Expansion in bioreactors of human progenitor populations from cord blood and mobilized peripheral blood

Umbilical cord blood (UCB) and mobilized peripheral blood (MPB) provide an alternate source to bone marrow for transplantation. Expansion in vitro of stem/progenitor cell populations from these sources may provide adult-sized grafts otherwise not attainable because of the limited cell numbers available in the case of UCB or because of numerous rounds of apheresis required for sufficient MPB cells. We asked whether continuous perfusion culture could be employed in ex vivo expansion to produce clinically relevant numbers of stem/progenitor cells from these sources. To evaluate MPB, 1-10 million leukocytes, from patients who had received either granulocyte colony-stimulating factor (G-CSF) or cyclophosphamide and granulocyte-macrophage colony-stimulating factor (GM-CSF), were inoculated into bioreactors, with or without irradiated, allogeneic stroma. The growth factor combination in the perfusion medium consisted of interleukin-3 (IL-3), stem cell factor (SCF), GM-CSF and erythropoietin (Epo). Under the best conditions tested, total cell numbers, granulocyte-macrophage colony-forming units (CFU-GM), and long-term culture-initiating cell (LTC-IC) populations were expanded by about 50-, 80-, and 20-fold, respectively, over 14 days. At low cell inocula (1 million), the presence of stroma enhanced the expansion of total cells and CFU-GM but not of LTC-IC. When SCF was not included in the medium, both total cells and CFU-GM expanded to a much lesser extent, but again the expansion of LTC-IC was not affected. At the higher cell inoculum (10 million), expansions of total cells and CFU-GM were equivalent with or without stroma. To evaluate UCB, cells were placed into bioreactors with or without irradiated, allogeneic stroma, and the bioreactors were perfused with medium containing the four standard growth factors. After 6-14 days, in several independent experiments, 20-24 million cells were harvested from bioreactors perfused with SCF-containing medium, irrespective of the presence or absence of preformed stroma. Similarly, in reactors perfused with SCF-containing medium (with or without stroma), an average 40- to 60-fold expansion of CFU-GM was obtained, yielding an average of 1.5-1.8 x 10(5) CFU-GM per reactor. Harvested cells were thus up to 40-fold enriched in CFU-GM in comparison to the inoculum. In the absence of SCF, cell expansions averaged 1.5- to 2-fold, and CFU-GM were expanded only 10- to 14-fold by day 14. As before, the presence of preformed stroma did not affect either cell or CFU-GM yields, provided the cell inoculum was at least 4.5 million cells.(ABSTRACT TRUNCATED AT 400 WORDS)     

Tissue culture surface characteristics influence the expansion of human bone marrow cells

Human cell therapy applications in tissue engineering, such as the ex vivo production of hematopoietic cells for transplantation, have recently entered the clinic. Although considerable effort has been focused on the development of biological processes to generate therapeutic cells, little has been published on the design and manufacture of devices for implementation of these processes in a robust and reproducible fashion at a clinical scale. In this study, the effect of tissue culture surface chemistry and texture was assessed in human bone marrow (BM) mononuclear cell (MNC) and CD34-enriched cell cultures. Growth and differentiation was assessed by total, progenitor (CFU-GM), stromal (CFU-F), and primitive (LTC-IC) cell output. Tissue culture treated (TCT) plastic significantly increased MNC culture output as compared with non-TCT plastic, whereas CD34-enriched cell cultures gave lower output (than MNC cultures) that was unaffected by TCT plastic. Interestingly, the level of MNC culture output was significantly different on four commercial TCT surfaces, with the best performing surface giving output that was 1.6- to 2.8-fold greater than the worst one. The surface giving the highest output was the best at supporting development of a distinct morphological feature in the adherent layer (i.e. cobblestone area) indicative of primitive cells, and X-ray photoelectron spectroscopy (XPS) was used to characterize this surface. For custom injection molding of culture devices, the use of three different resins resulted in MNC culture output that was equivalent to commercial cultureware controls, whereas CD34-enriched cell cultures were highly sensitive to resins containing additives. When the texture of molded parts was roughened by sandblasting of the tool, MNC culture output was significantly reduced and higher spikes of IL-6 and G-CSF production were observed, presumably due to macrophage activation. In conclusion, the manufacture of BM MNC culture devices for clinical applications was optimized by consideration of plastic resin, surface treatment, and texture of the culture substratum. Although CD34-enriched cells were insensitive to surface treatment, they were considerably more sensitive to biocompatibility issues related to resin selection. The development of robust systems for BM MNC expansion will enable clinical trials designed to test the safety and efficacy of cells produced in this novel tissue engineering application.     

The measurement of paste content in hardened concrete using automatic image analyzing technique

This paper describes a technique for the measurement of paste contents of mortars and concretes. The technique involves etching the polished surfaces of samples with a saturated solution of salicylic acid in 80:20 methyl alcohol: water mixture which does not affect limestone aggregates. The contrast difference between the exposed aggregates and the etched areas is then enhanced by painting the aggregates black and filling the etched areas with a white powder. An automatic image analyzer can then measure either the blackened aggregates or the whitened etched areas.     

Potentials and challenges of peer-to-peer based content distribution

Multimedia content currently accounts for over three quarters of all Internet traffic. This increase in traffic volume and content availability derives from a paradigm shift from the traditional text and picture based Web, to more resource demanding audio and video content. A controversial driver for this development is content distribution systems based on peer-to-peer overlay networks. Flooding the Internet with often illegal content, these networks now pose challenges to all actors in the value chain. However, if viewed as surmountable challenges in an evolutionary path, peer-to-peer technology has the potential of increasing efficiency in content distribution and unleashing resources to form scalable and resilient overlay networks of unprecedented dimensions.

In this paper we examine the potentials and challenges of peer-to-peer technology in content distribution, and analyse how, and under which circumstances, peer-to-peer technology can be used to increase the efficiency of multimedia services. The paper provides an up-to-date overview of the development of peer-to-peer networks as well as describing the economics laws governing their use. To conclude the study, the paper analyses Skype, a well known telecommunications service utilising the peer-to-peer technology, as well as demonstrating the benefits of peer-to-peer based content distribution using empirical data from the Danish Broadcasting Corporation.     

Retroviral infection is limited by Brownian motion

Replication-defective retroviruses are frequently used as gene carriers for gene transfer into target cells. Here we show that the short half-lives of retroviruses limit the distance that they can effectively travel in solution by Brownian motion, and thus the possibility of successful gene transfer. This physiochemical limitation can be overcome, and effective contact between the retroviral gene carrier and the target cell can be obtained, by using net convective flow of retrovirus-containing medium through a layer of target cells. Using model cell lines (NIH-3T3 and CV-1), it was shown that gene transfer rates can be increased by more than an order of magnitude using the same concentration infection medium. High transduction rates could be obtained even in the absence of polycations, such as Polybrene, which heretofore have been required to achieve reasonable transduction rates. This development may play an important role in realizing human gene therapy.     

Outdoor atmospheric corrosion of carbon steel and weathering steel exposed to the tropical-coastal climate of Thailand

This study focuses on atmospheric corrosion of carbon steel and weathering steel exposed at two different locations with dissimilar meteorological parameters and airborne pollutants in Thailand. The samples are subjected to an outdoor atmosphere for up to 36 months at Rayong, close to the Gulf of Thailand, and Phangnga, near the Andaman Sea. Thickness loss (µm), corrosion rate (µm/year) together with corrosion product morphology and composition are systematically analyzed using X-ray diffraction and a scanning electron microscope. Corrosion resistance of the tested steels exposed at both locations is discussed based on the above-mentioned parameters and calculated corrosion kinetics. The results indicate that the total time of wetness, amount of rainfall and chloride deposition rate play an important role in corrosion behavior of the tested steels. Alloying elements, copper, chromium and nickel, are shown to improve corrosion resistance of the samples when exposed at the location with a higher chloride concentration.     

Is rectal pain sensitivity a biological marker for irritable bowel syndrome: psychological influences on pain perception

BACKGROUND & AIMS: Rectal pain sensitivity has been called a biological marker for irritable bowel syndrome, but this conclusion may be premature. This article is a critical review of the evidence for psychological influences on perception. METHODS: The world literature accessible through Index Medicus from 1973 to 1997 was systematically reviewed. RESULTS: Evidence favoring a biological basis for pain sensitivity is that two thirds of patients report pain at abnormally low thresholds of rectal distention despite normal somatic pain thresholds. Pain thresholds are not correlated with anxiety or depression. Evidence favoring psychological influences on perception is that patients with the irritable bowel syndrome rate even sham distentions as more painful, and when perception tests that minimize psychological influences are used, they have normal sensory thresholds. Also, stress alters sensory thresholds. Sensitization by repeated distention has been cited as evidence of a biological basis for hyperalgesia, but it is not unique to patients with irritable bowel. Brain imaging shows that different regions are activated by painful distention in patients with irritable bowel syndrome, but this is consistent with psychological influences on perception. CONCLUSIONS: Psychological factors influence pain thresholds in patients with the irritable bowel syndrome. Two cognitive traits, selective attention to gastrointestinal sensations and disease attribution, may account for increased pain sensitivity.     

A Macintosh software package for simulation of human red blood cell metabolism.

We have developed a computer software package for Macintosh to simulate the metabolism and hemoglobin binding affinity of human red blood cell. The model is capable of simulating hemoglobin binding of ligands, metabolite concentrations, and metabolic fluxes at physiological steady state and in response to extracellular parameter variations, such as pH, osmolarity, glucose, and adenine concentrations. The kinetic parameters of enzymes, extracellular conditions, and initial intracellular metabolite concentrations can be specified by the user in order to model a particular situation. The software is use friendly, utilizing menu, window, and mouse to interact with the user. It also provides a pathway map of the red cell, which allows a direct access to enzyme kinetics by clicking the enzymes in the map.     

Glycerol monocaprate (monocaprin) reduces contamination by Escherichia coli and Salmonella enteritidis on hard surfaces

Emulsions of glycerol monocaprate (monocaprin) kill a variety of pathogenic bacteria and viruses in suspension. In this study the microbicidal activity of monocaprin against enterobacteria was tested on contaminated hard surfaces. Surfaces were contaminated with nutrient broth or meat juice containing large numbers of Escherichia coli or Salmonella enteritidis. They were then treated with acidified monocaprin emulsions and the surviving bacteria counted. Monocaprin killed S. enteritidis in chicken meat juice on plastic cutting boards and reduced the number of viable E. coli and S. enteritidis by more than 5 log₁₀ in 2 min on a laminated plastic kitchen counter contaminated with nutrient broth. Monocaprin rapidly killed E. coli on glass, stainless steel, laminated plastic, glazed ceramic tiles and polypropylene boards. It was most effective on glass and stainless steel and more effective on dry than on wet surfaces. It was concluded that acidified monocaprin emulsions reduce contamination by pathogenic enterobacteria on hard surfaces. They may be useful as sanitizers in the home, and possibly in public places, where contaminated surfaces are a potential source of transmission of pathogens to humans. Cleaning with monocaprin emulsions may therefore be a means to improve hygiene and infection control.     

Single,twin and triple buried heating pipes: on potential savings in heat losses and costs

In order to make district heating systems competitive in areas with single family houses or in other areas with low heat demands it is necessary to reduce the heat losses from the pipes. In recent years the twin pipe has become popular in the Nordic countries. In the article we describe how the heat loss and the heat loss coefficients can be calculated. We introduce the triple pipe with three media pipes (two supply pipes and one return pipe). The temperature dependency and the ageing of polyurethane insulation are briefly discussed. A comparison is made for different 80 mm distribution pipes and for different service pipes with respect to heat losses and to resources, i.e. materials needed for the casing and polyurethane insulation and the gravel in the excavations. For the distribution pipe we found that an egg‐shaped twin pipe can reduce the heat loss by 37% and the investments by 12% compared with a pair of single pipes. For the service pipes we found that the triple pipe reduces the heat loss by 45% compared with a common pair of single pipes and by 24% compared with circular twin pipes. The reduction in investment index is 21%. The article also addresses the question of the heat exchange between the two media pipes in a twin pipe. Copyright © 2005 John Wiley & Sons, Ltd.