Adipose-tissue-derived therapeutic cells in their natural environment as an autologous cell therapy strategy: the microtissue-stromal vascular fraction.

The prerequisite for a successful clinical use of autologous adipose-tissue-derived cells is the highest possible regenerative potential of the applied cell population, the stromal vascular fraction (SVF). Current isolation methods depend on high enzyme concentration, lysis buffer, long incubation steps and mechanical stress, resulting in single cell dissociation. The aim of the study was to limit cell manipulation and obtain a derivative comprising therapeutic cells (microtissue-SVF) without dissociation from their natural extracellular matrix, by employing a gentle good manufacturing practice (GMP)-grade isolation. The microtissue-SVF yielded larger numbers of viable cells as compared to the improved standard-SVF, both with low enzyme concentration and minimal dead cell content. It comprised stromal tissue compounds (collagen, glycosaminoglycans, fibroblasts), capillaries and vessel structures (CD31+, smooth muscle actin+). A broad range of cell types was identified by surface-marker characterisation, including mesenchymal, haematopoietic, pericytic, blood and lymphatic vascular and epithelial cells. Subpopulations such as supra-adventitial adipose-derived stromal/stem cells and endothelial progenitor cells were significantly more abundant in the microtissue-SVF, corroborated by significantly higher potency for angiogenic tube-like structure formation in vitro. The microtissue-SVF showed the characteristic phenotype and tri-lineage mesenchymal differentiation potential in vitro and an immunomodulatory and pro-angiogenic secretome. In vivo implantation of the microtissue-SVF combined with fat demonstrated successful graft integration in nude mice. The present study demonstrated a fast and gentle isolation by minor manipulation of liposuction material, achieving a therapeutically relevant cell population with high vascularisation potential and immunomodulatory properties still embedded in a fraction of its original matrix.

[Health indicator-based cluster analysis of districts and urban districts in North Rhine-Westphalia]. / Sozialräumliche Clusteranalyse der Kreise und kreisfreien Städte und Gesundheitsindikatoren in NRW.

OBJECTIVE: North Rhine-Westphalia (NRW's) indicator set for health reporting activities comprises more than 70 regional health indicators, which means that these data are available for health reporting purposes for all 54 districts and urban districts. Morbidity and mortality indicators differ in part quite considerably and require further interpretation. With the help of selected indicators, the authors of the following article try to explain the relation between social status and health status. METHODOLOGY: Ten years ago, NRW, as part of its health reporting activities, started to carry out multivariate analyses to classify socio-demographically different types of regions, leading to the establishment of six types of regions which can be linked to health-related data. Social structure indicators are part of a first step submitted to a main component analysis and grouped together by a small number of features and/or factors which clearly reflect differences in living conditions. As a result, two factors were extracted: an economic prosperity factor which is mainly determined by the disposable income and a so-called A-factor which mainly describes the fact that poorer, elderly, unemployed and foreign population groups live concentrated in regions with a declining population but high population density. These factors are, in a second step, used for a cluster analysis aimed at classifying the 54 districts and urban districts and at establishing different types of regions. In a subsequent step, the cluster method is used to explain regional variations of selected health indicators. RESULTS: It is a proven fact that morbidity and mortality are influenced by social status. With the help of selected indicators, six clusters with a different socio-economic structure influencing the health status of the population can be established for NRW. Special attention should be paid to the cluster of the Ruhr area with its below-average social situation. With 90% NRW's population primarily living within the other 5 clusters which are differently structured but increasingly adjusting their living conditions to each other. The authors of this publication assign four health status indicators to predefined clusters and analyse the relation between the social and health status: female and male life expectancy, the proportion of underweight live births, infant mortality and avoidable deaths.In regions with high A-factor values (poverty pole), i. e., in several ways socially deprived regions, male and female average life expectancy is significantly lower than in regions with a clearly less pronounced accumulation of problems. Moreover, a significantly higher life expectancy for male live births can be observed in regions with a high disposable income. The model fails to establish a convincing correlation between social status and infant mortality and breast cancer. CONCLUSIONS: Knowledge about socio-demographic differences in the health status of the population is particularly important for prevention measures in order to be able to react appropriately to health risks in districts and urban districts. The analysis shows that an intense regional accumulation of problems will have a negative influence on health status, an influence which is more significant than the positive influence of prosperous regions on the health status.

Measurement of hydrocarbon-degrading microbial populations by a 96-well plate most-probable-number procedure.

A 96-well microtiter plate most-probable-number (MPN) procedure was developed to enumerate hydrocarbon-degrading microorganisms. The performance of this method, which uses number 2 fuel oil (F2) as the selective growth substrate and reduction of iodonitrotetrazolium violet (INT) to detect positive wells, was evaluated by comparison with an established 24-well microtiter plate MPN procedure (the Sheen Screen), which uses weathered North Slope crude oil as the selective substrate and detects positive wells by emulsification or dispersion of the oil. Both procedures gave similar estimates of the hydrocarbon-degrader population densities in several oil-degrading enrichment cultures and sand samples from a variety of coastal sites. Although several oils were effective substrates for the 96-well procedure, the combination of F2 with INT was best, because the color change associated with INT reduction was more easily detected in the small wells than was disruption of the crude oil slick. The method's accuracy was evaluated by comparing hydrocarbon-degrader MPNs with heterotrophic plate counts for several pure and mixed cultures. For some organisms, it seems likely that a single cell cannot initiate sufficient growth to produce a positive result. Thus, this and other hydrocarbon-degrader MPN procedures might underestimate the hydrocarbon-degrading population, even for culturable organisms.

[Family formation in North Rhine-Westphalia: models and microsimulation using panel data]. / Familienentwicklung in Nordrhein-Westfalen: Modellierung und Mikrosimulation mit Paneldaten.

PIP: This publication is the last in a series presenting results from a longitudinal study of reproductive behavior in North Rhine-Westphalia, Federal Republic of Germany. The study, conducted between 1981 and 1987, involved three surveys of women aged 18-30 at the beginning of the period; 1,054 women participated in all three surveys. The focus of the present paper is on the use of panel data and microsimulation techniques to develop models for forecasting processes of family formation, particularly the birth of children. Results of the simulation models are compared with actual findings from the survey.^ieng

Induction of neutralizing antibodies and immunity in vaccinated guinea pigs by cyanogen bromide-peptides of VP3 of foot-and-mouth disease virus.

The specificity of guinea pig antisera against large cyanogen bromide-cleaved peptides of the virus capsid protein VP3 of foot-and-mouth disease virus type O1, strain Kaufbeuren has been characterized by double immunodiffusion, virus neutralization and protection tests. Antibodies to purified 146S particles and the cleavage peptides of VP3 showed an incomplete cross-section against VP3 peptide antigen when reacted in immunodiffusion tests, indicating that new antigenic determinants are exhibited by the peptides which are not recognized by the antiserum against the native virus proteins. The immune response against the reduced, unfolded chain constituents of VP3 was lower in comparison to that of native virus particles but still some immunological determinants remained actively capable of inducing virus-neutralizing antibodies in immunized guinea pigs.