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Sapere aude!
(2022)
Simultaneous Barcode Sequencing of Diverse Museum Collection Specimens Using a Mixed RNA Bait Set
(2022)
A growing number of publications presenting results from sequencing natural history collection specimens reflect the importance of DNA sequence information from such samples. Ancient DNA extraction and library preparation methods in combination with target gene capture are a way of unlocking archival DNA, including from formalin-fixed wet-collection material. Here we report on an experiment, in which we used an RNA bait set containing baits from a wide taxonomic range of species for DNA hybridisation capture of nuclear and mitochondrial targets for analysing natural history collection specimens. The bait set used consists of 2,492 mitochondrial and 530 nuclear RNA baits and comprises specific barcode loci of diverse animal groups including both invertebrates and vertebrates. The baits allowed to capture DNA sequence information of target barcode loci from 84% of the 37 samples tested, with nuclear markers being captured more frequently and consensus sequences of these being more complete compared to mitochondrial markers. Samples from dry material had a higher rate of success than wet-collection specimens, although target sequence information could be captured from 50% of formalin-fixed samples. Our study illustrates how efforts to obtain barcode sequence information from natural history collection specimens may be combined and are a way of implementing barcoding inventories of scientific collection material.
Etmopteridae (lantern sharks) is the most species-rich family of sharks, comprising more than 50 species.
Many species are described from few individuals, and re-collection of specimens is often hindered by the remoteness of their sampling sites.
For taxonomic studies, comparative morphological analysis of type specimens housed in natural history collections has been the main source of evidence.
In contrast, DNA sequence information has rarely been used.
Most lantern shark collection specimens, including the types, were formalin fixed before long-term storage in ethanol solutions.
The DNA damage caused by both fixation and preservation of specimens has excluded these specimens from DNA sequence-based phylogenetic analyses so far.
However, recent advances in the field of ancient DNA have allowed recovery of wet-collection specimen DNA sequence data.
Here we analyse archival mitochondrial DNA sequences, obtained using ancient DNA approaches, of two wet-collection lantern shark paratype specimens, namely Etmopterus litvinovi and E. pycnolepis, for which the type series represent the only known individuals.
Target capture of mitochondrial markers from single-stranded DNA libraries allows for phylogenetic placement of both species.
Our results suggest synonymy of E. benchleyi with E. litvinovi but support the species status of E. pycnolepis. This revised taxonomy is helpful for future conservation and management efforts, as our results indicate a larger distribution range of E. litvinovi. This study further demonstrates the importance of wet-collection type specimens as genetic resource for taxonomic research.
The degree of detrimental effects inflicted on mankind by the COVID-19 pandemic increased the need to develop ASSURED (Affordable, Sensitive, Specific, User-friendly, Rapid and Robust, Equipment-free, and Deliverable) POCT (point of care testing) to overcome the current and any future pandemics. Much effort in research and development is currently advancing the progress to overcome the diagnostic pressure built up by emerging new pathogens. LAMP (loop-mediated isothermal amplification) is a well-researched isothermal technique for specific nucleic acid amplification which can be combined with a highly sensitive immunochromatographic readout via lateral flow assays (LFA). Here we discuss LAMP-LFA robustness, sensitivity, and specificity for SARS-CoV-2 N-gene detection in cDNA and clinical swab-extracted RNA samples. The LFA readout is designed to produce highly specific results by incorporation of biotin and FITC labels to 11-dUTP and LF (loop forming forward) primer, respectively. The LAMP-LFA assay was established using cDNA for N-gene with an accuracy of 95.65%. To validate the study, 82 SARS-CoV-2-positive RNA samples were tested. Reverse transcriptase (RT)-LAMP-LFA was positive for the RNA samples with an accuracy of 81.66%; SARS-CoV-2 viral RNA was detected by RT-LAMP-LFA for as low as CT-33. Our method reduced the detection time to 15 min and indicates therefore that RT-LAMP in combination with LFA represents a promising nucleic acid biosensing POCT platform that combines with smartphone based semi-quantitative data analysis.
This study focuses on three key aspects: (a) crude throat swab samples in a viral transport medium (VTM) as templates for RT-LAMP reactions; (b) a biotinylated DNA probe with enhanced specificity for LFA readouts; and (c) a digital semi-quantification of LFA readouts. Throat swab samples from SARS-CoV-2 positive and negative patients were used in their crude (no cleaning or pre-treatment) forms for the RT-LAMP reaction. The samples were heat-inactivated but not treated for any kind of nucleic acid extraction or purification. The RT-LAMP (20 min processing time) product was read out by an LFA approach using two labels: FITC and biotin. FITC was enzymatically incorporated into the RT-LAMP amplicon with the LF-LAMP primer, and biotin was introduced using biotinylated DNA probes, specifically for the amplicon region after RT-LAMP amplification. This assay setup with biotinylated DNA probe-based LFA readouts of the RT-LAMP amplicon was 98.11% sensitive and 96.15% specific. The LFA result was further analysed by a smartphone-based IVD device, wherein the T-line intensity was recorded. The LFA T-line intensity was then correlated with the qRT-PCR Ct value of the positive swab samples. A digital semi-quantification of RT-LAMP-LFA was reported with a correlation coefficient of R2 = 0.702. The overall RT-LAMP-LFA assay time was recorded to be 35 min with a LoD of three RNA copies/µL (Ct-33). With these three advancements, the nucleic acid testing-point of care technique (NAT-POCT) is exemplified as a versatile biosensor platform with great potential and applicability for the detection of pathogens without the need for sample storage, transportation, or pre-processing.
Hydrometric networks play a vital role in providing information for decision-making in water resource management. They should be set up optimally to provide as much information as possible that is as accurate as possible and, at the same time, be cost-effective. Although the design of hydrometric networks is a well-identified problem in hydrometeorology and has received considerable attention, there is still scope for further advancement. In this study, we use complex network analysis, defined as a collection of nodes interconnected by links, to propose a new measure that identifies critical nodes of station networks. The approach can support the design and redesign of hydrometric station networks. The science of complex networks is a relatively young field and has gained significant momentum over the last few years in different areas such as brain networks, social networks, technological networks, or climate networks. The identification of influential nodes in complex networks is an important field of research. We propose a new node-ranking measure – the weighted degree–betweenness (WDB) measure – to evaluate the importance of nodes in a network. It is compared to previously proposed measures used on synthetic sample networks and then applied to a real-world rain gauge network comprising 1229 stations across Germany to demonstrate its applicability. The proposed measure is evaluated using the decline rate of the network efficiency and the kriging error. The results suggest that WDB effectively quantifies the importance of rain gauges, although the benefits of the method need to be investigated in more detail.
The temporal dynamics of climate processes are spread across different timescales and, as such, the study of these processes at only one selected timescale might not reveal the complete mechanisms and interactions within and between the (sub-) processes. To capture the non-linear interactions between climatic events, the method of event synchronization has found increasing attention recently. The main drawback with the present estimation of event synchronization is its restriction to analysing the time series at one reference timescale only. The study of event synchronization at multiple scales would be of great interest to comprehend the dynamics of the investigated climate processes. In this paper, the wavelet-based multi-scale event synchronization (MSES) method is proposed by combining the wavelet transform and event synchronization. Wavelets are used extensively to comprehend multi-scale processes and the dynamics of processes across various timescales. The proposed method allows the study of spatio-temporal patterns across different timescales. The method is tested on synthetic and real-world time series in order to check its replicability and applicability. The results indicate that MSES is able to capture relationships that exist between processes at different timescales.
Quantifying the roles of single stations within homogeneous regions using complex network analysis
(2018)
Regionalization and pooling stations to form homogeneous regions or communities are essential for reliable parameter transfer, prediction in ungauged basins, and estimation of missing information. Over the years, several clustering methods have been proposed for regional analysis. Most of these methods are able to quantify the study region in terms of homogeneity but fail to provide microscopic information about the interaction between communities, as well as about each station within the communities. We propose a complex network-based approach to extract this valuable information and demonstrate the potential of our approach using a rainfall network constructed from the Indian gridded daily precipitation data. The communities were identified using the network-theoretical community detection algorithm for maximizing the modularity. Further, the grid points (nodes) were classified into universal roles according to their pattern of within- and between-community connections. The method thus yields zoomed-in details of individual rainfall grids within each community.
In recent years, complex network analysis facilitated the identification of universal and unexpected patterns in complex climate systems. However, the analysis and representation of a multiscale complex relationship that exists in the global climate system are limited. A logical first step in addressing this issue is to construct multiple networks over different timescales. Therefore, we propose to apply the wavelet multiscale correlation (WMC) similarity measure, which is a combination of two state-of-the-art methods, viz. wavelet and Pearson’s correlation, for investigating multiscale processes through complex networks. Firstly we decompose the data over different timescales using the wavelet approach and subsequently construct a corresponding network by Pearson’s correlation. The proposed approach is illustrated and tested on two synthetics and one real-world example. The first synthetic case study shows the efficacy of the proposed approach to unravel scale-specific connections, which are often undiscovered at a single scale. The second synthetic case study illustrates that by dividing and constructing a separate network for each time window we can detect significant changes in the signal structure. The real-world example investigates the behavior of the global sea surface temperature (SST) network at different timescales. Intriguingly, we notice that spatial dependent structure in SST evolves temporally. Overall, the proposed measure has an immense potential to provide essential insights on understanding and extending complex multivariate process studies at multiple scales.
Hydrologic regionalization deals with the investigation of homogeneity in watersheds and provides a classification of watersheds for regional analysis. The classification thus obtained can be used as a basis for mapping data from gauged to ungauged sites and can improve extreme event prediction. This paper proposes a wavelet power spectrum (WPS) coupled with the self-organizing map method for clustering hydrologic catchments. The application of this technique is implemented for gauged catchments. As a test case study, monthly streamflow records observed at 117 selected catchments throughout the western United States from 1951 through 2002. Further, based on WPS of each station, catchments are classified into homogeneous clusters, which provides a representative WPS pattern for the streamflow stations in each cluster.
The quantification of spatial propagation of extreme precipitation events is vital in water resources planning and disaster mitigation. However, quantifying these extreme events has always been challenging as many traditional methods are insufficient to capture the nonlinear interrelationships between extreme event time series. Therefore, it is crucial to develop suitable methods for analyzing the dynamics of extreme events over a river basin with a diverse climate and complicated topography. Over the last decade, complex network analysis emerged as a powerful tool to study the intricate spatiotemporal relationship between many variables in a compact way. In this study, we employ two nonlinear concepts of event synchronization and edit distance to investigate the extreme precipitation pattern in the Ganga river basin. We use the network degree to understand the spatial synchronization pattern of extreme rainfall and identify essential sites in the river basin with respect to potential prediction skills. The study also attempts to quantify the influence of precipitation seasonality and topography on extreme events. The findings of the study reveal that (1) the network degree is decreased in the southwest to northwest direction, (2) the timing of 50th percentile precipitation within a year influences the spatial distribution of degree, (3) the timing is inversely related to elevation, and (4) the lower elevation greatly influences connectivity of the sites. The study highlights that edit distance could be a promising alternative to analyze event-like data by incorporating event time and amplitude and constructing complex networks of climate extremes.
Sea surface temperature (SST) patterns can – as surface climate forcing – affect weather and climate at large distances. One example is El Niño-Southern Oscillation (ENSO) that causes climate anomalies around the globe via teleconnections. Although several studies identified and characterized these teleconnections, our understanding of climate processes remains incomplete, since interactions and feedbacks are typically exhibited at unique or multiple temporal and spatial scales. This study characterizes the interactions between the cells of a global SST data set at different temporal and spatial scales using climate networks. These networks are constructed using wavelet multi-scale correlation that investigate the correlation between the SST time series at a range of scales allowing instantaneously deeper insights into the correlation patterns compared to traditional methods like empirical orthogonal functions or classical correlation analysis. This allows us to identify and visualise regions of – at a certain timescale – similarly evolving SSTs and distinguish them from those with long-range teleconnections to other ocean regions. Our findings re-confirm accepted knowledge about known highly linked SST patterns like ENSO and the Pacific Decadal Oscillation, but also suggest new insights into the characteristics and origins of long-range teleconnections like the connection between ENSO and Indian Ocean Dipole.
Numerical simulation of fluid-flow processes in a 3D high-resolution carbonate reservoir analogue
(2014)
A high-resolution three-dimensional (3D) outcrop model of a Jurassic carbonate ramp was used in order to perform a series of detailed and systematic flow simulations. The aim of this study was to test the impact of small- and large-scale geological features on reservoir performance and oil recovery. The digital outcrop model contains a wide range of sedimentological, diagenetic and structural features, including discontinuity surfaces, shoal bodies, mud mounds, oyster bioherms and fractures. Flow simulations are performed for numerical well testing and secondary oil recovery. Numerical well testing enables synthetic but systematic pressure responses to be generated for different geological features observed in the outcrops. This allows us to assess and rank the relative impact of specific geological features on reservoir performance. The outcome documents that, owing to the realistic representation of matrix heterogeneity, most diagenetic and structural features cannot be linked to a unique pressure signature. Instead, reservoir performance is controlled by subseismic faults and oyster bioherms acting as thief zones. Numerical simulations of secondary recovery processes reveal strong channelling of fluid flow into high-permeability layers as the primary control for oil recovery. However, appropriate reservoir-engineering solutions, such as optimizing well placement and injection fluid, can reduce channelling and increase oil recovery.
Pancreatic steatosis associates with beta-cell failure and may participate in the development of type-2-diabetes. Our previous studies have shown that diabetes-susceptible mice accumulate more adipocytes in the pancreas than diabetes-resistant mice. In addition, we have demonstrated that the co-culture of pancreatic islets and adipocytes affect insulin secretion. The aim of this current study was to elucidate if and to what extent pancreas-resident mesenchymal stromal cells (MSCs) with adipogenic progenitor potential differ from the corresponding stromal-type cells of the inguinal white adipose tissue (iWAT). miRNA (miRNome) and mRNA expression (transcriptome) analyses of MSCs isolated by flow cytometry of both tissues revealed 121 differentially expressed miRNAs and 1227 differentially expressed genes (DEGs). Target prediction analysis estimated 510 DEGs to be regulated by 58 differentially expressed miRNAs. Pathway analyses of DEGs and miRNA target genes showed unique transcriptional and miRNA signatures in pancreas (pMSCs) and iWAT MSCs (iwatMSCs), for instance fibrogenic and adipogenic differentiation, respectively. Accordingly, iwatMSCs revealed a higher adipogenic lineage commitment, whereas pMSCs showed an elevated fibrogenesis. As a low degree of adipogenesis was also observed in pMSCs of diabetes-susceptible mice, we conclude that the development of pancreatic steatosis has to be induced by other factors not related to cell-autonomous transcriptomic changes and miRNA-based signals.
Type 2 diabetes (T2D) is a complex metabolic disease regulated by an interaction of genetic predisposition and environmental factors. To understand the genetic contribution in the development of diabetes, mice varying in their disease susceptibility were crossed with the obese and diabetes-prone New Zealand obese (NZO) mouse. Subsequent whole-genome sequence scans revealed one major quantitative trait loci (QTL),Nidd/DBAon chromosome 4, linked to elevated blood glucose and reduced plasma insulin and low levels of pancreatic insulin. Phenotypical characterization of congenic mice carrying 13.6 Mbp of the critical fragment of DBA mice displayed severe hyperglycemia and impaired glucose clearance at week 10, decreased glucose response in week 13, and loss of beta-cells and pancreatic insulin in week 16. To identify the responsible gene variant(s), further congenic mice were generated and phenotyped, which resulted in a fragment of 3.3 Mbp that was sufficient to induce hyperglycemia. By combining transcriptome analysis and haplotype mapping, the number of putative responsible variant(s) was narrowed from initial 284 to 18 genes, including gene models and non-coding RNAs. Consideration of haplotype blocks reduced the number of candidate genes to four (Kti12,Osbpl9,Ttc39a, andCalr4) as potential T2D candidates as they display a differential expression in pancreatic islets and/or sequence variation. In conclusion, the integration of comparative analysis of multiple inbred populations such as haplotype mapping, transcriptomics, and sequence data substantially improved the mapping resolution of the diabetes QTLNidd/DBA. Future studies are necessary to understand the exact role of the different candidates in beta-cell function and their contribution in maintaining glycemic control.
Information on structural features of a fracture network at early stages of Enhanced Geothermal System development is mostly restricted to borehole images and, if available, outcrop data. However, using this information to image discontinuities in deep reservoirs is difficult. Wellbore failure data provides only some information on components of the in situ stress state and its heterogeneity. Our working hypothesis is that slip on natural fractures primarily controls these stress heterogeneities. Based on this, we introduce stress-based tomography in a Bayesian framework to characterize the fracture network and its heterogeneity in potential Enhanced Geothermal System reservoirs. In this procedure, first a random initial discrete fracture network (DFN) realization is generated based on prior information about the network. The observations needed to calibrate the DFN are based on local variations of the orientation and magnitude of at least one principal stress component along boreholes. A Markov Chain Monte Carlo sequence is employed to update the DFN iteratively by a fracture translation within the domain. The Markov sequence compares the simulated stress profile with the observed stress profiles in the borehole, evaluates each iteration with Metropolis-Hastings acceptance criteria, and stores acceptable DFN realizations in an ensemble. Finally, this obtained ensemble is used to visualize the potential occurrence of fractures in a probability map, indicating possible fracture locations and lengths. We test this methodology to reconstruct simple synthetic and more complex outcrop-based fracture networks and successfully image the significant fractures in the domain.
Parsing of argumentative structures has become a very active line of research in recent years. Like discourse parsing or any other natural language task that requires prediction of linguistic structures, most approaches choose to learn a local model and then perform global decoding over the local probability distributions, often imposing constraints that are specific to the task at hand. Specifically for argumentation parsing, two decoding approaches have been recently proposed: Minimum Spanning Trees (MST) and Integer Linear Programming (ILP), following similar trends in discourse parsing. In contrast to discourse parsing though, where trees are not always used as underlying annotation schemes, argumentation structures so far have always been represented with trees. Using the ‘argumentative microtext corpus’ [in: Argumentation and Reasoned Action: Proceedings of the 1st European Conference on Argumentation, Lisbon 2015 / Vol. 2, College Publications, London, 2016, pp. 801–815] as underlying data and replicating three different decoding mechanisms, in this paper we propose a novel ILP decoder and an extension to our earlier MST work, and then thoroughly compare the approaches. The result is that our new decoder outperforms related work in important respects, and that in general, ILP and MST yield very similar performance.
The behaviour of individuals, businesses, and government entities before, during, and immediately after a disaster can dramatically affect the impact and recovery time. However, existing risk-assessment methods rarely include this critical factor. In this Perspective, we show why this is a concern, and demonstrate that although initial efforts have inevitably represented human behaviour in limited terms, innovations in flood-risk assessment that integrate societal behaviour and behavioural adaptation dynamics into such quantifications may lead to more accurate characterization of risks and improved assessment of the effectiveness of risk-management strategies and investments. Such multidisciplinary approaches can inform flood-risk management policy development.
Im Artikel werden von Frauen verfasste Filmdrehbücher der 1910er Jahre im Russischen Kaiserreich chronologisch untersucht. Zunächst werden die ersten Drehbuchautorinnen Makarova und Tat’jana Suchotina-Tolstaja, die am Anfang der 1910er Jahre in Koautorschaft mit den bekannten Autoren (Makarova mit den Regisseur Vladimir Gončarov; Suchotina-Tolstaja mit ihrem Vater Leo Tolstoj) arbeiteten, und ihre Filme in Betracht gezogen. Dann wird der Film Ključi sčastʹja / Schlüssel zum Glück (Vladimir Gardin, Jakov Protazanov, 1913, Russisches Kaiserreich) nach dem Roman von Anastasija Verbickaja näher behandelt. Verbickajas Film demonstrierte, dass eine Drehbuchautorin eine selbständige Autorin sein kann und diente als Impuls für die Entwicklung der Frauenfilmdramaturgie im Russischen Kaiserreich, deren Aufschwung in der zweiten Hälfte der 1910er Jahre begann, und prägte bestimmte Erwartungen von auf weiblichen Drehbüchern basierenden Filmen. Maria Kallaš, die an den Drehbüchern zu den Verfilmungen des russischen literarischen Kanons 1913 arbeitete, kritisierte Verbickajas Text als pseudofeministisch und behauptete in ihrem Essay „Ženskie kabare“ („Frauenkabarett“), dass Frauenliteratur noch „keine eigene Sprache“ habe (1916). Anna Mar begann ihre Arbeit im Kino 1914, parallel zu Verbiсkajas Nachfolgerinnen, und konzentrierte sich in ihren Filmen auf die soziale Problematik – die Stellung moderner Frauen in der Gesellschaft. Damit eröffnete Mar eine neue Entwicklungsperspektive für das weibliche Drehbuchschreiben.
Accurately predicting total electron content (TEC) during geomagnetic storms is still a challenging task for ionospheric models. In this work, a neural-network (NN)-based model is proposed which predicts relative TEC with respect to the preceding 27-day median TEC, during storm time for the European region (with longitudes 30 degrees W-50 degrees E and latitudes 32.5 degrees N-70 degrees N). The 27-day median TEC (referred to as median TEC), latitude, longitude, universal time, storm time, solar radio flux index F10.7, global storm index SYM-H and geomagnetic activity index Hp30 are used as inputs and the output of the network is the relative TEC. The relative TEC can be converted to the actual TEC knowing the median TEC. The median TEC is calculated at each grid point over the European region considering data from the last 27 days before the storm using global ionosphere maps (GIMs) from international GNSS service (IGS) sources. A storm event is defined when the storm time disturbance index Dst drops below 50 nanotesla. The model was trained with storm-time relative TEC data from the time period of 1998 until 2019 (2015 is excluded) and contains 365 storms. Unseen storm data from 33 storm events during 2015 and 2020 were used to test the model. The UQRG GIMs were used because of their high temporal resolution (15 min) compared to other products from different analysis centers. The NN-based model predictions show the seasonal behavior of the storms including positive and negative storm phases during winter and summer, respectively, and show a mixture of both phases during equinoxes. The model's performance was also compared with the Neustrelitz TEC model (NTCM) and the NN-based quiet-time TEC model, both developed at the German Aerospace Agency (DLR). The storm model has a root mean squared error (RMSE) of 3.38 TEC units (TECU), which is an improvement by 1.87 TECU compared to the NTCM, where an RMSE of 5.25 TECU was found. This improvement corresponds to a performance increase by 35.6%. The storm-time model outperforms the quiet-time model by 1.34 TECU, which corresponds to a performance increase by 28.4% from 4.72 to 3.38 TECU. The quiet-time model was trained with Carrington averaged TEC and, therefore, is ideal to be used as an input instead of the GIM derived 27-day median. We found an improvement by 0.8 TECU which corresponds to a performance increase by 17% from 4.72 to 3.92 TECU for the storm-time model using the quiet-time-model predicted TEC as an input compared to solely using the quiet-time model.
In clinical settings, significant resources are spent on data collection and monitoring patients' health parameters to improve decision-making and provide better care. With increased digitization, the healthcare sector is shifting towards implementing digital technologies for data management and in administration. New technologies offer better treatment opportunities and streamline clinical workflow, but the complexity can cause ineffectiveness, frustration, and errors. To address this, we believe digital solutions alone are not sufficient. Therefore, we take a human-centred design approach for AI development, and apply systems engineering methods to identify system leverage points. We demonstrate how automation enables monitoring clinical parameters, using existing non-intrusive sensor technology, resulting in more resources toward patient care. Furthermore, we provide a framework on digitization of clinical data for integration with data management.
The subsurface harbors a large fraction of Earth's living biomass, forming complex microbial ecosystems. Without a profound knowledge of the ongoing biologically mediated processes and their reaction to anthropogenic changes it is difficult to assess the long-term stability and feasibility of any type of geotechnical utilization, as these influence subsurface ecosystems. Despite recent advances in many areas of subsurface microbiology, the direct quantification of turnover processes is still in its infancy, mainly due to the extremely low cell abundances. We provide an overview of the currently available techniques for the quantification of microbial turnover processes and discuss their specific strengths and limitations. Most techniques employed so far have focused on specific processes, e.g. sulfate reduction or methanogenesis. Recent studies show that processes that were previously thought to exclude each other can occur simultaneously, albeit at very low rates. Without the identification of the respective processes it is impossible to quantify total microbial activity. Even in cases where all simultaneously occurring processes can be identified, the typically very low rates prevent quantification. In many cases a simple measure of total microbial activity would be a better and more robust measure than assays for several specific processes. Enzyme or molecular assays provide a more general approach as they target key metabolic compounds. Depending on the compound targeted a broader spectrum of microbial processes can be quantified. The two most promising compounds are ATP and hydrogenase, as both are ubiquitous in microbes. Technical constraints limit the applicability of currently available ATP-assays for subsurface samples. A recently developed hydrogenase radiotracer assay has the potential to become a key tool for the quantification of subsurface microbial activity.
Subsurface microbial communities undertake many terminal electron-accepting processes, often simultaneously. Using a tritium-based assay, we measured the potential hydrogen oxidation catalyzed by hydrogenase enzymes in several subsurface sedimentary environments (Lake Van, Barents Sea, Equatorial Pacific, and Gulf of Mexico) with different predominant electron-acceptors. Hydrogenases constitute a diverse family of enzymes expressed by microorganisms that utilize molecular hydrogen as a metabolic substrate, product, or intermediate. The assay reveals the potential for utilizing molecular hydrogen and allows qualitative detection of microbial activity irrespective of the predominant electron-accepting process. Because the method only requires samples frozen immediately after recovery, the assay can be used for identifying microbial activity in subsurface ecosystems without the need to preserve live material. We measured potential hydrogen oxidation rates in all samples from multiple depths at several sites that collectively span a wide range of environmental conditions and biogeochemical zones. Potential activity normalized to total cell abundance ranges over five orders of magnitude and varies, dependent upon the predominant terminal electron acceptor. Lowest per-cell potential rates characterize the zone of nitrate reduction and highest per-cell potential rates occur in the methanogenic zone. Possible reasons for this relationship to predominant electron acceptor include (i) increasing importance of fermentation in successively deeper biogeochemical zones and (ii) adaptation of H(2)ases to successively higher concentrations of H-2 in successively deeper zones.
Efficient Removal of Tetracycline and Bisphenol A from Water with a New Hybrid Clay/TiO2 Composite
(2023)
New TiO2 hybrid composites were prepared fromkaolinclay, predried and carbonized biomass, and titanium tetraisopropoxideand explored for tetracycline (TET) and bisphenol A (BPA) removalfrom water. Overall, the removal rate is 84% for TET and 51% for BPA.The maximum adsorption capacities (q (m))are 30 and 23 mg/g for TET and BPA, respectively. These capacitiesare far greater than those obtained for unmodified TiO2. Increasing the ionic strength of the solution does not change theadsorption capacity of the adsorbent. pH changes only slightly changeBPA adsorption, while a pH > 7 significantly reduces the adsorptionof TET on the material. The Brouers-Sotolongo fractal modelbest describes the kinetic data for both TET and BPA adsorption, predictingthat the adsorption process occurs via a complex mechanism involvingvarious forces of attraction. Temkin and Freundlich isotherms, whichbest fit the equilibrium adsorption data for TET and BPA, respectively,suggest that adsorption sites are heterogeneous in nature. Overall,the composite materials are much more effective for TET removal fromaqueous solution than for BPA. This phenomenon is assigned to a differencein the TET/adsorbent interactions vs the BPA/adsorbent interactions:the decisive factor appears to be favorable electrostatic interactionsfor TET yielding a more effective TET removal.
Background: While incidences of cancer are continuously increasing, drug resistance of malignant cells is observed towards almost all pharmaceuticals. Several isoflavonoids and flavonoids are known for their cytotoxicity towards various cancer cells. Methods: The cytotoxicity of compounds was determined based on the resazurin reduction assay. Caspases activation was evaluated using the caspase-Glo assay. Flow cytometry was used to analyze the cell cycle (propodium iodide (PI) staining), apoptosis (annexin V/PI staining), mitochondrial membrane potential (MMP) (JC-1) and reactive oxygen species (ROS) (H2DCFH-DA). CCRF-CEM leukemia cells were used as model cells for mechanistic studies. Results: Compounds 1, 2 and 4 displayed IC50 values below 20 mu M towards CCRF-CEM and CEM/ADR5000 leukemia cells, and were further tested towards a panel of 7 carcinoma cells. The IC50 values of the compounds against carcinoma cells varied from 16.90 mu M (in resistant U87MG.Delta EGFR glioblastoma cells) to 48.67 mu M (against HepG2 hepatocarcinoma cells) for 1, from 7.85 mu M (in U87MG.Delta EGFR cells) to 14.44 mu M (in resistant MDA-MB231/BCRP breast adenocarcinoma cells) for 2, from 4.96 mu M (towards U87MG.Delta EGFRcells) to 7.76 mu M (against MDA-MB231/BCRP cells) for 4, and from 0.07 mu M (against MDA-MB231 cells) to 2.15 mu M (against HepG2 cells) for doxorubicin. Compounds 2 and 4 induced apoptosis in CCRF-CEM cells mediated by MMP alteration and increased ROS production. Conclusion: The present report indicates that isoflavones and biflavonoids from Ormocarpum kirkii are cytotoxic compounds with the potential of being exploited in cancer chemotherapy. Compounds 2 and 4 deserve further studies to develop new anticancer drugs to fight sensitive and resistant cancer cell lines.
Chromatographic separation of the extract of the roots of Dorstenia kameruniana (family Moraceae) led to the isolation of three new benzylbenzofuran derivatives, 2-(p-hydroxybenzyl)benzofuran-6-ol (1), 2-(p-hydroxybenzyl)-7-methoxybenzofuran-6-ol (2) and 2-(p-hydroxy)-3-(3-methylbut-2-en-1-yl)benzyl)benzofuran-6-ol (3) (named dorsmerunin A, B and C, respectively), along with the known furanocoumarin, bergapten (4). The twigs of Dorstenia kameruniana also produced compounds 1-4 as well as the known chalcone licoagrochalcone A (5). The structures were elucidated by NMR spectroscopy and mass spectrometry. The isolated compounds displayed cytotoxicity against the sensitive CCRF-CEM and multidrug-resistant CEM/ADR5000 leukemia cells, where compounds 4 and 5 had the highest activities (IC50 values of 7.17 mu M and 5.16 mu M, respectively) against CCRF-CEM leukemia cells. Compound 5 also showed cytotoxicity against 7 sensitive or drug-resistant solid tumor cell lines (breast carcinoma, colon carcinoma, glioblastoma), with IC50 below 50 mu M, whilst 4 showed selective activity.
A new isoflavone, 4′-prenyloxyvigvexin A (1) and a new pterocarpan, (6aR,11aR)-3,8-dimethoxybitucarpin B (2) were isolated from the leaves of Lonchocarpus bussei and the stem bark of Lonchocarpus eriocalyx, respectively. The extract of L. bussei also gave four known isoflavones, maximaisoflavone H, 7,2′-dimethoxy-3′,4′-methylenedioxyisoflavone, 6,7,3′-trimethoxy-4′,5′-methylenedioxyisoflavone, durmillone; a chalcone, 4-hydroxylonchocarpin; a geranylated phenylpropanol, colenemol; and two known pterocarpans, (6aR,11aR)-maackiain and (6aR,11aR)-edunol. (6aR,11aR)-Edunol was also isolated from the stem bark of L. eriocalyx. The structures of the isolated compounds were elucidated by spectroscopy. The cytotoxicity of the compounds was tested by resazurin assay using drug-sensitive and multidrug-resistant cancer cell lines. Significant antiproliferative effects with IC50 values below 10 μM were observed for the isoflavones 6,7,3′-trimethoxy-4′,5′-methylenedioxyisoflavone and durmillone against leukemia CCRF-CEM cells; for the chalcone, 4-hydroxylonchocarpin and durmillone against its resistant counterpart CEM/ADR5000 cells; as well as for durmillone against the resistant breast adenocarcinoma MDA-MB231/BCRP cells and resistant gliobastoma U87MG.ΔEGFR cells.
The cross-linguistic finding of greater demands in processing object relatives as compared to subject relatives in individuals with aphasia and non-brain-damaged speakers has been explained within the Relativized Minimality approach. Based on this account, the asymmetry is attributed to an element intervening between the moved element and its extraction site in object relatives, but not in subject relatives. Moreover, it has been proposed that processing of object relatives is facilitated if the intervening and the moved elements differ in their internal feature structure. The present study investigates these predictions in German-speaking individuals with aphasia and a group of control participants by combining the visual world eye-tracking methodology with an auditory referent identification task. Our results provide support for the Relativized Minimality approach. Particularly, the degree of featural distinctness was shown to modulate the occurrence of the effects in aphasia. We claim that, due to reduced processing capacities, individuals with aphasia need a higher degree of featural dissimilarity to distinguish the moved from the intervening element in object relatives to overcome their syntactic deficit. (C) 2017 Elsevier Ltd. All rights reserved.
Exploring generalisation following treatment of language deficits in aphasia can provide insights into the functional relation of the cognitive processing systems involved. In the present study, we first review treatment outcomes of interventions targeting sentence processing deficits and, second report a treatment study examining the occurrence of practice effects and generalisation in sentence comprehension and production. In order to explore the potential linkage between processing systems involved in comprehending and producing sentences, we investigated whether improvements generalise within (i.e., uni-modal generalisation in comprehension or in production) and/or across modalities (i.e., cross-modal generalisation from comprehension to production or vice versa). Two individuals with aphasia displaying co-occurring deficits in sentence comprehension and production were trained on complex, non-canonical sentences in both modalities. Two evidence-based treatment protocols were applied in a crossover intervention study with sequence of treatment phases being randomly allocated. Both participants benefited significantly from treatment, leading to uni-modal generalisation in both comprehension and production. However, cross-modal generalisation did not occur. The magnitude of uni-modal generalisation in sentence production was related to participants’ sentence comprehension performance prior to treatment. These findings support the assumption of modality-specific sub-systems for sentence comprehension and production, being linked uni-directionally from comprehension to production.
Structural changes at the intra- as well as intermicellar level were induced by the LCST-type collapse transition of poly(N-isopropyl acrylamide) in ABA triblock copolymer micelles in water. The distinct process kinetics was followed in situ and in real-time using time-resolved small-angle neutron scattering (SANS), while a micellar solution of a triblock copolymer, consisting of two short deuterated polystyrene endblocks and a long thermoresponsive poly(N-isopropyl acrylamide) middle block, was heated rapidly above its cloud point. A very fast collapse together with a multistep aggregation behavior is observed. The findings of the transition occurring at several size and time levels may have implications for the design and application of such thermoresponsive self-assembled systems.
We investigate concentrated solutions of poly(styrene-b-N-isopropyl acrylamide) (P(S-b-NIPAM)) diblock copolymers in deuterated water (D2O). Both structural changes and the changes of the segmental dynamics occurring upon heating through the lower critical solution temperature (LCST) of PNIPAM are studied using small-angle neutron scattering and neutron spin-echo spectroscopy. The collapse of the micellar shell and the cluster formation of collapsed micelles at the LCST as well as an increase of the segmental diffusion coefficient after crossing the LCST are detected. Comparing to our recent results on a triblock copolymer P(S-b-NIPAM-b-S) [25], we observe that the collapse transition of P(S-b-NIPAM) is more complex and that the PNIPAM segmental dynamics are faster than in P(S-b-NIPAM-b-S).
We have studied I lie thermal behavior of amphiphilic, symmetric triblock copolymers having short, deuterated polystyrene (PS) end blocks and a large poly(N-isopropylacrylarnicle) (PNIPAM) middle block exhibiting a lower critical solution temperature (LCST) in aqueous solution. A wide range of concentrations (0.1-300 mg/mL) is investigated using it number of analytical methods such as fluorescence correlation spectroscopy (FCS), turbidimetry, dynamic light scattering (DLS), small-angle neutron scattering (SANS), and neutron spin-echo spectroscopy (NSE). The critical micelle concentration is determined using FCS to be 1 mu M or less. The collapse of the micelles at the LCST is investigated using turbidimetry and DLS and shows a weak dependence on the degree of polymerization of the PNIPAM block. SANS with contrast matching allows its to reveal the core-shell Structure of the micelles as well as their correlation as a function of temperature. The segmental dynamics of the PNIPAM shell are studied as a function of temperature and arc found to be faster in the collapsed state than in the swollen state. The mode detected has a linear dispersion in q(2) and is found to be faster in the collapsed state as compared to the swollen state. We attribute this result to the averaging over mobile and immobilized segments.
In aqueous solution, symmetric triblock copolymers with a thermoresponsive middle block and hydrophobic end blocks form flower-like core-shell micelles which collapse and aggregate upon heating through the cloud point (CP). The collapse of the micellar shell and the intermicellar aggregation are followed in situ and in real-time using time-resolved small-angle neutron scattering (SANS), while heating micellar solutions of a poly((styrene-d(8))-b-(N-isopropyl acrylamide)-b-(styrene-d(8))) triblock copolymer in D2O rapidly through their CP. The influence of polymer concentration as well as of the start and target temperatures is addressed. In all cases, the micellar collapse is very fast. The collapsed micelles immediately form small clusters which contain voids. They densify which slows down or even stops their growth. For low concentrations and target temperatures just above the CP, i.e. shallow temperature jumps, the subsequent growth of the clusters is described by diffusion-limited aggregation. In contrast, for higher concentrations and/or higher target temperatures, i.e. deep temperature jumps, intermicellar bridges dominate the growth. Eventually, in all cases, the clusters coagulate which results in macroscopic phase separation. For shallow temperature jumps, the cluster surfaces stay rough; whereas for deep temperature jumps, a concentration gradient develops at late stages. These results are important for the development of conditions for thermal switching in applications, e.g. for the use of thermoresponsive micellar systems for transport and delivery purposes.
A concentrated solution of a symmetric triblock copolymer with a thermoresponsive poly(methoxy diethylene glycol acrylate) (PMDEGA) middle block and short hydrophobic, fully deuterated polystyrene end blocks is investigated in D2O where it undergoes a lower critical solution temperature-type phase transition at ca. 36 A degrees C. Small-angle neutron scattering (SANS) in a wide temperature range (15-50 A degrees C) is used to characterize the size and inner structure of the micelles as well as the correlation between the micelles and the formation of aggregates by the micelles above the cloud point (CP). A model featuring spherical core-shell micelles, which are correlated by a hard-sphere potential or a sticky hard-sphere potential together with a Guinier form factor describing aggregates formed by the micelles above the CP, fits the SANS curves well in the entire temperature range. The thickness of the thermoresponsive micellar PMDEGA shell as well as the hard-sphere radius increase slightly already below the cloud point. Whereas the thickness of the thermoresponsive micellar shell hardly shrinks when heating through the CP and up to 50 A degrees C, the hard-sphere radius decreases within 3.5 K at the CP. The volume fraction decreases already significantly below the CP, which may be at the origin of the previously observed gel-sol transition far below the CP (Miasnikova et al., Langmuir 28: 4479-4490, 2012). Above the CP, small, and at higher temperatures, large aggregates are formed by the micelles.
The central rift of the Red Sea has 25 brine pools with different physical and geochemical characteristics. Atlantis II (ATIID), Discovery Deeps (DD) and Chain Deep (CD) are characterized by high salinity, temperature and metal content. Several studies reported microbial communities in these brine pools, but few studies addressed the brine pool sediments. Therefore, sediment cores were collected from ATIID, DD, CD brine pools and an adjacent brine-influenced site. Sixteen different lithologic sediment sections were subjected to shotgun DNA pyrosequencing to generate 1.47 billion base pairs (1.47 x 10(9) bp). We generated sediment-specific reads and attempted to annotate all reads. We report the phylogenetic and biochemical uniqueness of the deepest ATIID sulfur-rich brine pool sediments. In contrary to all other sediment sections, bacteria dominate the deepest ATIID sulfur-rich brine pool sediments. This decrease in virus-to-bacteria ratio in selected sections and depth coincided with an overrepresentation of mobile genetic elements. Skewing in the composition of viruses-to-mobile genetic elements may uniquely contribute to the distinct microbial consortium in sediments in proximity to hydrothermally active vents of the Red Sea and possibly in their surroundings, through differential horizontal gene transfer.
Quality attributes of fruit determine its acceptability by the retailer and consumer. The objective of this work was to investigate the potential of absorption (μa) and reduced scattering (μs’) coefficients of European pear to analyze its fruit flesh firmness and soluble solids content (SSC). The absolute reference values, μa* (cm−1) and μs’* (cm−1), of pear were invasively measured, employing multi-spectral photon density wave (PDW) spectroscopy at preselected wavelengths of 515, 690, and 940 nm considering two batches of unripe and overripe fruit. On eight measuring dates during fruit development, μa and μs’ were analyzed non-destructively by means of laser light backscattering imaging (LLBI) at similar wavelengths of 532, 660, and 830 nm by means of fitting according to Farrell’s diffusion theory, using fix reference values of either μa* or μs’*. Both, the μa* and the μa as well as μs’* and μs’ showed similar trends. Considering the non-destructively measured data during fruit development, μa at 660 nm decreased 91 till 141 days after full bloom (dafb) from 1.49 cm−1 to 0.74 cm−1 due to chlorophyll degradation. At 830 nm, μa only slightly decreased from 0.41 cm−1 to 0.35 cm−1. The μs’ at all wavelengths revealed a decreasing trend as the fruit developed. The difference measured at 532 nm was most pronounced decreasing from 24 cm−1 to 10 cm−1, while at 660 nm and 830 nm values decreased from 15 cm−1 to 13 cm−1 and from 10 cm−1 to 8 cm−1, respectively. When building calibration models with partial least-squares regression analysis on the optical properties for non-destructive analysis of the fruit SSC, μa at 532 nm and 830 nm resulted in a correlation coefficient of R = 0.66, however, showing high measuring uncertainty. The combination of all three wavelengths gave an enhanced, encouraging R = 0.89 for firmness analysis using μs’ in the freshly picked fruit.
The Relativized Minimality approach to A'-dependencies (Friedmann et al., 2009) predicts that headed object relative clauses (RCs) and which questions are the most difficult, due to the presence of a lexical restriction on both the subject and the object DP which creates intervention. We investigated comprehension of center-embedded headed object RCs with Italian children, where Number and Gender feature values on subject and object DPs are manipulated. We found that. Number conditions are always more accurate than Gender ones, showing that intervention is sensitive to DP-internal structure. We propose a finer definition of the lexical restriction where external and syntactically active features (such as Number) reduce intervention whereas internal and (possibly) lexicalized features (such as Gender) do so to a lesser extent. Our results are also compatible with a memory interference approach in which the human parser is sensitive to highly specific properties of the linguistic input, such as the cue-based model (Van Dyke, 2007).
The predictions of two contrasting approaches to the acquisition of transitive relative clauses were tested within the same groups of German-speaking participants aged from 3 to 5 years old. The input frequency approach predicts that object relative clauses with inanimate heads (e.g., the pullover that the man is scratching) are comprehended earlier and more accurately than those with an animate head (e.g., the man that the boy is scratching). In contrast, the structural intervention approach predicts that object relative clauses with two full NP arguments mismatching in number (e.g., the man that the boys are scratching) are comprehended earlier and more accurately than those with number-matching NPs (e.g., the man that the boy is scratching). These approaches were tested in two steps. First, we ran a corpus analysis to ensure that object relative clauses with number-mismatching NPs are not more frequent than object relative clauses with number-matching NPs in child directed speech. Next, the comprehension of these structures was tested experimentally in 3-, 4-, and 5-year-olds respectively by means of a color naming task. By comparing the predictions of the two approaches within the same participant groups, we were able to uncover that the effects predicted by the input frequency and by the structural intervention approaches co-exist and that they both influence the performance of children on transitive relative clauses, but in a manner that is modulated by age. These results reveal a sensitivity to animacy mismatch already being demonstrated by 3-year-olds and show that animacy is initially deployed more reliably than number to interpret relative clauses correctly. In all age groups, the animacy mismatch appears to explain the performance of children, thus, showing that the comprehension of frequent object relative clauses is enhanced compared to the other conditions. Starting with 4-year-olds but especially in 5-year-olds, the number mismatch supported comprehension—a facilitation that is unlikely to be driven by input frequency. Once children fine-tune their sensitivity to verb agreement information around the age of four, they are also able to deploy number marking to overcome the intervention effects. This study highlights the importance of testing experimentally contrasting theoretical approaches in order to characterize the multifaceted, developmental nature of language acquisition.
We elicited the production of various types of relative clauses in a group of German-speaking children with specific language impairment (SLI) and typically developing controls in order to test the movement optionality account of grammatical difficulty in SLI. The results show that German-speaking children with SLI are impaired in relative clause production compared to typically developing children. The alternative structures that they produce consist of simple main clauses, as well as nominal and prepositional phrases produced in isolation, sometimes contextually appropriate, and sometimes not. Crucially for evaluating the movement optionality account, children with SLI produce very few instances of embedded clauses where the relative clause head noun is pronounced in situ; in fact, such responses are more common among the typically developing child controls. These results underscore the difficulty German-speaking children with SLI have with structures involving movement, but provide no specific support for the movement optionality account.
The predictions of two contrasting approaches to the acquisition of transitive relative clauses were tested within the same groups of German-speaking participants aged from 3 to 5 years old. The input frequency approach predicts that object relative clauses with inanimate heads (e.g., the pullover that the man is scratching) are comprehended earlier and more accurately than those with an animate head (e.g., the man that the boy is scratching). In contrast, the structural intervention approach predicts that object relative clauses with two full NP arguments mismatching in number (e.g., the man that the boys are scratching) are comprehended earlier and more accurately than those with number-matching NPs (e.g., the man that the boy is scratching). These approaches were tested in two steps. First, we ran a corpus analysis to ensure that object relative clauses with number-mismatching NPs are not more frequent than object relative clauses with number-matching NPs in child directed speech. Next, the comprehension of these structures was tested experimentally in 3-, 4-, and 5-year-olds respectively by means of a color naming task. By comparing the predictions of the two approaches within the same participant groups, we were able to uncover that the effects predicted by the input frequency and by the structural intervention approaches co-exist and that they both influence the performance of children on transitive relative clauses, but in a manner that is modulated by age. These results reveal a sensitivity to animacy mismatch already being demonstrated by 3-year-olds and show that animacy is initially deployed more reliably than number to interpret relative clauses correctly. In all age groups, the animacy mismatch appears to explain the performance of children, thus, showing that the comprehension of frequent object relative clauses is enhanced compared to the other conditions. Starting with 4-year-olds but especially in 5-year-olds, the number mismatch supported comprehension-a facilitation that is unlikely to be driven by input frequency. Once children fine-tune their sensitivity to verb agreement information around the age of four, they are also able to deploy number marking to overcome the intervention effects. This study highlights the importance of testing experimentally contrasting theoretical approaches in order to characterize the multifaceted, developmental nature of language acquisition.
This study investigates whether number dissimilarities on subject and object DPs facilitate the comprehension of subject-and object-extracted centre-embedded relative clauses in children with Grammatical Specific Language Impairment (G-SLI). We compared the performance of a group of English-speaking children with G-SLI (mean age: 12; 11) with that of two groups of younger typically developing (TD) children, matched on grammar and receptive vocabulary, respectively. All groups were more accurate on subject-extracted relative clauses than object-extracted ones and, crucially, they all showed greater accuracy for sentences with dissimilar number features (i.e., one singular, one plural) on the head noun and the embedded DP. These findings are interpreted in the light of current psycholinguistic models of sentence comprehension in TD children and provide further insight into the linguistic nature of G-SLI.