LeukoCatch, a quick and efficient tool for the preparation of leukocyte extracts from blood

<p>Abstract</p> <p>Background</p> <p>Whole-protein extracts from peripheral blood leukocytes are ideal for basic and clinical research. However, lack of a simple preparation technique has limited the use of such extracts. The aim of this study is to develop a simple and easy system that can selectively obtain leukocyte extracts without hemoglobin.</p> <p>Methods</p> <p>A filter that captures the leukocytes but not RBCs was set at the bottom of a 10-mL medical syringe by sandwiching it between plastic stoppers. The capturing efficiency of leukocytes with this tool, called LeukoCatch, was examined using human macrophage cells (MONO-MAC-6). The abilities of LeukoCatch system to capture the leukocyte proteins and to remove the hemoglobin from RBCs were tested by western blot analysis using human blood samples.</p> <p>Results</p> <p>This study presents the development of LeukoCatch, a novel tool that allows the preparation of leukocyte extracts from blood samples within 3 min without centrifugation. Tissue-cultured human macrophage cells were tested to determine the optimal filter numbers and pass-through frequencies of LeukoCatch, which was then applied to 2-mL blood samples. Samples were passed 2~5 times through a LeukoCatch equipped with 5 filters, washed twice with phosphate-buffered saline for red cell removal, and leukocyte proteins were extracted with 0.5 mL of elution buffer. Western blot analysis of the purified extract indicated that more than 90% of hemoglobin was removed by the LeukoCatch and that the protein recovery rate of leukocytes was at least 4 times better than that of the conventional centrifugation method.</p> <p>Conclusion</p> <p>We conclude that LeukoCatch is useful not only for diagnosis at the bedside but also for basic research using blood samples or tissue culture cells.</p

Metformin Prevents and Reverses Inflammation in a Non-Diabetic Mouse Model of Nonalcoholic Steatohepatitis

Background: Optimal treatment for nonalcoholic steatohepatitis (NASH) has not yet been established, particularly for individuals without diabetes. We examined the effects of metformin, commonly used to treat patients with type 2 diabetes, on liver pathology in a non-diabetic NASH mouse model. Methodology/Principal Findings: Eight-week-old C57BL/6 mice were fed a methionine- and choline-deficient plus high fat (MCD+HF) diet with or without 0.1% metformin for 8 weeks. Co-administration of metformin significantly decreased fasting plasma glucose levels, but did not affect glucose tolerance or peripheral insulin sensitivity. Metformin ameliorated MCD+HF diet-induced hepatic steatosis, inflammation, and fibrosis. Furthermore, metformin significantly reversed hepatic steatosis and inflammation when administered after the development of experimental NASH. Conclusions/Significance: These histological changes were accompanied by reduced hepatic triglyceride content, suppressed hepatic stellate cell activation, and the downregulation of genes involved in fatty acid metabolism, inflammation, and fibrogenesis. Metformin prevented and reversed steatosis and inflammation of NASH in an experimental non-diabetic model without affecting peripheral insulin resistance. © 2012 Kita et al

Eco-Friendly Cotton/Linen Fabric Treatment Using Aqueous Ozone and Ultraviolet Photolysis

Chemicals for the scouring and bleaching of fabrics have a high environmental load. In addition, in recent years, the high consumption of these products has become a problem in the manufacture of natural fabric products. Therefore, environmentally friendly, low-waste processes for fabric treatment are required. In this paper, we discuss the bleaching of fabrics using advanced oxidation processes (AOP). These processes use electrochemically generated aqueous ozone and ultraviolet (UV) irradiation to achieve bleaching. However, colour reversion often occurs. In this study, we suppressed unwanted colour reversion by treatment with rongalite. After treatment, changes in fabric colour were determined by measuring the colour difference and reflectance spectra. The best bleaching effect was obtained when ozone and UV irradiation treatments were combined, achieving results similar to those of a conventional bleaching method after 60 min of UV irradiation. In addition, the AOP treatment resulted in the simultaneous scouring of the fabric, as shown by the increased hydrophilicity of the fabric after AOP treatment. Thus, this AOP process represents a new fabric bleaching process that has an extremely low environmental impact

The McGill Pain Questionnaire, Japanese Version, Reconsidered: Confirming the Theoretical Structure

BACKGROUND: Based on a tripartite theoretical model of pain, the Pain Rating Index (PRI) of the McGill Pain Questionnaire (MPQ) continues to be one of the most frequently used instruments to measure clinical pain. However, language and cultural barriers have hindered its wide use and standardization in Japan. Although a number of exploratory factor analysis studies have failed to support consistently the theoretical structure of the MPQ, a few previous confirmatory factor analysis studies did statistically support the a priori model