Cryoelectron Microscopy of Vitreous Sections: A Step Further Towards the Native State

Author(s):  
Cedric Bouchet-Marquis ◽  
Stanislav Fakan
Author(s):  
Masako Osumi ◽  
Misuzu Nagano ◽  
Hiroko Kazama

We have found that microbodies appeared profusely together with a remarkable increase in catalase activity in normal alkane-grown cells of hydrocarbon-utilizing Candida yeasts, and that the microbodies multiplied by division in these cells. These features of Candida yeasts seem to provide a useful model system for studies on the biogenesis of the microbody. Subsequently, we have succeeded in isolation of Candida microbodies in an apparently native state, as judged biochemically and morphologically. The presence of DNA in the purified microbody fraction thus obtained was proved by the diphenylamine method. DNA molecule of about 15 urn in contour length was released from an isolated microbody. The physicochemical analyses of the microbody DNA revealed that its buoyant density differed from nuclear and mitochondrial DNAs. All these results lead us to the possibility that there is a novel type of DNA in microbodies.


Author(s):  
Tokio Nei ◽  
Haruo Yotsumoto ◽  
Yoichi Hasegawa ◽  
Yuji Nagasawa

In order to observe biological specimens in their native state, that is, still containing their water content, various methods of specimen preparation have been used, the principal two of which are the chamber method and the freeze method.Using its recently developed cold stage for installation in the pre-evacuation chamber of a scanning electron microscope, we have succeeded in directly observing a biological specimen in its frozen state without the need for such conventional specimen preparation techniques as drying and metallic vacuum evaporation. (Echlin, too, has reported on the observation of surface structures using the same freeze method.)In the experiment referred to herein, a small sliced specimen was place in the specimen holder. After it was rapidly frozen by freon cooled with liquid nitrogen, it was inserted into the cold stage of the specimen chamber.


2017 ◽  
Vol 14 (6) ◽  
pp. 778-784 ◽  
Author(s):  
Joanna Brzeska

Background: Cross-linking structure of polyurethanes determines no degradability of these materials. However, introducing the hydrolysable substrates (of natural or synthetic origin) into the cross-linked polyurethanes structure makes them biodegradable. Moreover compounds (such as polycaprolactone triol, glycerin, lysine triisocyanate, etc.) that are used for polyurethane cross-linking are degraded in non-toxic products. All these kinds of compounds can be introduced into soft or hard segments via urethane bonds. Objective: The review focuses on kind of multifunctional polyols and isocyanates, and low molecular crosslinkers used for cross-linked polyurethanes obtaining. These compounds are natural substrates (in the native state or after modification) or are synthetic compounds with degradable linkages. They belong to polyesters, plant oils, proteins, saccharides, and others (e.g. lignocellulosic materials), and they are synthesized chemically or via biosynthesis by algae, plants, microorganisms, and by animals. Conclusion: Incorporation of degradable groups (such as ester moieties) into the polymer structure, and using of substrates with the structure known and metabolized by microorganisms for soft or hard segments building, facilitate degradation of cross-linked polyurethanes.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Teodora Bavaro ◽  
Sara Tengattini ◽  
Refaya Rezwan ◽  
Enrica Chiesa ◽  
Caterina Temporini ◽  
...  

AbstractExogenous application of human epidermal growth factor (hEGF) stimulates epidermal wound healing. The aim of this study was to develop bioconjugates based on hEGF mimicking the protein in its native state and thus suitable for tissue engineering applications, in particular for treating skin-related disorders as burns. Ribonuclease A (RNase A) was used to investigate a number of different activated-agarose carriers: cyanogen bromide (CNBr)-activated-agarose and glyoxyl-agarose showed to preserve the appropriate orientation of the protein for receptor binding. EGF was immobilized on these carriers and immobilization yield was evaluated (100% and 12%, respectively). A peptide mapping of unbound protein regions was carried out by LC–MS to take evidence of the residues involved in the immobilization and, consequently, the flexibility and surface accessibility of immobilized EGF. To assess cell proliferative activities, 10, 25, 50, and 100 ng/mL of each immobilized EGF sample were seeded on fibroblast cells and incubated for 24, 48 and 72 h. The immobilized growth factor showed significantly high cell proliferative activity at 50 and 100 ng/mL compared to control and soluble EGF. Although both of the immobilized samples show dose-dependency when seeded with high number of fibroblast cells, CNBr-agarose-EGF showed a significantly high activity at 100 ng/mL and 72 h incubation, compared to glyoxyl-agarose-EGF.


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