Functional and phenotypical comparison of myofibroblasts derived from biopsies and bronchoalveolar lavage in mild asthma and scleroderma

BACKGROUND: Activated fibroblasts, which have previously been obtained from bronchoalveolar lavage fluid (BALF), are proposed to be important cells in the fibrotic processes of asthma and scleroderma (SSc). We have studied the motility for BALF derived fibroblasts in patients with SSc that may explain the presence of these cells in the airway lumen. Furthermore, we have compared phenotypic alterations in activated fibroblasts from BALF and bronchial biopsies from patients with mild asthma and SSc that may account for the distinct fibrotic responses. METHODS: Fibroblasts were cultured from BALF and bronchial biopsies from patients with mild asthma and SSc. The motility was studied using a cell migration assay. Western Blotting was used to study the expression of alpha-smooth muscle actin (α-SMA), ED-A fibronectin, and serine arginine splicing factor 20 (SRp20). The protein expression pattern was analyzed to reveal potential biomarkers using two-dimensional electrophoresis (2-DE) and sequencing dual matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-TOF). The Mann-Whitney method was used to calculate statistical significance. RESULTS: Increased migration and levels of ED-A fibronectin were observed in BALF fibroblasts from both groups of patients, supported by increased expression of RhoA, Rac1, and the splicing factor SRp20. However, these observations were exclusively accompanied by increased expression of α-SMA in patients with mild asthma. Compared to BALF fibroblasts in mild asthma, fibroblasts in SSc displayed a differential protein expression pattern of cytoskeletal- and scavenger proteins. These identified proteins facilitate cell migration, oxidative stress, and the excessive deposition of extracellular matrix observed in patients with SSc. CONCLUSION: This study demonstrates a possible origin for fibroblasts in the airway lumen in patients with SSc and important differences between fibroblast phenotypes in mild asthma and SSc. The findings may explain the distinct fibrotic processes and highlight the motile BALF fibroblast as a potential target cell in these disorders

Towards an anti-fibrotic therapy for scleroderma: targeting myofibroblast differentiation and recruitment

BACKGROUND: In response to normal tissue injury, fibroblasts migrate into the wound where they synthesize and remodel new extracellular matrix. The fibroblast responsible for this process is called the myofibroblast, which expresses the highly contractile protein alpha-smooth muscle actin (alpha-SMA). In normal tissue repair, the myofibroblast disappears. Conversely, abnormal myofibroblast persistence is a key feature of fibrotic dieases, including scleroderma (systemic sclerosis, SSc). Myofibroblasts can be derived from differentiation of local resident fibroblasts or by recruitment of microvascular pericytes. CLINICAL PROBLEM ADDRESSED: Controlling myofibroblast differentiation and persistence is crucial for developing anti-fibrotic therapies targeting SSc. BASIC SCIENCE ADVANCES: Insights have been recently generated into how the proteins transforming growth factor beta (TGFbeta), endothelin-1 (ET-1), connective tissue growth factor (CCN2/CTGF) and platelet derived growth factor (PDGF) contribute to myofibroblast differentiation and pericyte recruitment in general and to the persistent myofibroblast phenotype of lesional SSc fibroblast, specifically. RELEVANCE TO CLINICAL CARE: This minireview summarizes recent findings pertinent to the origin of myofibroblasts in SSc and how this knowledge might be used to control the fibrosis in this disease. CONCLUSIONS: TGFbeta, ET-1, CCN2 and PDGF are likely to cooperate in driving tissue repair and fibrogenic responses in fibroblasts. TGFbeta, ET-1 and CCN2 appear to contribute to myofibroblast differentiation; PDGF appears to be involved with pericyte recruitment. Thus, different therapeutic strategies may exist for targeting the multisystem fibrotic disorder SSc

A methodology for exploring biomarker – phenotype associations: application to flow cytometry data and systemic sclerosis clinical manifestations

BACKGROUND: This work seeks to develop a methodology for identifying reliable biomarkers of disease activity, progression and outcome through the identification of significant associations between high-throughput flow cytometry (FC) data and interstitial lung disease (ILD) - a systemic sclerosis (SSc, or scleroderma) clinical phenotype which is the leading cause of morbidity and mortality in SSc. A specific aim of the work involves developing a clinically useful screening tool that could yield accurate assessments of disease state such as the risk or presence of SSc-ILD, the activity of lung involvement and the likelihood to respond to therapeutic intervention. Ultimately this instrument could facilitate a refined stratification of SSc patients into clinically relevant subsets at the time of diagnosis and subsequently during the course of the disease and thus help in preventing bad outcomes from disease progression or unnecessary treatment side effects. The methods utilized in the work involve: (1) clinical and peripheral blood flow cytometry data (Immune Response In Scleroderma, IRIS) from consented patients followed at the Johns Hopkins Scleroderma Center. (2) machine learning (Conditional Random Forests - CRF) coupled with Gene Set Enrichment Analysis (GSEA) to identify subsets of FC variables that are highly effective in classifying ILD patients; and (3) stochastic simulation to design, train and validate ILD risk screening tools. RESULTS: Our hybrid analysis approach (CRF-GSEA) proved successful in predicting SSc patient ILD status with a high degree of success (>82 % correct classification in validation; 79 patients in the training data set, 40 patients in the validation data set). CONCLUSIONS: IRIS flow cytometry data provides useful information in assessing the ILD status of SSc patients. Our new approach combining Conditional Random Forests and Gene Set Enrichment Analysis was successful in identifying a subset of flow cytometry variables to create a screening tool that proved effective in correctly identifying ILD patients in the training and validation data sets. From a somewhat broader perspective, the identification of subsets of flow cytometry variables that exhibit coordinated movement (i.e., multi-variable up or down regulation) may lead to insights into possible effector pathways and thereby improve the state of knowledge of systemic sclerosis pathogenesis. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12859-015-0722-x) contains supplementary material, which is available to authorized users

The effect of energy crops on the emission of greenhouse gases

Podjęto próbę wstępnego oszacowania wpływu upraw energetycznych na atmosferę. Rozważono cztery źródła emisji związane z produkcją biomasy roślinnej. Analiza pozwoliła na obliczenie poziomu emisji gazów cieplarnianych na przestrzeni 14 lat użytkowania plantacji. Emisja, szczególnie N2O, z gleby w znacznym stopniu wpływa na bilans uprawy. Jest ona wielokrotnie wyższa niż emisja gazów cieplarnianych związana ze zużyciem paliwa w czasie pracy maszyn rolniczych. Jednocześnie emisja z gleby na plantacji wierzby jest wyższa w porównaniu z topolą. Szczególnie widoczna różnica jest przy dawce nawożenia 75 kg N. Na plantacji wierzby podczas 14 lat jej użytkowania emisja gazów cieplarnianych wyniosła 10444,08 kg CO2 equ/ha, przy tych samych założeniach dla plantacji topoli kształtowała się ona na poziomie 8.666,72 kg CO2 equ/ha.An attempt was made to introductory evaluating the impact of growing energy crops on the atmosphere. Four emission sources connected with plant biomass production were considered. Analysis enabled to calculate the level of greenhouse gas emission during 14 years of plantation use. Emissions, particularly N2O from the soil, considerably affected the balance of cultivated crops. They are many times higher than greenhouse gas emissions connected with fuel consumption during operation of agricultural machines. At the same time the emissions from soil under willow plantation are much higher as compared with the poplar plantation. The difference was particularly distinct at fertilization rate of 75 kg N. During 14 years' using of the willow plantation the greenhouse gas emission amounted to 10444.08 kg CO2 equ/ha, while for the poplar plantation emission reached the level of 8666.72 kg CO2 equ/ha

Production development of biofuels in the European Union

Przedstawiono zmiany w produkcji biopaliw w państwach członkowskich Unii Europejskiej na przestrzeni ostatnich lat. Omówiono kluczowe czynniki sukcesu Niemiec i Szwecji w zwiększaniu wielkości produkcji biopaliw. Niemcy są liderem europejskim na rynku biodiesla, natomiast Szwecja dynamicznie rozwija produkcję bioetanolu. W produkcji bioetanolu istotną rolę odgrywa także Słowenia z produkcją 320 tys. t w 2006 r. Największym producentem i konsumentem biopaliw w UE są jednak Niemcy. W 2004 r. produkcja biodiesla osiągnęła poziom 1035 tys. t, a do 2006 r. została podwojona (2662 tys. t). Ważnym producentem biodiesla jest także Francja z produkcją 743 tys. t w 2006 r. Przegląd osiągnięć poszczególnych państw UE w zakresie produkcji i wykorzystania biopaliw nie pozwala jednak sądzić, że cel Unii - 5,75% udziału biopaliw w 2010 r. - ma szanse zostać realizowany.Paper discussed the changes in production of biofuels in Member Countries of the EU within last years. The key success factors for biofuels' production development in Germany and Sweden were mentioned. Germany is a leader on European biodiesel market, while Sweden dynamically increases bioethanol production. Important role in bioethanol production plays also Slovenia (320000 t in 2006). The biggest producer and consumer of biofuels in the EU is still Germany. Biodiesel production in 2004 reached there the level of 1035000 t and was doubled in 2006 (2662000 t). Important producer of biodiesel is also France at production of 743000 t (2006). However, the review of achievements on - 5.75% share of biofuels in 2010 - is going to be realized

Balance of the agricultural biomass (straw) for energy purposes

Jednym z celów strategicznych Polski jest zwiększenie udziału energii ze źródeł odnawialnych w finalnym zużyciu energii, co najmniej do 15% w 2020 r. (Polityka Energetyczna Polski do 2030 roku). Polska energetyka systemowa jest zobligowana do wykorzystywania biomasy rolnej w procesie produkcji energii. Celem niniejszej pracy jest określenie bilansu biomasy rolnej, która może być przeznaczona do energetycznego wykorzystania. Na podstawie przyjętych założeń oszacowano nadwyżkę słomy produkowanej w Polsce. Obliczono, że w ciągu ostatnich trzech lat nadwyżka zwiększyła się z ok. 1,3 mln t w 2007 r. do 2,6 mln t w 2009 r.One of the strategic purposes of Poland is to increase the share of energy from renewable sources in total energy consumption at least to 15% in 2020 (Energetic Policy of Poland until 2030). Polish systemic energetics has been obliged to use the agricultural biomass in process of energy generation. The study undertaken aimed at evaluating the balance of agricultural biomass which may be provided for energetic usage. On the basis of assumed rules, the surplus of straw produced in Poland was estimated. It was calculated that during last three years this surplus increased from about 1.3 million ton in 2007, up to 2.6 million ton in 2009