Agent based modelling helps in understanding the rules by which fibroblasts support keratinocyte colony formation

Background: Autologous keratincoytes are routinely expanded using irradiated mouse fibroblasts and bovine serum for clinical use. With growing concerns about the safety of these xenobiotic materials, it is desirable to culture keratinocytes in media without animal derived products. An improved understanding of epithelial/mesenchymal interactions could assist in this. Methodology/Principal Findings: A keratincyte/fibroblast o-culture model was developed by extending an agent-based keratinocyte colony formation model to include the response of keratinocytes to both fibroblasts and serum. The model was validated by comparison of the in virtuo and in vitro multicellular behaviour of keratinocytes and fibroblasts in single and co-culture in Greens medium. To test the robustness of the model, several properties of the fibroblasts were changed to investigate their influence on the multicellular morphogenesis of keratinocyes and fibroblasts. The model was then used to generate hypotheses to explore the interactions of both proliferative and growth arrested fibroblasts with keratinocytes. The key predictions arising from the model which were confirmed by in vitro experiments were that 1) the ratio of fibroblasts to keratinocytes would critically influence keratinocyte colony expansion, 2) this ratio needed to be optimum at the beginning of the co-culture, 3) proliferative fibroblasts would be more effective than irradiated cells in expanding keratinocytes and 4) in the presence of an adequate number of fibroblasts, keratinocyte expansion would be independent of serum. Conclusions: A closely associated computational and biological approach is a powerful tool for understanding complex biological systems such as the interactions between keratinocytes and fibroblasts. The key outcome of this study is the finding that the early addition of a critical ratio of proliferative fibroblasts can give rapid keratinocyte expansion without the use of irradiated mouse fibroblasts and bovine serum

VKORC1 Pharmacogenetics and Pharmacoproteomics in Patients on Warfarin Anticoagulant Therapy: Transthyretin Precursor as a Potential Biomarker

Recognizing specific protein changes in response to drug administration in humans has the potential for the development of personalized medicine. Such changes can be identified by pharmacoproteomics approach based on proteomic technologies. It can also be helpful in matching a particular target-based therapy to a particular marker in a subgroup of patients, in addition to the profile of genetic polymorphism. Warfarin is a commonly prescribed oral anticoagulant in patients with prosthetic valve disease, venous thromboembolism and stroke.We used a combined pharmacogenetics and iTRAQ-coupled LC-MS/MS pharmacoproteomics approach to analyze plasma protein profiles of 53 patients, and identified significantly upregulated level of transthyretin precursor in patients receiving low dose of warfarin but not in those on high dose of warfarin. In addition, real-time RT-PCR, western blotting, human IL-6 ELISA assay were done for the results validation.This combined pharmacogenomics and pharmacoproteomics approach may be applied for other target-based therapies, in matching a particular marker in a subgroup of patients, in addition to the profile of genetic polymorphism

A user's guide to the Encyclopedia of DNA elements (ENCODE)

The mission of the Encyclopedia of DNA Elements (ENCODE) Project is to enable the scientific and medical communities to interpret the human genome sequence and apply it to understand human biology and improve health. The ENCODE Consortium is integrating multiple technologies and approaches in a collective effort to discover and define the functional elements encoded in the human genome, including genes, transcripts, and transcriptional regulatory regions, together with their attendant chromatin states and DNA methylation patterns. In the process, standards to ensure high-quality data have been implemented, and novel algorithms have been developed to facilitate analysis. Data and derived results are made available through a freely accessible database. Here we provide an overview of the project and the resources it is generating and illustrate the application of ENCODE data to interpret the human genome

Growth and reproduction in the Antarctic brooding bivalve Adacnarca nitens (Philobryidae) from the Ross Sea

We present information on the reproductive biology, population structure, and growth of the brooding Antarctic bivalve Adacnarca nitens Pelseneer 1903, from the Ross Sea, Antarctica. Individuals ranging from 0.85 - 6.00 mm were found attached to a hydrozoan colony. This species shows low fecundity and large egg size, common to other brooding species. The minimum size at which oogenesis was detected was 2.3 mm and the minimum size at which brooding was evident was 3.9 mm. Embryos of a full range of developmental stages were brooded simultaneously in females. The population showed a log-normal distribution and results suggest non-periodic reproduction with continuous embryonic development. The reproductive traits of A. nitens are discussed in the context of circum-Antarctic species distribution and limitations to dispersal in brooding benthic invertebrates

Caracterização imunofenotípica das subpopulações de linfócitos do lavado broncoalveolar de pacientes com silicose Phenotypic characterization of lymphocyte subsets in bronchoalveolar lavage of patients with silicosis

A lavagem broncoalveolar é um procedimento simples e seguro, na avaliação das pneumopatias relacionadas à exposição a poeiras minerais. O objetivo do estudo foi caracterizar as subpopulações celulares no lavado broncoalveolar (LBA) de pacientes silicóticos. A lavagem broncoalveolar foi realizada em 26 trabalhadores com diferentes formas de silicose: forma simples (n = 12), complicada (n = 13) e um paciente com a forma aguda da doença. Como grupo controle, foram incluídos sete indivíduos sadios. Os pacientes com silicose apresentaram intensa pleocitose com predomínio de macrófagos alveolares e tendência à linfocitose. As subpopulações de linfócitos presentes no lavado broncoalveolar (LBA) dos indivíduos sadios apresentaram fenótipo de células maduras. A grande maioria era constituída por células CD2+TCR<FONT FACE=Symbol>ab</FONT> (87,3%) e somente 2,9% das células T apresentaram marcação CD2+TCR<FONT FACE=Symbol>gd</FONT>. A relação CD4/CD8 foi de 1,8, com poucas (16%) células T imaturas duplo-negativas CD4-CD8-. Em contraste, pacientes com silicose apresentaram redução acentuada das subpopulações dos linfócitos maduros CD2+CD4+, CD2+CD8+ e aumento marcante (47%) de células imaturas (DN) duplo-negativas (CD4-CD8-). Não foi observado aumento das células NK (CD56+). A análise do conteúdo protéico e a determinação da relação Ig/albumina permitiram caracterizar produção local de imunoglobulinas no microambiente pulmonar. Como não foi observado aumento percentual de plasmócitos e linfócitos B (CD19+) no LBA desses pacientes, é possível concluir que as células produtoras de imunoglobulinas estão possivelmente localizadas no interstício pulmonar. Estes resultados sugerem que, durante a evolução da silicose, ocorre o desenvolvimento de linfopoese extratímica e surgimento de órgão linfóide terciário, no microambiente pulmonar desses pacientes.<br>Bronchoalveolar lavage is a safe and simple technique to evaluate lung disease related to exposure to mineral dusts. The aim of this study was to characterize the lymphocyte subsets in bronchoalveolar lavage of patients with silicosis. Bronchoalveolar lavage was carried out in 26 workers with different forms of silicosis: simple form (n = 12), complicated (n = 13) and 1 patient with acute form of the disease. As a control group, 7 healthy individuals were included. Compared to the control group, silicotic patients showed intense pleocytosis constituted mainly by alveolar macrophages with slight lymphocytosis. Lymphocyte subsets present in the bronchoalveolar fluid (BAL) of normal individuals were mature lymphocytes with phenotype CD2+TCR<FONT FACE=Symbol>ab</FONT> (87.3%) and only 2.9% were CD2+TCR<FONT FACE=Symbol>gd</FONT>. CD4/CD8 ratio was 1.8 with few (16%) immature double negative T cells subsets (CD4-CD8-). In contrast, silicotic patients showed reduction of the more mature lymphocyte subset CD2+CD4+, CD2+CD8+ and a great increase (47%) of immature (CD4-CD8-) T cell subsets. No increase in the NK (CD56+) cell population was observed. Biochemical analysis of protein contents and determination of the Ig/albumin ratio characterized local immunoglobulin production within the pulmonary microenvironment. Furthermore, lack of increase of plasma cells, as well as the maintenance of the percentage of B lymphocyte population (CD19+) in the BAL of silicotic patients, favors the hypothesis that the cells responsible for Ig production are possibly located in the interstitial space. Altogether the results suggest development of lymphopoiesis and tertiary lymphoid tissue within the pulmonary microenvironment during the clinical course of silicosis

New records of shelled marine molluscs at Bouvet Island and preliminary assessment of their biogeographic affinities

Bouvet Island is one of the most isolated islands in the Southern Ocean and its marine benthic fauna has rarely been investigated and so is poorly known. This study adds 30 molluscan species to the previous faunal record of 16 species. During the expedition ANT XXI-2 of PFS Polarstern 25 species of shelled gastropods, 12 species of bivalves, 3 species of polyplacophorans and 1 species of solenogaster (Aplacophora) were collected at four sites around Bouvet Island in depths between 120 m and 550 m. High percentages of the species representing the shelled molluscan fauna of Bouvet Island are known from sub and high (continental) Antarctica