3 research outputs found
Enhancing Kojic Acid Production in Aspergillus oryzae: Leveraging Crude Cellulase from Achatina fulica for Strain Improvement via Protoplasting and UV Mutagenesis
No full textThis study aims to prove the ability of crude cellulase enzymes from snails for protoplasting Aspergillus oryzae cells and its application for strain improvement with UV mutagenesis. Snail enzyme was obtained from Achatina fulica by dissolving its digestion track and fractionating it with ammonium sulfate. The activity of fractions was measured Spectrophotometrically and used for cell protoplasting for 2 hours, then irradiated with UV for 10, 15, and 20 minutes, respectively, with 5 cm in the distance. Screening of mutants is carried out with 1% FeCl3, and the potential mutant strain was tested for kojic acid production in an aerobic state and determined by Spectrophotometry at 268 nm. The cellulase activity in crude snail enzyme was 11.5807 U/ml and increased to 16.3984 U/ml after fractionation. The best protoplast formation was obtained with a 60% fraction, which showed transparent performance under the microscope. The UV mutagenesis of protoplasts showed that the highest number of potential mutants was obtained from UV treatment for 15 minutes (41.67%). The potential mutants look dark brown (DBC), such as strain 10H3, and produced higher kojic acid concentration than the parent strain. In conclusion, UV mutagenesis of Aspergillus oryzae through protoplasting by crude cellulase of snail enzyme was effective and improved kojic acid concentration
Whipworms in humans and pigs: origins and demography
No full text© 2016 Hawash et al. Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and
reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to
the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver
(http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. The attached file is the published version of the article.NHM Repositor
Translational research in rheumatoid arthritis: Exploiting melanocortin receptors
No full textThe copyright of this thesis rests with the author and no quotation from it or information derived from it may be published without the prior written consent of the authorRheumatoid arthritis (RA) is a chronic inflammatory disease affecting 1% of the population. The aetiology of rheumatoid arthritis is unknown, although there are multiple postulated theories. In 1950, Philip Hench won the Nobel prize for treating patients with rheumatoid arthritis with cortisone. He also treated 6 patients with adrenocorticotropic hormone (ACTH) with good results. ACTH is a melanocortin. The melanocortin system describes the five melanocortin receptors, their ligands, agonists and antagonists and the accessory proteins. The aim of this study was to explore the melanocortin receptors in rheumatoid arthritis synovium.
Methods
HA-tagged stable cell lines were created for MC1R, MC3R and MC5R. Multiple antibodies were tested for their utility using Western Blot, immunohistochemistry and flow cytometry. Samples of synovium from 28 patients with RA were tested using RTPCR for the presence of MC1R and MC3R. Gene expression was correlated with clinical characteristics, cytokine (RTPCR) expression and immunohistochemical score.
Results
The stable cell lines expressed MC1R, MC3R and MC5R respectively. Of the antibodies tested none were found to be of utility in detecting MC1R or MC3R .The MC1R RQ values in rheumatoid synovium appear to split into two groups, high and low. The medians of the two groups are significantly different (p=0.0005). There is almost a 5 cycle, or 64 fold, difference in gene expression between the medians of the two groups (1.59 v 6.23). Of note no MC3R positive samples were CD138 high (i.e. no MC3R positive samples had a significant plasma cell infiltrate) (p=0.006). Categorical analysis using Fishers Exact test revealed an association between MC1R high samples and CD68 lining high scores, (i.e. MC1R high samples also had a high macrophage score in the lining of the sample) (p=0.02). MC1R low samples were associated with not being on combination therapy,
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this did not quite reach significance (p=0.07). Linear regression analysis confirmed these associations for MC1R. PCA analysis did not show any grouping of samples according to any of the variables tested, likely due to sample size.
Conclusion
MC1R and MC3R are found in human synovium. Current commercial antibodies are not of utility in detecting MC1R or MC3R. Synovial samples can be split into high and low MC1R gene expression groups. MC3R was either present or absent. High expression of MC1R was associated with a high macrophage score and MC3R expression was associated with a low plasma cell score. MC1R and MC3R expression in RA synovium could be used as biomarkers of disease state or severity as well as a target for therapy
