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Nowadays, a challenge in wildlife management and nature conservation is to reach a state of human-wildlife coexistence, integrating wildlife into the human-dominated landscape. Achieving a state of coexistence is urgent as human-wildlife conflicts increase over time. Thus a "route guide" for researchers and conservation practitioners will be needed to identify if a human-wildlife interaction is heading towards conflict or coexistence, enabling them to conduct management activities, when possible, to achieve human-wildlife coexistence. Researchers have used different individual-based attributes as a proxy to measure support towards wildlife species by the general public. Different operationalizations from Environmental Economics and Environmental and Conservation Psychology research fields have been used to measure support. Examples of operationalization are the willingness-to-pay and Likert-type scale, or rating scale, from the first and second research fields. In the first, participants must indicate how much they would be willing to pay to protect a specific wildlife species population in a particular area and time. In the second, participants are asked to rate statements through, e.g., a five-point ordinal rating scale with opposite alternatives between, e.g., strongly agree and strongly disagree. In the human dimension of natural resources management research, variations of these methodologies have been used to measure support, not only for one wildlife species but for a set. For the willingness-to-pay variation, i.e., money allocation, participants must distribute a constant sum of money among a set of wildlife species. For the rating scale variation, each of the wildlife species in the set corresponds to a statement to be rated. The thesis aims to contrast these two variations, i.e., money allocation and rating scale, in their capacity to assess support changes towards a set of 12 native wildlife species from different taxa.
A survey was applied in 2018 (n: 368) and replicated in 2019 (n: 359) among urban dwellers who cohabit with the wildlife species set, in Valdivia, south of Chile. The surveys were applied before and after information disclosure and exposure in an experimental and longitudinal research design structure, respectively. As information disclosure, the threatened and endemic status of the wildlife species was presented to the participants. On the other hand, mass media coverage of a human-wildlife conflict involving one of the species included in study, the South American Sea Lion, was used for information exposure. The results indicate that the money allocation method identified support changes among the wildlife species to a greater extent than the rating scale for both types of information (Chapters 2, 3, and 4). The money allocation in the experimental design structure grouped the wildlife species based on their threatened and endemic status, while the rating scale did not come with the same results (Chapter 3). In the longitudinal design structure, the South American Sea Lion support decreased based on the average values of the money allocation and rating scale after the information exposure (Chapter 4). Differently, when the South American Sea Lion position support is compared with the other wildlife species, based on the money allocation, there was a descent, while the rating scale presented an ascent after the mass media coverage of the human-wildlife conflict (Chapter 4). This difference between the results of the two methods, in both research design structures, can be explained to a certain extent due to their scaling technique characteristics. The money allocation is a comparative scale; therefore, the support given to one wildlife species will affect the possible support given to the other species. In contrast, the rating scale is a non-comparative scale, i.e., the support given to a wildlife species is independent of the support given to the other wildlife species in the set. In the experimental research design structure (Chapters 2 and 3), to give or increase the support to a threatened or endemic wildlife species, a bill should be taken from another wildlife species, usually not threatened nor endemic. On the contrary, in the rating scale, there was no need to choose; the support could be increased for a wildlife species without decreasing the support for other wildlife species. In the longitudinal study design structure, the money allocation allows direct comparison between wildlife species from one year to another, while the rating scale does not. For the money allocation, the possible amount of support to be given to a wildlife species, i.e., 12 bills of 1,000 CLP each, did not vary from 2018 to 2019. For the rating scale, the values received among the wildlife species can vary within the rating scale from one year to another, misleading to incorrect interpretations. The money allocation method can be suitable for monitoring human-wildlife interactions, i.e., to position and visualize support shifts. The money allocation could be used as an overview of human-wildlife interactions in a specific area, working as a first assessment.
Pentathiepins are cyclic polysulfides that exert antiproliferative and cytotoxic activity in cancer cells, induce oxidative stress and apoptosis, and potently inhibit GPx1. These properties render this class of compounds promising candidates for the development of anticancer drugs. However, the biological effects and how they intertwine to promote high cytotoxicity have not been systematically assessed throughout a panel of cancer cell lines from distinct tissues of origin. In this thesis, six novel pentathiepins were analyzed and constitute the second generation of compounds with additional properties such as fluorescence or improved water solubility to facilitate cellular testing. All compounds underwent extensive biological evaluation in 14 human cancer cell lines. These studies included investigations of the inhibitory potential with regards to GPx1 and cell proliferation, examined the cytotoxicity in human cancer cell lines, as well as the induction of oxidative stress and DNA strand breaks. Furthermore, selected hallmarks of apoptosis, ferroptosis, and autophagy were studied. Experimental approaches regarding these cellular mechanisms included observing morphological changes, detecting phosphatidyl serine exposure and caspase activity, and quantifying cleaved PARP1 and levels of LC3B II. In addition, the analysis of the cell cycle aimed to identify aberrations or arrests in cell division.
Five of the six tested pentathiepins proved to be potent inhibitors of the GPx1, while all six exerted high cytotoxic and antiproliferative activity, although to different extents. There was a clear connection observed between the potential to provoke oxidative stress and damage to DNA in the form of single- and double-strand breaks both extra- and intracellularly. Furthermore, various experiments supported apoptosis but not ferroptosis as the mechanism of cell death in four different cell lines. In particular, the externalization of PS, the detection of activated caspases, and the cleavage of PARP1 corroborated this conclusion. Additionally, indications for autophagy were found, but more investigations are required to verify the current data. The findings of this dissertation are mainly in line with the postulated mechanism of action proposed for pentathiepins and a previous publication from our group that described their biological activity. However, the influence of modulators such as oxygen and GSH on the biological effects was ambiguous and dependent on the compound. The expression profile of the cell lines concerning GPx1 and CAT did not influence the cellular response toward the treatment, whereas the cell doubling time correlated with the cytotoxicity.
As the various pentathiepins give rise to different biological responses, modulation of the biological effects depends on the distinct chemical structures fused to the sulfur ring. This may allow for future optimization of the anticancer activity of pentathiepins. An analysis of the structure-activity relationships revealed that the piperazine scaffold was associated with superior biological activity compared to the pyrrolo-pyrazine backbone. Furthermore, substituents with electron-withdrawing properties or those providing a free electron pair, such as fluorine or morpholine, were advantageous. These findings should help design and synthesize the next generation of pentathiepins, thereby expanding the library of compounds, allowing for the further deduction of structure-activity relationships and an improved understanding of their mechanism of action.
Background
The Federal Ministry of Education and Research of Germany (BMBF) funds a network of university medicines (NUM) to support COVID-19 and pandemic research at national level. The “COVID-19 Data Exchange Platform” (CODEX) as part of NUM establishes a harmonised infrastructure that supports research use of COVID-19 datasets. The broad consent (BC) of the Medical Informatics Initiative (MII) is agreed by all German federal states and forms the legal base for data processing. All 34 participating university hospitals (NUM sites) work upon a harmonised infrastructural as well as legal basis for their data protection-compliant collection and transfer of their research dataset to the central CODEX platform. Each NUM site ensures that the exchanged consent information conforms to the already-balloted HL7 FHIR consent profiles and the interoperability concept of the MII Task Force “Consent Implementation” (TFCI). The Independent Trusted Third-Party (TTP) of the University Medicine Greifswald supports data protection-compliant data processing and provides the consent management solutions gICS.
Methods
Based on a stakeholder dialogue a required set of FHIR-functionalities was identified and technically specified supported by official FHIR experts. Next, a “TTP-FHIR Gateway” for the HL7 FHIR-compliant exchange of consent information using gICS was implemented. A last step included external integration tests and the development of a pre-configured consent template for the BC for the NUM sites.
Results
A FHIR-compliant gICS-release and a corresponding consent template for the BC were provided to all NUM sites in June 2021. All FHIR functionalities comply with the already-balloted FHIR consent profiles of the HL7 Working Group Consent Management. The consent template simplifies the technical BC rollout and the corresponding implementation of the TFCI interoperability concept at the NUM sites.
Conclusions
This article shows that a HL7 FHIR-compliant and interoperable nationwide exchange of consent information could be built using of the consent management software gICS and the provided TTP-FHIR Gateway. The initial functional scope of the solution covers the requirements identified in the NUM-CODEX setting. The semantic correctness of these functionalities was validated by project-partners from the Ludwig-Maximilian University in Munich. The production rollout of the solution package to all NUM sites has started successfully.
Diagenetic illite growth in porous sandstones leads to significant modifications of the initial pore system which result in tight reservoirs. Understanding and quantifying these changes provides insight into the porosity-permeability history of the reservoir and improves predictions on petrophysical behavior. To characterize the various stages of diagenetic alteration, a focused ion beam – scanning electron microscopy (FIB-SEM) study was undertaken on aeolian sandstones from the Bebertal outcrop of the Parchim Formation (Early Permian Upper Rotliegend group). Based on 3D microscopic reconstructions, three different textural types of illite crystals occur, common to many tight Rotliegend sandstones, namely (1) feldspar grain alterations and associated illite meshworks, (2) tangential grain coats, and (3) pore-filling laths and fibers. Reaction textures, pore structure quantifications, and numerical simulations of fluid transport have revealed that different generations of nano-porosity are connected to the diagenetic alteration of feldspars and the authigenic growth of pore-filling illites. The latter leads to the formation of microstructures that range from authigenic compact tangential grain coatings to highly porous, pore-filling structures. K-feldspar replacement and initial grain coatings of illite are composed primarily of disordered 1Md illite whereas the epitaxially grown illite lath- and fiber-shaped crystals occurring as pore-filling structures are of the trans-vacant 1Mtv polytype. Although all analyzed 3D structures offer connected pathways, the largest reduction in sandstone permeability occurred during the initial formation of the tangential illite coatings that sealed altered feldspars and the subsequent growth of pore-filling laths and fibrous illites. Analyses of both illite pore-size and crystallite-size distributions indicate that crystal growth occurred by a continuous nucleation and growth mechanism probably controlled by the multiple influx of potassium-rich fluids during late Triassic and Jurassic times. The detailed insight into the textural varieties of illite crystal growth and its calculated permeabilities provides important constraints for understanding the complexities of fluid-flow in tight reservoir sandstones.
Lacewings (Neuroptera) have predatory larvae with highly specialised mouthparts. Larvae of many groups within Neuroptera are well represented as fossils preserved in ambers; however, larvae of some groups are less often reported in the literature. Here we report such a rare case, a larva of the group Hemerobiidae, an aphidlion, preserved in a piece of Eocene Baltic amber (about 40 million years old). It is preserved together with three possible prey items, wingless aphids, most likely representatives of Germaraphis (or at least closely related to this group). The aphidlion can be identified based on the morphology of the antennae, simple curved and toothless stylets, well developed labial palps, and the absence of other mouth-part structures such as a protruding labrum or maxillary palps. A long, club-shaped distal element of the labial palps identifies the specimen as a larva of Hemerobiidae. The aphids can be identified based on their very long, beak-like mouth parts. This find is, to our knowledge, the first example of a lacewing larva preserved together with its potential prey. We briefly discuss other cases in which fossils preserved in amber allow us to reconstruct aspects of behaviour and interactions of fossil lacewing larvae.
Forest ecosystems around the world and especially boreal forests, are facing
drastically changing climatic conditions. It is known that these changes could
challenge their functionality and vitality. Still, the exact impact is not fully
understood, as tree growth is a complex process and depends on countless
environmental and genetic factors. To estimate the effects of climate change
on tree growth and forest development precisely, we must learn more about
tree growth itself. A comprehensive approach is needed where trees and
forests are investigated on different scales and levels of detail, ranging from
global studies to studies on single individuals.
In this dissertation, I follow such a comprehensive approach, using the
North American conifer white spruce as an example. I present three papers
in the form of three chapters in which my co-authors and I studied the
growth and anatomy of white spruce (Picea glauca [Moench] Voss) and how
it is influenced by environmental, climatic, and genetic factors.
We used diverse approaches and methods on different spatial scales, ranging from
investigations on the landscape to the local scale. We established three paired
plots with forest and treeline sites (two cold-limited and one drought-limited).
as well as one additional forest site. In the first chapter, we concentrated
on the genetic diversity of white spruce within and between populations at
all study sites throughout Alaska. The genetic investigations were combined
with analyses on the individual growth response of trees to climatic conditions
to find whether genetic similarities or spatial proximity caused similarities
in growth and climatic sensitivity. In the second chapter, we studied the
direct and indirect effects of environmental conditions on the xylem tissue
of white spruce. We analyzed the impact of precipitation, temperature, and
tree height on four xylem anatomical traits in trees growing at the three
treelines. The investigated traits represented the main functions of xylem
tissue (i.e., water transport and structural support). In the third chapter,
we investigated similar xylem anatomical traits at one cold-limited treeline.
We compared xylem anatomy and annual increment between genetic groups
and individuals and between spatial groups to investigate whether spatial or
genetic grouping influenced the anatomy and growth of white spruce.
We found an overall high gene flow and high genetic diversity in white
spruce. However, the sensitivity of the growth and anatomical traits of white
spruce was driven mainly by spatial rather than genetic effects and differed
between study sites. Trees from the drought-limited site were more sensitive
towards precipitation and a moisture index, while trees from the cold-limited
sites were more sensitive towards temperature. A strong direct effect of tem-
perature was primarily found in latewood traits related to the structural sup-
port of the tree. Earlywood traits related to water transport, however, were
influenced mainly by tree height. Tree height itself was potentially affected
by diverse abiotic and biotic factors (e.g., (micro)climate, soil conditions,
and competition). Thus, traits related to water transport were indirectly
influenced by environmental conditions. Genetic effects in xylem anatomical
traits were found in the earlywood hydraulic diameter and latewood den-
sity, whereas in general, primarily spatial rather than genetic grouping was
influencing the anatomy of white spruce.
Overall, white spruce showed to be a genetically diverse species with a
high gene flow. The effects of spatial proximity and spatial grouping on the
sensitivity and anatomy of white spruce indicate high phenotypic plastic-
ity. This high phenotypic plasticity combined with the vast genetic diversity
translates into an immense potential for the species to adjust (phenotypically)
and possibly adapt (genetically) to changing conditions. Thus, in terms of
climate change, white spruce may be a rather persistent species that manages
to cope with the drastic changes. Though additional work might be needed to
draw a more solid conclusion, the presented work shows how a comprehensive
study approach can help to interpret and understand the growth and ecology
of a tree species. It may be an inspiration for future studies to broaden their
approaches and to use comprehensive methods on different levels of detail to
not only observe trees but to explore and understand them.
Fast screening of enzyme variants is crucial for tailoring biocatalysts for the asymmetric synthesis of non-natural chiral chemicals, such as amines. However, most existing screening methods either are limited by the throughput or require specialized equipment. Herein, we report a simple, high-throughput, low-equipment dependent, and generally applicable growth selection system for engineering amine-forming or converting enzymes and apply it to improve biocatalysts belonging to three different enzyme classes. This results in (i) an amine transaminase variant with 110-fold increased specific activity for the asymmetric synthesis of the chiral amine intermediate of Linagliptin; (ii) a 270-fold improved monoamine oxidase to prepare the chiral amine intermediate of Cinacalcet by deracemization; and (iii) an ammonia lyase variant with a 26-fold increased activity in the asymmetric synthesis of a non-natural amino acid. Our growth selection system is adaptable to different enzyme classes, varying levels of enzyme activities, and thus a flexible tool for various stages of an engineering campaign.
Target proteins in biotechnological applications are highly diverse. Therefore, versatile flexible expression systems for their functional overproduction are required. In order to find the right heterologous gene expression strategy, suitable host-vector systems, which combine different genetic circuits, are useful. In this study, we designed a novel Bacillus subtilis expression toolbox, which allows the overproduction and secretion of potentially toxic enzymes. This toolbox comprises a set of 60 expression vectors, which combine two promoter variants, four strong secretion signals, a translation-enhancing downstream box, and three plasmid backbones. This B. subtilis toolbox is based on a tailor-made, clean deletion mutant strain, which is protease and sporulation deficient and exhibits reduced autolysis and secondary metabolism. The appropriateness of this alternative expression platform was tested for the overproduction of two difficult-to-produce eukaryotic model proteins. These included the sulfhydryl oxidase Sox from Saccharomyces cerevisiae, which forms reactive hydrogen peroxide and undesired cross-linking of functional proteins, and the human interleukin-1β, a pro-inflammatory cytokine. For the best performing Sox and interleukin, overproducing and secreting variants of these new B. subtilis toolbox fermentation strategies were developed and tested. This study demonstrates the suitability of the prokaryotic B. subtilis host-vector system for the extracellular production of two eukaryotic proteins with biotechnological relevance.
Introduction
Neurofilament light (NfL) can be detected in blood of healthy individuals and at elevated levels in those with different neurological diseases. We investigated if the choice of biological matrix can affect results when using NfL as biomarker in epidemiological studies.
Method
We obtained paired serum and EDTA-plasma samples of 299 individuals aged 37–67 years (BiDirect study) and serum samples of 373 individuals aged 65–83 years (MEMO study). In BiDirect, Passing–Bablok analyses were performed to assess proportional and systematic differences between biological matrices. Associations between serum or EDTA-plasma NfL and renal function (serum creatinine, serum cystatin C, glomerular filtration rate, and kidney disease) were investigated using linear or logistic regression, respectively. All regression coefficients were estimated (1) per one ng/L increase and (2) per one standard deviation increase (standardization using z-scores). In MEMO, regression coefficients were estimated (1) per one ng/L increase of serum or calculated EDTA-plasma NfL and (2) per one standard deviation increase providing a comparison to the results from BiDirect.
Results
We found proportional and systematic differences between paired NfL measurements in BiDirect, i.e., serum NfL [ng/L] = −0.33 [ng/L] + 1.11 × EDTA-plasma NfL [ng/L]. Linear regression coefficients for the associations between NfL and renal function did not vary between the different NfL measurements. In MEMO, one standard deviation increase in serum NfL was associated with greater changes in the outcomes than in BiDirect.
Conclusion
Although there are differences between serum and EDTA-plasma NfL, results can be used interchangeably if standardized values are used.
Background
Pycnogonida (sea spiders) is the sister group of all other extant chelicerates (spiders, scorpions and relatives) and thus represents an important taxon to inform early chelicerate evolution. Notably, phylogenetic analyses have challenged traditional hypotheses on the relationships of the major pycnogonid lineages (families), indicating external morphological traits previously used to deduce inter-familial affinities to be highly homoplastic. This erodes some of the support for phylogenetic information content in external morphology and calls for the study of additional data classes to test and underpin in-group relationships advocated in molecular analyses. In this regard, pycnogonid internal anatomy remains largely unexplored and taxon coverage in the studies available is limited.
Results
Based on micro-computed X-ray tomography and 3D reconstruction, we created a comprehensive atlas of in-situ representations of the central nervous system and midgut layout in all pycnogonid families. Beyond that, immunolabeling for tubulin and synapsin was used to reveal selected details of ganglionic architecture. The ventral nerve cord consistently features an array of separate ganglia, but some lineages exhibit extended composite ganglia, due to neuromere fusion. Further, inter-ganglionic distances and ganglion positions relative to segment borders vary, with an anterior shift in several families. Intersegmental nerves target longitudinal muscles and are lacking if the latter are reduced. Across families, the midgut displays linear leg diverticula. In Pycnogonidae, however, complex multi-branching diverticula occur, which may be evolutionarily correlated with a reduction of the heart.
Conclusions
Several gross neuroanatomical features are linked to external morphology, including intersegmental nerve reduction in concert with trunk segment fusion, or antero-posterior ganglion shifts in partial correlation to trunk elongation/compaction. Mapping on a recent phylogenomic phylogeny shows disjunct distributions of these traits. Other characters show no such dependency and help to underpin closer affinities in sub-branches of the pycnogonid tree, as exemplified by the tripartite subesophageal ganglion of Pycnogonidae and Rhynchothoracidae. Building on this gross anatomical atlas, future studies should now aim to leverage the full potential of neuroanatomy for phylogenetic interrogation by deciphering pycnogonid nervous system architecture in more detail, given that pioneering work on neuron subsets revealed complex character sets with unequivocal homologies across some families.
Liquid chromatography-mass spectrometry (LC-MS)-based untargeted metabolomics experiments have become increasingly popular because of the wide range of metabolites that can be analyzed and the possibility to measure novel compounds. LC-MS instrumentation and analysis conditions can differ substantially among laboratories and experiments, thus resulting in non-standardized datasets demanding customized annotation workflows. We present an ecosystem of R packages, centered around the MetaboCoreUtils, MetaboAnnotation and CompoundDb packages that together provide a modular infrastructure for the annotation of untargeted metabolomics data. Initial annotation can be performed based on MS1 properties such as m/z and retention times, followed by an MS2-based annotation in which experimental fragment spectra are compared against a reference library. Such reference databases can be created and managed with the CompoundDb package. The ecosystem supports data from a variety of formats, including, but not limited to, MSP, MGF, mzML, mzXML, netCDF as well as MassBank text files and SQL databases. Through its highly customizable functionality, the presented infrastructure allows to build reproducible annotation workflows tailored for and adapted to most untargeted LC-MS-based datasets. All core functionality, which supports base R data types, is exported, also facilitating its re-use in other R packages. Finally, all packages are thoroughly unit-tested and documented and are available on GitHub and through Bioconductor.
Background
Microvascular decompression (MVD) success rates exceed 90% in hemifacial spasm (HFS). However, postoperative recovery patterns and durations are variable.
Objective
We aim to study factors that might influence the postoperative patterns and duration needed until final recovery.
Method
Only patients following de-novo MVD with a minimum follow-up of 6 months were included. Overall trend of recovery was modeled. Patients were grouped according to recognizable clinical recovery patterns. Uni- and multivariable analyses were used to identify the factors affecting allocation to the identified patterns and time needed to final recovery.
Results
A total of 323 (92.6%) patients had > 90% symptom improvement, and 269 (77.1%) patients had complete resolution at the last follow-up. The overall trend of recovery showed steep remission within the first 6 months, followed by relapse peaking around 8 months with a second remission ~ 16 months. Five main recovery patterns were identified.
Pattern analysis showed that evident proximal indentation of the facial nerve at root exit zone (REZ), males and facial palsy are associated with earlier recovery at multivariable and univariable levels. anterior inferior cerebellar artery (AICA), AICA/vertebral artery compressions and shorter disease durations are related to immediate resolution of the symptoms only on the univariable level. Time analysis showed that proximal indentation (vs. distal indentation), males and facial palsy witnessed significantly earlier recoveries.
Conclusion
Our main finding is that in contrast to peripheral indentation, proximal indentation of the facial nerve at REZ is associated with earlier recovery. Postoperative facial palsy and AICA compressions are associated with earlier recoveries. We recommend a minimum of 1 year before evaluating the final outcome of MVD for HFS.
Background:
Microvascular decompression (MVD) success rates exceed 90% in hemifacial spasm (HFS).
However, postoperative recovery patterns and durations are variable.
Objective:
We aim to study factors that might influence the postoperative patterns and duration needed until
final recovery.
Method:
Only patients following de-novo MVD with a minimum follow-up of 6 months were included.
Overall trend of recovery was modeled. Patients were grouped according to recognizable clinical
recovery patterns. Uni- and multivariable analyses were used to identify the factors affecting
allocation to the identified patterns and time needed to final recovery.
Results:
323(92.6%) patients had >90% symptom improvement and 269(77.1%) patients had complete
resolution at the last follow–up. The overall trend of recovery showed steep remission within the
first 6 months, followed by relapse peaking around 8 months with a second remission ~16
months. Five main recovery patterns were identified.
Patterns analysis showed that evident proximal indentation of the facial nerve at REZ, males and
facial palsy are associated with earlier recovery at multivariable and univariable levels. AICA,
AICA/VA compressions and shorter disease durations are related to immediate resolution of the
symptoms only on the univariable level. Time analysis showed that proximal indentation (vs.
distal indentation), males and facial palsy witnessed significantly earlier recoveries.
Conclusion:
Our main finding is that in contrast to peripheral indentation, proximal indentation of the facial
nerve at REZ is associated with earlier recovery. Postoperative facial palsy and AICA
compressions are associated with earlier recoveries. We recommend a minimum of 1 year before
evaluating the final outcome of MVD for HFS.
A New Laboratory Workflow Integrating the Free Light Chains Kappa Quotient into Routine CSF Analysis
(2022)
We performed this cohort study to test whether further analysis of intrathecal inflammation can be omitted if the free light chain kappa (FLCκ) quotient is within the reference range in the corresponding quotient diagram. FLCκ concentrations were measured in serum and cerebrospinal fluid (CSF) samples. The intrathecal fraction (IF) of FLCκ was calculated in relation to the hyperbolic reference range. 679 patient samples were used as a discovery cohort (DC). The sensitivity and negative predictive value (NPV) of the FLCκ-IF for the detection of an intrathecal humoral immune response (CSF-specific OCB and/or IF IgG/A/M > 0%) was determined. Based on these data, a diagnostic algorithm was developed and prospectively validated in an independent validation cohort (VC, n = 278). The sensitivity of the FLCκ-IF was 98% in the DC and 97% in the VC with a corresponding NPV of 99%. The use of the FLCκ-IF as a first line analysis would have reduced the Ig and OCB analysis by 62% in the DC and 74% in the VC. The absence of a FLCκ-IF predicts the absence of a humoral intrathecal immune response with a very high NPV of 99%. Thus, integration of our proposed algorithm into routine CSF laboratory analysis could help to reduce analytical efforts.
The present study covers the synthesis, purification and evaluation of a novel aminomethacrylate-based copolymer in terms of its suitability for improving the solubility and in vitro release of poorly water-soluble drug compounds. The new copolymer was synthesized by solvent polymerization with radical initiation and by use of a chain transfer agent. Based on its composition, it can be considered as a modified type of dimethylaminoethyl methacrylate-butyl methacrylate-methyl methacrylate “EUDRAGIT® E PO” (ModE). ModE was specifically developed to provide a copolymer with processing and application properties that exceed those of commercially available (co-)polymers in solubility enhancement technologies where possible. By varying the concentration of the chain transfer agent in the radical polymerization process, the molecular weight of ModE was varied in a range of 173–305 kDa. To evaluate the solubility-enhancing properties of ModE, a series of drug-loaded extrudates were prepared by hot melt extrusion using the novel—as well as several commercially available—(co-)polymers. These extrudates were then subjected to comparative tests for amorphousness, solubility-enhancing properties, storage stability, and drug release. Celecoxib, efavirenz, and fenofibrate were used as model drugs in all experiments. Of all the (co-)polymers included in the study, ModE with a molecular weight of 173 kDa showed the best performance in terms of desired properties and was shown to be particularly suitable for preparing amorphous solid dispersions (ASDs) of the three model drugs, which in a first set of dissolution experiments showed better release behavior under pH conditions of the fasting stomach than higher molecular weight ModE types, as well as a variety of commercially available (co-)polymers. Therefore, the results demonstrate the successful synthesis of a new copolymer, which in future studies will be investigated in more detail for universal application in the field of solubility enhancement.
Four aerobic bacteria with bacteriolytic capabilities were isolated from the brackish water site Strait Uzynaral of Lake Balkhash in Kazakhstan. The morphology and physiology of the bacterial isolates have subsequently been analyzed. Using matrix assisted laser desorption ionization-time of flight mass spectrum and partial 16S rRNA gene sequence analyses, three of the isolates have been identified as Pseudomonas veronii and one as Paenibacillus apiarius. We determined the capability of both species to lyse pre-grown cells of the Gram-negative strains Pseudomonas putida SBUG 24 and Escherichia coli SBUG 13 as well as the Gram-positive strains Micrococcus luteus SBUG 16 and Arthrobacter citreus SBUG 321 on solid media. The bacteriolysis process was analyzed by creating growth curves and electron micrographs of co-cultures with the bacteriolytic isolates and the lysis sensitive strain Arthrobacter citreus SBUG 321 in nutrient-poor liquid media. One metabolite of Paenibacillus apiarius was isolated and structurally characterized by various chemical structure determination methods. It is a novel antibiotic substance.
The GATA1 transcription factor is essential for normal erythropoiesis and megakaryocytic differentiation. Germline GATA1 pathogenic variants in the N-terminal zinc finger (N-ZF) are typically associated with X-linked thrombocytopenia, platelet dysfunction, and dyserythropoietic anemia. A few variants in the C-terminal ZF (C-ZF) domain are described with normal platelet count but altered platelet function as the main characteristic. Independently performed molecular genetic analysis identified a novel hemizygous variant (c.865C>T, p.H289Y) in the C-ZF region of GATA1 in a German patient and in a Spanish patient. We characterized the bleeding and platelet phenotype of these patients and compared these findings with the parameters of two German siblings carrying the likely pathogenic variant p.D218N in the GATA1 N-ZF domain. The main difference was profound thrombocytopenia in the brothers carrying the p.D218N variant compared to a normal platelet count in patients carrying the p.H289Y variant; only the Spanish patient occasionally developed mild thrombocytopenia. A functional platelet defect affecting αIIbβ3 integrin activation and α-granule secretion was present in all patients. Additionally, mild anemia, anisocytosis, and poikilocytosis were observed in the patients with the C-ZF variant. Our data support the concept that GATA1 variants located in the different ZF regions can lead to clinically diverse manifestations.
Structured examination and treatment are essential in medicine. For dental students, a structured approach to the assessment of oral mucosal lesions is missing thus far. To validate an approach, a structured questionnaire was compared with the habitually used free description of oral lesions (white lesions, ulcers, hyperplasia). Thirty-three dental students were divided into two groups (Group 1 (n = 17) used the free description; Group 2 (n = 16) used a guided questionnaire) to characterize mucosal lesions in patients and make a tentative diagnosis. Although no difference was found between the groups regarding the suspected diagnosis or the histopathological findings, there was a significant advantage of the structured questionnaire in all aspects of the description compared to the free description (p = 0.000018). Thus, a structured description is an important aspect in the evaluation of oral mucosal changes, and a guided questionnaire should be implemented in the study of dentistry.
The thesis develops a scholarly-artistic modular method of analysis for literary studies, film studies and comic studies. Artistic modules of analysis are combined with established research methods used by the humanities on order to deepen the level of understanding of the analysed comic/literary/cineastic work. Martin Rowson's comic adaptation of Laurence Sterne's novel "The Life and Opinions of Tristram Shandy, Gentleman" is analysed with the scholartistic method and treated as an addative literary adaptation.
Socio-cognitive abilities and challenges change across the healthy lifespan and are essential for successful human interaction. Identifying effective socio-cognitive training approaches for healthy individuals may prevent development of mental or physical disease and reduced quality of life. A systematic search was conducted in MEDLINE Ovid, Web of Science Core Collection, CENTRAL, and PsycInfo databases. Studies that investigated different socio-cognitive trainings for healthy individuals across the human lifespan assessing effects on theory of mind, emotion recognition, perspective taking, and social decision making were included. A random-effects pairwise meta-analysis was conducted. Risk-of-Bias was assessed using the Cochrane Risk-of-Bias-2-Tool. Twenty-three intervention studies with N = 1835 participants were included in the systematic review; twelve randomized controlled trials in the meta-analysis (N = 875). Socio-cognitive trainings differed regarding duration and content in different age groups, with theory of mind being the domain most frequently trained. Results of the meta-analysis showed that trainings were highly effective for improving theory of mind in children aged 3–5 years (SMD = 2.51 (95%CI: 0.48–4.53)), children aged 7–9 years (SMD = 2.71 (95%CI: − 0.28 to 5.71)), and older adults (SMD = 5.90 (95%CI: 2.77–9.02). Theory of mind training was highly effective in all investigated age-groups for improving theory of mind, yet, more research on transfer effects to other socio-cognitive processes and further investigation of training effects in other socio-cognitive domains (e.g., emotion recognition, visual perspective taking, social decision making) is needed. Identified characteristics of successful socio-cognitive trainings in different age groups may help designing future training studies for other populations.
To reduce global greenhouse gas emissions in order to limit global warming to 1.5°C, individuals and households play a key role. Behavior change interventions to promote pro-environmental behavior in individuals are needed to reduce emissions globally. This systematic literature review aims to assess the a) evidence-based effectiveness of such interventions and b) the content of very successful interventions without limiting the results to specific emitting sectors or countries. Based on the “PICOS” mnemonic and PRISMA statement, a search strategy was developed, and eligibility criteria were defined. Three databases (Embase, PsycInfo, and Web of Science) were searched to retrieve and review potential literature. As a result, 54 publications from 2010 to 2021 were included in the analysis. The results show that most interventions only have small positive effects or none at all. A total of 15 very successful interventions focused on the sectors of mobility, energy, and waste and incorporated improved (infra-) structures, education, feedback, enablement or made the sustainable option the default. Six evidence-based recommendations for content, timing, and setting are deducted and given for interventions on enhancing pro-environmental behavior (PEB). In summary, although the various interventions and intervention types to promote PEB differ in their effectiveness, very successful interventions have common elements. Future research should focus on high-/low-impact and high-/low-cost behavior to develop interventions that aim at high-impact but low-cost behavior changes, or avoid low-impact but high-cost behavior.
MicroRNAs (miRNAs) and long non-coding RNAs (lncRNAs) are involved in the modulation of the DNA-damage response (DDR) and upon exposure to ionizing radiation (IR), their expression fluctuates. In this study, we propose a workflow that enables the creation of regulatory networks by integrating transcriptomics data as well as regulatory data in order to better understand the interplay between genes, transcription factors (TFs), miRNAs, and lncRNAs in the cellular response to IR. We preprocessed and analyzed publicly available gene expression profiles and then applied our consensus and integration approach using open source data and tools. To exemplify the benefits of our proposed workflow, we identified a total of 32 differentially expressed transcripts corresponding to 20 unique differentially expressed genes (DEGs) and using these DEGs, we constructed a regulatory network consisting of 106 interactions and 100 nodes (11 DEGs, 78 miRNAs, 1 DEG acting as a TF, and 10 lncRNAs). Overrepresentation analyses (ORAs) furthermore linked our DEGs and miRNAs to annotations pertaining to the DDR and to IR. Our results show that MDM2 and E2F7 function as network hubs, and E2F7, miR-25-3p, let-7a-5p, and miR-497-5p are the four nodes with the highest betweenness centrality. In brief, our workflow, that is based on open source data and tools, and that generates a regulatory network, provides novel insights into the regulatory mechanisms involving miRNAs and lncRNAs in the cellular response to IR.
Introduction: Adiposity and excessive weight are on the rise in western industrialized countries. In cases where conservative measures fail and surgical interventions are not (yet) desired, gastric balloon therapy has proven to be a safe and reversible endoscopic method. Methods: Aside from weight progression under gastric balloon therapy and by using MRI, our research paper describes the behavior of different abdominal body fat compartments at the beginning and at the end of the gastric balloon therapy. Additionally, the volume of the left liver lobe as well as the fill volume and performance of the gastric balloon were analyzed over the duration of treatment. For assessing potential impacts of weight reduction on the muscle mass, we determined the area of the m. psoas on a comparable cross-sectional area at the beginning and at the end of the therapy. Results: We were able to verify a significant reduction of the layer of subcutaneous fat, adipose capsule of the kidney, and intra-abdominal fatty tissue during the therapy. The volume of the left liver lobe was shrinking in addition to a muscle loss during the balloon therapy. The volume of the gastric balloon remained stable (not hyperinflation). There were variable gas bubbles in the gastric balloon. Conclusion: The gastric balloon is a temporary and successful option for weight reduction by reducing body fat, liver volume, but also muscle mass.
In this work, spatial distributions for reactive stable and transient species that are involved
in the reaction cycle of H2O2, a key species for biomedical applications, were
determined directly in the effluent of a kINPen-sci plasma jet. The small diameter
of cold atmospheric pressure plasma jets and their operation at atmospheric pressure
that causes strong quenching reactions make diagnostics challenging. Here, various diagnostic
techniques have been employed and adapted for the use in the effluent of a
cold atmospheric pressure plasma jet, which were laser atomic absorption spectroscopy
(LAAS) at 811.5 nm for the detection of Ar(3P2), picosecond two-photon absorption
laser-induced fluorescence spectroscopy (ps-TALIF) at 225 nm and 205 nm for the
detection of O and H atoms, respectively, and continuous wave cavity ring-down spectroscopy
(cw-CRDS) at 1.506 µm for the detection of HO2, and cw-CRDS at 8000 µm
for the detection of H2O2. All these methods provide absolute number densities. In
this work, spatial distributions within the small diameter of the effluent of a CAPJ
were obtained, which have not been reported so far literature. In order to overcome the
line-of-sight limitations of CRDS, radial scans were performed and transformed into a
spatial distribution by using Abel inversion.
Based on the determined spatial density distributions for H atoms, O atoms, HO2
radicals, and H2O2 molecules, together with the investigated impact of humidity in the
feed gas on the excitation dynamics and the production of Ar(3P2), and finally on a
comparison of the experimental results to a plasma chemical and reacting flow model,
three different zones with varying reaction kinetics were identified. The densities close
to the nozzle of the kINPen-sci plasma jet were dominated by reactions within the
plasma zone including the dissociation of H2O added to the Ar feed gas and O2 that
was presumably transferred into the plasma zone by counter-propagating ionisation
waves. Notably, also the larger molecules, such as HO2 and H2O2 were mainly formed
within the plasma zone of the plasma jet. Between 1.5 mm and 5 mm below the nozzle,
the atomic species and molecular radicals generated in the plasma zone were consumed
by chemical reactions with the surrounding gas, whose composition was controlled by
applying a gas curtain. At further distances from the nozzle, where typically biological
samples are positioned, only H2O2 and HO2 were observed.
With this work, it is successfully demonstrated that even for the small diameters of
cold atmospheric pressure plasma jets the determination of spatial profiles for reactive
transient and stable species is possible within the effluent. By combining the experimental
results, important insights into the formation and consumption of H2O2 and its
precursors were gained, which are essential for the understanding of use of plasmas in
biomedical applications.
Active Middle Ear Implant Evoked Auditory Brainstem Response Intensity-Latency Characteristics
(2022)
Objective
To analyze intensity-latency functions of intraoperative auditory evoked brainstem responses (ABRs) to stimulation by the Vibrant Soundbridge (VSB) active middle ear implant with respect to coupling efficiency, VSB evoked ABR thresholds, and coupling modality [oval window (OW) placement vs. Incus placement and vs. round window (RW) placement].
Study Design
Exploratory study.
Setting
Bi-centric study at tertiary referral centers.
Patients
Twenty-four patients (10 female, 14 male, mean age: 58 years) who received a VSB.
Outcome Measures
Wave-V intensity-latency functions of intraoperative VSB evoked ABRs using a modified audio processor programmed to preoperative bone conduction thresholds for stimulation. Threshold level correction to coupling efficiency and ABR thresholds. Individual plots and exponential function fits.
Results
After ABR threshold level correction, the latency functions could be aligned. A large variance of latencies was observed at individual threshold level. Wave-V latency was longest in the Incus placement subgroup (9.73 ms, SD: 1.04) as compared to OW placement subgroup (9.47 ms, SD: 1.05), with the shortest latency in the RW placement subgroup (8.99 ms, SD: 0.68). For increasing stimulation levels, the variance decreased with intensity-latency function slopes converging toward a steady-state (saturation) latency caused by saturation of audio processor (stimulation) gain. Latency saturation was reached at a stimulation level of 50 dB nHL for the OW placement subgroup, 35 dB nHL for the Incus placement subgroup, and 30 dB nHL for the RW placement subgroup. The latency and saturation results indicated decreased dynamic range for RW placement, i.e., reverse stimulation.
Conclusions
VSB evoked ABR wave-V intensity-latency function slopes were similar to acoustic stimulation at high stimulation levels with a shift toward longer latencies caused by audio processor signal delay. Saturation of latencies occurred for higher stimulation levels due to saturation of audio processor gain. Thus, the analysis of VSB evoked intensity-latency functions appears to allow for the objective assessment of a patient's individual dynamic range. This can further improve diagnostics as well as intraoperative and postoperative quality control.
Abstract
Amine transaminases (ATAs) are biocatalysts for the synthesis of chiral amines and can be identified in sequence databases by specific sequence motifs. This study shows that the activity level towards the model substrate 1‐phenylethylamine can be predicted solely from the sequence. To demonstrate this, 15 putative ATAs with a different distribution of hydrophobic or hydrophilic amino acid side chains near the active site were characterized. Hydrophobic side chains were associated with a high activity level and were a better predictor of activity than global sequence identity to known ATAs with high or low activities. Enzyme stability investigations revealed that four out of the 15 ATAs showed a good operational stability.
Genetic variants in α-actinin-2 (ACTN2) are associated with several forms of (cardio)myopathy. We previously reported a heterozygous missense (c.740C>T) ACTN2 gene variant, associated with hypertrophic cardiomyopathy, and characterized by an electro-mechanical phenotype in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Here, we created with CRISPR/Cas9 genetic tools two heterozygous functional knock-out hiPSC lines with a second wild-type (ACTN2wt) and missense ACTN2 (ACTN2mut) allele, respectively. We evaluated their impact on cardiomyocyte structure and function, using a combination of different technologies, including immunofluorescence and live cell imaging, RNA-seq, and mass spectrometry. This study showed that ACTN2mut presents a higher percentage of multinucleation, protein aggregation, hypertrophy, myofibrillar disarray, and activation of both the ubiquitin-proteasome system and the autophagy-lysosomal pathway as compared to ACTN2wt in 2D-cultured hiPSC-CMs. Furthermore, the expression of ACTN2mut was associated with a marked reduction of sarcomere-associated protein levels in 2D-cultured hiPSC-CMs and force impairment in engineered heart tissues. In conclusion, our study highlights the activation of proteolytic systems in ACTN2mut hiPSC-CMs likely to cope with ACTN2 aggregation and therefore directs towards proteopathy as an additional cellular pathology caused by this ACTN2 variant, which may contribute to human ACTN2-associated cardiomyopathies.
Dementia is a leading cause of disability and dependency in older people worldwide. As the number of people affected increases, so does the need for innovative care models. Dementia care management (DCM) is an empirically validated approach for improving the care and quality of life for people with dementia (PwD) and caregivers. The aim of this study is to investigate the influencing factors and critical pathways for the implementation of a regionally adapted DCM standard in the existing primary care structures in the German region of Siegen-Wittgenstein (SW). Utilizing participatory research methods, five local health care experts as co-researchers conducted N = 13 semi-structured interviews with 22 local professionals and one caregiver as peer reviewers. Data collection and analysis were based on the Consolidated Framework for Implementation Research (CFIR). Our results show that among the most mentioned influencing factors, three CFIR constructs can be identified as both barriers and facilitators: Patients’ needs and resources, Relative advantage, and Cosmopolitanism. The insufficient involvement of relevant stakeholders is the major barrier and the comprehensive consideration of patient needs through dementia care managers is the strongest facilitating factor. The study underlines the vital role of barrier analysis in site-specific DCM implementation.
Objective
To identify and summarise evaluated interventions aiming to improve the communication of palliative care (PC) and end-of-life (EoL) issues in physicians caring for cancer patients. Such interventions are needed with regard to the aim of an earlier communication of those issues in oncology daily practice, which is associated with a range of benefits for patients and caregivers but is often impeded by physicians’ communication insecurities.
Design
Systematic review based on the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines.Data sourcesRelevant publications were systematically searched in MEDLINE, PsycINFO, CINAHL and Web of Science databases in September 2020 with an update in July 2021.
Eligibility criteria
We included publications reporting a quantitative evaluation of a communication intervention on one or more PC/EoL issues with a communication-related main outcome. Target group had to be physicians caring for cancer patients non-specialist in PC.
Data extraction and synthesis
Two independent raters extracted intervention characteristics, publication characteristics and publication quality. Results were narratively synthesised.
Results
24 publications reporting 22 interventions were included. 13 publications reported randomised controlled trials. A majority of the interventions addressed one specific PC/EoL issue, most often breaking bad news. Teaching strategies mostly involved role-plays. Target group were mainly oncologists. In addition to self-reported outcome measurements for evaluation, most publications also reported the use of external rating data. All but one publication reported significant intervention effects on at least one outcome parameter. Publication quality was overall moderate.
Conclusions
The empirically tested communication interventions on PC/EoL issues seem to effectively improve physicians’ communication. Future interventions should focus on other issues than breaking bad news, such as preparing for the future. Target group should also be organ-specific oncologists, as all primary caring physicians are responsible for timely communication. Our risk-of-bias assessment revealed some weaknesses, indicating that more high-quality studies for evaluation are needed.
PROSPERO registration number CRD42020191054.
Abstract
Introduction
Using the situated expectancy–value theory, it becomes possible to explain motivational functioning across alternating learning conditions not only at a particular moment but also over time. The situated expectancy–value theory provides evidence for the critical role of context. The present research examines how adolescents' success expectancies, task values, and effort develop when the conditions of the academic environment change. It also evaluates whether adolescents adopt more adaptive expectancy–value profiles in response to the need‐based nature of self‐directed learning as an extension of regular instruction. Within the self‐directed learning approach under investigation, adolescents take responsibility for their own learning processes.
Methods
The present research offers insights into the expectancy–value profiles of 754 German adolescents (Mage = 13.56; SD = 1.2; 49.4% female). A four‐wave study was used to examine perceptions of self‐efficacy, intrinsic value, utility value, and effort. Latent profile analyses and latent transition analyses were employed.
Results
Notably, the results provide evidence that expanding instruction via self‐directed learning intervals that occur for 1 week per semester contributes to more favorable expectancy–value profiles within a student's favorite subject. A mixed profile (highly confident, hardly interested) disappeared. Instead, success expectations aligned more closely with adolescents' task values and effort. All profiles settled at a higher level.
Conclusions
The findings indicate that instruction that is expanded via self‐directed learning intervals has positive consequences for motivational profiles over time. An initial self‐directed learning episode led to a shift to more interested profile groups. The continuation of a positive trend even after a self‐directed learning interval indicates that adolescents are able to continually adapt their learning to their needs during teacher‐directed instruction. This study provides clues about how to design curricula in a way that counteracts the downward trend in students' motivation to learn.
Agriculture in the populated islands of the Galapagos Archipelago, a protected area due to its unique biodiversity, has been detrimental to its conservation but highly required to meet food necessities. A potential solution to make agricultural farming more sustainable is adopting water-saving technologies (WSTs). Therefore, this study aimed to test the effectiveness of using WSTs such as Groasis Waterboxx® in three of the most valuable crops in the islands through participatory research with the involvement of a group of farmers from the Floreana and Santa Cruz islands and explore a possible transition to more sustainable agricultural practices. Capsicum annuum, Cucumis sativus and Solanum lycopersicum were cultivated using Groasis Waterboxx® and compared to conventional irrigation practices (drip-irrigated controls) to assess the variability of productivity, the number of fruits and individual fruit weight (IFW). In addition, differences in plant traits were analyzed by crop, and island. Results suggested that WSTs such as Groasis Waterboxx® may provide on-farm benefits regarding the yields of the studied traits. From this study, it is difficult to determine whether participation in such a research study will permanently change irrigation practices. However, the participant’s responses to the study suggest an increase in their understanding of the use and benefits of WST.
Background
A redistribution of tasks between specialized nurses and primary care physicians, i.e., models of advanced nursing practice, has the potential to improve the treatment and care of the growing number of people with dementia (PwD). Especially in rural areas with limited access to primary care physicians and specialists, these models might improve PwD’s quality of life and well-being. However, such care models are not available in Germany in regular healthcare. This study examines the acceptance, safety, efficacy, and health economic efficiency of an advanced nursing practice model for PwD in the primary care setting in Germany.
Methods
InDePendent is a two-arm, multi-center, cluster-randomized controlled intervention study. Inclusion criteria are age ≥70 years, cognitively impaired (DemTect ≤8) or formally diagnosed with dementia, and living in the own home. Patients will be recruited by general practitioners or specialists. Randomization is carried out at the physicians’ level in a ratio of 1:2 (intervention vs. waiting-control group). After study inclusion, all participants will receive a baseline assessment and a follow-up assessment after 6 months. Patients of the intervention group will receive advanced dementia care management for 6 months, carried out by specialized nurses, who will conduct certain tasks, usually carried out by primary care physicians. This includes a standardized assessment of the patients’ unmet needs, the generation and implementation of an individualized care plan to address the patients’ needs in close coordination with the GP. PwD in the waiting-control group will receive routine care for 6 months and subsequently become part of the intervention group. The primary outcome is the number of unmet needs after 6 months measured by the Camberwell Assessment of Need for the Elderly (CANE). The primary analysis after 6 months is carried out using multilevel models and will be based on the intention-to-treat principle. Secondary outcomes are quality of life, caregiver burden, acceptance, and cost-effectiveness. In total, n=465 participants are needed to assess significant differences in the number of unmet needs between the intervention and control groups.
Discussion
The study will provide evidence about the acceptance, efficacy, and cost-effectiveness of an innovative interprofessional concept based on advanced nursing care. Results will contribute to the implementation of such models in the German healthcare system. The goal is to improve the current treatment and care situation for PwD and their caregivers and to expand nursing roles.
Advancing Radiation-Detected Resonance Ionization towards Heavier Elements and More Exotic Nuclides
(2022)
RAdiation-Detected Resonance Ionization Spectroscopy (RADRIS) is a versatile method for highly sensitive laser spectroscopy studies of the heaviest actinides. Most of these nuclides need to be produced at accelerator facilities in fusion-evaporation reactions and are studied immediately after their production and separation from the primary beam due to their short half-lives and low production rates of only a few atoms per second or less. Only recently, the first laser spectroscopic investigation of nobelium (Z=102) was performed by applying the RADRIS technique in a buffer-gas-filled stopping cell at the GSI in Darmstadt, Germany. To expand this technique to other nobelium isotopes and for the search for atomic levels in the heaviest actinide element, lawrencium (Z=103), the sensitivity of the RADRIS setup needed to be further improved. Therefore, a new movable double-detector setup was developed, which enhances the overall efficiency by approximately 65% compared to the previously used single-detector setup. Further development work was performed to enable the study of longer-lived (t1/2>1 h) and shorter-lived nuclides (t1/2<1 s) with the RADRIS method. With a new rotatable multi-detector design, the long-lived isotope 254Fm (t1/2=3.2 h) becomes within reach for laser spectroscopy. Upcoming experiments will also tackle the short-lived isotope 251No (t1/2=0.8 s) by applying a newly implemented short RADRIS measurement cycle.
We are currently facing an antimicrobial resistance crisis, which means that a lot of bacterial pathogens have developed resistance to common antibiotics. Hence, novel and innovative solutions are urgently needed to combat resistant human pathogens. A new source of antimicrobial compounds could be bacterial volatiles. Volatiles are ubiquitous produced, chemically divers and playing essential roles in intra- and interspecies interactions like communication and antimicrobial defense. In the last years, an increasing number of studies showed bioactivities of bacterial volatiles, including antibacterial, antifungal and anti-oomycete activities, indicating bacterial volatiles as an exciting source for novel antimicrobial compounds. In this review we introduce the chemical diversity of bacterial volatiles, their antimicrobial activities and methods for testing this activity. Concluding, we discuss the possibility of using antimicrobial volatiles to antagonize the antimicrobial resistance crisis.
The experience of abuse in the period of childhood and youth is a key stressor that has con-sequences on the developing brain and is associated with the genesis of mental disorders. Childhood abuse and depression often cooccur together and have both been associated with cortical thickness resulting in a difficulty to detangle the influence of each factor. In prior studies, childhood abuse and depression were inconsistently related to whole-brain cortical thickness. Thus, this thesis aims to investigate the link between childhood abuse, depres-sive symptoms, and alterations of the cortex.
Therefore, this study analyses 1,551 individuals of the general population. A significant in-teraction effect of childhood abuse and depressive symptoms is observed for whole-brain cortical thickness. Yet, the results indicate no influence of childhood abuse or depression alone. A thinner cortex was associated with more severe depressive symptoms in the abused, but not in the non-abused group. In non-depressed participants, an increased whole-brain cortex was found in the abused, compared to the non-abused group. Similar interaction effects were observed in 12 out of 34 cortical regions.
The results suggest, in line with prior findings, that depressed individuals with a history of childhood abuse are a specific ecophenotype which is also reflected in specific brain altera-tions. Cortical regions that are distinct associated with the interaction of depressive symp-toms and childhood abuse are involved in various fields such as sensory processing, self-conception, and memory. Greater cortical thickness in subjects with childhood abuse and without depressive symptoms might act compensatory and thus reflect resilience against depressive symptoms.
Practical implications concern the treatment and diagnostic system as well as the im-portance of early prevention programs. An individualised treatment is necessary as various studies found a less favourable outcome in depressive patients with a history of maltreat-ment. Therefore, it seems urgent to assess experiences of childhood abuse at the beginning of psychiatric and psychotherapeutic treatment. In addition, early prevention programs are in need to support vulnerable family systems and thereby strengthening the economic, health and social system.
Organic cation transporter 1 (OCT1) is a membrane transporter that affects hepatic uptake of cationic and weakly basic drugs. OCT1 transports structurally highly diverse substrates. The mechanisms conferring this polyspecificity are unknown. Here, we analyzed differences in transport kinetics between human and mouse OCT1 orthologs to identify amino acids that contribute to the polyspecificity of OCT1. Following stable transfection of HEK293 cells, we observed more than twofold differences in the transport kinetics of 22 out of 28 tested substrates. We found that the β2-adrenergic drug fenoterol was transported with eightfold higher affinity but at ninefold lower capacity by human OCT1. In contrast, the anticholinergic drug trospium was transported with 11-fold higher affinity but at ninefold lower capacity by mouse Oct1. Using human–mouse chimeric constructs and site-directed mutagenesis, we identified nonconserved amino acids Cys36 and Phe32 as responsible for the species-specific differences in fenoterol and trospium uptake. Substitution of Cys36 (human) to Tyr36 (mouse) caused a reversal of the affinity and capacity of fenoterol but not trospium uptake. Substitution of Phe32 to Leu32 caused reversal of trospium but not fenoterol uptake kinetics. Comparison of the uptake of structurally similar β2-adrenergics and molecular docking analyses indicated the second phenol ring, 3.3 to 4.8 Å from the protonated amino group, as essential for the affinity for fenoterol conferred by Cys36. This is the first study to report single amino acids as determinants of OCT1 polyspecificity. Our findings suggest that structure–function data of OCT1 is not directly transferrable between substrates or species.
In recent years, the colon has become a hot topic in biopharmaceutical research as several in vitro models of the human colon have been presented. A major focus is on the characterization of the microbiota and its capabilities. The aim of the present study was to further develop the MimiCol, preserving its properties and accelerating data acquisition. Emphasis was placed on the simplicity of its design and easy scalability. To prove the viability of the concept, degradation of sulfasalazine was investigated, and the bacterial composition during the experiment was assessed by 16S rRNA sequencing. The transfer of the experimental conditions to the new model was successful. Commercially available components were implemented in the setup. The model MimiCol3 represented the colon ascendens satisfactorily in its properties regarding volume, pH value, and redox potential. 16S rRNA sequencing led to further insights into the bacterial composition in the vessels. Degradation of sulfasalazine was in good agreement with in vivo data. The new model of the colon ascendens MimiCol3 enabled us to collect more reliable data, as three experiments were conducted simultaneously under the same conditions.
An Enzyme Cascade Reaction for the Recovery of Hydroxytyrosol Dervatives from Olive Mill Wastewater
(2022)
Abstract
The valorization of olive mill wastewaters (OMWW), a by‐product of the olive milling, is getting rising attention. Lipophilization of the main phenolic compound 3‐hydroxytyrosol (HT) could facilitate its extraction. An immobilized variant of the promiscuous hydrolase/acyltransferase from Pyrobaculum calidifontis VA1 (PestE) was used to perform acetylation in water using ethyl acetate as acyl donor. PestE was used in a segmented flow setting to allow continuous operation. Additionally, HT precursors were made accessible by pretreatment with almond β‐glucosidase and the hydrolytic activity of PestE_I208A_L209F_N288A.
Background
Approaching epidemiological data with flexible machine learning algorithms is of great value for understanding disease-specific association patterns. However, it can be difficult to correctly extract and understand those patterns due to the lack of model interpretability.
Method
We here propose a machine learning workflow that combines random forests with Bayesian network surrogate models to allow for a deeper level of interpretation of complex association patterns. We first evaluate the proposed workflow on synthetic data. We then apply it to data from the large population-based Study of Health in Pomerania (SHIP). Based on this combination, we discover and interpret broad patterns of individual serum TSH concentrations, an important marker of thyroid functionality.
Results
Evaluations using simulated data show that feature associations can be correctly recovered by combining random forests and Bayesian networks. The presented model achieves predictive accuracy that is similar to state-of-the-art models (root mean square error of 0.66, mean absolute error of 0.55, coefficient of determination of R2 = 0.15). We identify 62 relevant features from the final random forest model, ranging from general health variables over dietary and genetic factors to physiological, hematological and hemostasis parameters. The Bayesian network model is used to put these features into context and make the black-box random forest model more understandable.
Conclusion
We demonstrate that the combination of random forest and Bayesian network analysis is helpful to reveal and interpret broad association patterns of individual TSH concentrations. The discovered patterns are in line with state-of-the-art literature. They may be useful for future thyroid research and improved dosing of therapeutics.
Purpose
Postapproval reports of intraocular inflammation (IOI) and occlusive retinal vasculitis following intravitreal brolucizumab are accumulating. A role of anti-drug antibodies (ADAs) to brolucizumab is under current scientific discussion. The purpose of the present study was to measure brolucizumab ADAs in a cross-sectional ophthalmic patient population and to compare the occurrence of brolucizumab ADAs with that of ranibizumab ADAs.
Methods
One hundred and ninety-two serum samples and 54 vitreous samples were collected from patients with a range of eye diseases including neovascular age-related macular degeneration (AMD), diabetic retinopathy, retinal vein occlusion, cataract, glaucoma, dry eye disease, macular hole, epiretinal membranes and intraocular lens (IOL) dislocation. Serum and vitreous samples were analysed for immune globuline (Ig) G ADAs to brolucizumab and ranibizumab using indirect enzyme-linked immunosorbent assay (ELISA). Optical Density (OD) was read at 450 nm (wavelength correction at 550 nm) for ADA level measurements.
Results
Presence of brolucizumab ADAs was observed in patients with and without prior brolucizumab exposure. Both the frequency of notable ADA signals (OD > 0.1) and the mean ADA signal in serum samples were higher for brolucizumab than for ranibizumab. Two patients who experienced severe IOI and occlusive retinal vasculitis following intravitreal brolucizumab had high brolucizumab ADA serum levels. In one of these two patients, high brolucizumab ADA levels were also found in vitreous. Another patient developed moderate IOI without retinal vasculitis in the presence of low brolucizumab ADA serum levels. Overall, notable brolucizumab ADA levels were less frequent in vitreous than in the corresponding serum samples but with a tendency for higher prevalence in vitreous from patients with diabetic retinopathy.
Conclusion
Brolucizumab ADAs occur with significant prevalence in a typical ophthalmic patient population and may represent a risk factor for IOI and occlusive retinal vasculitis following brolucizumab.
Background
Understanding how SARS-CoV-2 affects respiratory centres in the brainstem may help to preclude assisted ventilation for patients in intensive care setting. Viral invasion appears unlikely, although autoimmunity has been implicated, the responsible antigens remain unknown. We previously predicted the involvement of three epitopes within distinct brainstem proteins: disabled homolog 1 (DAB1), apoptosis-inducing-factor-1 (AIFM1), and surfeit-locus-protein-1 (SURF1).
Methods
Here, we used microarrays to screen serum from COVID-19 patients admitted to intensive care and compared those with controls who experienced mild course of the disease.
Findings
The results confirm the occurrence of IgG and IgM antibodies against the hypothesised epitopes in COVID-19 patients. Importantly, while IgM levels were similar in both groups, IgG levels were significantly elevated in severely ill patients compared to controls, suggesting a pathogenic role of IgG.
Interpretation
The newly discovered anti-neuronal antibodies might be promising markers of severe disease and the targeted peptide epitopes might be used for targeted immunomodulation. Further work is needed to determine whether these antibodies may play a role in long-COVID.
Funding
AF, CF and PR received support from the German Research Foundation (grants FL 379/22-1, 327654276-SFB 1315, FR 4479/1-1, PR 1274/8-1). SH, DR, and DB received support from the Ministry of Economy, State of Mecklenburg Western Pomerania, Germany (grant COVIDPROTECT: “Optimisation of diagnostic and therapeutic pathways for COVID-19 patients in MV”). SH received support from the Research Group Molecular Medicine University of Greifswald (FVMM, seed funding FOVB-2021-01). AV received support from the Else Kröner Fresenius Foundation and the Alzheimer Research Initiative.
Species of the genus Drosera, known for carnivorous plants, such as sundew, have been traditionally used for centuries as medicinal plants. Efficacy-determining compounds are naphthoquinones and flavonoids. Flavonoids possess a broad spectrum of bioactive properties, including biofilm inhibitory activity. Biofilms render antibiotics ineffective, contributing to the current rise in antimicrobial resistance. In this study, the biofilm inhibitory activity of two European sundew species (Drosera rotundifolia and Drosera intermedia) grown agriculturally in Germany and four commercial sundew products (declared as Drosera longifolia, Drosera sp. and Drosera planta trit.) against three multidrug-resistant Escherichia coli strains was tested. The aim of the study was to comparatively investigate the biofilm inhibitory potential of sundew species extracts grown locally in northern Germany and commercial sundew products. The minimum biofilm inhibitory concentration of the European sundew species was approx. 35 µg mL−1. In comparison, commercial sundew products ranged in concentration from 75 to 140 µg mL−1. Additionally, individual compounds isolated from European sundew were tested. Among these compounds, biofilm inhibitory activity was determined for four of the eight substances, with 2″-O-galloyl hyperoside standing out for its activity (38 µg mL−1). The whole plant extracts of Drosera rotundifolia and Drosera intermedia proved to be more effective than the commercial products and the single compounds in its biofilm inhibition activity against Escherichia coli strains. Sundew extracts may serve as a potential therapeutic approach for targeting biofilm production.
APOE ε4 in Depression-Associated Memory Impairment—Evidence from Genetic and MicroRNA Analyses
(2022)
(1) Background: The aim of this study was to replicate a reported interaction between APOE ε4 status and depression on memory function in two independent, nondemented samples from the general population and to examine the potential role of circulating plasma miRNAs. (2) Methods: The impact of the APOE ε4 allele on verbal memory and the interaction with depression is investigated in two large general-population cohorts from the Study of Health in Pomerania (SHIP, total n = 6286). Additionally, biological insights are gained by examining the potential role of circulating plasma miRNAs as potential epigenetic regulators. Analyses are performed using linear regression models adjusted for relevant biological and environmental covariates. (3) Results: Current depression as well as carrying the APOE ε4 allele were associated with impaired memory performance, with increasing effect for subjects with both risk factors. In a subcohort with available miRNA data subjects with current depressive symptoms and carrying APOE e4 revealed reduced levels of hsa-miR-107, a prominent risk marker for early Alzheimer’s Disease. (4) Conclusions: Our results confirm the effect of depressive symptoms and APOE ε4 status on memory performance. Additionally, miRNA analysis identified hsa-miR-107 as a possible biological link between APOE ε4, depressive symptoms, and cognitive impairment.
Influenza A Virus (IAV) infection followed by bacterial pneumonia often leads to hospitalization and death in individuals from high risk groups. Following infection, IAV triggers the process of viral RNA replication which in turn disrupts healthy gut microbial community, while the gut microbiota plays an instrumental role in protecting the host by evolving colonization resistance. Although the underlying mechanisms of IAV infection have been unraveled, the underlying complex mechanisms evolved by gut microbiota in order to induce host immune response following IAV infection remain evasive. In this work, we developed a novel Maximal-Clique based Community Detection algorithm for Weighted undirected Networks (MCCD-WN) and compared its performance with other existing algorithms using three sets of benchmark networks. Moreover, we applied our algorithm to gut microbiome data derived from fecal samples of both healthy and IAV-infected pigs over a sequence of time-points. The results we obtained from the real-life IAV dataset unveil the role of the microbial families Ruminococcaceae, Lachnospiraceae, Spirochaetaceae and Prevotellaceae in the gut microbiome of the IAV-infected cohort. Furthermore, the additional integration of metaproteomic data enabled not only the identification of microbial biomarkers, but also the elucidation of their functional roles in protecting the host following IAV infection. Our network analysis reveals a fast recovery of the infected cohort after the second IAV infection and provides insights into crucial roles of Desulfovibrionaceae and Lactobacillaceae families in combating Influenza A Virus infection. Source code of the community detection algorithm can be downloaded from https://github.com/AniBhar84/MCCD-WN.
The biomechanical (Young's modulus, adhesion force, deformability) properties of platelets depend on the cytoskeleton and have an undisputed influence on physiological and pathological processes such as hemostasis and thrombosis. The alterations of these biomechanical properties can be used as label-free diagnostic markers in initiation or progressive diseases such as MYH9-inherited disease. Therefore, the focus of my thesis was to investigate the relationship between the changes in platelet cytoskeleton proteins and the resulting biomechanical properties using biophysical methods.
In the first chapter of my thesis I focused on my review of the biophysical methods that are most commonly used to assess and quantify the biomechanical properties of platelets. In this review, I provide an in-depth insight into the governing principles and instrumentation setup and discuss relevant examples applied to platelet mechanics. In addition, my review also summarizes the limitations of these biophysical methods and highlight latest improvements. The review covers the following techniques: micropipette aspiration, atomic force microscopy (AFM), scanning ion conductance microscopy (SICM), tensile force microscopy on hydrogel substrates, microcolumns, and deformable 3D substrates, and real-time deformability cytometry (RT-DC). This review is directed toward clinician scientists who are interested in exploring applications of single-cell based biophysical approaches in unraveling the role of platelet biomechanics in hemostasis and thrombosis research.
In the second chapter of my thesis, I present my research paper on the influence of commonly used ex vivo anticoagulants on the intrinsic biomechanical properties and functional parameters (e.g. activation profils) of human platelets. To comprehensively assess this, platelets obtained in different ex vivo anticoagulants such as ACD-A, Na-Citrate, K2-EDTA, Li-Heparin, and r-Hirudin were used, and their biomechanical properties were determined by real-time fluorescence and deformability cytometry (RT-FDC). Flow cytometry, and confocal laser scanning fluorescence microscopy were used to determine platelet function properties. K2-EDTA and Li-Heparin were found to affect platelet biomechanics by increasing actin polymerization of non-stimulated human platelets. This increased actin polymerization results in decreased platelet deformation. It is recommended that an ex vivo anticoagulant such as ACD-A, Na-Citrate, or r-Hirudin be chosen for the study of the cytoskeleton of human platelets and, if possible, that it not be exchanged, because comparability of results is not assured. Furthermore, I demonstrate the significance of choosing correct ex vivo anticoagulants in RT-FDC by showing that platelets from a healthy donor and a MYH9 patient with the E1841K point mutation differ in their deformation. This paper is the first comprehensive investigation at the single platelet level to establish the relevance of preanalytical standardization in platelet sample preparation for biomechanical studies.
The third chapter of my thesis is focused on the biomechanical analyses of platelets and thrombi from MYH9-related disease. Here I studied three Myh9 mouse lines with a point mutation in the Myh9 gene at positions 702, 1424, or 1841. Furthermore, two MYH9 patients (MYH9 p.D1424N, MYH9 p.E1841K) were examined. MYH9-related disease (MYH9-RD) presents with macrothrombocytopenia with a moderate bleeding tendency. It is caused by mutations in the MYH9 gene that lead to alteration of non-muscle myosin heavy chains type IIA (NMMHC IIA), resulting in disruption of the platelet cytoskeleton. Western blot analysis, flow cytometry, in vitro aggregometry, and transmission electron microscopy demonstrated that Myh9 point mutant mice have comparable primary function compared to the control group. The heterozygous point mutations in the Myh9 gene resulted in decreased platelet deformation (RT-FDC), decreased platelet adhesion to collagen (single platelet force spectroscopy-SPFS), and decreased platelet-platelet interaction forces (SPFS). Decreased platelet force (Micropost Arrays) results in softer thrombi (colloidal probe Spectroscopy), impaired clot retraction, and thus prolonged bleeding time. The R702C, D1424N, and E1841K mutations have a similar effect on platelet biomechanical functions, although the E1841K mutation had less impact on thrombus formation and stiffness. MYH9-RD patients have an increased risk of bleeding, and the antifibrinolytic drug tranexamic acid (TXA) is one way to control bleeding complications in these patients. It was shown that TXA treatment significantly reduced bleeding time in the three Myh9 mouse models, confirming that the enhanced bleeding phenotype due to decreased platelet forces in Myh9 mutant mice can be compensated by the addition of TXA.
With the biophysical methods and research results presented in my thesis, it is clear that it is essential to study the altered response of the platelet cytoskeleton by cytoskeletal mutations, biochemical, physical stimuli, or by pharmacological aspects. This will provide us with an opportunity to better understand the underlying mechanisms and thus contribute to better clinical treatment.
Application of In Vivo Imaging Techniques and Diagnostic Tools in Oral Drug Delivery Research
(2022)
Drug absorption following oral administration is determined by complex and dynamic interactions between gastrointestinal (GI) physiology, the drug, and its formulation. Since many of these interactions are not fully understood, the COST action on “Understanding Gastrointestinal Absorption-related Processes (UNGAP)” was initiated in 2017, with the aim to improve the current comprehension of intestinal drug absorption and foster future developments in this field. In this regard, in vivo techniques used for the characterization of human GI physiology and the intraluminal behavior of orally administered dosage forms in the GI tract are fundamental to gaining deeper mechanistic understanding of the interplay between human GI physiology and drug product performance. In this review, the potential applications, advantages, and limitations of the most important in vivo techniques relevant to oral biopharmaceutics are presented from the perspectives of different research fields.
Introduction
Privacy concerns are an important barrier to adoption and continued use of digital technologies, particularly in the health sector. With the introduction of mobile health applications (mHealth apps), the construct of app information privacy concerns has received increased attention. However, few validated measures exist to capture said concerns in population samples, although they can help to improve public health efforts.
Methods
Using a cross-sectional survey of German adults (mean age = 35.62; 63.5% female), this study examined psychometric properties of the app information privacy concerns scale (AIPC). Analyses comprised confirmatory factor analysis, factorial validity (exploratory factor analysis), internal consistency, convergent validity (i.e., correlations with privacy victimhood, and app privacy concerns), and discriminant validity (i.e., daily app use, adoption intentions, and attitudes toward COVID-19 contact tracing app use).
Results
The analysis did not support the proposed three-factor structure of the AIPC (i.e., anxiety, personal attitude, and requirements). Instead, a four-factor model was preferable that differentiated requirements regarding disclosure policies, and personal control. In addition, factors mirroring anxiety and personal attitude were extracted, but shared a significant overlap. However, these factors showed good reliability, convergent and discriminant validity.
Discussion
The findings underline the role of app information privacy concerns as a significant barrier to mHealth app use. In this context, anxiety and personal attitudes seemed particularly relevant, which has implications for health communication. Moreover, the observed differentiation of external (disclosure) and internal (control) requirements aligns with health behavior change models and thus is a promising area for future research.
(1) Background: In Germany, new recommendations for dental examinations of children and the use of fluorides have been introduced. The pediatrician (PA) should refer the patient to the dentist for dental examinations and check-ups (DEs) from the sixth month of age. Therefore, our aim was to determine with a questionnaire the extent to which PAs find DE useful, make referrals for DE and recommend fluoride. (2) Methods: The nationwide empirical survey was conducted with a self-developed and validated standardized online questionnaire. In addition to personal information, 16 items were collected. Agreement with the items was recorded using Likert scales. The data were primarily analysed descriptively. (3) Results: 696 PAs participated in the survey (age: 51.7 (8.4) years, women/men: 428/286 (61.5/38.5%). A total of 11% of PAs found referral by eruption of first tooth very important (important/neutral/unimportant: 13.8/32/43.2%), compared to 70% for complete deciduous teeth (21.3/7.3/1.4%). A total of 48.8% of PAs always recommended fluoridated toothpaste from the first tooth (often/occasionally/rarely/never: 18.3/7.8/8/17.1%) and 50.6% completely refused to recommend fluoride-free toothpaste (always/often/occasionally/rarely: 9.8/9/14.7/15.9%). A total of 44.8% never recommended the use of fluoridated toothpaste if the child cannot yet spit (always/often/occasionally/rarely: 19.2/13.9/7.8/14.3%). (4) Conclusions: Among PAs, referral to DEs was increasingly implemented as children grew older. Specific fluoride recommendations were accepted.
In addition to the classical oestrogen receptors, ERα and ERβ, a G protein-coupled oestrogen receptor (GPER) has been identified that primarily mediates the rapid, non-genomic signalling of oestrogens. Data on GPER expression at the protein level are contradictory; therefore, the present study was conducted to re-evaluate GPER expression by immunohistochemistry to obtain broad GPER expression profiles in human non-neoplastic and neoplastic tissues, especially those not investigated in this respect so far. We developed and thoroughly characterised a novel rabbit monoclonal anti-human GPER antibody, 20H15L21, using Western blot analyses and immunocytochemistry. The antibody was then applied to a large series of formalin-fixed, paraffin-embedded human tissue samples. In normal tissue, GPER was identified in distinct cell populations of the cortex and the anterior pituitary; islets and pancreatic ducts; fundic glands of the stomach; the epithelium of the duodenum and gallbladder; hepatocytes; proximal tubules of the kidney; the adrenal medulla; and syncytiotrophoblasts and decidua cells of the placenta. GPER was also expressed in hepatocellular, pancreatic, renal, and endometrial cancers, pancreatic neuroendocrine tumours, and pheochromocytomas. The novel antibody 20H15L21 will serve as a valuable tool for basic research and the identification of GPER-expressing tumours during histopathological examinations.
A massive shift in agricultural practices over the past decades, to support exceptionally high yields and productivities involving intensive agriculture, have led to unsustainable agriculture practices across the globe. Sustenance of such high yields and productivities demand high use of organic and industrial fertilizers. This acts as a negative pressure on the environment. Excessive use of fertilizers leads to nutrient surplus in the fields, which, as a part of catchment runoff, flows into the water bodies as diffuse pollution. These nutrients through rivers are eventually passed into seas. High nutrients ending up into water bodies cause eutrophication. The situation is worsened when such unsustainable agricultural activities are carried out on drained peatlands. As a result, the nutrients that were not part of the nutrient cycle in the landscape for years begin to leach out due to mineralization of peatlands, thereby putting an additional load of nutrients on the environment, that was already under the negative impact of nutrient surplus. In view of the above, a small lowland catchment of the Ryck river in northeast Germany was assessed for its nitrogen losses from agricultural lands through empirical modelling. Initial empirical modelling resulted in an average annual total nitrogen loss of 14.7 kg ha−1 year−1. After a comparative analysis of these results with procured data, the empirical equation was modified to suit the catchment, yielding more accurate results. The study showed that 75.6% of peatlands in the catchment are under agricultural use. Subsequently, a proposal was made for potential wetland buffer zones in the Ryck catchment. Altogether, 13 peatland sites across 8 sub-catchments were recommended for mitigation of high nutrient runoff. In the end, nutrient efficiency of proposed WBZs in one of the sub-catchments of Ryck has been discussed. The results show that (i) the modified empirical equation can act as a key tool in application-based future strategies for nitrogen reduction in the Ryck catchment, (ii) restoration of peatlands and introduction of WBZs can help in mitigating the nutrient runoff for improved water quality of Ryck, and subsequently (ii) contribute to efficient reduction of riverine loads of nutrients into the Baltic Sea.