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Many uraemic patients die throughout the world because of expensive renal replacement therapy and reduced number of renal grafts. The recent in vitro generation of a histocompatible renal tissue using nuclear transplantation may open new opportunities in renal transplantation and may help to solve the problem of chronic kidney shortage.
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AIM: N-Glycans in recombinant human erythropoietin (EPO) are essential to in vivo biological activity. This paper is to develop a method for mapping sialyated or asialyated N-glycans of EPO. METHODS: At first, N-glycans linked to asparagines in glycoprotein EPO were released by peptide N-glycosidase F. To map asialyated N-glycans, sialic acid in N-glycans were removed by incubating N-glycans with sialidase. Oligosaccharides were labeled with a sensitive fluorescent dye 8-aminopyrene-1,3,6-trisulfonate (APTS), and all of the labeled oligosaccharides released from EPO were mapped by capillary gel electrophoresis with laser-induced fluorescence. The relationship between N-glycans and bioactivity of EPO was investigated on the basis of N-glycan mapping spectra. RESULTS: N-Glycans of seven different batches of EPO were mapped. Each sample was analysed twice, with and without sialidase treatment. The results showed that N-glycans of each sample were approximately the same. But when the expression vector was different, the types of N-glycans and their relative content were quite different. In case of asialyated N-glycan mapping, the retention time of each oligosaccharide delayed greatly, and most importantly, the resulted sialic acid peak can be used as a quantitative standard to determine sialic acid content in N-glycans of EPO. In addition, the difference of N-glycan mapping was observed when the in vivo biological activity of EPO was different. CONCLUSION: The approach in this article for determining N-glycan mapping can be applied to determine the source of EPO and the difference between each batch. It is also a suitable tool for routinely controlling the inner quality of EPO by coupled with peptide mapping.
The Leonardo da Vinci training programme supports innovative transnational initiatives for promoting the knowledge, aptitudes and skills necessary for successful integration into working life. Biomedical engineering is an emerging interdisciplinary field that contributes to understand, define and solve problems in biomedical technology within industrial and health service contexts. Paper presents a Leonardo da Vinci pilot-project called Web-based learning and training in the field of biomedical and design engineering (WEBD). This project has started on 2001. The WEBD project proposes to use advanced learning technologies to provide education in the www. Project uses interactive 3D graphics and virtual reality tools. The WEBD distance training permits users to experience and interact with a life-like model or environment, in safety and at convenient times, while providing a degree of control over the simulation that is usually not possible in the real-life situation.
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Our objective is to determine the quality of tissue engineered human skin via immunostaining, RT-PCR and electron microscopy (SEM and TEM). Culture-expanded human keratinocytes and fibroblasts were used to construct bilayer tissue-engineered skin. The in vitro skin construct was cultured for 5 days and implanted on the dorsum of athymic mice for 30 days. Immunostaining of the in vivo skin construct appeared positive for monoclonal mouse anti-human cytokeratin, anti-human involucrin and anti-human collagen type I. RT-PCR analysis revealed loss of the expression for keratin type 1, 10 and 5 and re-expression of keratin type 14, the marker for basal keratinocytes cells in normal skin. SEM showed fibroblasts proliferating in the 5 days in vitro skin. TEM of the in vivo skin construct showed an active fibrocyte cell secreting dense collagen fibrils. We have successfully constructed bilayer tissue engineered human skin that has similar features to normal human skin.
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Restoration of disturbed functions of the organs and tissues is the main task of contemporary genetic and cellular biotechnology, including genetic and cellular therapy. Duchenne dystrophy, one of the most widespread human genetic diseases, is at the same time the most extensively studied from the viewpoint of both genetic and histological changes leading to muscle fiber degeneration. Although many studies carried out on models, recognized analogous to Duchenne dystrophy, gave hopeful results, clinical tests with the use of developed methods gave no expected success and the rate of mortality from this disease amounts to 100%. Based on the world experience and analysis of the authors' data, possible influence of the intensity of regeneration on success of genetic and cellular therapy has been considered.
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The management of type 1 diabetes is being advanced by innovations in glucose measurement, development of an artificial pancreas, and in alternate routes of insulin delivery. Emerging technologies will allow insulin to be delivered more effectively. In the future, patients with type 1 diabetes will receive insulin in optimal quantities (because of more information about blood glucose values) at optimal times (because of better integration of blood glucose values with appropriate insulin dosages) by way of optimal routes into the body (because of needle-free routes of administration) in order to achieve optimal blood glucose control. Emerging technologies for improved care of patients with type 1 diabetes include: 1. new methods for monitoring glucose, including noninvasive, minimally invasive, continuous, and alternate site measurement technologies; 2. new methods for integrating glucose values and insulin dosages, known as the artificial pancreas; and 3. new routes of insulin administration, including inhaled, oral, buccal, nasal, and transdermal drug delivery systems. These new technologies will facilitate proper treatment of type 1 diabetes and improve the lives of affected patients.
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