The recessive form of dystrophic epidermolysis bullosa (RDEB) is a crippling disease caused by impairments in the junctions of the dermis and the basement membrane of the epidermis. Using ectopic expression of hTERT/hTERT + BMI-1 in primary cells, we developed expansible cultures of RDEB fibroblasts and keratinocytes. We showed that they display the properties of their founders, including morphology, contraction ability and expression of the respective specific markers including reduced secretion of type VII collagen (C7). The immortalized keratinocytes retained normal stratification in 3D skin equivalents. The comparison of secreted protein patterns from immortalized RDEB and healthy keratinocytes revealed the differences in the contents of the extracellular matrix that were earlier observed specifically for RDEB. We demonstrated the possibility to reverse the genotype of immortalized cells to the state closer to the progenitors by the Cre-dependent hTERT switch off. Increased β-galactosidase activity and reduced proliferation of fibroblasts were shown after splitting out of transgenes. We anticipate our cell lines to be tractable models for studying RDEB from the level of single-cell changes to the evaluation of 3D skin equivalents. Our approach permits the creation of standardized and expandable models of RDEB that can be compared with the models based on primary cell cultures. 相似文献
Infrared and Raman spectra of atactic PMMA and infrared spectra of stereoregular PMMA and of its four deuterated derivatives , were measured. They were used to assign the bands of stretching and deformation vibrations of CH2, CCH3 and OCH3 groups in the infrared and Raman spectra and to discuss the effect of stereoregularity on these bands in the infrared spectra. 相似文献
In this work, the curing of «ED-20» epoxy resin with partially siloxy-substituted aluminum, iron, and zirconium siloxanes that we obtained previously was studied. The initial content of a metallosiloxane in the compositions was 5–50 wt% with respect to the resin. In all the cases, thermal curing was used to obtain a series of samples in the form of solid homogeneous materials. The fact of the epoxy ring opening in the resin was confirmed by IR spectroscopy. The catalytic properties of the metal atom in a metallosiloxane were found to affect the curing process. The samples demonstrate rather a high resistance to thermooxidative destruction, and in most cases, their glass transition temperatures are lower than those obtained upon standard curing of «ED-20» resin with triethylenetetramine. Partially siloxy-substituted metalloalkoxysiloxanes can be efficient agents for curing and formation of a hybrid material based on epoxy resins. 相似文献
This study focuses on hydrological and biotic variables in Lake Glubokoe, which is located in Thala Hills of Enderby Land (East Antarctica). Water and sediment samples and physical measurements were collected once a week in the austral summer (19 December 2010 – 6 February 2011). This lake exhibits perennial ice cover that reached a thickness of 2.5–2.7 m during the study period. A very low concentration of planktonic chlorophyll‐a (0.06–0.45 μg L?1) was measured in the lake, indicating its ultra‐oligotrophic status. The water was poorly populated by algae and metazoans, especially in upper waters below ice cover to a depth of 2 m. Small planktonic organisms (2–5 μm) were observed throughout the study period, but larger organisms (>8 μm) such as the cyanobacteria Planktolyngbya limnetica occurred only during the warmest period (January). Only few individuals of metazoans (rotifers) were found in planktonic samples. Due to deep light penetration (10–15% of incoming active solar radiation reached the depth of 30 m), thick cyanobacterial mats (30 cm) cover all the bottom surface (grey silts) in the lake. Abundant benthic biota associated with these mats was found (up to 1000 ind. m?2). Among the benthic metazoans, bdelloid rotifers and tardigrades were the dominating taxa. The results of this study suggest a typical ecological feature of most subglacial lakes in East Antarctica is that metazoans are very poor in the pelagic zone, preferring instead to occupy an area near the lake bottom because of a favourable constant temperature of 4 °C, good level of dissolved oxygen and available food resources as the bacterial detritus. 相似文献
This study was undertaken to evaluate the compatibility of Giemsa staining protocol with the comet assay. We showed, for the first time, that DNA comets can be visualized and analyzed using Giemsa staining. We generated DNA damage dose response curves for human peripheral blood lymphocytes exposed to X-ray radiation using the comet assay with either SybrGreen I or Giemsa stain. The dose response curves were fitted by linear regressions (R2 > 0.977). The SybrGreen I results showed only ~1.2-fold higher slope coefficient (method sensitivity) compared to the Giemsa results. The unexpectedly high sensitivity of Giemsa staining for the comet assay is due to the Romanowsky-Giemsa effect, the stain photo-stability and the higher resolution of bright-field imaging compared to fluorescence imaging. Our results demonstrate that Giemsa staining can effectively be used for measuring DNA damage by the comet assay. The low cost and availability of Giemsa stain makes this method affordable for any low budget research and will facilitate new applications of the comet assay in biology and medicine. 相似文献
Esters with acyl groups can be formed by the esterification of polyglycerol. The purpose of the present study was to produce fatty acid esters [hexanoic (caproic), octanoic (caprylic), decanoic (capric), dodecanoic (lauric), tetradecanoic (myristic), hexadecanoic (palmitic), octadecanoic (stearic)] and polyglycerol (average number‐of degrees of polymerization of 5) with varying degrees of esterification and to examine their emulsifying properties. A number of fundamental catalysts of polyglycerol acylation reactions by methyl esters of carboxylic acid were studied, and sodium methoxide was found to be the best choice. The temperature rate of transesterification increased from 180 to 220 °C with the fatty acid chain alkyl residue. Synthesized mono‐, di‐, tri‐, tetra‐, and heptaesters of various fatty acids and polyglycerol provided the highest hydroxyl values from 15 to 815 mg KOH g?1 and saponification values from 82 to 321 mg KOH g?1. The emulsifying properties were assessed for all polyglycerol and fatty acid esters, with results showing maximum emulsifying effect for tri‐ and tetraesters of capric, lauric, and caprylic acids. Regardless of the hydrophilic–lipophilic balance value (HLB) of polyglycerol esters and carboxylic acid, a 4:1 ratio of sunflower oil to water formed a water‐in‐oil type emulsion. When mixing oil and water in a 1:1 ratio, mono‐ and diesters of polyglycerol formed an oil‐in‐water type emulsion, heptaesters formed a water‐in‐oil type emulsion, and tri‐ and tetraesters formed both of types of emulsions, depending on the length of the acid hydrocarbon radicals. 相似文献