Leukocyte redistribution as immunological biomarker of corticosteroid resistance in severe asthma.

BACKGROUND: Earlier studies have suggested that the leukocyte redistribution can be considered as an immunological marker of the clinical response to corticosteroids (CS), representing an easy measurable potential biomarker in severe asthma. OBJECTIVE: The aim of this study was to determinate the utility of the leukocyte redistribution as a biomarker of disease heterogeneity in patients with severe asthma and as a bioindicator of potential CS resistance. METHODS: We developed an unbiased clustering approach based on the clinical data and the flow cytometry results of peripheral blood leukocyte phenotypes of 142 patients with severe asthma before and after systemic CS administration. RESULTS: Based on the differences in the blood count eosinophils, neutrophils and lymphocytes, together with the flow cytometry measurements of basic T cell, B cell and NK cell subpopulations before and after systemic CS administration, we identified two severe asthma clusters, which differed in the cell frequencies, response to CS and atopy status. Patients in cluster 1 had higher frequency of blood eosinophils at baseline, were sensitized to less allergens and had better steroid responsiveness, measured as the pronounced leukocyte redistribution after the administration of systemic CS. Patients in cluster 2 were determined by the higher frequency of B-cells and stronger IgE sensitization status to the multiple allergens. They also displayed higher steroid resistance, as the clinical correlate for the lower leukocyte redistribution after administration of systemic CS. CONCLUSION: The flow cytometry-based profiling of the basic populations of immune cells in the blood and its analysis before and after systemic corticosteroid administration could improve personalized treatment approaches in patients with severe asthma.

Development of integrated wetland change detection approach: In case of Erdos Larus Relictus National Nature Reserve, China.

Wetlands are threatened by the global warming and the human exploitation pressure, and have been shrinking quickly in recent years. Timely and accurate wetland area change detection is the primary task for wetland conservation and restoration. The objective of this study is to develop an integrated change detection approach which integrates the advantages of spectral mixture analysis (SMA) and change vector analysis (CVA) for the change identification of wetland dynamics. In the proposed approach, water, vegetation and soil fractions of wetlands were derived by SMA; then, the detailed change information (including change magnitude and 12 change direction categories) were calculated through CVA. The proposed approach was applied for the wetlands change in Erdos Larus Relictus National Nature Reserve (ELRNNR), China, using time-series Landsat images during 1977-2017. We found that the wetland faced serious degradation, with water fraction changed to soil (5.79 km2), to vegetation (1.35 km2) and to both soil and vegetation (3.53 km2). From 1977 to 2000, a slight degradation occurred in the northeast edge of Bojiang Lake and a marginal degradation in Bojiang and Houjia Lakes inside the ELRNNR, with water fraction changed to soil and vegetation. During 2000-2010, severe degradation occurred in ELRNNR, and from 2010 to 2017, the wetland was more susceptible to the precipitation change and human activities. Analysis of the result indicated that the long-term drought and effects of mismanagement as well as misuse by human beings were the driving factors of wetland degradation. The proposed approach in this study achieves a higher accuracy than the classification approach to detect wetland change, with the ability to obtain more detailed change information.

Human CD40 ligand-expressing type 3 innate lymphoid cells induce IL-10-producing immature transitional regulatory B cells.

BACKGROUND: Type 3 innate lymphoid cells (ILC3s) are involved in maintenance of mucosal homeostasis; however, their role in immunoregulation has been unknown. Immature transitional regulatory B (itBreg) cells are innate-like B cells with immunosuppressive properties, and the in vivo mechanisms by which they are induced have not been fully clarified. OBJECTIVE: We aimed to investigate the ILC3-B-cell interaction that probably takes place in human tonsils. METHODS: ILC3s were isolated from peripheral blood and palatine tonsils, expanded, and cocultured with naive B cells. Tonsillar ILC3s and regulatory B cells were visualized with immunofluorescence histology. ILC3 frequencies were measured in tonsil tissue of allergic and nonallergic patients and in peripheral blood of allergic asthmatic patients and healthy control subjects. RESULTS: A mutually beneficial relationship was revealed between ILC3s and B cells: ILC3s induced IL-15 production in B cells through B cell-activating factor receptor, whereas IL-15, a potent growth factor for ILC3s, induced CD40 ligand (CD40L) expression on circulating and tonsillar ILC3s. IL-15-activated CD40L+ ILC3s helped B-cell survival, proliferation, and differentiation of IL-10-secreting, PD-L1-expressing functional itBreg cells in a CD40L- and B cell-activating factor receptor-dependent manner. ILC3s and regulatory B cells were in close connection with each other in palatine tonsils. ILC3 frequency was reduced in tonsil tissue of allergic patients and in peripheral blood of allergic asthmatic patients. CONCLUSION: Human CD40L+ ILC3s provide innate B-cell help and are involved in an innate immunoregulatory mechanism through induction of itBreg cell differentiation, which takes place in palatine tonsils in vivo. This mechanism, which can contribute to maintenance of immune tolerance, becomes insufficient in allergic diseases.

Evaluating Crop Area Mapping from MODIS Time-Series as an Assessment Tool for Zimbabwe's "Fast Track Land Reform Programme".

Moderate Resolution Imaging Spectroradiometer (MODIS) data forms the basis for numerous land use and land cover (LULC) mapping and analysis frameworks at regional scale. Compared to other satellite sensors, the spatial, temporal and spectral specifications of MODIS are considered as highly suitable for LULC classifications which support many different aspects of social, environmental and developmental research. The LULC mapping of this study was carried out in the context of the development of an evaluation approach for Zimbabwe's land reform program. Within the discourse about the success of this program, a lack of spatially explicit methods to produce objective data, such as on the extent of agricultural area, is apparent. We therefore assessed the suitability of moderate spatial and high temporal resolution imagery and phenological parameters to retrieve regional figures about the extent of cropland area in former freehold tenure in a series of 13 years from 2001-2013. Time-series data was processed with TIMESAT and was stratified according to agro-ecological potential zoning of Zimbabwe. Random Forest (RF) classifications were used to produce annual binary crop/non crop maps which were evaluated with high spatial resolution data from other satellite sensors. We assessed the cropland products in former freehold tenure in terms of classification accuracy, inter-annual comparability and heterogeneity. Although general LULC patterns were depicted in classification results and an overall accuracy of over 80% was achieved, user accuracies for rainfed agriculture were limited to below 65%. We conclude that phenological analysis has to be treated with caution when rainfed agriculture and grassland in semi-humid tropical regions have to be separated based on MODIS spectral data and phenological parameters. Because classification results significantly underestimate redistributed commercial farmland in Zimbabwe, we argue that the method cannot be used to produce spatial information on land-use which could be linked to tenure change. Hence capabilities of moderate resolution data are limited to assess Zimbabwe's land reform. To make use of the unquestionable potential of MODIS time-series analysis, we propose an analysis of plant productivity which allows to link annual growth and production of vegetation to ownership after Zimbabwe's land reform.

Sensitivity of proxies on non-linear interactions in the climate system.

Recent climate change is affecting the earth system to an unprecedented extent and intensity and has the potential to cause severe ecological and socioeconomic consequences. To understand natural and anthropogenic induced processes, feedbacks, trends, and dynamics in the climate system, it is also essential to consider longer timescales. In this context, annually resolved tree-ring data are often used to reconstruct past temperature or precipitation variability as well as atmospheric or oceanic indices such as the North Atlantic Oscillation (NAO) or the Atlantic Multidecadal Oscillation (AMO). The aim of this study is to assess weather-type sensitivity across the Northern Atlantic region based on two tree-ring width networks. Our results indicate that nonstationarities in superordinate space and time scales of the climate system (here synoptic- to global scale, NAO, AMO) can affect the climate sensitivity of tree-rings in subordinate levels of the system (here meso- to synoptic scale, weather-types). This scale bias effect has the capability to impact even large multiproxy networks and the ability of these networks to provide information about past climate conditions. To avoid scale biases in climate reconstructions, interdependencies between the different scales in the climate system must be considered, especially internal ocean/atmosphere dynamics.

Defining adult asthma endotypes by clinical features and patterns of volatile organic compounds in exhaled air.

BACKGROUND: Several classifications of adult asthma patients using cluster analyses based on clinical and demographic information has resulted in clinical phenotypic clusters that do not address molecular mechanisms. Volatile organic compounds (VOC) in exhaled air are released during inflammation in response to oxidative stress as a result of activated leukocytes. VOC profiles in exhaled air could distinguish between asthma patients and healthy subjects. In this study, we aimed to classify new asthma endotypes by combining inflammatory mechanisms investigated by VOC profiles in exhaled air and clinical information of asthma patients. METHODS: Breath samples were analyzed for VOC profiles by gas chromatography-mass spectrometry from asthma patients (n = 195) and healthy controls (n = 40). A total of 945 determined compounds were subjected to discriminant analysis to find those that could discriminate healthy from asthmatic subjects. 2-step cluster analysis based on clinical information and VOCs in exhaled air were used to form asthma endotypes. RESULTS: We identified 16 VOCs, which could distinguish between healthy and asthma subjects with a sensitivity of 100% and a specificity of 91.1%. Cluster analysis based on VOCs in exhaled air and the clinical parameters FEV1, FEV1 change after 3 weeks of hospitalization, allergic sensitization, Junipers symptoms score and asthma medications resulted in the formation of 7 different asthma endotype clusters. We identified asthma clusters with different VOC profiles but similar clinical characteristics and endotypes with similar VOC profiles, but distinct clinical characteristics. CONCLUSION: This study demonstrates that both, clinical presentation of asthma and inflammatory mechanisms in the airways should be considered for classification of asthma subtypes.

Differential serum protein markers and the clinical severity of asthma.

BACKGROUND: Asthma is a heterogeneous disease characterized by different clinical phenotypes and the involvement of multiple inflammatory pathways. During airway inflammation, many cytokines and chemokines are released and some are detectable in the sera. OBJECTIVE: Serum chemokines and cytokines, involved in airway inflammation in asthma patients, were investigated. METHODS: A total of 191 asthma patients were classified by hierarchical cluster analysis, including the following parameters: forced expiratory volume in 1 second (FEV1), eosinophil cationic protein (ECP) serum levels, blood eosinophils, Junipers asthma symptom score, and the change in FEV1, ECP serum levels, and blood eosinophils after 3 weeks of asthma therapy. Serum proteins were measured by multiplex analysis. Receiver operating characteristic (ROC) curves were used to evaluate the validity of serum proteins for discriminating between asthma clusters. RESULTS: Classification of asthma patients identified one cluster with high ECP serum levels, increased blood eosinophils, low FEV1 values, and good FEV1 improvement in response to asthma therapy (n=60) and one cluster with low ECP serum levels, low numbers of blood eosinophils, higher FEV1 values, and no FEV1 improvement in response to asthma therapy (n=131). Serum interleukin (IL)-8, eotaxin, vascular endothelial growth factor (VEGF), cutaneous T-cell-attracting chemokine (CTACK), growth-related oncogene (GRO)-α, and hepatocyte growth factor (HGF) were significantly different between the two clusters of asthma patients. ROC analysis for serum proteins calculated a sensitivity of 55.9% and specificity of 75.8% for discriminating between them. CONCLUSION: Serum cytokine and chemokine levels might be predictors for the severity of asthmatic inflammation, asthma control, and response to therapy, and therefore might be useful for treatment optimization.

Spatio-temporal analyses of cropland degradation in the irrigated lowlands of Uzbekistan using remote-sensing and logistic regression modeling.

Advancing land degradation in the irrigated areas of Central Asia hinders sustainable development of this predominantly agricultural region. To support decisions on mitigating cropland degradation, this study combines linear trend analysis and spatial logistic regression modeling to expose a land degradation trend in the Khorezm region, Uzbekistan, and to analyze the causes. Time series of the 250-m MODIS NDVI, summed over the growing seasons of 2000-2010, were used to derive areas with an apparent negative vegetation trend; this was interpreted as an indicator of land degradation. About one third (161,000 ha) of the region's area experienced negative trends of different magnitude. The vegetation decline was particularly evident on the low-fertility lands bordering on the natural sandy desert, suggesting that these areas should be prioritized in mitigation planning. The results of logistic modeling indicate that the spatial pattern of the observed trend is mainly associated with the level of the groundwater table (odds = 330 %), land-use intensity (odds = 103 %), low soil quality (odds = 49 %), slope (odds = 29 %), and salinity of the groundwater (odds = 26 %). Areas, threatened by land degradation, were mapped by fitting the estimated model parameters to available data. The elaborated approach, combining remote-sensing and GIS, can form the basis for developing a common tool for monitoring land degradation trends in irrigated croplands of Central Asia.

Inhibition of angiogenesis by IL-32: possible role in asthma.

BACKGROUND: IL-32 is a proinflammatory cytokine involved in various chronic inflammatory diseases. Chronic airway inflammation in asthmatic patients results in structural airway changes, including angiogenesis. Vascular endothelial growth factor (VEGF) is a key inducer of angiogenesis in the airways of asthmatic patients. OBJECTIVE: The aim of the study was to investigate the expression and function of IL-32 in patients with angiogenesis and asthma. METHODS: The expression and regulation of IL-32 in normal human bronchial epithelial (NHBE) cells was analyzed by using RT-PCR, ELISA, Western blotting, immunofluorescent staining, and flow cytometry. After knockdown of IL-32 in NHBE cells by small interfering RNA (siRNA) transfections, VEGF secretion was quantified by means of ELISA. New blood vessel formation was determined with human umbilical vein endothelial cells by culturing with supernatants from IL-32 siRNA-transfected NHBE cells. IL-32 was determined in serum and induced sputum samples of asthmatic patients and healthy control subjects by means of ELISA. RESULTS: IL-32 is expressed in NHBE cells on stimulation with IFN-γ, TNF-α, T(H)1 cells, and rhinovirus. Inhibition of IL-32 expression resulted in significantly increased secretion of the proangiogenic factors VEGF and platelet-derived growth factor by NHBE cells. Human umbilical vein endothelial cells cultured in supernatants from IL-32 siRNA-transfected NHBE cells showed enhanced in vitro angiogenesis. IL-32 is detectable in induced sputum from asthmatic patients. IL-32 serum levels were significantly higher in asthmatic patients compared with those seen in healthy control subjects and correlated with response to asthma treatment. CONCLUSION: IL-32 is induced by IFN-γ, TNF-α, T(H)1 cells, and rhinovirus in bronchial epithelial cells. It inhibits angiogenesis, and its serum levels are associated with a good treatment response in asthmatic patients.

IL-32 is expressed by human primary keratinocytes and modulates keratinocyte apoptosis in atopic dermatitis.

BACKGROUND: Keratinocyte (KC) apoptosis is an important mechanism of eczema and spongiosis in patients with atopic dermatitis (AD) and is mediated by IFN-gamma, which is secreted by T(H)1 cells. IL-32 is a proinflammatory cytokine that is involved in the inflammatory processes of rheumatoid arthritis, chronic obstructive pulmonary disease, and Crohn disease. Recently, it was shown that upregulation of IL-32 induces apoptosis. OBJECTIVE: The aim of the study was to investigate the expression and function of IL-32 in patients with AD. METHODS: The expression of IL-32 in KCs was analyzed by means of RT-PCR, ELISA, and flow cytometry. Transfections of small interfering RNA were performed in primary KCs, and apoptosis was analyzed by means of terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling, annexin-V, and 7-amino actinomycin D stainings. Immunofluorescence stainings were used to detect IL-32 in skin biopsy specimens, and serum levels of IL-32 were analyzed by means of ELISA. RESULTS: We report that IL-32 is expressed in human primary KCs on stimulation with IFN-gamma, TNF-alpha, and T(H)1 cells in contrast to T(H)2, regulatory T (Treg), or T(H)17 cells, which showed no effect. Transfection of primary KCs and artificial skin equivalents with small interfering RNA to IL-32, which resulted in a clear decrease in IL-32 expression, significantly reduced KC apoptosis. Immunofluorescence staining demonstrated that IL-32 was expressed in AD lesional skin, whereas it was present in neither skin biopsy specimens from healthy donors nor in lesional skin from patients with psoriasis. Serum levels of IL-32 from patients with AD correlated with disease severity, but increased serum levels of IL-32 were also detected in asthmatic patients. CONCLUSION: The present study demonstrates KCs as a source of IL-32, which modulates KC apoptosis and contributes to the pathophysiology of AD.

Geostatistical analysis of the spatiotemporal dynamics of powdery mildew and leaf rust in wheat.

Plant diseases are dynamic systems that progress or regress in spatial and temporal dimensions. Site-specific or temporally optimized disease control requires profound knowledge about the development of each stressor. The spatiotemporal dynamics of leaf rust (Puccinia recondite f. sp. tritici) and powdery mildew (Blumeria graminis f. sp. tritici) in wheat was analyzed in order to evaluate typical species-dependent characteristics of disease spread. During two growing seasons, severity data and other relevant plant growth parameters were collected in wheat fields. Spatial characteristics of both diseases were assessed by cluster analyses using spatial analysis by distance indices, whereas the temporal epidemic trends were assessed using statistical parameters. Multivariate statistics were used to identify parameters suitable for characterizing disease trends into four classes of temporal dynamics. The results of the spatial analysis showed that both diseases generally occurred in patches but a differentiation between the diseases by their spatial patterns and spread was not possible. In contrast, temporal characteristics allowed for a differentiation of the diseases, due to the fact that a typical trend was found for leaf rust which differed from the trend of powdery mildew. Therefore, these trends suggested a high potential for temporally optimized disease control. Precise powdery mildew control would be more complicated due to the observed high variability in spatial and temporal dynamics. The general results suggest that, in spite of the high variability in spatiotemporal dynamics, disease control that is optimized in space and time is generally possible but requires consideration of disease- and case-dependent characteristics.

Effect of sustained high altitude on asthma patients.

Due to factors such as allergen avoidance and the decrease of air pollution, sustained stays in a high-altitude climate have been recommended for asthma patients for a long time. There are also documented effects and favorable influence on the health of permanent residents at high altitude; for example, the frequency of allergic sensitization to house dust mite in asymptomatic subjects is much lower than at sea level. In the context of this article, 'high altitude' means 1500-2500 m above sea level. The aim of the review is to summarize the available data on the effects of a sustained stay of asthmatic patient data between 1500-1800 m above sea level in alpine altitudes (Europe). Climatic conditions in South America or in Africa are completely different from the altitudes discussed in this review. We conclude that the available evidence suggests a significant benefit of high altitude for asthmatic patients, particularly in steroid-dependent patients.

Activin A is an acute allergen-responsive cytokine and provides a link to TGF-beta-mediated airway remodeling in asthma.

BACKGROUND: Allergic asthma typically shows activated, allergen-specific CD4(+) T cells in the early phase and airway remodeling in the late phase of the disease. Although TGF-beta plays a crucial role in airway remodeling, it is only marginally induced in CD4(+) T cells in the early allergen-dependent activation of the immune system. OBJECTIVE: To elucidate the transition between early- and late-phase events, we investigated the role of activin A, a close family member of TGF-beta. METHODS: Activin A and TGF-beta(1) levels were measured systemically in the serum and in CD4(+) T cells of asthmatic patients, as well as locally in the lung. RESULTS: Activin A serum levels were increased in patients with severe asthma compared with levels in patients with moderate asthma and healthy control subjects, whereas all patients showed significantly increased TGF-beta(1) serum levels independent of disease severity. In T cells only patients with moderate asthma showed increased activin A mRNA expression, whereas TGF-beta(1) expression was equal to that seen in healthy subjects. Accordingly, ovalbumin sensitization in a mouse model of allergic asthma could induce activin A mRNA expression, but not TGF-beta(1) expression, in the lung. Immunohistochemistry of mice and human specimens revealed an abundant expression of activin A by infiltrating lymphocytes and structural cells of the lung. Although TGF-beta(1) more potently enhanced proliferation and Smad 2/3-dependent reporter genes in fibroblasts, activin A was capable of inducing TGF-beta(1) and vice versa. CONCLUSION: Activin A provides a link between acute allergen-specific T-cell responses and chronic TGF-beta(1)-mediated airway remodeling in asthma.

Native Art v 1 and recombinant Art v 1 are able to induce humoral and T cell-mediated in vitro and in vivo responses in mugwort allergy.

BACKGROUND: Mugwort pollen is an important allergen source in hay fever and pollen-related food allergy. Little is known about the clinical relevance of the major mugwort allergen Art v 1 and its importance in allergy. OBJECTIVE: In this study we aimed to investigate the allergenicity of mugwort extract compared with the allergenicity of native (n)Art v 1 and recombinant (r)Art v 1, one major allergen of mugwort, in vivo and in vitro. METHODS: Thirty-two patients allergic to mugwort and 10 control subjects were investigated by means of skin prick and nasal provocation testing with different concentrations of mugwort extract, nArt v 1, and rArt v 1. nArt v 1 was purified from aqueous mugwort extract, and rArt v 1 was cloned, expressed in Escherichia coli, and then purified. The in vitro allergenicity was measured by means of ImmunoCAP, ELISA, ELISA-inhibition experiments, and T-cell proliferation assays. RESULTS: nArt v 1 and rArt v 1 were able to elicit positive in vivo and in vitro reactions. The IgE-binding capacity, as determined by means of ELISA, was slightly higher for nArt v 1 than for rArt v 1, and both allergens were able to induce T-cell proliferation in sensitized patients. However, rArt v 1 elicited a reduced response in skin and nasal provocation tests compared with nArt v 1. Compared with mugwort extract, both nArt v 1 and rArt v 1 showed lower sensitivity in patients with mugwort allergy in vivo. CONCLUSIONS: Art v 1, either in its native or recombinant form, is able to induce allergic reactions in patients with mugwort allergy. rArt v 1 induced comparable humoral and cell-mediated responses in vitro but showed reduced in vivo allergenicity compared with biochemically purified nArt v 1.

Interleukin-2 primes eosinophil degranulation in hypereosinophilia and Wells' syndrome.

Patients with hypereosinophilia frequently suffer from eosinophil-mediated damages of the heart, lungs, skin, and other organs, while some do not. The reason(s) for this difference is not known. We observed that eosinophils from most patients with hypereosinophilia express the alpha-chain of the IL-2 receptor (CD25), and that IL-2 enhances platelet-activating factor-stimulated release of eosinophil cationic protein from CD25-expressing but not from CD25-negative eosinophils. Such a "priming" effect has previously been described for eosinophil hematopoietins. These data suggest that patients with increased eosinophil surface CD25 expression are at higher risk of eosinophil degranulation and subsequent tissue damage when IL-2 is present at inflammatory sites.

Eosinophils express functional IL-13 in eosinophilic inflammatory diseases.

IL-13 is an immunoregulatory and effector cytokine in allergic diseases such as bronchial asthma. A variety of immune and non-immune cells are known as IL-13 producers. In this study we investigated whether and under what conditions human eosinophils generate IL-13. Freshly isolated highly purified peripheral blood eosinophils from patients with several eosinophilic inflammatory diseases and from normal control individuals were investigated. We observed that blood eosinophils from patients suffering from bronchial asthma, atopic dermatitis, parasitic infections, hypereosinophilic syndrome, and idiopathic eosinophilic esophagitis expressed IL-13, as assessed by ELISA, ELISPOT assay, flow cytometry, and immunocytochemistry. By using nasal polyp tissues and immunohistochemistry, we demonstrated IL-13 expression in eosinophils under in vivo conditions. In contrast, blood eosinophils from control individuals as well as blood neutrophils from both eosinophilic and control patients did not produce detectable IL-13 levels. However, when blood eosinophils from control individuals were stimulated with GM-CSF or IL-5 in vitro, they generated IL-13 mRNA and protein, suggesting that IL-13 expression by eosinophils under inflammatory conditions is a cytokine-driven process. Stimulation of blood eosinophils containing IL-13 by eotaxin resulted in a rapid release of this cytokine. Eosinophil-derived IL-13 was functional, as it increased the surface expression of the low affinity IgE receptor (CD23) on purified B cells. In conclusion, human eosinophils are able to produce and release functional IL-13 in eosinophilic inflammatory responses.