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In diesem Artikel bewerten wir die bisherigen Ergebnisse der Standardisierungsanstrengungen der Plattform Industrie 4.0 im Hinblick auf das Verfolgen einheitlicher und in sich konsistenter Konzepte. Die von uns betrachteten Ergebnisse sind das Referenzarchitekturmodell Industrie 4.0 (RAMI4.0) sowie das Konzept der Verwaltungsschale (VWS). Um seinen Zweck zu erfullen, den Diskurs zur Standardisierung von Industrie 4.0 (I4.0)-relevanten Aspekten im Sinne einer Schichtung zu vereinfachen, mussten die vom RAMI4.0 eingefuhrten Begriffsachsen ein klares Ordnungskriterium aufweisen. Dies ist zumindest bei der Architekturachse (Business, Functional, Information, Communication, Integration, Asset) nicht der Fall, weswegen diese Begriffsachse nicht die gewunschte „Schichtung“ erreicht. Stattdessen reprasentieren die Werte der Architekturachse eher verschiedene, sich uberlappende Aspekte. Das Konzept der Verwaltungsschale analysieren wir mit Hilfe der drei grundsatzlich verschiedenartigen Konzepte der „administrativen Prozesse“, des „Komponentenmodells“ und des „Datenmodells“. Wir stellen fest, dass sich im Laufe der Zeit Elemente aller drei Konzepte in unterschiedlicher Zusammensetzung wiederfinden und insbesondere die ursprungliche Idee der VWS als eine „schmale informatische Hulle“ in sich widerspruchlich ist. Diese konzeptuelle Unscharfe hat unseres Erachtens erheblich dazu beigetragen, dass die Ergebnisse der Standardisierung der VWS bis jetzt hinter den ursprunglichen Planen zuruckgeblieben sind.

This article presents a novel methodology to the underwater documentation of pile fields in archaeological lakeside settlement sites using Structure from Motion (SfM). Mapping the piles of such sites is an indispensable basis to the exploitation of the high resolution absolute chronological data gained through dendrochronology. In a case study at the underwater site of Ploča, Mičov Grad at Lake Ohrid, North Macedonia, nine consecutive 10 m2 strips and a 6 m2 excavation section were uncovered, the situation documented, and the wood piles sampled. The gained data was vectorized in a geographic information system. During two field campaigns, a total of 794 wooden elements on a surface of 96 m2 could be documented three-dimensionally with a residual error of less than 2 cm. The exceptionally high number of fishes in the 5 m deep water resulted in a significant covering of potentially important information on the relevant photos. We present a machine learning approach, especially developed and successfully applied to the automatic detection and masking of these fishes in order to eliminate them from the images. The discussed documentation workflow enables an efficient, cost-effective, accurate and reproducible mapping of pile fields. So far, no other method applied to the recording of pile fields has allowed for a comparably high resolution of spatial information., Journal of Archaeological Science: Reports, 39

In this article, we introduce a classification of system interactions to guide the discourse on their interfaces and interoperability. It is based on a simple, but nevertheless complete classification of system interactions with respect to information transport and processing. Information transport can only be uni- or bidirectional and information processing is subclassified along the binary dimensions of state, determinism and synchronicity. For interactions with bidirectional information flow we are able to define a criterion for a layered structure of systems: we name a bidirectional interaction ”horizontal” if all interacting systems behave the same with respect to state, determinism and synchronicity and we name it “vertical” — providing a semantic direction — if there is a behavioral asymmetry between the interacting systems with respect to these properties. It is shown that horizontal interactions are essentially stateful, asynchronous and nondeterministic and are described by protocols. Vertical interactions are essentially top-down-usage, described by object models or operations, and bottom-up-observation, described by anonymous events. The interaction classification thereby helps to better understand the significant relationships that are created between interacting discrete systems by their interactions and guides us on how to talk about discrete systems, their interfaces and interoperability. To show its conceptual power, we apply the interaction classification to assess several other architectural models, communication technologies and so called software design or architectural styles like SOA and REST.

We quantify confinement effects on hindered transport in mesoporous silica particles through using physical reconstructions of their mesopore space morphology obtained by electron tomography as geometric models in direct diffusion simulations for passive, finite-size tracers. Studied are fully porous particles with mean mesopore sizes of dmeso = 16.0 and 23.9 nm, prepared by classical sol–gel processing, and solid core–porous shell particles (dmeso = 9.4 and 16.8 nm) originating from a layer-by-layer assembly of sol particles around a solid, impermeable core followed by thermal consolidation of the porous shell. Because shell thickness and core size are independently adjustable, core–shell particles allow to decouple the intraparticle diffusion distance in a fixed-bed reactor or chromatographic column from the external surface area of the particles and the hydraulic permeability of the bed, impossible with fully porous particles. Effective diffusivity, accessible porosity, and pore network connectivity recorded in the four reconstructions as a function of λ, the ratio of tracer to mean mesopore size, demonstrate an unfavorable shell morphology for the core–shell particles that opposes their design advantage over fully porous particles. The reconstructions reveal that core–shell particles contain an increased number of narrow and constricted as well as closed pores. These structural features reflect compacted and sintered packings and are most likely formed during shell consolidation. The presented expressions for hindered diffusion and accessible porosity can be used to optimize mesopore space morphologies in sensitive applications, e.g., catalysis under confinement and controlled drug release.

The blue blood of the horseshoe crab is a natural, irreplaceable, and precious resource that is highly valued by the biomedical industry. The Limulus amebocyte lysate (LAL) obtained from horseshoe crab blood cells functions as a surprisingly sophisticated sensing system that allows for the extremely sensitive detection of bacterial and fungal cell-wall components. Notably, LAL tests have markedly contributed to the quality control of pharmaceutical drugs and medical devices as successful alternatives to the rabbit pyrogen test. Furthermore, LAL-based endotoxin and (1→3)-β-D-glucan (β-glucan) assay techniques are expected to have optimal use as effective biomarkers, serving as adjuncts in the diagnosis of bacterial sepsis and fungal infections. The innovative β-glucan assay has substantially contributed to the early diagnosis and management of invasive fungal diseases; however, the clinical significance of the endotoxin assay remains unclear and is challenging to elucidate. Many obstacles need to be overcome to enhance the analytical sensitivity and clinical performance of the LAL assay in detecting circulating levels of endotoxin in human blood. Additionally, there are complex interactions between endotoxin molecules and blood components that are attributable to the unique physicochemical properties of lipopolysaccharide (LPS). In this regard, while exploring the potential of new LPS-sensing technologies, a novel platform for the ultrasensitive detection of blood endotoxin will enable a reappraisal of the LAL assay for the highly sensitive and reliable detection of endotoxemia.

Endotoxins are a highly pyrogenic and immunogenic contaminant of bacterial origin that must be avoided during the manufacturing of biopharmaceutical products to ensure safety and efficacy. Low endotoxin recovery, also known as a masking effect, is defined as the ability to detect50% [21] of the expected endotoxin in an endotoxin assay. Masking can be caused by the ability of endotoxins to build aggregates, bind to the protein or organise in micelles or vesicles that in turn inhibit detection of the endotoxin in the solution being tested. Therefore, a masking effect can result from physical parameters of the molecule being tested or from the buffer/environmental conditions of the solution the molecule is in. This can subsequently lead to the underestimation of endotoxin contaminations and lead to a potential false negative test. Tight control over the effectiveness of the downstream process and the use of well-characterised endotoxin testing assays are needed to ensure optimal endotoxin removal. This manuscript demonstrates the capacity to remove the endotoxins within a proven acceptable range by also controlling and evaluating the potential masking effects during downstream process at ambient temperature and also during sample storage condition until the analyse was performed. The endotoxin removal study (ERS) is divided in the initial part to evaluate the process buffers and the conditions of the molecule to avoid the underestimation of endotoxins in process samples in advance. This pre-study is a necessary prerequisite to evaluate the results after the endotoxin spiked downstream unit operations. With those aspects, the removal capacity can be demonstrated. A study was carried out to characterise the endotoxin removal capability of the purification process including controlling of masking effects. The endotoxin removal capacity on ion exchange chromatography and during ultrafiltration/diafiltration unit operations of the downstream processing of an immunoglobulin G1 antibody was conducted using various process parameters to understand their impact on endotoxin removal. In the small-scale study, the processing steps from each tested unit operation were spiked with Escherichia coli endotoxins. The potential masking effect during purification was addressed by controlling the hold time by spiking studies of the different generated pools at ambient temperature. By conducting a masking study, all generated protein pools (flow-through/wash, eluate and regeneration pools) had no masking effect caused through sample handling prior to analysis. Overall, this study showed that endotoxins could be successfully removed by anion exchange chromatography. A partial removal could be achieved by cation exchange chromatography and endotoxins could not be removed with ultrafiltration/diafiltration.

Specialized and systematic underwater fieldwork at the prehistoric site of Ploča Mičov Grad at Gradište (North Macedonia) on the eastern shore of Lake Ohrid was undertaken in 2018 and 2019. It has substantiated the archeological site's outstanding preservation condition, and furthermore proven the numerous construction timbers’ suitability for dendrochronological analysis. Dendrochronological analysis on archaeological timbers was applied, combined with radiocarbon dating. Bayesian radiocarbon modeling allowed to ‘wiggle match’ the dendrochronological mean curves, i.e. allowed the precise chronological anchoring of ‘floating’ tree-ring sequences. Furthermore, radiocarbon dates of plant remains from the site's main archaeological layer are statistically evaluated. Based on the new findings, the strikingly high density of wooden piles at the site can be attributed to several construction phases of Neolithic (middle of 5th millennium BC) and Bronze Age (2nd millennium BC: 1800, 1400 and 1300 BCE) settlements. Intense settlement activity is furthermore evidenced by a cultural layer of mainly organic material under the lakebed up to 1.7 m in thickness, which accumulated during the Neolithic occupation of the bay in the middle of the 5th millennium BC. The presented research enables precise absolute dating of a series of settlement phases at Ploča Mičov Grad from the Neolithic and the Bronze Age, and hence provides important reference points for an absolute chronological framework for the prehistory of the southwestern Balkans. The investigations underline the potential of future research on waterlogged prehistoric settlements in the region., Journal of Archaeological Science: Reports, 38

For release of parenteral drug products, bacterial endotoxin testing is one of a panel of necessary tests. In order to ensure the validity of such tests, various controls are performed, including demonstration of compendial method suitability or method qualification. In addition to compendial suitability testing, quality control (QC) sample hold-time studies are requested by authorities like the Food and Drug Administration (FDA) as described in &ldquo, Guidance for Industry: Pyrogen and Endotoxins Testing.&rdquo, It is requested to be determine whether the ability to detect endotoxins can be affected by storage and handling of the sample to be tested. To accomplish these studies, endotoxin is introduced or spiked into the undiluted product and held for a certain period of time in process-representative containers. This time period reflects procedural maximum QC sample hold time from sampling until analysis. Inadequate detection of endotoxin can be caused by adsorption of endotoxin to container surfaces or molecular masking effects, in which the binding sites on the endotoxin molecules are prevented from triggering the enzymatic cascade necessary in the assay, are obscured. The endotoxin may form macromolecular structures, such as sheets or blebs, or the binding sites may otherwise be rendered unavailable due to the sample matrix composition. In either case, the endotoxin assay may yield falsely low results if and when masking occurs. In this work, the QC sample hold times of different in-process controls within the production process of a biopharmaceutical product were analyzed. One out of eight different samples showed a strong masking of endotoxin. Analysis of the sample composition revealed that either kifunensine, mycophenolic acid (MPA), or lauryl-N, N-dimethylamine oxide (LDAO) was responsible for masking. Further analysis clearly identified LDAO as the root cause for masking. A novel one-step mechanism for LDAO-induced endotoxin masking is proposed. The principle is similar to an already-proposed two-step mechanism for endotoxin masking, but the LDAO case combines these two steps: the disturbance of the salt bridges and hydrophobic interactions with LPS in one molecule. These molecular interactions occur quickly when both endotoxin and LDAO are present in the same matrix. Thus, depending on the masking agents, low endotoxin recovery (LER) can occur regardless of the QC sample hold duration.

Among the mechanisms put-up by the host to defend against invading microorganisms, antimicrobial peptides represent the first line. In different species of mammals, the cathelicidin family of antimicrobial peptides AMPs has been identified, and in humans, LL-37 is the only type of cathelicidin identified. LL-37 has many different biological activities, such as regulation of responses to inflammation, besides its lipopolysaccharide (LPS)-neutralizing and antimicrobial and activities. Recently, employing a murine septic model that involves cecal ligation and puncture (CLP), we examined the effect of LL-37. The results indicated that LL-37 exhibits multiple protective actions on septic mice; firstly, the survival of CLP mice was found to be improved by LL-37 by the suppression of the macrophage pyroptosis that induces the release of pro-inflammatory cytokines (such as IL-1β) and augments inflammatory reactions in sepsis; secondly, the release of neutrophil extracellular traps (NETs), which have potent bactericidal activity, is enhanced by LL-37, and protects mice from CLP-induced sepsis; thirdly, LL-37 stimulates neutrophils to release antimicrobial microvesicles (ectosomes), which improve the pathological condition of sepsis. These findings indicate that LL-37 protects CLP septic mice through at least three mechanisms, i.e., the suppression of pro-inflammatory macrophage pyroptosis and the release of antimicrobial NETs (induction of NETosis) and ectosomes from neutrophils. Thus, LL-37 can be a potential therapeutic candidate for sepsis due to its multiple properties, including the modulation of cell death (pyroptosis and NETosis) and the release of antimicrobial NETs and ectosomes as well as its own bactericidal and LPS-neutralizing activities.

Sepsis is a life-threatening disease caused by systemic dys-regulated inflammatory response to infection. We previously revealed that LL-37, a human cathelicidin antimicrobial peptide, improves the survival of cecal ligation and puncture septic mice. Ectosomes, microvesicles released from neutrophils, are reported to be elevated in sepsis survivors; however, the functions of ectosomes in sepsis remain largely unknown. Therefore, we herein elucidated the protective action of LL-37 on sepsis, by focusing on LL-37-induced ectosome release in a cecal ligation and puncture model. The results demonstrated the enhancement of ectosome levels by LL-37 administration, accompanied by a reduction of bacterial load. Importantly, ectosomes isolated from LL-37-injected cecal ligation and puncture mice contained higher amounts of antimicrobial proteins/peptides and exhibited higher antibacterial activity, compared with those from PBS-injected cecal ligation and puncture mice, suggesting that LL-37 induces the release of ectosomes with antibacterial potential in vivo. Actually, LL-37 stimulated mouse bone-marrow neutrophils to release ectosomes ex vivo, and the LL-37-induced ectosomes possessed antibacterial potential. Furthermore, administration of LL-37-induced ectosomes reduced the bacterial load and improved the survival of cecal ligation and puncture mice. Together these observations suggest LL-37 induces the release of antimicrobial ectosomes in cecal ligation and puncture mice, thereby reducing the bacterial load and protecting mice from lethal septic conditions.

Endotoxin (lipopolysaccharide) testing of drugs is routinely required in pharmaceutical industries. Suitable compendial assays are defined by national pharmacopoeias. At this time, Limulus Amoebocyte Lysate (LAL) assays are the gold standard. LAL is used in vitro for specific detection of endotoxin based on endotoxin-activated Factor C-mediated clotting cascade. However, alternative mediated pathways (e.g., Factor G), impurities, and further factors may influence test results. Some of these influencing factors are eliminated by recombinant Factor C (rFC) test, which represents a promising alternative. rFC not only enables highly specific endotoxin testing, as interfering Horseshoe Crab blood components are eliminated, but also offers ethical and ecological advantages compared to classical LAL assays. However, the question remains whether rFC-based tests are robust test systems, equivalent or superior to LAL and suitable for routine bacterial endotoxin testing. Pharmaceutical test users have validated the test successfully for their specific products, but no long-term studies have been published that combine testing of unknown samples, inter-laboratory, -operator, and -lot changes. Thus, it was of great interest to investigate rFC test performance in a routine setting within a proficiency test program set-up. Over a period of six years comparative endotoxin testing was conducted with one kinetic chromogenic LAL assay and two rFC-based assays. Results of this study demonstrate that both rFC-based assays were comparable to LAL. All results met acceptance criteria defined by compendial bacterial endotoxin testing. RFC-based methods generated results with even better endotoxin recovery rates compared to LAL. Therefore, rFC-based tests were found to represent reliable methods, as equivalent or even superior to LAL assays and suitable for routine bacterial endotoxin testing.

We have witnessed a sharp rise in the quest for a methodologically sound analysis of the “migration,”1 “transfer,”2 and “borrowing”3 of concepts between different legal systems.4 At the time of its foundation as a modern federal state in 1848, the Swiss Confederation (Switzerland) formed a republican island in an ocean of monarchies in the heart of Europe. The foundation of the United States of America, in turn, may be viewed as the first large-scale experiment in federal republicanism in the modern era, having been initiated at a time when what today constitutes Switzerland was a rather loose confederation of...

The performance of SBA-15 and KIT-6 silicas as ordered mesoporous supports for heterogeneous catalysis, selective adsorption, and controlled drug release relies on their properties for diffusive transport of finite-size solutes or particles. We investigate this issue through a reconstruction–simulation approach, applied to purely mesoporous SBA-15 and KIT-6 silica samples with a mean pore size of 9.4 nm. Physical reconstruction by electron tomography confirms a highly interconnected, three-dimensional mesopore network for both samples but also reveals constrictions (both samples) and dead ends (SBA-15 only) in the supposedly uniform cylinders of the primary pore system. Pore-scale simulations of the diffusion of pointlike and finite-size tracers in the reconstructions show that a small number of bottlenecks suffices for observing a dramatic decline in the effective diffusion coefficient, accessible porosity, and pore interconnectivity with increasing tracer size. When the tracer size approaches one-third ...

Hindered diffusion of solutes is the rate-limiting step in many processes for which random porous media play a central role as providers of adsorbing or reactive interfaces. The key to an optimized layout of these processes is the knowledge of the overall diffusive hindrance factor H(λ) = Deff,H(λ)/Dm, which quantifies the degree to which diffusion through a material (represented by the effective diffusion coefficient Deff,H) is hindered compared with diffusion in the bulk liquid (represented by Dm) in dependence of λ, the ratio of solute size to mean pore size. To arrive at an adequate hindrance factor expression for random mesoporous silica, we use electron tomography to physically reconstruct the mesopore space of three macro-mesoporous silica monoliths. The samples share the same general mesopore shape and topology at varied mean feature size, as established by morphological analysis, and serve as realistic models in pore-scale simulations of hindered diffusion. From a large set of Deff,H(λ) values fo...