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Open AccessProtocol
Turning the World Upside-Down in Cellulose for Improved Culturing and Imaging of Respiratory Challenges within a Human 3D Model
Cells 2019, 8(10), 1292; https://doi.org/10.3390/cells8101292 (registering DOI) - 21 Oct 2019
Abstract
Polarized growth of human-derived respiratory epithelial cells on hydrogel-coated filters offers big advantages concerning detailed experiments with respect to drug screening or host pathogen interactions. Different microscopic approaches, such as confocal analyses and high content screening, help to examine such 3D respiratory samples, [...] Read more.
Polarized growth of human-derived respiratory epithelial cells on hydrogel-coated filters offers big advantages concerning detailed experiments with respect to drug screening or host pathogen interactions. Different microscopic approaches, such as confocal analyses and high content screening, help to examine such 3D respiratory samples, resulting in high-resolution pictures and enabling quantitative analyses of high cell numbers. A major problem employing these techniques relates to single-use instead of multiple-use of Transwell filters and difficulties in the digestion of collagen if subsequent analyses are needed. Up to date, cells are seeded in collagen-based matrices to the inner field of Transwell inserts, which makes it impossible to image due to the design of the inserts and hard to perform other analyses since digestion of the collagen matrix also affects Transwell grown cells. To overcome these problems, we optimized culturing conditions for monitoring cell differentiation or repeated dose experiments over a long time period. For this, cells are seeded upside-down to the bottom side of filters within an animal-free cellulose hydrogel. These cells were then grown inverted under static conditions and were differentiated in air-liquid interphase (ALI). Full differentiation of goblet (Normal Human Bronchial Epithelial (NHBE))/Club (small airway epithelia (SAE)) cells and ciliated cells was detected after 12 days in ALI. Inverted cell cultures could then be used for ‘follow-up’ live cell imaging experiments, as well as, flow-cytometric analyses due to easy digestion of the cellulose compared to classical collagen matrices. Additionally, this culture technique also enables easy addition of immune cells, such as dendritic cells (DCs), macrophages, neutrophils, T or B cells alone or in combination, to the inner field of the Transwell to monitor immune cell behavior after repeated respiratory challenge. Our detailed protocol offers the possibility of culturing human primary polarized cells into a fully differentiated, thick epithelium without any animal components over >700 days. Furthermore, this animal-free, inverted system allows investigation of the same inserts, because the complete Transwell can be readily transferred to glass-bottom dishes for live cell imaging analyses and then returned to its original plate for further cultivation. Full article
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Open AccessFeature PaperArticle
Comparison of Temperature Control and Temperature Difference Control for a Kaibel Dividing Wall Column
Processes 2019, 7(10), 773; https://doi.org/10.3390/pr7100773 (registering DOI) - 21 Oct 2019
Abstract
A dividing wall column (DWC) effectively intensifies the distillation process with a reduced energy consumption, capital investment, and space. The three-product DWC has been investigated intensively and extensively; however, the four-product Kaibel DWC has received scarce attention. This study aimed to propose feasible [...] Read more.
A dividing wall column (DWC) effectively intensifies the distillation process with a reduced energy consumption, capital investment, and space. The three-product DWC has been investigated intensively and extensively; however, the four-product Kaibel DWC has received scarce attention. This study aimed to propose feasible control structures for the Kaibel DWC using only temperature sensors in order to promote its industrialization. Two temperature control structures, two temperature difference control structures, and two double temperature difference control structures were studied. The feasibility of the six proposed control structures was verified with a wide variety of feed disturbances. In most cases, temperature difference control was better than temperature control to maintain product purities. The dynamic performances proved that the inserted feed disturbances were handled well. These results help to promote the industrialization of the Kaibel DWC. Full article
(This article belongs to the Special Issue Process Optimization and Control)
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Open AccessTechnical Note
Man-Machine-Interface Software Design of a Cotton Harvester Yield Monitor Calibration System
AgriEngineering 2019, 1(4), 511-522; https://doi.org/10.3390/agriengineering1040037 (registering DOI) - 21 Oct 2019
Abstract
Several yield monitors are available for use on cotton harvesters, but none are able to maintain yield measurement accuracy across cultivars and field conditions that vary spatially and/or temporally. Thus, the utility of yield monitors as tools for on-farm research is limited unless [...] Read more.
Several yield monitors are available for use on cotton harvesters, but none are able to maintain yield measurement accuracy across cultivars and field conditions that vary spatially and/or temporally. Thus, the utility of yield monitors as tools for on-farm research is limited unless steps are taken to calibrate the systems as cultivars and conditions change. This technical note details the man-machine-interface software system design portion of a harvester-based yield monitor calibration system for basket-type cotton strippers. The system was based upon the use of pressure sensors to measure the weight of the basket by monitoring the static pressure in the hydraulic lift cylinder circuit. To ensure accurate weighing, the system automatically lifted the basket to a target lift height, allowed basket time to settle, then weighed the contents of the basket. The software running the system was split into two parts that were run on an embedded low-level micro-controller, and a mobile computer located in the harvester cab. The system was field tested under commercial conditions and found to measure basket load weights within 2.5% of the reference scale. As such, the system was proven to be capable of providing an on-board auto-correction to a yield monitor for use in multi-variety field trials. Full article
(This article belongs to the Special Issue Robotics and Automation Engineering in Agriculture)
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Open AccessPerspective
The Neuroprotective Effects of Melatonin: Possible Role in the Pathophysiology of Neuropsychiatric Disease
Brain Sci. 2019, 9(10), 285; https://doi.org/10.3390/brainsci9100285 (registering DOI) - 21 Oct 2019
Abstract
Melatonin is a hormone that is secreted by the pineal gland. To date, melatonin is known to regulate the sleep cycle by controlling the circadian rhythm. However, recent advances in neuroscience and molecular biology have led to the discovery of new actions and [...] Read more.
Melatonin is a hormone that is secreted by the pineal gland. To date, melatonin is known to regulate the sleep cycle by controlling the circadian rhythm. However, recent advances in neuroscience and molecular biology have led to the discovery of new actions and effects of melatonin. In recent studies, melatonin was shown to have antioxidant activity and, possibly, to affect the development of Alzheimer’s disease (AD). In addition, melatonin has neuroprotective effects and affects neuroplasticity, thus indicating potential antidepressant properties. In the present review, the new functions of melatonin are summarized and a therapeutic target for the development of new drugs based on the mechanism of action of melatonin is proposed. Full article
(This article belongs to the Special Issue Neurogenesis and Gliogenesis in Health and Disease)
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Open AccessFeature PaperArticle
The Effect of Vitamin D Supplementation on its Metabolism and the Vitamin D Metabolite Ratio
Nutrients 2019, 11(10), 2539; https://doi.org/10.3390/nu11102539 (registering DOI) - 21 Oct 2019
Abstract
25-hydroxyvitamin D (25(OH)D) is commonly measured to assess vitamin D status. Other vitamin D metabolites such as 24,25-dihydroxyvitamin D (24,25(OH)2D) provide additional insights into vitamin D status or metabolism. Earlier studies suggested that the vitamin D metabolite ratio (VMR), calculated as [...] Read more.
25-hydroxyvitamin D (25(OH)D) is commonly measured to assess vitamin D status. Other vitamin D metabolites such as 24,25-dihydroxyvitamin D (24,25(OH)2D) provide additional insights into vitamin D status or metabolism. Earlier studies suggested that the vitamin D metabolite ratio (VMR), calculated as 24,25(OH)2D/25(OH)D, could predict the 25(OH)D increase after vitamin D supplementation. However, the evidence for this additional value is inconclusive. Therefore, our aim was to assess whether the increase in 25(OH)D after supplementation was predicted by the VMR better than baseline 25(OH)D. Plasma samples of 106 individuals (25(OH)D < 75 nmol/L) with hypertension who completed the Styrian Vitamin D Hypertension Trial (NC.T.02136771) were analyzed. Participants received vitamin D (2800 IU daily) or placebo for 8 weeks. The treatment effect (ANCOVA) for 25(OH)D3, 24,25(OH)2D3 and the VMR was 32 nmol/L, 3.3 nmol/L and 0.015 (all p < 0.001), respectively. Baseline 25(OH)D3 and 24,25(OH)2D3 predicted the change in 25(OH)D3 with comparable strength and magnitude. Correlation and regression analysis showed that the VMR did not predict the change in 25(OH)D3. Therefore, our data do not support routine measurement of 24,25(OH)2D3 in order to individually optimize the dosage of vitamin D supplementation. Our data also suggest that activity of 24-hydroxylase increases after vitamin D supplementation. Full article
(This article belongs to the Special Issue Calcium, Vitamin D and Health)
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Open AccessArticle
Impact of Cold versus Hot Brewing on the Phenolic Profile and Antioxidant Capacity of Rooibos (Aspalathus linearis) Herbal Tea
Antioxidants 2019, 8(10), 499; https://doi.org/10.3390/antiox8100499 (registering DOI) - 21 Oct 2019
Abstract
Consumption of rooibos (Aspalathus linearis) as herbal tea is growing in popularity worldwide and its health-promoting attributes are mainly ascribed to its phenolic composition, which may be affected by the brewing conditions used. An aspect so far overlooked is the impact [...] Read more.
Consumption of rooibos (Aspalathus linearis) as herbal tea is growing in popularity worldwide and its health-promoting attributes are mainly ascribed to its phenolic composition, which may be affected by the brewing conditions used. An aspect so far overlooked is the impact of cold brewing vs regular brewing and microwave boiling on the poly(phenolic) profile and in vitro antioxidant capacity of infusions prepared from red (‘fermented’, oxidized) and green (‘unfermented’, unoxidized) rooibos, the purpose of the present study. By using an untargeted metabolomics-based approach (UHPLC-QTOF mass spectrometry), 187 phenolic compounds were putatively annotated in both rooibos types, with flavonoids, tyrosols, and phenolic acids the most represented type of phenolic classes. Multivariate statistics (OPLS-DA) highlighted the phenolic classes most affected by the brewing conditions. Similar antioxidant capacities (ORAC and ABTS assays) were observed between cold- and regular-brewed green rooibos and boiled-brewed red rooibos. However, boiling green and red rooibos delivered infusions with the highest antioxidant capacities and total polyphenol content. The polyphenol content strongly correlated with the in vitro antioxidant capacities, especially for flavonoids and phenolic acids. These results contribute to a better understanding of the impact of the preparation method on the potential health benefits of rooibos tea. Full article
(This article belongs to the Special Issue Phenolic Profiling and Antioxidant Capacity in Plants)
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Open AccessArticle
Influence of Bioactive Compounds Incorporated in a Nanoemulsion as Coating on Avocado Fruits (Persea americana) during Postharvest Storage: Antioxidant Activity, Physicochemical Changes and Structural Evaluation
Antioxidants 2019, 8(10), 500; https://doi.org/10.3390/antiox8100500 (registering DOI) - 21 Oct 2019
Abstract
The objective of the present study was to determine the effect of the application of a nanoemulsion made of orange essential oil and Opuntia oligacantha extract on avocado quality during postharvest. The nanoemulsion was applied as a coating in whole fruits, and the [...] Read more.
The objective of the present study was to determine the effect of the application of a nanoemulsion made of orange essential oil and Opuntia oligacantha extract on avocado quality during postharvest. The nanoemulsion was applied as a coating in whole fruits, and the following treatments were assessed: concentrated nanoemulsion (CN), 50% nanoemulsion (N50), 25% nanoemulsion (N25) and control (C). Weight loss, firmness, polyphenol oxidase (PPO) activity, total soluble solids, pH, external and internal colour, total phenols, total flavonoids, antioxidant activity by 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH), while the structural evaluation of the epicarp was assessed through histological cuts. Significant differences were found (p < 0.05) among the treatments in all the response variables. The best results were with the N50 and N25 treatments for firmness and weight loss, finding that the activity of the PPO was diminished, and a delay in the darkening was observed in the coated fruits. Furthermore, the nanoemulsion treatments maintained the total phenol and total flavonoid contents and potentiated antioxidant activity at 60 days. This histological study showed that the nanoemulsion has a delaying effect on the maturation of the epicarp. The results indicate that using this nanoemulsion as a coating is an effective alternative to improve the postharvest life of avocado. Full article
(This article belongs to the Special Issue Natural Phenolic Compounds for Health, Food and Cosmetic Applications)
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Open AccessArticle
Deep Learning of Fuzzy Weighted Multi-Resolution Depth Motion Maps with Spatial Feature Fusion for Action Recognition
J. Imaging 2019, 5(10), 82; https://doi.org/10.3390/jimaging5100082 (registering DOI) - 21 Oct 2019
Abstract
Human action recognition (HAR) is an important yet challenging task. This paper presents a novel method. First, fuzzy weight functions are used in computations of depth motion maps (DMMs). Multiple length motion information is also used. These features are referred to as fuzzy [...] Read more.
Human action recognition (HAR) is an important yet challenging task. This paper presents a novel method. First, fuzzy weight functions are used in computations of depth motion maps (DMMs). Multiple length motion information is also used. These features are referred to as fuzzy weighted multi-resolution DMMs (FWMDMMs). This formulation allows for various aspects of individual actions to be emphasized. It also helps to characterise the importance of the temporal dimension. This is important to help overcome, e.g., variations in time over which a single type of action might be performed. A deep convolutional neural network (CNN) motion model is created and trained to extract discriminative and compact features. Transfer learning is also used to extract spatial information from RGB and depth data using the AlexNet network. Different late fusion techniques are then investigated to fuse the deep motion model with the spatial network. The result is a spatial temporal HAR model. The developed approach is capable of recognising both human action and human–object interaction. Three public domain datasets are used to evaluate the proposed solution. The experimental results demonstrate the robustness of this approach compared with state-of-the art algorithms. Full article
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Open AccessFeature PaperReview
Erythroxylum in Focus: An Interdisciplinary Review of an Overlooked Genus
Molecules 2019, 24(20), 3788; https://doi.org/10.3390/molecules24203788 (registering DOI) - 21 Oct 2019
Abstract
The genus Erythroxylum contains species used by indigenous people of South America long before the domestication of plants. Two species, E. coca and E. novogranatense, have been utilized for thousands of years specifically for their tropane alkaloid content. While abuse of the narcotic [...] Read more.
The genus Erythroxylum contains species used by indigenous people of South America long before the domestication of plants. Two species, E. coca and E. novogranatense, have been utilized for thousands of years specifically for their tropane alkaloid content. While abuse of the narcotic cocaine has impacted society on many levels, these species and their wild relatives contain untapped resources for the benefit of mankind in the form of foods, pharmaceuticals, phytotherapeutic products, and other high-value plant-derived metabolites. In this review, we describe the current state of knowledge of members within the genus and the recent advances in the realm of molecular biology and biochemistry. Full article
(This article belongs to the Special Issue Advances in Plant Alkaloid Research)
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Open AccessFeature PaperReview
The Microenvironment of Pituitary Tumors—Biological and Therapeutic Implications
Cancers 2019, 11(10), 1605; https://doi.org/10.3390/cancers11101605 (registering DOI) - 21 Oct 2019
Abstract
The tumor microenvironment (TME) includes resident and infiltrative non-tumor cells, as well as blood and lymph vessels, extracellular matrix molecules, and numerous soluble factors, such as cytokines and chemokines. While the TME is now considered to be a prognostic tool and a therapeutic [...] Read more.
The tumor microenvironment (TME) includes resident and infiltrative non-tumor cells, as well as blood and lymph vessels, extracellular matrix molecules, and numerous soluble factors, such as cytokines and chemokines. While the TME is now considered to be a prognostic tool and a therapeutic target for many cancers, little is known about its composition in pituitary tumors. This review summarizes our current knowledge of the TME within pituitary tumors and the strong interest in TME as a therapeutic target. While we cover the importance of angiogenesis and immune infiltrating cells, we also address the role of the elusive folliculostellate cells, the emerging literature on pituitary tumor-associated fibroblasts, and the contribution of extracellular matrix components in these tumors. The cases of human pituitary tumors treated with TME-targeting therapies are reviewed and emerging concepts of vascular normalization and combined therapies are presented. Together, this snapshot overview of the current literature pinpoints not only the underestimated role of TME components in pituitary tumor biology, but also the major promise it may offer for both prognosis and targeted therapeutics. Full article
Open AccessConcept Paper
About Chirality in Minkowski Spacetime
Symmetry 2019, 11(10), 1320; https://doi.org/10.3390/sym11101320 (registering DOI) - 21 Oct 2019
Abstract
In this paper, we show that Lorentz boosts are direct isometries according to the recent mathematical definitions of direct and indirect isometries and of chirality, working for any metric space. Here, these definitions are extended to the Minkowski spacetime. We also show that [...] Read more.
In this paper, we show that Lorentz boosts are direct isometries according to the recent mathematical definitions of direct and indirect isometries and of chirality, working for any metric space. Here, these definitions are extended to the Minkowski spacetime. We also show that the composition of parity inversion and time reversal is an indirect isometry, which is the opposite of what could be expected in Euclidean spaces. It is expected that the extended mathematical definition of chirality presented here can contribute to the unification of several definitions of chirality in space and in spacetime, and that it helps clarify the ubiquitous concept of chirality. Full article
Open AccessArticle
Helicobacter pylori Induces IL-33 Production and Recruits ST-2 to Lipid Rafts to Exacerbate Inflammation
Cells 2019, 8(10), 1290; https://doi.org/10.3390/cells8101290 (registering DOI) - 21 Oct 2019
Abstract
Helicobacter pylori colonizes human gastric epithelial cells and contributes to the development of several gastrointestinal disorders. Interleukin (IL)-33 is involved in various immune responses, with reported proinflammatory and anti-inflammatory effects, which may be associated with colitis and colitis-associated cancer. IL-33 induces the inflammatory [...] Read more.
Helicobacter pylori colonizes human gastric epithelial cells and contributes to the development of several gastrointestinal disorders. Interleukin (IL)-33 is involved in various immune responses, with reported proinflammatory and anti-inflammatory effects, which may be associated with colitis and colitis-associated cancer. IL-33 induces the inflammatory cascade through its receptor, suppression of tumorigenicity-2 (ST-2). Binding of IL-33 to membrane-bound ST-2 (mST-2) recruits the IL-1 receptor accessory protein (IL-1RAcP) and activates intracellular signaling pathways. However, whether IL-33/ST-2 is triggered by H. pylori infection and whether this interaction occurs in lipid rafts remain unclear. Our study showed that both IL-33 and ST-2 expression levels were significantly elevated in H. pylori-infected cells. Confocal microscopy showed that ST-2 mobilized into the membrane lipid rafts during infection. Depletion of membrane cholesterol dampened H. pylori-induced IL-33 and IL-8 production. Furthermore, in vivo studies revealed IL-33/ST-2 upregulation, and severe leukocyte infiltration was observed in gastric tissues infected with H. pylori. Together, these results demonstrate that ST-2 recruitment into the lipid rafts serves as a platform for IL-33-dependent H. pylori infection, which aggravates inflammation in the stomach. Full article
(This article belongs to the Special Issue Immunomodulatory Factors in Host Defense)
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Open AccessArticle
Atomic Force Microscope Guided SERS Spectra Observation for [email protected]@PVP Plasmonic Nanoparticles
Molecules 2019, 24(20), 3789; https://doi.org/10.3390/molecules24203789 (registering DOI) - 21 Oct 2019
Abstract
Recently polymer encapsulated surface-enhanced-Raman-scattering (SERS) probes with internal noble metal core–shell structure has found growing applications in biomedical applications. Here we studied the SERS spectra of [email protected][email protected] (4MBA: 4-mercaptobenzoic acid; PVP: polyvinylpyrrolidone) plasmonic nanoparticles produced from a chemical reduction method. By linking the [...] Read more.
Recently polymer encapsulated surface-enhanced-Raman-scattering (SERS) probes with internal noble metal core–shell structure has found growing applications in biomedical applications. Here we studied the SERS spectra of [email protected][email protected] (4MBA: 4-mercaptobenzoic acid; PVP: polyvinylpyrrolidone) plasmonic nanoparticles produced from a chemical reduction method. By linking the atomic force microscope (AFM) with the homebuilt confocal Raman spectrometer thus to use AFM images as guidance, we realized the measurement of the SERS spectra from separated nanoparticles. We investigated the cases for single nanoparticles and for dimer structures and report several observed results including SERS spectra linearly scaled with laser power, abrupt boosting and abnormal shape changing of SERS spectra for dimer structures. Based on the finite element method simulation, we explained the observed ratio of SERS signals between the dimer structure and the single nanoparticle, and attributed the observed abnormal spectra to the photothermal effect of these plasmonic nanoparticles. Our study provides valuable guidance for choosing appropriate laser power when applying similar SERS probes to image biological cells. Full article
(This article belongs to the Special Issue Optical Properties of Nanomaterials)
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Open AccessArticle
GSKIP-Mediated Anchoring Increases Phosphorylation of Tau by PKA but Not by GSK3beta via cAMP/PKA/GSKIP/GSK3/Tau Axis Signaling in Cerebrospinal Fluid and iPS Cells in Alzheimer Disease
J. Clin. Med. 2019, 8(10), 1751; https://doi.org/10.3390/jcm8101751 (registering DOI) - 21 Oct 2019
Abstract
Based on the protein kinase A (PKA)/GSK3β interaction protein (GSKIP)/glycogen synthase kinase 3β (GSK3β) axis, we hypothesized that these might play a role in Tau phosphorylation. Here, we report that the phosphorylation of Tau Ser409 in SHSY5Y cells was increased by overexpression of [...] Read more.
Based on the protein kinase A (PKA)/GSK3β interaction protein (GSKIP)/glycogen synthase kinase 3β (GSK3β) axis, we hypothesized that these might play a role in Tau phosphorylation. Here, we report that the phosphorylation of Tau Ser409 in SHSY5Y cells was increased by overexpression of GSKIP WT more than by PKA- and GSK3β-binding defective mutants (V41/L45 and L130, respectively). We conducted in vitro assays of various kinase combinations to show that a combination of GSK3β with PKA but not Ca2+/calmodulin-dependent protein kinase II (CaMK II) might provide a conformational shelter to harbor Tau Ser409. Cerebrospinal fluid (CSF) was evaluated to extend the clinical significance of Tau phosphorylation status in Alzheimer’s disease (AD), neurological disorders (NAD), and mild cognitive impairment (MCI). We found higher levels of different PKA–Tau phosphorylation sites (Ser214, Ser262, and Ser409) in AD than in NAD, MCI, and normal groups. Moreover, we used the CRISPR/Cas9 system to produce amyloid precursor protein (APPWT/D678H) isogenic mutants. These results demonstrated an enhanced level of phosphorylation by PKA but not by the control. This study is the first to demonstrate a transient increase in phosphor-Tau caused by PKA, but not GSK3β, in the CSF and induced pluripotent stem cells (iPSCs) of AD, implying that both GSKIP and GSK3β function as anchoring proteins to strengthen the cAMP/PKA/Tau axis signaling during AD pathogenesis. Full article
(This article belongs to the Special Issue Energetic Metabolism Impairment in Brain Dysfunction)
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Open AccessArticle
A Toolbox for Functional Analysis and the Systematic Identification of Diagnostic and Prognostic Gene Expression Signatures Combining Meta-Analysis and Machine Learning
Cancers 2019, 11(10), 1606; https://doi.org/10.3390/cancers11101606 (registering DOI) - 21 Oct 2019
Abstract
The identification of biomarker signatures is important for cancer diagnosis and prognosis. However, the detection of clinical reliable signatures is influenced by limited data availability, which may restrict statistical power. Moreover, methods for integration of large sample cohorts and signature identification are limited. [...] Read more.
The identification of biomarker signatures is important for cancer diagnosis and prognosis. However, the detection of clinical reliable signatures is influenced by limited data availability, which may restrict statistical power. Moreover, methods for integration of large sample cohorts and signature identification are limited. We present a step-by-step computational protocol for functional gene expression analysis and the identification of diagnostic and prognostic signatures by combining meta-analysis with machine learning and survival analysis. The novelty of the toolbox lies in its all-in-one functionality, generic design, and modularity. It is exemplified for lung cancer, including a comprehensive evaluation using different validation strategies. However, the protocol is not restricted to specific disease types and can therefore be used by a broad community. The accompanying R package vignette runs in ~1 h and describes the workflow in detail for use by researchers with limited bioinformatics training. Full article
(This article belongs to the Special Issue Application of Bioinformatics in Cancers)

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