Mixture Of Fragments Research Articles

Orientation, thermal characteristics and structural dimensions of Mangrove Kingfisher Halcyon senegaloides nest cavities in arboreal termitaria in central Mozambique

The breeding ecology of the Mangrove Kingfisher Halcyon senegaloides (family Alcedinidae) remains poorly known. While the few nests recorded in southern and East Africa have been in tree hollows, a seasonal population in central Mozambique woodland breeds in cavities in arboreal termitaria. Despite this breeding strategy being widely recorded in many tropical kingfisher species, it has received little research attention in the Afrotropical realm. We report on the siting and structural characteristics of arboreal termitaria used as nesting sites by Mangrove Kingfishers in central Mozambique and document the thermal dynamics of a nesting cavity in an arboreal termitarium. Arboreal termitaria (average volume 40.1 ± 23.8 l) occurred at a density of 0.52 ± 0.22 termitaria ha−1 in large trees or shrubs, mostly of the families Fabaceae and Malvaceae. Termitaria containing nest cavities were situated 10.8 ± 3.1 m above the ground, and most nests (78.6%) faced north. Nest cavity temperatures averaged 31.4 ± 2.6 °C, which is warmer and more stable than the average ambient temperatures of 28.7 ± 4.0 °C. ‘Carton’ termitaria (comprising a mixture of faecal matter and wood fragments) are thus believed to confer a thermal advantage for summer-nesting Mangrove Kingfishers.

Increasing microplastic exposure had minimal effects on fatty acid composition in zooplankton and yellow perch in a large, in-lake mesocosm experiment

Using 10 m diameter mesocosms in a Canadian boreal lake, we investigated the effects of microplastic (MP) exposure on the body weight and diet of yellow perch ( Perca flavescens) and the fatty acid composition of yellow perch and zooplankton. We exposed the aquatic ecosystem within seven mesocosms for 10 weeks to a mixture of polyethylene, polystyrene, and polyethylene terephthalate fragments, ranging in nominal addition concentrations from 6 to 29 240 particles L−1 (although realized water column concentrations were lower), as well as two negative controls. Increasing MP exposure did not affect yellow perch body weight (growth) or diet, or the overall fatty acid composition of yellow perch muscle or zooplankton. Results were highly variable across mesocosms. Despite high levels of MP ingestion by yellow perch, we did not find evidence of MPs leading to food dilution or any other effect where we could anticipate impacts on food web structure.

An ultrasensitive DNA-enhanced amplification method for detecting cfDNA drug-resistant mutations in non-small cell lung cancer with selective FEN-assisted degradation of dominant somatic fragments.

Blood cell-free DNA (cfDNA) can be a new reliable tool for detecting epidermal growth factor receptor (EGFR) mutations in non-small cell lung cancer (NSCLC) patients. However, the currently reported cfDNA assays have a limited role in detecting drug-resistant mutations due to their deficiencies in sensitivity, stability, or mutation detection rate. We developed an Archaeoglobus fulgidus-derived flap endonuclease (Afu FEN)-based DNA-enhanced amplification system of mutated cfDNA by designing a pair of hairpin probes to anneal with wild-type cfDNA to form two 5'-flaps, allowing for the specific cleavage of wild-type cfDNA by Afu FEN. When the dominant wild-type somatic cfDNA fragments were cleaved by structure-recognition-specific Afu FEN, the proportion of mutated cfDNA in the reaction system was greatly enriched. As the amount of mutated cfDNA in the system was further increased by PCR amplification, the mutation status could be easily detected through first-generation sequencing. In a mixture of synthetic wild-type and T790M EGFR DNA fragments, our new assay still could detect T790M mutation at the fg level with remarkably high sensitivity. We also tested its performance in detecting low variant allele frequency (VAF) mutations in clinical samples from NSCLC patients. The plasma cfDNA samples with low VAF (0.1 and 0.5 %) could be easily detected by DNA-enhanced amplification. This system with enhanced amplification of mutated cfDNA is an effective tool used for the early screening and individualized targeted therapy of NSCLC by providing arapid, sensitive, and economical way for the detection of drug-resistant mutations in tumors.

The “suevite” conundrum, Part 2: Re‐examining the type locality at the Ries impact structure, Germany

AbstractOne of the most common types of allochthonous impactite produced in hypervelocity impact events is impact breccia that contains melt particles. In numerous terrestrial hypervelocity impact structures such melt‐bearing breccias have been termed “suevite,” after the type locality at the Ries impact structure, Germany. Despite its widespread occurrence, the origin, emplacement, and classification of suevite remains debated. In this contribution, we re‐examine the nature and origin of suevite at the Ries impact structure. The results of new field and laboratory investigations, when combined and synthesized with results from previous studies, lead to a multi‐stage model for the origin and emplacement of allochthonous impactites during the Ries impact event. Following the creation of a transient cavity the so‐called Bunte Breccia and “megablocks” were emplaced via ballistic sedimentation and subsequent radial flow during the excavation stage to form a continuous ejecta blanket. At the end of the excavation stage, a mixture of melt and lithic fragments formed a lining to the transient cavity and it is this material that later became the crater, dike, and outer suevite (OS) units. The crater suevite represents the material from the displaced zone of the transient cavity that was transported and mixed but never left the cavity. The emplacement of dike suevite occurred during the modification stage as the crater suevite was intruded into fractures in the underlying crater floor. The OS and rare impact melt rocks overlying the ballistic (Bunte Breccia) ejecta deposits were emplaced as outwards‐directed ground‐hugging flows largely during the modification stage of crater formation. The OS flows varied both spatially and temporally in terms of the flow characteristics, from being dominated by solid particles and gas (cf. pyroclastic density currents) to a mixture of solid particles, liquid (impact melt), and minor gases (i.e., particulate impact melt‐rich flows). These particulate impact melt‐rich flows dominated by far. Minor “fallback” of material from an ejecta plume is evidenced by accretionary lapilli in the Nördlingen 1973 core. In summary, allochthonous impactites at the Ries impact structure are not unusual but are consistent with observations from other terrestrial and planetary craters, where melt‐rich impactites overly ballistic ejecta deposits both outside and inside crater rims and where melt‐rich impactites occur in crater interiors.

Mechanical properties of soils in internal waste bank of lignite mines and their primary characteristics

The modern development of urban areas is related to, among others, the location of industrial facilities on the outskirts of cities. More and more often, commercial buildings are founded on areas that have not been used for construction so far. Such areas include, among others: reclaimed lignite mine banks in the Konin region. The man-made soil is a chaotic mixture of fragments of glacial tills and Pliocene clays, often exceeding 20 m in thickness, which is naturally consolidated over time. Due to the method of formation of the embankment, despite the fact that banks are made of natural soil, their strength and deformation characteristics clearly differ from those characteristic of lithologically similar soils deposited as a result of geological processes. In this case, the use of a standard test approach may overestimate the strength and stiffness of the soil. Due to the complex structure of the bank in-situ tests were used for geotechnical exploration: CPTU and FVT, as well as laboratory tests in a triaxial apparatus and an oedometer. The results were compared with the results of studies conducted in similar naturally deposited soils. The obtained results provide valuable geotechnical characteristics of the embankment soil, which in its large fragments is built of natural soil clasts. The obtained results indicate a relatively small change in the geotechnical properties of the soils incorporated into the embankment within individual clasts of the natural soil (on a local scale) and a clear deterioration in the scale of the entire embankment, treated as the impact zone of building structures.

Ligand Screening of Membrane Proteins Embedded in Nanodiscs: How to Manage Non-Specific Interactions in Weak Affinity Chromatography?

Miniaturized weak affinity chromatography is emerging as an interesting alternative to conventional biophysical tools for performing fragment-screening studies in the context of fragment-based drug discovery. In order to push back the analytical limits, it is necessary not only to control non-specific interactions with chromatographic support, but also to adapt this methodology by comparing the results obtained on an affinity column to a control column. The work presented in this study focused on fragment screening that targets a model membrane protein, the adenosine A2A receptor, embedded in nanodiscs (NDs) as biomimetic membranes. By studying the retention behavior of test fragment mixtures on supports modified with different types of NDs, we were able to determine the contribution of ND-related non-specific interactions, in particular the electrostatic effect of anionic phospholipids and the hydrophobic effect of neutral phospholipids. Different strategies for the preparation of control columns (empty NDs, orthosteric site blocking) were investigated and are presented for the first time. With these two types of control columns, the screening enabled the identification of two new fragments of AA2AR, which were confirmed by competition experiments and whose Kd values, estimated directly during the screening or after the competition experiments in frontal mode, were in good agreement.

Optimizing NMR fragment-based drug screening for membrane protein targets

NMR spectroscopy has played a pivotal role in fragment-based drug discovery by coupling detection of weak ligand-target binding with structural mapping of the binding site. Fragment-based screening by NMR has been successfully applied to many soluble protein targets, but only to a limited number of membrane proteins, despite the fact that many drug targets are membrane proteins. This is partly because of difficulties preparing membrane proteins for NMR—especially human membrane proteins—and because of the inherent complexity associated with solution NMR spectroscopy on membrane protein samples, which require the inclusion of membrane-mimetic agents such as micelles, nanodiscs, or bicelles. Here, we developed a generalizable protocol for fragment-based screening of membrane proteins using NMR. We employed two human membrane protein targets, both in fully protonated detergent micelles: the single-pass C-terminal domain of the amyloid precursor protein, C99, and the tetraspan peripheral myelin protein 22 (PMP22). For both we determined the optimal NMR acquisition parameters, protein concentration, protein-to-micelle ratio, and upper limit to the concentration of D6-DMSO in screening samples. Furthermore, we conducted preliminary screens of a plate-format molecular fragment mixture library using our optimized conditions and were able to identify hit compounds that selectively bound to the respective target proteins. It is hoped that the approaches presented here will be useful in complementing existing methods for discovering lead compounds that target membrane proteins.

Quarter century of mercury litterfall at a coniferous forest responding to climate change, Central Europe.

This work evaluated the 25-year-long trends (1994-2018) in mercury (Hg) concentrations and fluxes in spruce litterfall at a forest research plot Načetín (NAC) recovering from acidic deposition in the Ore Mountains, Czech Republic. The mean litterfall Hg deposition averaged 51 ± 18µgm-2year-1, which has been the highest litterfall Hg deposition reported up to date on the European continent. In contrast, the wet deposition (2017-2019) was an order of magnitude lower averaging at 2.5 ± 1.5µgm-2year-1. All the spruce litterfall components bark, twigs, needles, cones, and a mixture of unidentified fragments had elevated mean Hg concentrations relative to background sites averaging 256 ± 77, 234 ± 62, 119 ± 23, 95 ± 14, and 44 ± 15µg kg-1, respectively. Elevated litterfall Hg deposition and concentrations were attributed to the nearby local Hg emission source-coal-fired power plants. Temporally, the decrease of Czech Hg emissions since the 1990s was reflected by the decreasing trend of Hg concentrations in litterfall bark, cones, and twigs, while in needles and other material, Hg increased but insignificantly. Total litterfall ratios of Hg/C, Hg/N, and Hg/S were lower than those in soil O horizons averaging at 0.23 ± 0.04, 9.5 ± 2.0, and 170 ± 37μgg-1, respectively. Since the beginning of monitoring, total litterfall Hg/C exhibited no trend, Hg/N decreased, and Hg/S increased. The litterfall biomass deposition averaging at 469 ± 176gm-2year-1 increased through time resulting in an increased Hg litterfall deposition at NAC by 1.1µgm-2year-1 despite the decreases in Czech Hg emissions. Peaks of annual litterfall Hg deposition up to 96µgm-2year-1 at NAC during the 25years of monitoring resulted from weather extremes such as rime-snow accumulation, wind gusts, droughts, and insect infestation, which all significantly affected the annual biomass deposition. Based on our observations, further increases in biomass and litterfall Hg deposition rates can be expected due to the onset of bark beetle infestation and the increasing number of droughts caused by climate change.

Dispersal of high-burnup fuel fragment surrogate particles during and after loss-of-coolant accident tests

The issue of fuel fragmentation, relocation, and dispersal is critical in the licensing and use of high-burnup (>62GWd/MTU) nuclear fuel in light water reactors (LWRs). In this work, two test series are reported that examine the fragment dispersal during a burst event and an additional dispersal following the burst due to vibrations in the rod, such as those induced by accident recovery systems. To examine dispersal during the balloon and burst portion of a Loss-of-Coolant Accident event, HfO2 fragments and yttria-stabilized zirconia pellets were filled into an as-fabricated cladding tube, which was then pressurized and subjected to loss-of-coolant accident testing in steam. Results from this test were found to be highly non-prototypic, and dispersal was both significantly more violent and significantly greater in magnitude than identified for actual fuel tests. These findings were attributed to the conservative (more dispersive) nature of the particles chosen for dispersal and to the details of the test conditions used that led to particularly wide bursts. The second set of tests examined dispersal following the burst when vibrations were induced in the rod, primarily via recovery activities such as Emergency Core Cooling System actuation leading to rapid water addition. Post-burst dispersal testing was performed by inducing sinusoidal oscillations with 2–25 nm peak-to-peak amplitude and 2–5 Hz frequencies in pre-burst rods that had been refilled with HfO2 fragments or high-burnup fragment surrogate mixture of HfO2 fragments and yttria-stabilized zirconia sands. Testing revealed that rods with large burst openings (7 mm wide in this work) led to unmitigated dispersal from above the burst zone but that smaller bursts (5 mm wide), although still much larger than the mean fragment size of 3 mm, led to effectively no dispersal because of interparticle locking. Additionally, mixture and moisture were found to impact the amount of dispersal: mixtures increased dispersal, and moisture drastically reduced it. The implications of these findings on likely dispersal from actual fuel are discussed.

Non-isochoric stable granular models taking into account fluidisation by pore gas pressure

In this paper, we study non-isochoric models for mixtures of solid particles, at high volume concentration, and a gas. One of the motivations for this work concerns geophysics and more particularly the pyroclastic density currents, whose dense basal parts are precisely mixtures of pyroclasts and lithic fragments and air. They are extremely destructive phenomena, capable of devastating urbanised areas, and are known to propagate over long distances, even over almost flat topography. Fluidisation of these dense granular flows by pore gas pressure is one response that could explain this behaviour and must therefore be taken into account in the models. Starting from a solid–gas mixing model and invoking the compressibility of the gas through a law of state, we rewrite the conservation of mass equation of the gas phase into an equation of the pore gas pressure whose net effect is to reduce the friction between the particles. The momentum equation of the solid phase is completed by generic constitutive laws, specified as in Schaeffer et al. (J. Fluid Mech., vol. 874, 2019, pp. 926–951) by a yield function and a dilatancy function. Therefore, the divergence of the velocity field, which reflects the ability of the granular flow to expand or compress, depends on the volume fraction, pressure, strain rate and inertial number. In addition, we require the dilatancy function to describe the rate of volume change of the granular material near an isochoric equilibrium state, i.e. at constant volume. This property ensures that the volume fraction, which is the solution to the conservation of mass equation, is positive and finite at all times. We also require that the non-isochoric fluidised model be linearly stable and dissipate energy (over time). In this theoretical framework, we derive the dilatancy models corresponding to classical rheologies such as Drucker–Prager and $\mu (I)$ (with or without expansion effects). The main result of this work is to show that it is possible to obtain non-isochoric and fluidised granular models satisfying all the properties necessary to correctly account for the physics of granular flows and which are well posed, at least linearly stable.

IPEV: identification of prokaryotic and eukaryotic virus-derived sequences in virome using deep learning.

The virome obtained through virus-like particle enrichment contains a mixture of prokaryotic and eukaryotic virus-derived fragments. Accurate identification and classification of these elements are crucial to understanding their roles and functions in microbial communities. However, the rapid mutation rates of viral genomes pose challenges in developing high-performance tools for classification, potentially limiting downstream analyses. We present IPEV, a novel method to distinguish prokaryotic and eukaryotic viruses in viromes, with a 2-dimensional convolutional neural network combining trinucleotide pair relative distance and frequency. Cross-validation assessments of IPEV demonstrate its state-of-the-art precision, significantly improving the F1-score by approximately 22% on an independent test set compared to existing methods when query viruses share less than 30% sequence similarity with known viruses. Furthermore, IPEV outperforms other methods in accuracy on marine and gut virome samples based on annotations by sequence alignments. IPEV reduces runtime by at most 1,225 times compared to existing methods under the same computing configuration. We also utilized IPEV to analyze longitudinal samples and found that the gut virome exhibits a higher degree of temporal stability than previously observed in persistent personal viromes, providing novel insights into the resilience of the gut virome in individuals. IPEV is a high-performance, user-friendly tool that assists biologists in identifying and classifying prokaryotic and eukaryotic viruses within viromes. The tool is available at https://github.com/basehc/IPEV.

Extracellular vesicle small RNA cargo discriminates non-cancer donors from pediatric B-lymphoblastic leukemia patients.

Pediatric B-acute lymphoblastic leukemia (B-ALL) is a disease of abnormally growing B lymphoblasts. Here we hypothesized that extracellular vesicles (EVs), which are nanosized particles released by all cells (including cancer cells), could be used to monitor B-ALL severity and progression by sampling plasma instead of bone marrow. EVs are especially attractive as they are present throughout the circulation regardless of the location of the originating cell. First, we used nanoparticle tracking analysis to compare EVs between non-cancer donor (NCD) and B-ALL blood plasma; we found that B-ALL plasma contains more EVs than NCD plasma. We then isolated EVs from NCD and pediatric B-ALL peripheral blood plasma using a synthetic peptide-based isolation technique (Vn96), which is clinically amenable and isolates a broad spectrum of EVs. RNA-seq analysis of small RNAs contained within the isolated EVs revealed a signature of differentially packaged and exclusively packaged RNAs that distinguish NCD from B-ALL. The plasma EVs contain a heterogenous mixture of miRNAs and fragments of long non-coding RNA (lncRNA) and messenger RNA (mRNA). Transcripts packaged in B-ALL EVs include those involved in negative cell cycle regulation, potentially suggesting that B-ALL cells may use EVs to discard gene sequences that control growth. In contrast, NCD EVs carry sequences representative of multiple organs, including brain, muscle, and epithelial cells. This signature could potentially be used to monitor B-ALL disease burden in pediatric B-ALL patients via blood draws instead of invasive bone marrow aspirates.

Heterobimetallic Complexes of Bi- or Polydentate N-Heterocyclic Carbene Ligands and Their Catalytic Properties

This review summarizes developments in the synthesis and catalytic applications of those heterobimetallic carbene complexes in which at least two different metals are bound to the same ligand by at least one M-C(carbene) bond each. Several new synthetic methods for such complexes yielding well-defined and thoroughly characterized compounds are presented. The new complexes were found to be catalytically active in several (most often tandem) reactions. In certain cases, the incorporation of two different metals into the same imidazole- or triazol-based NHC-carbene complex resulted in the substantially higher catalytic activity of the heterobimetallic complex compared either to its homobimetallic analogs or to mixtures of comparable mononuclear complex fragments containing the two metals independently. This is a clear demonstration of advantageous metal–metal cooperation within the catalyst. Opposite examples are also discussed, where the heterobimetallic carbene complex proved inferior in relation to its homobimetallic analogs or to mixtures of homonuclear fragments.

Evidence for postglacial seismicity in lacustrine records in the western Kola Peninsula (north-western Russia)

In this article we describe and interpret disturbances in the sedimentary records of four lakes on the western flank of Lake Imandra (NE Fennoscandia, Kola Peninsula). The research framework comprises sedimentological and textural criteria for a visual description of sedimentary structures sediment core data, chronological (radiocarbon dating) data, and ground-penetrating radar (GPR) data. Disturbances preserved in lake sequences contain a mixture of chaotic fragments of sand, silt, polychromatic gyttja, peat, and wood fragments embedded in the organogenic matrix. The synchronicity of disturbances, fast sediment accumulation in lakes, relationship with Quaternary faults, the observed mass movements in lake sediments are interpreted as potential consequences of the earthquake shaking. A seismic event with a magnitude Mw no less than 4.5–5 and intensity of shaking I0 = IV–VI took place in the Middle Holocene, 6400–6100 cal. yr BP. Our studies show that although this area is not seismically active today, some of the main fault zones experienced short periods of reactivation also in the postglacial time, when glacioisostatic rebound after the ice retreat was already very low.

Electron-beam synthesis of thin magneto-dielectric coatings with mixed nickel-iron and upper alumina layers

We have synthesized magneto-dielectric coatings with thicknesses 290, 550, 5460 and 6063 nm with an upper protective and electrically insulating layer of alumina ceramic and a lower magnetic iron-nickel layer. The coatings were formed in a helium atmosphere at forevacuum pressures (from 1.5 to 15 Pa) by electron-beam evaporation of a mixture of iron and nickel fragments, and of a solid alumina ceramic target placed in a separate crucible. Magnetic properties of the coatings have been studied by a ferromagnetic resonance technique with estimation of the effective saturation magnetization together with evaluation of the signal-to-noise ratio and relative broadening of the resonance lines. Owing to the layer of alumina ceramic, the values of relative dielectric permittivity (5.6) and electrical conductivity (0.78–3.00 mS/m) remain typical for thin-film dielectrics.

Indeks Kesesuaian Wisata Di Pantai Pemuteran Buleleng, Bali


 
 
 
 Marine ecotourism is one way of developing tourism in coastal areas that pays attention to ecology and maintains environmental stability. This study was conducted to determine the value of the Tourism Suitability Index (IKW) at Pemuteran Beach, Grokgak, Buleleng by considering 10 parameters such as water depth, beach type, beach width, water base material, water current velocity, beach slope, water brightness, coastal land cover, hazardous biota, and freshwater availability. The method of determining the location using the purposive sampling method is by taking an intentional and determined sample so that what you want to study can be represented by the number of stations, as many as 4 station points. The IKW analysis in this study refers to the suitability classification of coastal tourism and the tourism suitability matrix. Based on the results of the IKW analysis at Pemuteran Beach, each station, namely Station I has a value of 77%, station II is 79%, station III is 83%, and Station IV is 82%. It has the type and basic material of the beach with a black sand substrate with a mixture of coral fragments with a depth of 1.55-1.7 m; including shallow waters, the average width of the beach is quite wide, which is 11.5 m. In addition to this, Pemuteran Beach has a coastal slope of less than 10o with an average current speed of 0.27 m/s and has 100% water clarity. Coastal land cover is generally in the form of resident settlements.
 
 
 

Experimental and Theoretical Exploration of the Kinetics and Thermodynamics of the Nucleophile-Induced Fragmentation of Ylidenenorbornadiene Carboxylates.

Ylidenenorbornadienes (YNDs), prepared by [4 + 2] cycloadditions between fulvenes and acetylene carboxylates, react with thiol nucleophiles to yield mixtures of four to eight diastereomers depending on the symmetry of the YND substrate. The mixtures of diastereomers fragment via a retro-[4 + 2] cycloaddition with a large variation in rate, with half-lives ranging from 16 to 11,000 min at 80 °C. The diastereomer-enriched samples of propane thiol adducts [YND-propanethiol (PTs)] were isolated and identified by nuclear Overhauser effect spectroscopy (NOESY) correlations. Simulated kinetics were used to extrapolate the rate constants of individual diastereomers from the observed rate data, and it correlated well with rate constants measured directly and from isolated diastereomer-enriched samples. The individual diastereomers of a model system fragment at differing rates with half-lives ranging from 5 to 44 min in CDCl3. Density functional theory calculations were performed to investigate the mechanism of fragmentation and support an asynchronous retro-[4 + 2] cycloaddition transition state. The computations generally correlated well with the observed free energies of activation for four diastereomers of the model system as a whole, within 2.6 kcal/mol. However, the observed order of the fragmentation rates across the set of diastereomers deviated from the computational results. YNDs display wide variability in the rate of fragmentation, dependent on the stereoelectronics of the ylidene substituents. A Hammett study showed that the electron-rich aromatic rings attached to the ylidene bridge increase the fragmentation rate, while electron-deficient systems slow fragmentation rates.

New dating approach based on the petrographical, mineralogical and chemical characterization of ancient lime mortar: case study of the archaeological site of Hippo, Annaba city, Algeria

This work presents the results of a multidisciplinary study on the characterization of the composition of certain joint mortars from the ancient city of Hippo (Algeria), one of the most important North African cities in antiquity. Twenty mortar samples were analysed by X-Ray Fluorescence (XRF), powder X-ray diffraction (XRPD), optical microscopy (OM), scanning electron microscopy (SEM) with energy dispersive X-ray analysis (EDS) and thermogravimetric analysis (TGA). Their main physical properties, like solid and dry density and porosity, have been measured by geotechnical procedures. The typological observation by OM showed the existence of four types of sand used as aggregates that ranged from fine to coarse and were mixed with a white to russet natural lime binder. These mortars consisted mainly of mixtures of calcite and rock fragments, and sometimes pieces of red tile. It was recognized that the origins of the natural granules were sands produced by marine erosion of the Edough Mountains. The analysis by XRPD and TGA indicates that the mortars consisted of a mixture of lime/aggregates of low to medium hydraulicity. The analysis of the main chemical components by XRF allowed for the confirmation of the dating of certain monuments and suggested a new dating for other monuments.

Quantification of the mobility potential of antibiotic resistance genes through multiplexed ddPCR linkage analysis.

There is a clear need for global monitoring initiatives to evaluate the risks of antibiotic resistance genes (ARGs) towards human health. Therefore, not only ARG abundances within a given environment, but also their potential mobility, hence their ability to spread to human pathogenic bacteria needs to be quantified. We developed a novel, sequencing-independent method for assessing the linkage of an ARG to a mobile genetic element by statistical analysis of multiplexed droplet digital PCR (ddPCR) carried out on environmental DNA sheared into defined, short fragments. This allows quantifying the physical linkage between specific ARGs and mobile genetic elements, here demonstrated for the sulfonamide ARG sul1 and the Class 1 integron integrase gene intI1. The method's efficiency is demonstrated using mixtures of model DNA fragments with either linked and unlinked target genes: Linkage of the two target genes can be accurately quantified based on high correlation coefficients between observed and expected values (R2) as well as low mean absolute errors (MAE) for both target genes, sul1 (R2 = 0.9997, MAE = 0.71%, n = 24) and intI1 (R2 = 0.9991, MAE = 1.14%, n = 24). Furthermore, we demonstrate that adjusting the fragmentation length of DNA during shearing allows controlling rates of false positives and false negative detection of linkage. The presented method allows rapidly obtaining reliable results in a labor- and cost-efficient manner.

Naturally Occurring N-Terminal Fragments of Bovine Milk Osteopontin Are Transported across Models of the Intestinal Barrier

Osteopontin (OPN) is a bioactive integrin-binding protein found in high concentrations in milk, where it is present both as a full-length protein and as several N-terminally derived fragments. OPN resists gastric digestion, and via interaction with receptors in the gut or by crossing the intestinal barrier into circulation, ingested milk OPN may influence physiological processes. The aim of this study was to investigate OPN interaction with intestinal cells and its transport across models of the intestinal barrier. Immunodetection of OPN incubated with Caco-2 cells at 4 °C and 37 °C showed that OPN binds to the intestinal cells, but it is not internalised. Transepithelial transport was studied using mono- and co-cultures of Caco-2 cells and mucus-producing HT29-MTX cells in transwell membranes. OPN was shown to cross the barrier models in a time-, temperature-, and energy-dependent process inhibited by wortmannin, indicating that the transport takes place via the transcytosis pathway. Analyses of the naturally occurring milk mixture of full-length and N-terminal fragments showed that the N-terminal fragments of OPN bound intestinal cells most effectively and that the fragments were transported across the intestinal membrane models. This suggests that proteolytic processing of OPN increases its biological activity after ingestion.