Polysphaeroides filiformis, a proterozoic cyanobacterial microfossil and implications for cyanobacteria evolution.

Deciphering the fossil record of cyanobacteria is crucial to understand their role in the chemical and biological evolution of the early Earth. They profoundly modified the redox conditions of early ecosystems more than 2.4 Ga ago, the age of the Great Oxidation Event (GOE), and provided the ancestor of the chloroplast by endosymbiosis, leading the diversification of photosynthetic eukaryotes. Here, we analyze the morphology, ultrastructure, chemical composition, and metals distribution of Polysphaeroides filiformis from the 1040-1006 Ma Mbuji-Mayi Supergroup (DR Congo). We evidence trilaminar and bilayered ultrastructures for the sheath and the cell wall, respectively, and the preservation of Ni-tetrapyrrole moieties derived from chlorophyll in intracellular inclusions. This approach allows an unambiguous interpretation of P. filiformis as a branched and multiseriate photosynthetic cyanobacterium belonging to the family of Stigonemataceae. It also provides a possible minimum age for the emergence of multiseriate true branching nitrogen-fixing and probably heterocytous cyanobacteria.

[How could forensic psychiatry be made more attractive?] / Hogyan lehetne az igazságügyi pszichiátriát vonzóbbá tenni?

INTRODUCTION: Decrease in the number of forensic psychiatric experts recently reached a critical level. Shortage of forensic experts caused difficulties in the health care as well as in the justice system. OBJECTIVE: Surveying of how the field of forensic psychiatry and the forensic psychiatric expert work can be made more attractive. METHOD: We performed an online survey among forensic psychiatric experts and specialists in psychiatry. We complied a questionnaire that beside demographic data contained questions in 4 areas. The first 2 areas of the questionnaire - simplifying the forensic psychiatry training (10 items), decreasing the cost of the forensic psychiatry training (5 items) - consisted of simple tatements. Responders had to indicate on a 10-point Likert scale their level of agreement. In the last 2 areas, participants were asked to describe in unstructured format whether they see any circumstance that makes forensic psychiatry attractive or unattractive. The questionnaire was sent out in electronic form to the forensic psychiatric experts on the mailing list of the Hungarian Forensic Expert Chamber, and all specialists in psychiatry on the mailing list of the Hungarian Medical Chamber. RESULTS: Altogether 171 persons filled in the questionnaire with a mean age of 57.26 ± 11.57 years. There were 122 (71.3%) females among the participants. The following proposals received the highest ratings from the forensic psychiatric experts as well as from the specialists in psychiatry: increasing the number of the training institutes; decreasing the costs of the training; making the Hungarian Forensic Expert Chamber course free of charge; and introducing a stipend to cover the costs of the training. DISCUSSION: Results of the survey indicate that active forensic psychiatric experts and specialists in psychiatry representing a potential professional resource, concur that forensic psychiatry training can be made more attractive mainly with increasing the number of training institutes and decreasing the total costs of the training which is compiled from different constituents. CONCLUSION: Due to the critical lack of necessary professionals, urgent action is needed to make forensic psychiatry more popular and the forensic psychiatry qualification easier to obtain. Orv Hetil. 2023; 164(35): 1373-1380.

Blooming of a microbial community in an Ediacaran extreme volcanic lake system.

Ancient aquatic sediments are critical archives for studying early microbial life and the types of environments in which they thrived. The recently characterized Amane Tazgart microbialites in the Anti-Atlas, Morocco, are a rare and well-preserved non-marine deposit that evolved in an alkaline volcanic lake setting during the Ediacaran Period. A multiproxy geochemical toolbox reveals evidence pointing to spatio-temporal ecosystem organization and succession related to changing lake water chemistry. This is marked by secular transition from a cold/dry climate, hypersaline alkaline thermophilic and anoxic-oxic community, to a stable state warm/wet climate fully oxygenated fresh to brackish water ecosystem, predominated by oxygenic stromatolites. Extreme dissolved Arsenic concentrations suggest that these polyextremophiles required robust detoxification mechanisms to circumvent arsenic toxicity and phosphate deficiency. We propose that self-sustaining and versatile anoxic to oxic microbial ecosystems thrived in aquatic continental settings during the Ediacaran Period, when complex life co-evolved with a rise in atmospheric oxygen content.

Therapy for Temporomandibular Disorders: 3D-Printed Splints from Planning to Evaluation.

INTRODUCTION: This article describes the authors' digital workflow-based method for fabricating intraoral occlusal splints, from planning to the evaluation phase. MATERIALS AND METHODS: In our protocol, first, we had a registration phase. This included taking digital impressions, determining the centric relation (CR) position with the deprogrammer Luci Jig, and using the digital facebow for measuring the individual values. The laboratory phase was next, which included planning and manufacturing with a 3D printer. The last phase was delivery, when we checked the stability of the splint and adjusted the occlusal part. RESULT: The average cost is lower for a fully digital splint than for conventional methods. In terms of time, there was also a significant difference between the classic and digital routes. From a dental technical point of view, the execution was much more predictable. The printed material was very rigid and, therefore, fragile. Compared to the analog method, the retention was much weaker. CONCLUSION: The presented method permits time-efficient laboratory production, and may also be performed chairside in a dental office. The technology is perfectly applicable to everyday life. In addition to its many beneficial properties, its negative properties must also be highlighted.

Unmasking pipefish otolith using synchrotron-based scanning X-ray fluorescence.

Scientists use otoliths to trace fish life history, especially fish migrations. Otoliths incorporate signatures of individual growth and environmental use. For many species, distinct increment patterns in the otolith are difficult to discern; thus, questions remain about crucial life history information. To unravel the history of such species, we use synchrotron-based scanning X-ray fluorescence. It allows the mapping of elements on the entire otolith at a high spatial resolution. It gives access to precise fish migration history by tagging landmark signature for environmental transition and it also characterises localised growth processes at a mineral level. Freshwater pipefish, which are of conservation concern, have otoliths that are small and fragile. Growth increments are impossible to identify and count; therefore, there is a major lack of knowledge about their life history. We confirm for the first time, by mapping strontium that the two tropical pipefish species studied are diadromous (transition freshwater/marine/freshwater). Mapping of other elements uncovered the existence of different migratory routes during the marine phase. Another major breakthrough is that we can chemically count growth increments solely based on sulphur signal as it is implicated in biomineralization processes. This novel method circumvents reader bias issues and enables age estimation even for otoliths with seemingly untraceable increments. The high spatial resolution elemental mapping methods push back limits of studies on life traits or stock characterisation.

Towards routine 3D characterization of intact mesoscale samples by multi-scale and multimodal scanning X-ray tomography.

Non-invasive multi-scale and multimodal 3D characterization of heterogeneous or hierarchically structured intact mesoscale samples is of paramount importance in tackling challenging scientific problems. Scanning hard X-ray tomography techniques providing simultaneous complementary 3D information are ideally suited to such studies. However, the implementation of a robust on-site workflow remains the bottleneck for the widespread application of these powerful multimodal tomography methods. In this paper, we describe the development and implementation of such a robust, holistic workflow, including semi-automatic data reconstruction. Due to its flexibility, our approach is especially well suited for on-the-fly tuning of the experiments to study features of interest progressively at different length scales. To demonstrate the performance of the method, we studied, across multiple length scales, the elemental abundances and morphology of two complex biological systems, Arabidopsis plant seeds and mouse renal papilla samples. The proposed approach opens the way towards routine multimodal 3D characterization of intact samples by providing relevant information from pertinent sample regions in a wide range of scientific fields such as biology, geology, and material sciences.

Intracellular bound chlorophyll residues identify 1 Gyr-old fossils as eukaryotic algae.

The acquisition of photosynthesis is a fundamental step in the evolution of eukaryotes. However, few phototrophic organisms are unambiguously recognized in the Precambrian record. The in situ detection of metabolic byproducts in individual microfossils is the key for the direct identification of their metabolisms. Here, we report a new integrative methodology using synchrotron-based X-ray fluorescence and absorption. We evidence bound nickel-geoporphyrins moieties in low-grade metamorphic rocks, preserved in situ within cells of a ~1 Gyr-old multicellular eukaryote, Arctacellularia tetragonala. We identify these moieties as chlorophyll derivatives, indicating that A. tetragonala was a phototrophic eukaryote, one of the first unambiguous algae. This new approach, applicable to overmature rocks, creates a strong new proxy to understand the evolution of phototrophy and diversification of early ecosystems.

Cellular Detection of a Mitochondria Targeted Brominated Vinyl Triphenylamine Optical Probe (TP-Br) by X-Ray Fluorescence Microscopy.

Triphenylamine (TP) derivatives such as two-branch cationic vinylbenzimidazolium triphenylamine TP-2Bzim are promising turn-on fluorescent probes suitable for two-photon imaging, labelling mitochondria in live cells. Here, we designed two TP-2Bzim derivatives as bimodal probes suitable for X-ray fluorescence imaging. The conjugation of the TP core with a rhenium tricarbonyl moiety in the TP-RePyta probe altered the localisation in live cells from mitochondria to lysosomes. The introduction of bromine on the TP core generated the TP-Br probe retaining good photophysical properties and mitochondria labelling in live cells. The influence of calcium channels in the uptake of TP-Br was studied. Synchrotron Radiation X-ray Fluorescence (SXRF) imaging of bromine enabled the detection of TP-Br and suggested a negligible presence of the probe in an unbound state in the incubated cells, a crucial point in the development of these probes. This study paves the way towards the development of TP probes as specific organelle stainers suitable for SXRF imaging.

Catabolism of lysosome-related organelles in color-changing spiders supports intracellular turnover of pigments.

Pigment organelles of vertebrates belong to the lysosome-related organelle (LRO) family, of which melanin-producing melanosomes are the prototypes. While their anabolism has been extensively unraveled through the study of melanosomes in skin melanocytes, their catabolism remains poorly known. Here, we tap into the unique ability of crab spiders to reversibly change body coloration to examine the catabolism of their pigment organelles. By combining ultrastructural and metal analyses on high-pressure frozen integuments, we first assess whether pigment organelles of crab spiders belong to the LRO family and second, how their catabolism is intracellularly processed. Using scanning transmission electron microscopy, electron tomography, and nanoscale Synchrotron-based scanning X-ray fluorescence, we show that pigment organelles possess ultrastructural and chemical hallmarks of LROs, including intraluminal vesicles and metal deposits, similar to melanosomes. Monitoring ultrastructural changes during bleaching suggests that the catabolism of pigment organelles involves the degradation and removal of their intraluminal content, possibly through lysosomal mechanisms. In contrast to skin melanosomes, anabolism and catabolism of pigments proceed within the same cell without requiring either cell death or secretion/phagocytosis. Our work hence provides support for the hypothesis that the endolysosomal system is fully functionalized for within-cell turnover of pigments, leading to functional maintenance under adverse conditions and phenotypic plasticity. First formulated for eye melanosomes in the context of human vision, the hypothesis of intracellular turnover of pigments gets unprecedented strong support from pigment organelles of spiders.

Correlative optical photothermal infrared and X-ray fluorescence for chemical imaging of trace elements and relevant molecular structures directly in neurons.

Alzheimer's disease (AD) is the most common cause of dementia, costing about 1% of the global economy. Failures of clinical trials targeting amyloid-ß protein (Aß), a key trigger of AD, have been explained by drug inefficiency regardless of the mechanisms of amyloid neurotoxicity, which are very difficult to address by available technologies. Here, we combine two imaging modalities that stand at opposite ends of the electromagnetic spectrum, and therefore, can be used as complementary tools to assess structural and chemical information directly in a single neuron. Combining label-free super-resolution microspectroscopy for sub-cellular imaging based on novel optical photothermal infrared (O-PTIR) and synchrotron-based X-ray fluorescence (S-XRF) nano-imaging techniques, we capture elemental distribution and fibrillary forms of amyloid-ß proteins in the same neurons at an unprecedented resolution. Our results reveal that in primary AD-like neurons, iron clusters co-localize with elevated amyloid ß-sheet structures and oxidized lipids. Overall, our O-PTIR/S-XRF results motivate using high-resolution multimodal microspectroscopic approaches to understand the role of molecular structures and trace elements within a single neuronal cell.

Cytoplasmic aggregation of uranium in human dopaminergic cells after continuous exposure to soluble uranyl at non-cytotoxic concentrations.

Uranium exposure can lead to neurobehavioral alterations in particular of the monoaminergic system, even at non-cytotoxic concentrations. However, the mechanisms of uranium neurotoxicity after non-cytotoxic exposure are still poorly understood. In particular, imaging uranium in neurons at low intracellular concentration is still very challenging. We investigated uranium intracellular localization by means of synchrotron X-ray fluorescence imaging with high spatial resolution (< 300 nm) and high analytical sensitivity (< 1 µg.g-1 per 300 nm pixel). Neuron-like SH-SY5Y human cells differentiated into a dopaminergic phenotype were continuously exposed, for seven days, to a non-cytotoxic concentration (10 µM) of soluble natural uranyl. Cytoplasmic submicron uranium aggregates were observed accounting on average for 62 % of the intracellular uranium content. In some aggregates, uranium and iron were co-localized suggesting common metabolic pathways between uranium and iron storage. Uranium aggregates contained no calcium or phosphorous indicating that detoxification mechanisms in neuron-like cells are different from those described in bone or kidney cells. Uranium intracellular distribution was compared to fluorescently labeled organelles (lysosomes, early and late endosomes) and to fetuin-A, a high affinity uranium-binding protein. A strict correlation could not be evidenced between uranium and the labeled organelles, or with vesicles containing fetuin-A. Our results indicate a new mechanism of uranium cytoplasmic aggregation after non-cytotoxic uranyl exposure that could be involved in neuronal defense through uranium sequestration into less reactive species. The remaining soluble fraction of uranium would be responsible for protein binding and for the resulting neurotoxic effects.

Intracellular location matters: rationalization of the anti-inflammatory activity of a manganese(ii) superoxide dismutase mimic complex.

A conjugate of a Mn-based superoxide dismutase mimic with a Re-based multimodal probe 1[combining low line] was studied in a cellular model of oxidative stress. Its speciation was investigated using Re and Mn X-fluorescence. Interestingly, 1[combining low line] shows a distribution different from its unconjugated analogue but a similar concentration in mitochondria and a similar bioactivity.

Anti-inflammatory activity of superoxide dismutase mimics functionalized with cell-penetrating peptides.

A superoxide dismutase mimic (Mn1) was functionalized with three positively charged-peptides: RRRRRRRRR (Mn1-R9), RRWWWRRWRR (Mn1-RW9) or Fx-r-Fx-K (Mn1-MPP). Characterization of the physico-chemical properties of the complexes show that they share similar binding affinity for Mn2+, apparent reduction potential and intrinsic superoxide dismutase activity. However, their accumulation in cells is different (Mn1-R9 < Mn1-MPP < Mn1-RW9 < Mn1), as well as their subcellular distribution. In addition, the three functionalized-complexes display a better anti-inflammatory activity than Mn1 when assayed at 10 µM. This improvement is due to a combination of an anti-inflammatory effect of the peptidyl moiety itself, and of the SOD mimic for Mn1-RW9 and Mn1-MPP. In contrast, the enhanced anti-inflammatory activity of Mn1-R9 is solely due to the SOD mimic.

Manganese Mapping Using a Fluorescent Mn2+ Sensor and Nanosynchrotron X-ray Fluorescence Reveals the Role of the Golgi Apparatus as a Manganese Storage Site.

Elucidating dynamics in transition-metal distribution and localization under physiological and pathophysiological conditions is central to our understanding of metal-ion regulation. In this Forum Article, we focus on manganese and specifically recent developments that point to the relevance of the Golgi apparatus in manganese detoxification when this essential metal ion is overaccumulated because of either environmental exposure or mutations in manganese efflux transporters. In order to further evaluate the role of the Golgi apparatus as a manganese-ion storage compartment under subcytotoxic manganese levels, we use a combination of confocal microscopy using a sensitive "turn-on" fluorescent manganese sensor, M1, and nanosynchrotron X-ray fluorescence imaging to show that manganese ions are stored in the Golgi apparatus under micromolar manganese exposure concentrations. Our results, along with previous reports on manganese accumulation, now indicate a central role of the Golgi apparatus in manganese storage and trafficking under subcytotoxic manganese levels and hint toward a possible role of the Golgi apparatus in manganese storage even under physiological conditions.

Microbially induced potassium enrichment in Paleoproterozoic shales and implications for reverse weathering on early Earth.

Illitisation requires potassium incorporation into a smectite precursor, a process akin to reverse weathering. However, it remains unclear whether microbes facilitate K+ uptake to the sediments and whether illitisation was important in the geological past. The 2.1 billion-year-old Francevillian Series of Gabon has been shown to host mat-related structures (MRS) and, in this regard, these rocks offer a unique opportunity to test whether ancient microbes induced illitisation. Here, we show high K content confined to illite particles that are abundant in the facies bearing MRS, but not in the host sandstone and black shale. This observation suggests that microbial biofilms trapped K+ from the seawater and released it into the pore-waters during respiration, resulting in illitisation. The K-rich illite developed exclusively in the fossilized MRS thus provides a new biosignature for metasediments derived from K-feldspar-depleted rocks that were abundant crustal components on ancient Earth.

Author Correction: Microbially induced potassium enrichment in Paleoproterozoic shales and implications for reverse weathering on early Earth.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Graftable SCoMPIs enable the labeling and X-ray fluorescence imaging of proteins.

Bio-imaging techniques alternative to fluorescence microscopy are gaining increasing interest as complementary tools to visualize and analyze biological systems. Among them, X-ray fluorescence microspectroscopy provides information on the local content and distribution of heavy elements (Z ≥ 14) in cells or biological samples. In this context, similar tools to those developed for fluorescence microscopy are desired, including chemical probes or tags. In this work, we study rhenium complexes as a convenient and sensitive probe for X-ray fluorescence microspectroscopy. We demonstrate their ability to label and sense exogenously incubated or endogenous proteins inside cells.

Rapid and reliable diagnosis of Wilson disease using X-ray fluorescence.

Wilson's disease (WD) is a rare autosomal recessive disease due to mutations of the gene encoding the copper-transporter ATP7B. The diagnosis is hampered by the variability of symptoms induced by copper accumulation, the inconstancy of the pathognomonic signs and the absence of a reliable diagnostic test. We investigated the diagnostic potential of X-ray fluorescence (XRF) that allows quantitative analysis of multiple elements. Studies were performed on animal models using Wistar rats (n = 10) and Long Evans Cinnamon (LEC) rats (n = 11), and on human samples including normal livers (n = 10), alcohol cirrhosis (n = 8), haemochromatosis (n = 10), cholestasis (n = 6) and WD (n = 22). XRF experiments were first performed using synchrotron radiation to address the elemental composition at the cellular level. High-resolution mapping of tissue sections allowed measurement of the intensity and the distribution of copper, iron and zinc while preserving the morphology. Investigations were further conducted using a laboratory X-ray source for irradiating whole pieces of tissue. The sensitivity of XRF was highlighted by the discrimination of LEC rats from wild type even under a regimen using copper deficient food. XRF on whole formalin-fixed paraffin embedded needle biopsies allowed profiling of the elements in a few minutes. The intensity of copper related to iron and zinc significantly discriminated WD from other genetic or chronic liver diseases with 97.6% specificity and 100% sensitivity. This study established a definite diagnosis of Wilson's disease based on XRF. This rapid and versatile method can be easily implemented in a clinical setting.

MMX-I: data-processing software for multimodal X-ray imaging and tomography.

A new multi-platform freeware has been developed for the processing and reconstruction of scanning multi-technique X-ray imaging and tomography datasets. The software platform aims to treat different scanning imaging techniques: X-ray fluorescence, phase, absorption and dark field and any of their combinations, thus providing an easy-to-use data processing tool for the X-ray imaging user community. A dedicated data input stream copes with the input and management of large datasets (several hundred GB) collected during a typical multi-technique fast scan at the Nanoscopium beamline and even on a standard PC. To the authors' knowledge, this is the first software tool that aims at treating all of the modalities of scanning multi-technique imaging and tomography experiments.

Fabrication and characterization of high-efficiency double-sided blazed x-ray optics.

The focusing efficiency of conventional diffractive x-ray lenses is fundamentally limited due to their symmetric binary structures and the corresponding symmetry of their focusing and defocusing diffraction orders. Fresnel zone plates with asymmetric structure profiles can break this limitation; yet existing implementations compromise either on resolution, ease of use, or stability. We present a new way for the fabrication of such blazed lenses by patterning two complementary binary Fresnel zone plates on the front and back sides of the same membrane chip to provide a compact, inherently stable, single-chip device. The presented blazed double-sided zone plates with 200 nm smallest half-pitch provide up to 54.7% focusing efficiency at 6.2 keV, which is clearly beyond the value obtainable by their binary counterparts.