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Sterically distorted donor-acceptor p‑systems, termed DA springs, can be progressively planarized under mechanical load causing a bathochromic shift of the photoluminescence (PL) spectrum. By combining theory and experiment, we here use a simple linear force calibration for two different conformational mechanochromophores to determine molecular forces in polymers from the mechanochromic shift in PL wavelength during multiple uniaxial tensile tests. Two systems are used, i) a highly entangled linear glassy polyphenylene and ii) a covalent elastomeric polydimethylsiloxane network. The mean forces estimated by this method are validated using known threshold forces for the mechanochemical ring-opening reactions of two different spiropyran force probes. The agreement between both approaches underlines that these DA springs provide the unique opportunity for the online monitoring of local molecular forces present in diverse polymer matrices., (© 2024 Wiley‐VCH GmbH.)

Magnonics is a research field that has gained an increasing interest in both the fundamental and applied sciences in recent years. This field aims to explore and functionalize collective spin excitations in magnetically ordered materials for modern information technologies, sensing applications and advanced computational schemes. Spin waves, also known as magnons, carry spin angular momenta that allow for the transmission, storage and processing of information without moving charges. In integrated circuits, magnons enable on-chip data processing at ultrahigh frequencies without the Joule heating, which currently limits clock frequencies in conventional data processors to a few GHz. Recent developments in the field indicate that functional magnonic building blocks for in-memory computation, neural networks and Ising machines are within reach. At the same time, the miniaturization of magnonic circuits advances continuously as the synergy of materials science, electrical engineering and nanotechnology allows for novel on-chip excitation and detection schemes. Such circuits can already enable magnon wavelengths of 50 nm at microwave frequencies in a 5G frequency band. Research into non-charge-based technologies is urgently needed in view of the rapid growth of machine learning and artificial intelligence applications, which consume substantial energy when implemented on conventional data processing units. In its first part, the 2024 Magnonics Roadmap provides an update on the recent developments and achievements in the field of nano-magnonics while defining its future avenues and challenges. In its second part, the Roadmap addresses the rapidly growing research endeavors on hybrid structures and magnonics-enabled quantum engineering. We anticipate that these directions will continue to attract researchers to the field and, in addition to showcasing intriguing science, will enable unprecedented functionalities that enhance the efficiency of alternative information technologies and computational schemes., (Creative Commons Attribution license.)

Background: Patient-specific aiming devices (PSAD) may improve precision and accuracy of glenoid component positioning in total shoulder arthroplasty, especially in degenerative glenoids. The aim of this study was to compare precision and accuracy of guide wire positioning into different glenoid models using a PSAD versus a standard guide., Methods: Three experienced shoulder surgeons inserted 2.5 mm K-wires into polyurethane cast glenoid models of type Walch A, B and C (in total 180 models). Every surgeon placed guide wires into 10 glenoids of each type with a standard guide by DePuy Synthes in group (I) and with a PSAD in group (II). Deviation from planned version, inclination and entry point was measured, as well as investigation of a possible learning curve., Results: Maximal deviation in version in B- and C-glenoids in (I) was 20.3° versus 4.8° in (II) (p < 0.001) and in inclination was 20.0° in (I) versus 3.7° in (II) (p < 0.001). For B-glenoid, more than 50% of the guide wires in (I) had a version deviation between 11.9° and 20.3° compared to ≤ 2.2° in (II) (p < 0.001). 50% of B- and C-glenoids in (I) showed a median inclination deviation of 4.6° (0.0°-20.0°; p < 0.001) versus 1.8° (0.0°-4.0°; p < 0.001) in (II). Deviation from the entry point was always less than 5.0 mm when using PSAD compared to a maximum of 7.7 mm with the standard guide and was most pronounced in type C (p < 0.001)., Conclusion: PSAD enhance precision and accuracy of guide wire placement particularly for deformed B and C type glenoids compared to a standard guide in vitro. There was no learning curve for PSAD. However, findings of this study cannot be directly translated to the clinical reality and require further corroboration., (© 2024. The Author(s).)

Prokaryotic antiviral defence systems are frequently toxic for host cells and stringent regulation is required to ensure survival and fitness. These systems must be readily available in case of infection but tightly controlled to prevent activation of an unnecessary cellular response. Here we investigate how the bacterial cyclic oligonucleotide-based antiphage signalling system (CBASS) uses its intrinsic protein modification system to regulate the nucleotide cyclase. By integrating a type II CBASS system from Bacillus cereus into the model organism Bacillus subtilis, we show that the protein-conjugating Cap2 (CBASS associated protein 2) enzyme links the cyclase exclusively to the conserved phage shock protein A (PspA) in the absence of phage. The cyclase-PspA conjugation is reversed by the deconjugating isopeptidase Cap3 (CBASS associated protein 3). We propose a model in which the cyclase is held in an inactive state by conjugation to PspA in the absence of phage, with conjugation released upon infection, priming the cyclase for activation., (© 2024. The Author(s).)

Expanding upon the burgeoning discipline of magnonics, this research elucidates the intricate dynamics of spin waves (SWs) within three-dimensional nanoenvironments. It marks a shift from traditionally used planar systems to exploration of magnetization configurations and the resulting dynamics within 3D nanostructures. This study deploys micromagnetic simulations alongside ferromagnetic resonance measurements to scrutinize magnetic gyroids, periodic chiral configurations composed of chiral triple junctions with a period in nanoscale. Our findings uncover distinctive attributes intrinsic to the gyroid network, most notably the localization of collective SW excitations and the sensitivity of the gyroid's ferromagnetic response to the orientation of the static magnetic field, a correlation closely tied to the crystallographic alignment of the structure. Furthermore, we show that for the ferromagnetic resonance, multidomain gyroid films can be treated as a magnonic material with effective magnetization scaled by its filling factor. The implications of our research carry the potential for practical uses such as an effective, metamaterial-like substitute for ferromagnetic parts and lay the groundwork for radio frequency filters. The growing areas of 3D magnonics and spintronics present exciting opportunities to investigate and utilize gyroid nanostructures for signal processing purposes.

Aminophosphonates, like glyphosate (GS) or metal chelators such as ethylenediaminetetra(methylenephosphonic acid) (EDTMP), are released on a large scale worldwide. Here, we have characterized a bacterial strain capable of degrading synthetic aminophosphonates. The strain was isolated from LC/MS standard solution. Genome sequencing indicated that the strain belongs to the genus Ochrobactrum. Whole-genome classification using pyANI software to compute a pairwise ANI and other metrics between Brucella assemblies and Ochrobactrum contigs revealed that the bacterial strain is designated as Ochrobactrum sp. BTU1. Degradation batch tests with Ochrobactrum sp. BTU1 and the selected aminophosphonates GS, EDTMP, aminomethylphosphonic acid (AMPA), iminodi(methylene-phosphonic) (IDMP) and ethylaminobis(methylenephosphonic) acid (EABMP) showed that the strain can use all phosphonates as sole phosphorus source during phosphorus starvation. The highest growth rate was achieved with AMPA, while EDTMP and GS were least supportive for growth. Proteome analysis revealed that GS degradation is promoted by C-P lyase via the sarcosine pathway, i.e., initial cleavage at the C-P bond. We also identified C-P lyase to be responsible for degradation of EDTMP, EABMP, IDMP and AMPA. However, the identification of the metabolite ethylenediaminetri(methylenephosphonic acid) via LC/MS analysis in the test medium during EDTMP degradation indicates a different initial cleavage step as compared to GS. For EDTMP, it is evident that the initial cleavage occurs at the C-N bond. The detection of different key enzymes at regulated levels, form the bacterial proteoms during EDTMP exposure, further supports this finding. This study illustrates that widely used and structurally more complex aminophosphonates can be degraded by Ochrobactrum sp. BTU1 via the well-known degradation pathways but with different initial cleavage strategy compared to GS., (Copyright © 2024 The Authors. Published by Elsevier GmbH.. All rights reserved.)

Glyphosate (GS) specifically inhibits the 5-enolpyruvyl-shikimate-3-phosphate (EPSP) synthase that converts phosphoenolpyruvate (PEP) and shikimate-3-phosphate to EPSP in the shikimate pathway of bacteria and other organisms. The inhibition of the EPSP synthase depletes the cell of the EPSP-derived aromatic amino acids as well as of folate and quinones. A variety of mechanisms (e.g., EPSP synthase modification) has been described that confer GS resistance to bacteria. Here, we show that the Burkholderia anthina strain DSM 16086 quickly evolves GS resistance by the acquisition of mutations in the ppsR gene. ppsR codes for the pyruvate/ortho-P i dikinase PpsR that physically interacts and regulates the activity of the PEP synthetase PpsA. The mutational inactivation of ppsR causes an increase in the cellular PEP concentration, thereby abolishing the inhibition of the EPSP synthase by GS that competes with PEP for binding to the enzyme. Since the overexpression of the Escherichia coli ppsA gene in Bacillus subtilis and E. coli did not increase GS resistance in these organisms, the mutational inactivation of the ppsR gene resulting in PpsA overactivity is a GS resistance mechanism that is probably unique to B. anthina., (© 2023 The Authors. Environmental Microbiology Reports published by Applied Microbiology International and John Wiley & Sons Ltd.)

Glyphosate (GS) inhibits the 5-enolpyruvyl-shikimate-3-phosphate (EPSP) synthase that is required for aromatic amino acid, folate and quinone biosynthesis in Bacillus subtilis and Escherichia coli. The inhibition of the EPSP synthase by GS depletes the cell of these metabolites, resulting in cell death. Here, we show that like the laboratory B. subtilis strains also environmental and undomesticated isolates adapt to GS by reducing herbicide uptake. Although B. subtilis possesses a GS-insensitive EPSP synthase, the enzyme is strongly inhibited by GS in the native environment. Moreover, the B. subtilis EPSP synthase mutant was only viable in rich medium containing menaquinone, indicating that the bacteria require a catalytically efficient EPSP synthase under nutrient-poor conditions. The dependency of B. subtilis on the EPSP synthase probably limits its evolvability. In contrast, E. coli rapidly acquires GS resistance by target modification. However, the evolution of a GS-resistant EPSP synthase under non-selective growth conditions indicates that GS resistance causes fitness costs. Therefore, in both model organisms, the proper function of the EPSP synthase is critical for the cellular viability. This study also revealed that the uptake systems for folate precursors, phenylalanine and tyrosine need to be identified and characterized in B. subtilis., (© 2023 The Authors. Environmental Microbiology published by Applied Microbiology International and John Wiley & Sons Ltd.)

Prophages control their lifestyle to either be maintained within the host genome or enter the lytic cycle. Bacillus subtilis contains the SPβ prophage whose lysogenic state depends on the MrpR (YopR) protein, a key component of the lysis-lysogeny decision system. Using a historic B. subtilis strain harboring the heat-sensitive SPβ c2 mutant, we demonstrate that the lytic cycle of SPβ c2 can be induced by heat due to a single nucleotide exchange in the mrpR gene, rendering the encoded MrpRG136E protein temperature-sensitive. Structural characterization revealed that MrpR is a DNA-binding protein resembling the overall fold of tyrosine recombinases. MrpR has lost its recombinase function and the G136E exchange impairs its higher-order structure and DNA binding activity. Genome-wide profiling of MrpR binding revealed its association with the previously identified SPbeta repeated element (SPBRE) in the SPβ genome. MrpR functions as a master repressor of SPβ that binds to this conserved element to maintain lysogeny. The heat-inducible excision of the SPβ c2 mutant remains reliant on the serine recombinase SprA. A suppressor mutant analysis identified a previously unknown component of the lysis-lysogeny management system that is crucial for the induction of the lytic cycle of SPβ., (© The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Objective: The purpose of this study was to determine medical-surgical nurse leaders' evidence-based practice (EBP) attributes, perceived barriers to EBP, and whether there were differences in leaders' EBP competencies and EBP implementation by demographic and organizational factors., Background: Leaders are crucial to the development of cultures that support EBP implementation, but little is known about medical-surgical nurse leaders' capacity to perform this aspect of their role., Methods: A cross-sectional design using survey methodology was used. The survey contained demographic/work setting questions and 3 instruments to measure EBP beliefs, implementation, and competencies., Results: Senior leaders self-reported higher EBP attributes compared with nurses in other roles; nurses with an MSN or higher reported greater frequency of EBP implementation. A regression revealed that EBP competencies, EBP beliefs, having a DNP degree, and working in an Academy of Medical-Surgical Nurses Premier Recognition In the Specialty of Med-surg unit or a Pathway to Excellence® organization had significant, positive effects on EBP implementation scores ( R2 = 0.37)., Conclusions: Findings demonstrate there is a range of EBP beliefs and competencies, and barriers to EBP among medical-surgical nurse leaders. Because medical-surgical nursing is the largest acute care practice specialty in the United States and many other countries, leaders fully integrating EBP into their practice would be a substantial contribution to advancing EBP in healthcare., (Copyright © 2023 Wolters Kluwer Health, Inc. All rights reserved.)

A new Vibrio strain, K08M4 T , was isolated from the broad-nosed pipefish Syngnathus typhle in the Kiel Fjord. Infection experiments revealed that K08M4 T was highly virulent for juvenile pipefish. Cells of strain K08M4 T were Gram-stain-negative, curved rod-shaped and motile by means of a single polar flagellum. The strain grew aerobically at 9-40° C, at pH 4-10.5 and it tolerated up to 12 % (w/v) NaCl. The most prevalent (>10 %) cellular fatty acids of K08M4 T were C 16 : 1 ω 7 c and C 16 : 0 . Whole-genome comparisons revealed that K08M4 T represents a separate evolutionary lineage that is distinct from other Vibrio species and falls within the Splendidus clade. The genome is 4,886,292 bp in size, consists of two circular chromosomes (3,298,328 and 1, 587,964 bp) and comprises 4,178 protein-coding genes and 175 RNA genes. In this study, we describe the phenotypic features of the new isolate and present the annotation and analysis of its complete genome sequence. Based on these data, the new isolate represents a new species for which we propose the name Vibrio syngnathi sp. nov. The type strain is K08M4 T (=DSM 109818 T =CECT 30086 T ).

Focusing on visible plaques for phage isolation leaves the question if we miss the diversity of non-plaque forming phages. We addressed this question through direct plaque-based isolation by employing the new hosts Brevundimonas pondensis LVF1 and Serratia marcescens LVF3 dsDNA, ssDNA, dsRNA, and ssRNA host-associated metavirome analysis. Of the 25 distinctive dsDNA phage isolates, 14 were associated with Brevundimonas and 11 with Serratia . TEM analysis revealed that 6 were myoviruses, 18 siphoviruses and 1 podovirus, while phages infecting Brevundimonas belonged all to siphoviruses. The associated viromes suggested a higher phage diversity in summer than in winter, and dsDNA phages were the dominant group. Isolation of vB&#95;SmaP-Kaonashi was possible after investigating the viromes associated with Serratia , demonstrating the great potential of accompanying host-associated metavirome analysis. The ssDNA virome analysis showed that the B. pondensis LVF1 host is associated with Microviridae and Inoviridae phages, although none of them were isolated. The results demonstrated that the classical isolation technique is not exhausted, leading to the isolation of new dsDNA phages. It can be further improved by combination with metavirome techniques, which revealed further diversity., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Friedrich, Neubauer, Kuritsyn, Bodenberger, Tskhay, Hartmann, Poehlein, Bömeke, Hoppert, Schneider, Hertel and Daniel.)

Ferroelectric domain boundaries are quasi-two-dimensional functional interfaces with high prospects for nanoelectronic applications. Despite their reduced dimensionality, they can exhibit complex non-Ising polarization configurations and unexpected physical properties. Here, the impact of the three-dimensional (3D) curvature on the polarization profile of nominally uncharged 180° domain walls in LiNbO 3 is studied using second-harmonic generation microscopy and 3D polarimetry analysis. Correlations between the domain-wall curvature and the variation of its internal polarization unfold in the form of modulations of the Néel-like character, which we attribute to the flexoelectric effect. While the Néel-like character originates mainly from the tilting of the domain wall, the internal polarization adjusts its orientation due to the synergetic upshot of dipolar and monopolar bound charges and their variation with the 3D curvature. Our results show that curved interfaces in solid crystals may offer a rich playground for tailoring nanoscale polar states.

Luteibacter is a genus of the Rhodanobacteraceae family. The present study describes a novel species within the genus Luteibacter (EIF3 T ). The strain was analyzed genomically, morphologically and physiologically. Average nucleotide identity analysis revealed that it is a new species of Luteibacter . In silico analysis indicated two putative prophages (one incomplete, one intact). EIF3 T cells form an elliptical morphotype with an average length of 2.0 µm and width of 0.7 µm and multiple flagella at one end. The bacterial strain is an aerobic Gram-negative with optimal growth at 30 °C. EIF3 T is resistant towards erythromycin, tetracycline and vancomycin. We propose the name Luteibacter flocculans sp. nov. with EIF3 T (=DSM 112537 T = LMG 32416 T ) as type strain. Further, we describe the first known Luteibacter -associated bacteriophage called vB&#95;LflM-Pluto.

Phages are viruses of bacteria and have been known for over a century. They do not have a metabolism or protein synthesis machinery and rely on host cells for replication. The model organism Bacillus subtilis has served as a host strain for decades and enabled the isolation of many unique viral strains. However, many viral species representatives remained orphans as no, or only a few, related phages were ever re-isolated.The presented protocol describes how a CRISPR-Cas9 system with an artificial CRISPR-array can be set up and used to discriminate abundant and well-known B. subtilis phage from a host-based metagenome enrichment. The obtained viral suspension can be used for metagenome sequencing and isolating new viral strains., (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Purpose: The majority of methods for measuring glenoid bone loss in shoulder instability use the best-fit circle following the inferior glenoid rim. However, there is no precise method on how to draw this circle, particularly in case of a missing rim segment. Defining the radius is a source of substantial error. It was hypothesized that there is a relationship between the best-fit inferior circle (inner circle), defined by Sugaya, and the circle tangent to the supra- and infra-glenoid tubercles (outer circle), defined by Itoi, thus allowing a more consistent appreciation of the paleo-glenoid., Methods: Ninety-five normal dry scapulae were examined. The specimens were digitally photographed obtaining perpendicular images of the glenoid cavity. Using HOROS® imaging software, a best-fit inferior circle (inner circle) and a second circle fitting the most inferior and superior points of the glenoid (outer circle) were drawn by two investigators. The diameters and areas of the circles were recorded. Two-way random-effects intra-class correlation coefficients (ICC) were used to measure intra- and inter-observer agreement. A Bayesian measurement-error regression model was used to determine the relationship between outer and inner circle measurements., Results: The mean glenoid height was 35.1 mm and the glenoid width 25.6 mm. The mean diameter of the outer circle was 35.7 ± 4.2 mm and the mean diameter of the inner circle was 26.8 ± 3.2 mm. ICC showed excellent inter- and intra-observer agreement for both the outer circle diameter (ICC ≥ 0.95) and inner circle diameter (ICC ≥ 0.93). The two diameters demonstrated a very strong significant Pearson correlation (0.92, p < 0.001) and the regression showed excellent model fit R 2  = 0.87. The areas of the two circles were also highly and significantly correlated (r = 0.94; p < 0.001). The ratio of inner circle to outer diameters was 0.74., Conclusion: There is a strong correlation between the inner and outer glenoid circle diameters. This study sets the base for the use the combined outer and inner circle and its ratio to better appreciate the paleo-glenoid morphology and thus obtain a more reliable bone loss estimation. Application of this method aids in a more reliable estimation bone loss with potential benefit in surgical decision-making., (© 2022. The Author(s) under exclusive licence to European Society of Sports Traumatology, Knee Surgery, Arthroscopy (ESSKA).)

Bacteriophages, also called phages, are viruses of bacteria. They are the most common and diverse biological entities on this planet. For metagenomic investigation, their diversity is also their biggest obstacle. The direct metagenomic sequence of environmental phage communities often leads to short genomic fragments limiting the investigation to a few individual aspects of phage biology and diversity.The presented protocol for generating a host-associated metagenome reduces the phage diversity to a concise and accessible size. Metagenome sequencing often leads to complete genomes, and the availability of a suitable host system ensures further experimental investigation., (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Mechanochromophores based on conformational changes of donor-acceptor-donor (DAD) springs allow sensing of forces acting on polymer chains by monotonic changes of absorbance or photoluminescence (PL) wavelength. Here, we identify a series of thiophene (D)-flanked quinoxalines (A) as molecular torsional springs for force sensing in bulk polymers at room temperature. The mode of DAD linkage to the polymer matrix and linker rigidity are key parameters that influence the efficacy of force transduction to the DAD spring and thus mechanochromic response, as probed by in situ PL spectroscopy of bulk films during stress-strain experiments. The largest shift of the PL maximum, and thus the highest sensitivity, is obtained from an ansa -DAD spring exhibiting bridged D units and a stiff A linker. Using detailed spectroscopy and density functional theory calculations, we reveal conformer redistribution in the form of a thiophene ring flip as the major part of the overall mechanochromic response. At forces as low as 27 pN at early stages of deformation, the ring flip precedes mechanically induced planarization of the ansa -DAD spring, the latter process producing a PL shift of 21 nm nN -1 . Within the stress-strain diagram, the thiophene ring flip and DAD planarization are thus two separated processes that also cause irreversible and reversible mechanochromic responses, respectively, upon sample failure. As the thiophene ring flip requires much smaller forces than planarization of the DAD spring, such micromechanical motion gives access to sensing of tiny forces and expands both sensitivity and the force range of conformational mechanochromophores.

Background: Patient-specific instrumentation (PSI) may potentially improve humeral osteotomy in shoulder arthroplasty. The purpose of this study was to compare the deviation between planned and postosteotomy humeral inclination, retrotorsion, and height in shoulder arthroplasty, using PSI vs. standard cutting guides (SCG)., Methods: Twenty fresh-frozen cadaveric specimens were allocated to undergo humeral osteotomy using either PSI or SCG, such that the 2 groups have similar age, gender, and side. Preosteotomy computed tomography (CT) scan was performed and used for the 3-dimensional (3D) planning. The osteotomy procedure was performed using a PSI designed for each specimen or an SCG depending on the group. A postosteotomy CT scan was performed. The preosteotomy and postosteotomy 3D CT scan reconstructions were superimposed to calculate the deviation between planned and postosteotomy inclination, retrotorsion, and height. Outliers were defined as cases with 1 or more of the following deviations: >5° inclination, >10° retrotorsion, and >3 mm height. The deviation and outliers in inclination, retrotorsion, and height were compared between the 2 groups., Results: The deviations between planned and postosteotomy parameters were similar among the PSI and SCG groups for inclination (P = .260), whereas they were significantly greater in the SCG group for retrotorsion (P < .001) and height (P = .003). There were 8 outliers in the SCG group, compared with only 1 outlier in the PSI group (P = .005). Most outliers in the SCG group were due to deviation >10° in retrotorsion., Conclusion: After 3D planning, PSI had less deviation between planned and postosteotomy humeral retrotorsion and height, relative to SCG., (Copyright © 2022 Journal of Shoulder and Elbow Surgery Board of Trustees. Published by Elsevier Inc. All rights reserved.)

Over the past year, the AI community has constructed several deep learning models for diagnosing COVID-19 based on the visual features of chest X-rays. While deep learning researchers have commonly focused much of their attention on designing deep learning classifiers, only a fraction of these same researchers have dedicated effort to including a segmentation module in their system. This is unfortunate since other applications in radiology typically require segmentation as a necessary prerequisite step in building truly deployable clinical models. Differentiating COVID-19 from other pulmonary diseases can be challenging as various lung diseases share common visual features with COVID-19. To help clarify the diagnosis of suspected COVID-19 patients, we have designed our deep learning pipeline with a segmentation module and ensemble classifier. Following a detailed description of our deep learning pipeline, we present the strengths and shortcomings of our approach and compare our model with other similarly constructed models. While doing so, we focus our attention on widely circulated public datasets and describe several fallacies we have noticed in the literature concerning them. After performing a thorough comparative analysis, we demonstrate that our best model can successfully obtain an accuracy of 91 percent and sensitivity of 92 percent., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Authors.)

The Bacillus phage SPβ has been known for about 50 years, but only a few strains are available. We isolated four new wild-type strains of the SPbeta species. Phage vB&#95;BsuS-Goe14 introduces its prophage into the spoVK locus, previously not observed to be used by SPβ-like phages. Sequence data revealed the genome replication strategy and the genome packaging mode of SPβ-like phages. We extracted 55 SPβ-like prophages from public Bacillus genomes, thereby discovering three more integration loci and one additional type of integrase. The identified prophages resemble four new species clusters and three species orphans in the genus Spbetavirus. The determined core proteome of all SPβ-like prophages consists of 38 proteins. The integration cassette proved to be not conserved, even though, present in all strains. It consists of distinct integrases. Analysis of SPβ transcriptomes revealed three conserved genes, yopQ, yopR, and yokI, to be transcribed from a dormant prophage. While yopQ and yokI could be deleted from the prophage without activating the prophage, damaging of yopR led to a clear-plaque phenotype. Under the applied laboratory conditions, the yokI mutant showed an elevated virion release implying the YokI protein being a component of the arbitrium system., (© 2022 The Authors. Environmental Microbiology published by Society for Applied Microbiology and John Wiley & Sons Ltd.)

Roundup® is the brand name for herbicide solutions containing glyphosate, which specifically inhibits the 5-enolpyruvyl-shikimate-3-phosphate (EPSP) synthase of the shikimate pathway. The inhibition of the EPSP synthase causes plant death because EPSP is required for biosynthesis of aromatic amino acids. Glyphosate also inhibits the growth of archaea, bacteria, Apicomplexa, algae and fungi possessing an EPSP synthase. Here, we have characterized two glyphosate-resistant bacteria from a Roundup solution. Taxonomic classification revealed that the isolates 1CH1 and 2CH1 are Burkholderia anthina and Burkholderia cenocepacia strains respectively. Both isolates cannot utilize glyphosate as a source of phosphorus and synthesize glyphosate-sensitive EPSP synthase variants. Burkholderia. anthina 1CH1 and B. cenocepacia 2CH1 tolerate high levels of glyphosate because the herbicide is not taken up by the bacteria. Previously, it has been observed that the exposure of soil bacteria to herbicides like glyphosate promotes the development of antibiotic resistances. Antibiotic sensitivity testing revealed that the only the B. cenocepacia 2CH1 isolate showed increased resistance to a variety of antibiotics. Thus, the adaptation of B. anthina 1CH1 and B. cenocepacia 2CH1 to glyphosate did not generally increase the antibiotic resistance of both bacteria. However, our study confirms the genomic adaptability of bacteria belonging to the genus Burkholderia., (© 2021 The Authors. Environmental Microbiology Reports published by Society for Applied Microbiology and John Wiley & Sons Ltd.)

Virulent bacterial viruses, also known as phages or bacteriophages, are considered as a potential option to fight antibiotic-resistant bacteria. However, their biology is still poorly understood, and only a fraction of phage genes is assigned with a function. To enable the first classification, we explored new options to test phage genes for their requirement on viral replication. As a model, we used the smallest known Bacillus subtilis phage Goe1, and the Cas9-based mutagenesis vector pRH030 as a genetic tool. All phage genes were specifically disrupted, and individual survival rates and mutant genotypes were investigated. Surviving phages relied on the genome integrity through host intrinsic non-homologues end joining system or a natural alteration of the Cas9 target sequence. Quantification of phage survivors and verifying the underlying genetic situation enables the classification of genes in essential or non-essential sets for viral replication. We also observed structural genes to hold more natural mutations than genes of the genome replication machinery., (© 2021 The Authors. Environmental Microbiology Reports published by Society for Applied Microbiology and John Wiley & Sons Ltd.)

Serratia marcescens is a species that belongs to the family of Yersiniaceae. This family comprises taxa representing opportunistic human- and phytopathogens but also plant growth-promoting rhizobacteria (PGPR). This study describes a novel Gram-negative strain (LVF3R) of the species Serratia marcescens. The strain was characterized genomically, morphologically, and physiologically. In addition, the potential of the isolate to act as a host strain to assess the diversity of Serratia associated phages in environmental samples was explored. Average nucleotide identity analysis revealed that LVF3R belongs to the species Serratia marcescens. In silico analysis and ProphageSeq data resulted in the identification of one prophage, which is capable of viral particle formation. Electron microscopy showed cells of a rod-shaped, flagellated morphotype. The cells revealed a length and width of 1-1.6 μm and 0.8 μm, respectively. LVF3R showed optimal growth at 30 C and in the presence of up to 2% (w/v) NaCl. It exhibited resistances to ampicillin, erythromycin, oxacillin, oxytetracycline, rifampicin, tetracycline, and vancomycin. Genome data indicate that strain S. marcescens LVF3R is a potential PGPR strain. It harbors genes coding for indole acetic acid (IAA) biosynthesis, siderophore production, plant polymer degradation enzymes, acetoin synthesis, flagellar proteins, type IV secretion system, chemotaxis, phosphorous solubilization, and biofilm formation., Competing Interests: The authors have declared that no competing interests exist.

Phages are viruses of bacteria and are the smallest and most common biological entities in the environment. They can reproduce immediately after infection or integrate as a prophage into their host genome. SPβ is a prophage of the Gram-positive model organism Bacillus subtilis 168, and it has been known for more than 50 years. It is sensitive to dsDNA damage and is induced through exposure to mitomycin C or UV radiation. When induced from the prophage, SPβ requires 90 min to produce and release about 30 virions. Genomes of sequenced related strains range between 128 and 140 kb, and particle-packed dsDNA exhibits terminal redundancy. Formed particles are of the Siphoviridae morphotype. Related isolates are known to infect other B. subtilis clade members. When infecting a new host, SPβ presumably follows a two-step strategy, adsorbing primarily to teichoic acid and secondarily to a yet unknown factor. Once in the host, SPβ-related phages pass through complex lysis-lysogeny decisions and either enter a lytic cycle or integrate as a dormant prophage. As prophages, SPβ-related phages integrate at the host chromosome's replication terminus, and frequently into the spsM or kamA gene. As a prophage, it imparts additional properties to its host via phage-encoded proteins. The most notable of these functional proteins is sublancin 168, which is used as a molecular weapon by the host and ensures prophage maintenance. In this review, we summarise the existing knowledge about the biology of the phage regarding its life cycle and discuss its potential as a research object., (© 2021. The Author(s).)

Glyphosate is a nonselective herbicide that kills weeds and other plants competing with crops. Glyphosate specifically inhibits the 5-enolpyruvyl-shikimate-3-phosphate (EPSP) synthase, thereby depleting the cell of EPSP serving as a precursor for biosynthesis of aromatic amino acids. Glyphosate is considered to be toxicologically safe for animals and humans. Therefore, it became the most-important herbicide in agriculture. However, its intensive application in agriculture is a serious environmental issue because it may negatively affect the biodiversity. A few years after the discovery of the mode of action of glyphosate, it has been observed that bacteria evolve glyphosate resistance by acquiring mutations in the EPSP synthase gene, rendering the encoded enzyme less sensitive to the herbicide. The identification of glyphosate-resistant EPSP synthase variants paved the way for engineering crops tolerating increased amounts of the herbicide. This review intends to summarize the molecular mechanisms underlying glyphosate resistance in bacteria. Bacteria can evolve glyphosate resistance by (i) reducing glyphosate sensitivity or elevating production of the EPSP synthase, by (ii) degrading or (iii) detoxifying glyphosate and by (iv) decreasing the uptake or increasing the export of the herbicide. The variety of glyphosate resistance mechanisms illustrates the adaptability of bacteria to anthropogenic substances due to genomic alterations., (© 2021 The Authors. Environmental Microbiology published by Society for Applied Microbiology and John Wiley & Sons Ltd.)

The Bacillus subtilis subsp. subtilis type strain DSM10 has been used as a reference in various studies. However, detailed information about the genome has not been available. Therefore, whole-genome sequencing was performed, and the sequence was compared with that of the related B. subtilis strain NCIB3610., (Copyright © 2021 Lilge et al.)

Kinneretia sp. strain DAIF2 was isolated from a eutrophic freshwater pond. The genome consists of a single chromosome (6,010,585 bp) with a GC content of 69.3%. The whole-genome-based phylogeny of DAIF2 revealed a closest relation to the genus Kinneretia ., (Copyright © 2021 Hollensteiner et al.)

We present the complete genome of Stenotrophomonas indicatrix DAIF1, which was isolated from an oligotrophic pond in a water protection area. Whole-genome alignments indicated that strain DAIF1 belongs to the species Stenotrophomonas indicatrix The whole genome (4,639,375 bp) harbors 4,108 protein-encoding genes, including 3,029 genes with assigned functions., (Copyright © 2021 Friedrich et al.)

The AI research community has recently been intensely focused on diagnosing COVID-19 by applying deep learning technology to the X-ray scans taken of COVID-19 patients. Differentiating COVID-19 from other pneumonia-inducing illnesses is a highly challenging task as it shares many of the same imaging characteristics as other pulmonary diseases. This is especially true given the small number of COVID-19 X-rays that are publicly available. Deep learning experts commonly use transfer learning to offset the small number of images typically available in medical imaging tasks. Our COV-SNET model is a deep neural network that was pretrained on over one hundred thousand X-ray images. In this paper, we designed two COV-SNET models with the purpose of diagnosing COVID-19. The experimental results demonstrate the robustness of our deep learning models, ultimately achieving sensitivities of 95% for our three-class and two-class models. We also discuss the strengths and weaknesses of such an approach, focusing mainly on the limitations of public X-ray datasets on current COVID-19 deep learning models. Finally, we conclude with possible future directions for this research., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2021 The Authors.)

Here, we present pRH030, a new CRISPR-Cas9 tool for the genetic engineering of Bacillus phages and beyond. It is based on the Streptococcus pyogenes cas9 with its native constitutive promoter, tracrRNA, and a gRNA precursor. The constitutive expression of Cas9 was conducive to the inactivation of viral attackers and enhanced phage mutagenesis efficiency up to 100%. The gRNA precursor can be built up to an artificial CRISPR array with up to 5 spacers (target sequences) assembled from ordinary oligonucleotides and directly cloned into pRH030. Required time and resources remain comparable to a single gRNA cloning. These properties make pRH030 an attractive new system for the modification of Bacillus phages and qualify it for research beyond genetic construction., (© 2020 The Authors. Letters in Applied Microbiology published by John Wiley & Sons Ltd on behalf of Society for Applied Microbiology.)

Many filamentous vibriophages encode virulence genes that lead to the emergence of pathogenic bacteria. Most genomes of filamentous vibriophages characterized up until today were isolated from human pathogens. Despite genome-based predictions that environmental Vibrios also contain filamentous phages that contribute to bacterial virulence, empirical evidence is scarce. This study aimed to characterize the bacteriophages of a marine pathogen, Vibrio alginolyticus (Kiel- alginolyticus ecotype) and to determine their role in bacterial virulence. To do so, we sequenced the phage-containing supernatant of eight different V. alginolyticus strains, characterized the phages therein and performed infection experiments on juvenile pipefish to assess their contribution to bacterial virulence. We were able to identify two actively replicating filamentous phages. Unique to this study was that all eight bacteria of the Kiel- alginolyticus ecotype have identical bacteriophages, supporting our previously established theory of a clonal expansion of the Kiel- alginolyticus ecotype. We further found that in one of the two filamentous phages, two phage-morphogenesis proteins (Zot and Ace) share high sequence similarity with putative toxins encoded on the Vibrio cholerae phage CTXΦ. The coverage of this filamentous phage correlated positively with virulence (measured in controlled infection experiments on the eukaryotic host), suggesting that this phage contributes to bacterial virulence.

A number of amines with three bulky alkyl groups at the nitrogen, which surpass the steric crowding of triisopropylamine considerably, were prepared by using different synthetic methods. It turned out that treatment of N -chlorodialkylamines with organometallic compounds, for example, Grignard reagents, in the presence of a major excess of tetramethylenediamine offered the most effective access to the target compounds. The limits of this method were also tested. The trialkylamines underwent a dealkylation reaction, depending on the degree of steric stress, even at ambient temperature. Because olefins were formed in this transformation, it showed some similarity with the Hofmann elimination. However, the thermal decay of sterically overcrowded tertiary amines was not promoted by bases. Instead, this reaction was strongly accelerated by protic conditions and even by trace amounts of water. Reaction mechanisms, which were analyzed with the help of quantum chemical calculations, are suggested to explain the experimental results.

We present the first two complete genomes of the Janthinobacterium lividum species, namely strains EIF1 and EIF2, which both possess the ability to synthesize violacein. The violet pigment violacein is a secondary metabolite with antibacterial, antifungal, antiviral, and antitumoral properties. Both strains were isolated from environmental oligotrophic water ponds in Göttingen. The strains were phylogenetically classified by average nucleotide identity (ANI) analysis and showed a species assignment to J. lividum with 97.72% (EIF1) and 97.66% (EIF2) identity. These are the first complete genome sequences of strains belonging to the species J. lividum. The genome of strain EIF1 consists of one circular chromosome (6,373,589 bp) with a GC-content of 61.98%. The genome contains 5,551 coding sequences, 122 rRNAs, 93 tRNAs, and 1 tm-RNA. The genome of EIF2 comprises one circular chromosome (6,399,352 bp) with a GC-content of 61.63% and a circular plasmid p356839 (356,839 bp) with a GC-content of 57.21%. The chromosome encodes 5,691 coding sequences, 122 rRNAs, 93 tRNAs, and 1 tm-RNA and the plasmid harbors 245 coding sequences. In addition to the highly conserved chromosomally encoded violacein operon, the plasmid comprises a nonribosomal peptide synthetase cluster with similarity to xenoamicin, which is a bioactive compound effective against protozoan parasites., (© The Author(s) 2020. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.)

Here, we present the complete genome sequence of the Bacillus subtilis strain SP1. This strain is a descendant of the laboratory strain 168. The strain is suitable for biotechnological applications because the prototrophy for tryptophan has been restored. Due to laboratory cultivation, the strain has acquired 24 additional sequence variations., (Copyright © 2020 Richts et al.)

Nitrilases are industrially important biocatalysts due to their ability to degrade nitriles to carboxylic acids and ammonia. In this study, a workflow for simple and fast recovery of nitrilase candidates from metagenomes is presented. For identification of active enzymes, a NADH-coupled high-throughput assay was established. Purification of enzymes could be omitted as the assay is based on crude extract containing the expressed putative nitrilases. In addition, long incubation times were avoided by combining nitrile and NADH conversion in a single reaction. This allowed the direct measurement of nitrile degradation and provided not only insights into substrate spectrum and specificity but also in degradation efficiency. The novel assay was used for investigation of candidate nitrilase-encoding genes. Seventy putative nitrilase-encoding gene and the corresponding deduced protein sequences identified during sequence-based screens of metagenomes derived from nitrile-treated microbial communities were analyzed. Subsequently, the assay was applied to 13 selected candidate genes and proteins. Six of the generated corresponding Escherichia coli clones produced nitrilases that showed activity and one unusual nitrilase was purified and analyzed. The activity of the novel arylacetonitrilase Nit09 exhibited a broad pH range and a high long-term stability. The enzyme showed high activity for arylacetonitriles with a K M of 1.29 mM and a V max of 13.85 U/mg protein for phenylacetonitrile. In conclusion, we provided a setup for simple and rapid analysis of putative nitrilase-encoding genes from sequence to function. The suitability was demonstrated by identification, isolation, and characterization of the arylacetonitrilase. KEY POINTS: • A simple and fast high-throughput nitrilase screening was developed. • A set of putative nitrilases was successfully screened with the assay. • A novel arylacetonitrilase was identified, purified, and characterized in detail.

Purpose: Os acromiale is a common entity in the middle-age group, in whom it is frequently associated with rotator cuff tears. However, it can be a cause of shoulder pain in the young athletes. We want to increase awareness of this pathology that may occultly affect the young athlete as well as to present the results of a perfusion-preserving arthrodesis., Methods: Four consecutive young patients (17-21 years old) with a history of at least 6 months of unrecognized shoulder pain were surgically treated for os acromiale. Through a superior approach, stabilization of the neo-joint by means of cannulated screws and autogenic graft augmentation was performed., Results: Union of the os acromiale was achieved in all the patients. They had an excellent functional outcome, reaching all the maximum Simple Shoulder Test (12) and Oxford shoulder Score (48) scores. All the patients were able to return to their previous sports level., Conclusion: Awareness of the os acromiale in the young athlete, appropriate clinical examination, and image studies are crucial to confirm diagnosis. Surgical treatment aiming at fusion in situ has shown excellent result., (© 2020 The Author(s).)

We present here the complete genome sequences of plant growth-promoting Klebsiella sp. strain MPUS7, Serratia sp. strain NGAS9, and Citrobacter sp. strain LUTT5, isolated from rhizosphere soils and tubers of potato ( Solanum tuberosum L.) plants growing in the northern and southern highlands of Tanzania., (Copyright © 2020 Aloo et al.)

Bacillus velezensis FZB42 is a Gram-positive, endospore-forming rhizobacterium that is associated with plant roots and promotes plant growth. It was used as host to isolate phage vB&#95;BveM-Goe7 (Goe7). Goe7 exhibits a Myoviridae morphology with a contractile tail and an icosahedral head. Its genome is 158,674 bp in size and contains 5137-bp-long terminal repeats (LTRs). It also contains five tRNA-encoding genes and 251 coding DNA sequences (CDS), of which 65 were annotated. The adsorption constant of Goe7 is 6.1 ± 0.24 × 10 -8  ml/min, with a latency period of 75 min and a burst size of 114 particles per burst. A BLASTn sequence comparison against the non-redundant nucleotide database of NCBI revealed that Goe7 is most similar to Bacillus subtilis phage vB&#95;BsuM-Goe3.

vB&#95;BmeM-Goe8 is a phage preying on Bacillus megaterium. Its genome has a GC content of 38.9%, is 161,583 bp in size, and has defined ends consisting of 7436-bp-long terminal repeats. It harbours 11 genes encoding tRNAs and 246 coding DNA sequences, 66 of which were annotated. The particle reveals Myoviridae morphology, and the formation of a double baseplate upon tail sheath contraction indicates morphological relatedness to the group of SPO1-like phages. BLASTn comparison against the NCBI non-redundant nucleotide database revealed that Bacillus phage Mater is the closest relative of vB&#95;BmeM-Goe8.

Bacillus thuringiensis (Bt) is non-pathogenic for humans and serves as a biological control agent in agriculture. Understanding its phages will help to prevent industrial production loss of Bt products and will lead to a better understanding of phages in general. The complete genome of the new B. thuringiensis phage isolate vB&#95;BthM-Goe5 (Goe5) was sequenced, revealing a linear 157,804-bp-long dsDNA chromosome flanked by 2579-bp-long terminal repeats. It contains two tRNAs and 272 protein coding regions, 69 of which could be assigned with an annotation. Morphological investigation, using transmission electron microscopy, revealed Myoviridae morphology. The formation of a double baseplate upon tail sheath contraction indicates a link to the group of SPO1-related phages. Comparative genomics with all Bacillus-related viral genomes available in the NCBI genome database during this investigation indicated that Goe5 was a unique isolate, with Bacillus phage Bastille as its closest relative.

We present the recently isolated virus vB&#95;BthP-Goe4 infecting Bacillus thuringiensis HD1. Morphological investigation via transmission electron microscopy revealed key characteristics of the genus Phi29virus , but with an elongated head resulting in larger virion particles of approximately 50 nm width and 120 nm height. Genome sequencing and analysis resulted in a linear phage chromosome of approximately 26 kb, harbouring 40 protein-encoding genes and a packaging RNA. Sequence comparison confirmed the relation to the Phi29virus genus and genomes of other related strains. A global average nucleotide identity analysis of all identified φ29-like viruses revealed the formation of several new groups previously not observed. The largest group includes Goe4 and may significantly expand the genus Phi29virus ( Salasvirus ) or the Picovirinae subfamily.

Phages are currently under discussion as a solution for the antibiotic crisis, as they may cure diseases caused by multi-drug-resistant pathogens. However, knowledge of phage biology and genetics is limited, which impedes risk assessment of therapeutic applications. In order to enable advances in phage genetic research, the aim of this work was to create a toolkit for simple and fast genetic engineering of phages recruiting Bacillus subtilis as host system. The model organism B. subtilis represents a non-pathogenic surrogate of its harmful relatives, such as Bacillus anthracis or Bacillus cereus . This toolkit comprises the application CutSPR, a bioinformatic tool for rapid primer design, and facilitates the cloning of specific CRISPR-Cas9-based mutagenesis plasmids. The employment of the prophage-free and super-competent B. subtilis TS01 strain enables an easy and fast introduction of specific constructs for in vivo phage mutagenesis. Clean gene deletions and a functional clean gene insertion into the genome of the model phage vB&#95;BsuP-Goe1 served as proof of concept and demonstrate reliability and high efficiency. The here presented toolkit allows comprehensive investigation of the diverse phage genetic pool, a better understanding of phage biology, and safe phage applications.

Background: Operative management of scapular body fractures, when indicated, typically involves extensive exposure through a posterior approach. We present our experience with a deltoid preserving approach that allows excellent exposure of the fracture lines for reduction and fixation while minimizing muscle detachment and overall tissue trauma., Technique: Exposure of the scapula was obtained through a posterior incision. The posterior deltoid was exposed and retracted superiorly while the arm was abducted in accordance with Brodsky et al. The scapula was exposed in the interval between infraspinatus and teres minor., Patients and Methods: Six patients were treated using this approach and were retrospectively reviewed. All were men with a mean age of 34 years (range 24-45 ± 6.7 years). The injuries involved two 14-A3.1 and four 14-A3.2 AO/OTA types of fractures. The mean follow-up after surgery was 28 months (range 21-36 ± 4.93 months)., Results: All fractures could be anatomically reduced and healed without compromise. The mean Constant score was 93.8 (range 91-97 ± 2.13), while range of motion and strength returned to levels equal to the uninjured shoulder. All patients returned to their previous level of activity. We did not observe atrophy of the posterior muscles or hardware complications, and none required hardware removal., Conclusion: The deltoid and external rotators preserving posterior approach permitted good visualization of the fractures while allowing reduction and fixation without extensive muscular dissection and provided excellent functional outcomes. We consider that it offers obvious advantages over more aggressive muscle detaching approaches., Level of Evidence: Therapeutic study, IV.

The new virus vB&#95;BveP-Goe6 was isolated on the host organism Bacillus velezensis FZB42. The virus morphology indicated its association with the genus Phi29virus The genome of vB&#95;BveP-Goe6 (19,105 bp) comprises a linear chromosome with a GC content of 39.99%. The genome harbors 26 putative protein-coding genes and a noncoding packaging RNA., (Copyright © 2018 Schilling et al.)

The species Bacillus licheniformis includes important strains that are used in industrial production processes. Currently the physiological model used to adapt these processes is based on the closely related model organism B. subtilis. In this study we found that both organisms reveal significant differences in the regulation of subtilisin, their main natural protease and a product of industrial fermentation processes. We identified and characterized a novel antisense sRNA AprAs, which represents an RNA based repressor of apr, the gene encoding for the industrial relevant subtilisin protease. Reduction of the AprAs level leads to an enhanced proteolytic activity and an increase of Apr protein expression in the mutant strain. A vector based complementation of the AprAs deficient mutant confirmed this effect and demonstrated the necessity of cis transcription for full efficiency. A comparative analysis of the corresponding genome loci from B. licheniformis and B. subtilis revealed the absence of an aprAs promoter in B. subtilis and indicates that AprAs is a B. licheniformis species specific phenomenon. The discovery of AprAs is of great biotechnological interest since subtilisin Carlsberg is one of the main products of industrial fermentation by B. licheniformis.

The Spounavirinae viruses are ubiquitous in nature and have an obligatory virulent lifestyle. They infect Firmicutes , a bacterial phylum containing an array of environmental non-pathogenic and pathogenic organisms. To expand the knowledge of this viral subfamily, new strains were isolated and investigated in this study. Here we present two new viruses, vB&#95;BsuM-Goe2 and vB&#95;BsuM-Goe3, isolated from raw sewage and infecting Bacillus species. Both were morphologically classified via transmission electron microscopy (TEM) as members of the Spounavirinae subfamily belonging to the Myoviridae family. Genomic sequencing and analyses allowed further affiliation of vB&#95;BsuM-Goe2 to the SPO1-like virus group and vB&#95;BsuM-Goe3 to the Bastille-like virus group. Experimentally determined adsorption constant, latency period, burst size and host range for both viruses revealed different survival strategies. Thus vB&#95;BsuM-Goe2 seemed to rely on fewer host species compared to vB&#95;BsuM-Goe3, but efficiently recruits those. Stability tests pointed out that both viruses are best preserved in LB-medium or TMK-buffer at 4 or 21 °C, whereas cryopreservation strongly reduced viability.

The properties of ferroelectric domain walls can significantly differ from those of their parent material. Elucidating their internal structure is essential for the design of advanced devices exploiting nanoscale ferroicity and such localized functional properties. Here, we probe the internal structure of 180° ferroelectric domain walls in lead zirconate titanate (PZT) thin films and lithium tantalate bulk crystals by means of second-harmonic generation microscopy. In both systems, we detect a pronounced second-harmonic signal at the walls. Local polarimetry analysis of this signal combined with numerical modelling reveals the existence of a planar polarization within the walls, with Néel and Bloch-like configurations in PZT and lithium tantalate, respectively. Moreover, we find domain wall chirality reversal at line defects crossing lithium tantalate crystals. Our results demonstrate a clear deviation from the ideal Ising configuration that is traditionally expected in uniaxial ferroelectrics, corroborating recent theoretical predictions of a more complex, often chiral structure.