance of NTR1 neuroblastoma cells is discussed.