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Production of applicable and homogeneous biochar for soil amendment purposes would benefit from knowledge on how feedstock heterogeneity impacts key biochar pore properties and how the properties are transformed due to pyrolysis. This study aimed (1) to quantify how clonal differences and within-tree heterogeneity of a hybrid aspen feedstock (wood) impact biochar pore properties and (2) to estimate how pore properties of non-pyrolysed wood materials are transformed when pyrolysed into biochar. The study was conducted by collecting samples from a hybrid aspen (Populus tremula L. × Populus tremuloides Michx.) clonal field trial. Key pore properties of non-pyrolysed and pyrolysed wood samples were quantified with 3D X-ray imaging and quantitative image analyses. The results demonstrated how pyrolysis shifted distinctively bi-modal pore size distributions of the wood samples towards smaller pore size regions. The bi-modal wood tissue structure controlled the pore structure also in the biochars. Due to decreasing cell wall thicknesses, the pyrolysis increased the porosity of the materials. While the thermal process homogenized differences in the wall thicknesses, the thicknesses of the feedstock were also shown to control the resulting thicknesses in the biochars. Mechanisms of biochar pore property formation can be considered important when designing applicable biochars for a chosen purpose. Clonal differences and within-tree heterogeneity had a direct impact only on the wall thicknesses and the pore diameters of vessels. These impacts can be of interest when planning feedstock utilization in biochar production. However, the results suggest that relatively homogeneous biochar can be produced from hybrid aspen feedstocks.

Funding Information: The work at Aalto University has been financially supported by the Academy of Finland (COOLCAT consortium, decision no. 329977 and 329978 ; ALDI consortium, decision no. 331082 ). This work made use of Aalto University Bioeconomy, OtaNano and RawMatters infrastructure. Hannu Revitzer (Aalto University) is thanked for the ICP-OES analysis, Aalto workshop people (especially Seppo Jääskeläinen) for working on the reactor modifications. The DFT calculations were made possible by computational resources provided by the CSC — IT Center for Science, Espoo, Finland ( https://www.csc.fi/en/ ) and computer capacity from the Finnish Grid and Cloud Infrastructure (urn:nbn:fi:research-infras-2016072533). The University of Helsinki acknowledges support from Academy of Finland (project 295696 ) as well as ESRF for beamtime and Blanka Detlefs and Christoph Sahle for expert support. Preliminary XANES measurements were performed using the Helsinki Center for X-ray Spectroscopy Hel-XAS instrument under theproposal number 2021–0011. The development of active catalysts for carbon dioxide (CO2) hydrogenation to methanol is intimately related to the creation of effective metal-oxide interfaces. In this work, we investigated how the order of addition of copper and zinc on zirconia influences the catalytic properties, the catalytic activity and selectivity toward methanol. Regarding the carbon dioxide conversion and methanol production, the catalysts on which the promoter (zinc) was atomically deposited after copper impregnation (i.e., ZnO/Cu/ZrO2 and ZnO/Cu/ZnO/ZrO2) were superior catalysts compared to the reverse copper-after-zinc catalyst (Cu/ZnO/ZrO2). Temperature-programmed experiments and in situ diffuse reflectance infrared Fourier transform-spectroscopy (DRIFTS) experiments allowed us to elucidate the benefits of the zinc-after-copper pair to store CO2 as carbonate species and further convert them into formate species, key intermediates in the formation of methanol. This research provides insights into the potential of atomic layer deposition in the development of tailored heterogeneous catalysts for efficient CO2 valorization to methanol.

In this article, we demonstrate the viability of highly monochromatic full-field X-ray absorption near edge structure based tomography using a laboratory-scale Johann-type X-ray absorption spectrometer utilising a conventional X-ray tube source. In this proof-of-concept, by using a phantom embedded with elemental Se, Na$$_2$$ 2 SeO$$_3$$ 3 , and Na$$_2$$ 2 SeO$$_4$$ 4 , we show that the three-dimensional distributions of Se in different oxidation states can be mapped and distinguished from the phantom matrix and each other with absorption edge contrast tomography. The presented method allows for volumetric analyses of chemical speciation in mm-scale samples using low-brilliance X-ray sources, and represents a new analytic tool for materials engineering and research in many fields including biology and chemistry.

Funding Information: This project had received funding from the Academy of Finland (Decision Nos. 265680, 294625, 306845, 308265, and 341361) and EraNet Neuron ACROBAT. Publisher Copyright: Copyright © 2022 Nieminen, Pospelov, Koponen, Yrjölä, Shulga, Khirug and Rivera. Transcranial magnetic stimulation (TMS) is widely applied on humans for research and clinical purposes. TMS studies on small animals, e.g., rodents, can provide valuable knowledge of the underlying neurophysiological mechanisms. Administering TMS on small animals is, however, prone to technical difficulties, mainly due to their small head size. In this study, we aimed to develop an energy-efficient coil and a compatible experimental set-up for administering TMS on rodents. We applied a convex optimization process to develop a minimum-energy coil for TMS on rats. As the coil windings of the optimized coil extend to a wide region, we designed and manufactured a holder on which the rat lies upside down, with its head supported by the coil. We used the set-up to record TMS–electromyography, with electromyography recorded from limb muscles with intramuscular electrodes. The upside-down placement of the rat allowed the operator to easily navigate the TMS without the coil blocking their field of view. With this paradigm, we obtained consistent motor evoked potentials from all tested animals.

Research conducted by the veterinary education community is critical to continual improvement of educational outcomes. Additionally, research productivity is one metric in promotion and tenure decisions. We sought to identify challenges encountered or anticipated when undertaking or planning veterinary educational research (VER), to learn how these challenges might be overcome, and to synthesize tips for success from those who have performed VER. A branching survey was developed and deployed along the authors’ worldwide veterinary education contacts in a cascading manner. The survey collected quantitative and qualitative information from participants who had performed VER and those who planned to perform VER in the future. The 258 participants represented 41 countries. Of the participants, 204 had performed VER (79%) and 54 planned to in the future (21%). The median time spent teaching was 14 years, and median time performing VER was 5 years. The most commonly reported challenges in performing VER were lack of funding, lack of time, and difficulties encountered when undertaking a study, including data collection, analysis, and publishing. When asked about overcoming the challenges, a major theme emerged around people, who provided expertise and mentoring. The most commonly reported tip for success was collaboration; 73% of experienced researchers reported people as most helpful upon beginning VER. Collaborators provided diverse help with ideas, study design, statistics, and other aspects. These results suggest that institutions can offer support to academics in the form of small grants, protected research time, writing workshops, and mentorship to assist with the production of meaningful VER.

Elevated states of brain plasticity typical for critical periods of early postnatal life can be reinstated in the adult brain through interventions, such as antidepressant treatment and environmental enrichment, and induced plasticity may be critical for the antidepressant action. Parvalbumin-positive (PV) interneurons regulate the closure of developmental critical periods and can alternate between high and low plasticity states in response to experience in adulthood. We now show that PV plasticity states and cortical networks are regulated through the activation of TrkB neurotrophin receptors. Visual cortical plasticity induced by fluoxetine, a widely prescribed selective serotonin reuptake inhibitor (SSRI) antidepressant, was lost in mice with reduced expression of TrkB in PV interneurons. Conversely, optogenetic gain-of-function studies revealed that activation of an optically activatable TrkB (optoTrkB) specifically in PV interneurons switches adult cortical networks into a state of elevated plasticity within minutes by decreasing the intrinsic excitability of PV interneurons, recapitulating the effects of fluoxetine. TrkB activation shifted cortical networks towards a low PV configuration, promoting oscillatory synchrony, increased excitatory-inhibitory balance, and ocular dominance plasticity. OptoTrkB activation promotes the phosphorylation of Kv3.1 channels and reduces the expression of Kv3.2 mRNA providing a mechanism for the lower excitability. In addition, decreased expression and puncta of Synaptotagmin2 (Syt2), a presynaptic marker of PV interneurons involved in Ca2+-dependent neurotransmitter release, suggests lower inputs onto pyramidal neurons suppressing feed-forward inhibition. Together, the results provide mechanistic insights into how TrkB activation in PV interneurons orchestrates the activity of cortical networks and mediating antidepressant responses in the adult brain.

BackgroundThe segmentation of 3D cell nuclei is essential in many tasks, such as targeted molecular radiotherapies (MRT) for metastatic tumours, toxicity screening, and the observation of proliferating cells. In recent years, one popular method for automatic segmentation of nuclei has been deep learning enhanced marker-controlled watershed transform. In this method, convolutional neural networks (CNNs) have been used to create nuclei masks and markers, and the watershed algorithm for the instance segmentation. We studied whether this method could be improved for the segmentation of densely cultivated 3D nuclei via developing multiple system configurations in which we studied the effect of edge emphasizing CNNs, and optimized H-minima transform for mask and marker generation, respectively.ResultsThe dataset used for training and evaluation consisted of twelve in vitro cultivated densely packed 3D human carcinoma cell spheroids imaged using a confocal microscope. With this dataset, the evaluation was performed using a cross-validation scheme. In addition, four independent datasets were used for evaluation. The datasets were resampled near isotropic for our experiments. The baseline deep learning enhanced marker-controlled watershed obtained an average of 0.69 Panoptic Quality (PQ) and 0.66 Aggregated Jaccard Index (AJI) over the twelve spheroids. Using a system configuration, which was otherwise the same but used 3D-based edge emphasizing CNNs and optimized H-minima transform, the scores increased to 0.76 and 0.77, respectively. When using the independent datasets for evaluation, the best performing system configuration was shown to outperform or equal the baseline and a set of well-known cell segmentation approaches.ConclusionsThe use of edge emphasizing U-Nets and optimized H-minima transform can improve the marker-controlled watershed transform for segmentation of densely cultivated 3D cell nuclei. A novel dataset of twelve spheroids was introduced to the public.

The use of Li-ion batteries is increasing rapidly. Understanding the processes behind active material aging helps to enhance the materials, and therefore, development of new in situ methods for structural studies is important. In addition, understanding the effect of different synthesis methods on the active material properties is necessary to optimize the material cycle life. In this work, the performance of LiCoO2 doped with Mg during the lithiation step is compared to LiCoO2 prepared using an Mg-doped Co3O4 precursor. In situ laboratory-scale X-ray absorption near-edge spectroscopy is used to analyze the Co valence changes in LiCoO2 to understand the electrochemical behavior of the investigated materials. The maximum reachable Co valence state is found to decrease upon aging, a small decrease indicating a good cycle-life, and this is attributed to the enhanced stacking order, better Mg distribution in the lattice, and fine primary particle size in the material. In the synthesis conditions used in this study, Mg doping during the lithiation step is shown to perform better compared to the precursor doping. Overlithiation is shown to reduce the electrochemical performance of nondoped and precursor-doped LiCoO2 materials but not to affect the cyclability of lithiation-doped LiCoO2. (c) 2022 The Author(s). Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

Background Computed tomography angiography (CTA) imaging is needed in current guideline-based stroke diagnosis, and infarct core size is one factor in guiding treatment decisions. We studied the efficacy of a convolutional neural network (CNN) in final infarct volume prediction from CTA and compared the results to a CT perfusion (CTP)-based commercially available software (RAPID, iSchemaView). Methods We retrospectively selected 83 consecutive stroke cases treated with thrombolytic therapy or receiving supportive care that presented to Helsinki University Hospital between January 2018 and July 2019. We compared CNN-derived ischaemic lesion volumes to final infarct volumes that were manually segmented from follow-up CT and to CTP-RAPID ischaemic core volumes. Results An overall correlation of r = 0.83 was found between CNN outputs and final infarct volumes. The strongest correlation was found in a subgroup of patients that presented more than 9 h of symptom onset (r = 0.90). A good correlation was found between the CNN outputs and CTP-RAPID ischaemic core volumes (r = 0.89) and the CNN was able to classify patients for thrombolytic therapy or supportive care with a 1.00 sensitivity and 0.94 specificity. Conclusions A CTA-based CNN software can provide good infarct core volume estimates as observed in follow-up imaging studies. CNN-derived infarct volumes had a good correlation to CTP-RAPID ischaemic core volumes.

In stroke imaging, CT angiography (CTA) is used for detecting arterial occlusions. These images could also provide information on the extent of ischemia. The study aim was to develop and evaluate a convolutional neural network (CNN)–based algorithm for detecting and segmenting acute ischemic lesions from CTA images of patients with suspected middle cerebral artery stroke. These results were compared to volumes reported by widely used CT perfusion–based RAPID software (IschemaView). A 42-layer-deep CNN was trained on 50 CTA volumes with manually delineated targets. The lower bound for predicted lesion size to reliably discern stroke from false positives was estimated. The severity of false positives and false negatives was reviewed visually to assess the clinical applicability and to further guide the method development. The CNN model corresponded to the manual segmentations with voxel-wise sensitivity 0.54 (95% confidence interval: 0.44–0.63), precision 0.69 (0.60–0.76), and Sørensen–Dice coefficient 0.61 (0.52–0.67). Stroke/nonstroke differentiation accuracy 0.88 (0.81–0.94) was achieved when only considering the predicted lesion size (i.e., regardless of location). By visual estimation, 46% of cases showed some false findings, such as CNN highlighting chronic periventricular white matter changes or beam hardening artifacts, but only in 9% the errors were severe, translating to 0.91 accuracy. The CNN model had a moderately strong correlation to RAPID-reported Tmax > 10 s volumes (Pearson’s r = 0.76 (0.58–0.86)). The results suggest that detecting anterior circulation ischemic strokes from CTA using a CNN-based algorithm can be feasible when accompanied with physiological knowledge to rule out false positives.

Alcohol use disorder (AUD) causes complex alterations in the brain that are poorly understood. The heterogeneity of drinking patterns and the high incidence of comorbid factors compromise mechanistic investigations in AUD patients. Here we used male Marchigian Sardinian alcohol-preferring (msP) rats, a well established animal model of chronic alcohol drinking, and a combination of longitudinal resting-state fMRI and manganese-enhanced MRI to provide objective measurements of brain connectivity and activity, respectively. We found that 1 month of chronic alcohol drinking changed the correlation between resting-state networks. The change was not homogeneous, resulting in the reorganization of pairwise interactions and a shift in the equilibrium of functional connections. We identified two fundamentally different forms of network reorganization. First is functional dedifferentiation, which is defined as a regional increase in neuronal activity and overall correlation, with a concomitant decrease in preferential connectivity between specific networks. Through this mechanism, occipital cortical areas lost their specific interaction with sensory-insular cortex, striatal, and sensorimotor networks. Second is functional narrowing, which is defined as an increase in neuronal activity and preferential connectivity between specific brain networks. Functional narrowing strengthened the interaction between striatal and prefrontocortical networks, involving the anterior insular, cingulate, orbitofrontal, prelimbic, and infralimbic cortices. Importantly, these two types of alterations persisted after alcohol discontinuation, suggesting that dedifferentiation and functional narrowing rendered persistent network states. Our results support the idea that chronic alcohol drinking, albeit at moderate intoxicating levels, induces an allostatic change in the brain functional connectivity that propagates into early abstinence., This research was supported by funding from the European Union Horizon 2020 research and innovation program under Grant 668863 (SyBil-AA); the ERA-Net NEURON framework with grants given to the project “Translational Neuroimaging in Alcoholism” (TRANSALC) by the Academy of Finland (Grant TRANSALC 01EW1112), the Bundesministerium für Bildung und Forschung (Grant FKZ01EW1112), and the Spanish Ministerio de Ciencia e Innovación (Grant PIM2010ERN-00679). S.C. and D.M. were funded by the Spanish Ministerio de Economía y Competitividad (MINECO) cofinanced by the European Regional Development Fund (ERDF) under Grants BFU2015-64380-C2-1-R and BFU2015-64380-C2-2-R, and the Generalitat Valenciana Government through the Prometeo Program (Grant PROMETEO/2019/015). S.C. was further supported by the Spanish Agency of Research (AEI) and ERDF under Grant PGC2018-101055-B-I00 and the Spanish Ministerio de Sanidad, Servicios Sociales e Igualdad (Grant 2017I065). Ú.P.-R. was supported by Grant FPU13/03537 and research stay internship EST15/00516 from the Spanish Ministerio de Educación, Cultura y Deporte (MECD). Further funding was obtained from the Spanish State Research Agency through the Severo Ochoa Program for Centres of Excellence in R&D (Grant SEV- 2017-0723). W.H.S. was further supported by the Deutsche Forschungsgemeinschaft (Center Grant TRR265-B08).

Aims Alcohol exposure during adolescence is associated with both increased risk for alcohol use disorders and anxiety in adulthood. Our present experiments examined this association using alcohol-preferring AA (Alko Alcohol) rats selected for high voluntary alcohol drinking. Methods Two groups of female AA rats acquired alcohol drinking at different ages. We gave the adolescent-onset group free choice to 10% alcohol and water for seven weeks, starting on post-natal day 42 (PND 42), whereas the adult-onset group started drinking alcohol on PND 112. After the 7-week drinking, we withdrew the adolescent group from alcohol for two weeks, followed by another voluntary 7-week drinking period, started at the same age as the adult-onset group. We assessed anxiety-like behaviour repeatedly during alcohol drinking with open field and elevated plus maze tests. At the end of alcohol drinking, we also tested the rats using the light/dark box, stress-induced body temperature test and social dominance test. Results During the first 7-week alcohol drinking, adolescent rats exhibited significantly slower acquisition of alcohol drinking and lower alcohol preference than the adult-onset group. However, when tested at the same age as the adult-onset rats, they displayed identical alcohol intake and preference. We found no alcohol-induced effects on anxiety- or stress-related behaviour in the experimental groups at any time points. Conclusions These data show that the genetically determined phenotype of high alcohol drinking of the female alcohol-preferring AA rats is not associated with a predisposition to develop anxiety-like behaviour following voluntary alcohol exposure, even when initiated during adolescence.

Background Conventional diagnostic methods have some limitations in diagnosing specific causes of canine hepatobiliary disorders. In the evaluation of the hepatobiliary system in dogs, ultrasonography (US) is the first imaging method of choice. Nonetheless, endoscopic retrograde cholangiopancreatography (ERCP) has also been proven to be a practicable technique for evaluating canine hepatobiliary (endoscopic retrograde cholangiography, ERC) and pancreatic duct (endoscopic retrograde pancreatography, ERP) disorders, providing additional therapeutic options by sphincterotomy (EST). To date, the efficacy and safety of diagnostic and therapeutic ERCP has not been evaluated in veterinary medicine literature. The present study sought to report complications and outcomes of dogs undergoing ERCP and EST, and to assess the usefulness of diagnostic ERCP by comparing the findings of US, ERCP and histopathological findings in liver and pancreas. Results This retrospective case series comprises data collected from 15 dogs that underwent successful ERC/ERCP. Nine dogs underwent EST following ERC. US and ERC were best in agreement when assessing the common bile duct. In case of disagreement between the modalities, the ERC findings of the ductal structures were in line with the available pathology findings more often than the US findings, whereas the opposite was noted for the gallbladder. The technical success rates were 88.2% for ERC, 66.7% for ERP, and 81.8% for EST, with no major complications during or immediately after the procedure. Immediate bile flow after EST was recorded in 7/9 dogs but only four showed coinciding clinical and laboratory improvement and four dogs were euthanized within 1-6 days after EST. Conclusions US remains a valuable initial diagnostic imaging method for hepatobiliary disorders and allows good assessment of the gallbladder. ERC can serve as a complementary procedure for diagnostic assessment of the hepatobiliary duct disorders. However, in order to improve the outcomes of EST, careful selection of patients for the procedure would require more advanced diagnostic imaging of the hepatobiliary area.

Porous silicon (PSi) is a biocompatible and biodegradable material, which can be utilized in biomedical applications. It has several favorable properties, which makes it an excellent material for building engineered nanosystems for drug delivery and diagnostic purposes. One significant hurdle for commercial applications of PSi is the lack of industrial scale production of nanosized PSi particles. Here, we report a novel two-step production method for PSi nanoparticles. The method is based on centrifuge chemical vapor deposition (cCVD) of elemental silicon in an industrial scale reactor followed by electrochemical post-processing to porous particles. Physical properties, biocompatibility and in vivo biodistribution of the cCVD produced nanoparticles were investigated and compared to PSi nanoparticles conventionally produced from silicon wafers by pulse electrochemical etching. Our results demonstrate that the cCVD production provides PSi nanoparticles with comparable physical and biological quality to the conventional method. This method may circumvent several limitations of the conventional method such as the requirements for high purity monocrystalline silicon substrates as starting material and the material losses during the top-down milling process of the pulse-etched films to porous nanoparticles. However, the electroless etching required for the porosification of cCVD-produced nanoparticles limited control over the pore size, but is amenable for scaling of the production to industrial requirements.

International audience; Different types of carbon materials are biocompatible with neural cells and can promote maturation. The mechanism of this effect is not clear. Here we have tested the capacity of a carbon material composed of amorphous sp 3 carbon backbone, embedded with a percolating network of sp 2 carbon domains to sustain neuronal cultures. We found that cortical neurons survive and develop faster on this novel carbon material. After 3 days in culture, there is a precocious increase in the frequency of neuronal activity and in the expression of maturation marker KCC2 on carbon films as compared to a commonly used glass surface. Accelerated development is accompanied by a dramatic increase in neuronal dendrite arborization. The mechanism for the precocious maturation involves the activation of intracellular calcium oscillations by the carbon material already after 1 day in culture. Carbon-induced oscillations are independent of network activity and reflect intrinsic spontaneous activation of developing neurons. Thus, these results reveal a novel mechanism for carbon material-induced neuronal survival and maturation. Brain trauma as well as neurodegenerative diseases are the leading cause of irreversible disability and low quality of life in the elderly population 1. A way to combat neurodegeneration is to promote reparation of neuronal networks, rewiring of neuronal connections, and eventual restoration or substitution of the lost functionality 2. One putative therapeutic avenue is providing scaffolds-special materials that support targeted differentiation of neuronal stem cells and neurite outgrowth of regenerating neurons. Carbon-derived materials possess numerous properties that make them usable as scaffolds 3. Carbon nanotubes (CNTs) and graphene are among the most studied carbon materials for biological applications. These materials enhance neuronal stem cell differentiation 4-6 , as well as promote neuronal survival, neuronal activity, and neuronal process outgrowth 7-13. Neurons cultured on CNTs have increased levels of neuronal K +-Cl − cotransporter KCC2, a key component in the functional maturation of inhibitory synaptic 14 and glutamatergic 15-17 transmission. Downregulation of this protein is also implicated in reactive plasticity following brain trauma 18. CNTs have been suggested to improve the electrical responsiveness of neurons by facilitating local electronic shortcuts between somas and dendrites 19. The ability of CNTs to form tight contacts with neurons is beneficial for neuron-electrode interfaces 20-24. 3D gra-phene substrates support growth and differentiation of neurons 25-27 that in combination with anti-inflammatory properties 28,29 makes graphene a next-generation neuronal tissue scaffold. Despite the importance of novel carbon materials for future engineering, we do not fully understand the mechanisms underlining the trophic action of carbon scaffolds. In this work, we propose a novel mechanism by which a new type of sputtered carbon material accelerates neuronal maturation. The carbon film material consists of conducting nanoscale sp 2 carbon islands embedded in a diamond-like sp 3 carbon matrix. We demonstrate OPEN

Publisher Copyright: © 2021 The Authors AIM: To report initial experiences of automatic detection of Crohn's disease (CD) using quantified motility in magnetic resonance enterography (MRE). MATERIALS AND METHODS: From 302 patients, three datasets with roughly equal proportions of CD and non-CD cases with various illnesses were drawn for testing and neural network training and validation. All datasets had unique MRE parameter configurations and were performed in free breathing. Nine neural networks were devised for automatic generation of three different regions of interests (ROI): small bowel, all bowel, and non-bowel. Additionally, a full-image ROI was tested. The motility in an MRE series was quantified via a registration procedure, which, accompanied with given ROIs, resulted in three motility indices (MI). A subset of the indices was used as an input for a binary logistic regression classifier, which predicted whether the MRE series represented CD. RESULTS: The highest mean area under the curve (AUC) score, 0.78, was reached using the full-image ROI and with the dataset with the highest cine series length. The best AUC scores for the other two datasets were only 0.54 and 0.49. CONCLUSION: The automatic system was able to detect CD in the group of MRE studies with lower temporal resolution and longer cine series showing potential in primary bowel disorder diagnostics. Larger ROI selections and utilising all available cine series for motility registration yielded slight performance improvements. (c) 2021 The Authors. Published by Elsevier Ltd on behalf of The Royal College of Radiologists. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/ 4.0/).

The Parkinson's disease (PD) risk gene GTP cyclohydrolase 1 (GCH1) catalyzes the rate-limiting step in tetrahydrobiopterin (BH4) synthesis, an essential cofactor in the synthesis of monoaminergic neurotransmitters. To investigate the mechanisms by which GCH1 deficiency may contribute to PD, we generated a loss of function zebrafishgch1mutant (gch1–/–), using CRISPR/Cas technology.gch1–/–zebrafish develop marked monoaminergic neurotransmitter deficiencies by 5 d postfertilization (dpf), movement deficits by 8 dpf and lethality by 12 dpf. Tyrosine hydroxylase (Th) protein levels were markedly reduced without loss of ascending dopaminergic (DAergic) neurons. L-DOPA treatment ofgch1–/–larvae improved survival without ameliorating the motor phenotype. RNAseq ofgch1–/–larval brain tissue identified highly upregulated transcripts involved in innate immune response. Subsequent experiments provided morphologic and functional evidence of microglial activation ingch1–/–. The results of our study suggest that GCH1 deficiency may unmask early, subclinical parkinsonism and only indirectly contribute to neuronal cell death via immune-mediated mechanisms. Our work highlights the importance of functional validation for genome-wide association studies (GWAS) risk factors and further emphasizes the important role of inflammation in the pathogenesis of PD.SIGNIFICANCE STATEMENTGenome-wide association studies have now identified at least 90 genetic risk factors for sporadic Parkinson's disease (PD). Zebrafish are an ideal tool to determine the mechanistic role of genome-wide association studies (GWAS) risk genes in a vertebrate animal model. The discovery of GTP cyclohydrolase 1 (GCH1) as a genetic risk factor for PD was counterintuitive, GCH1 is the rate-limiting enzyme in the synthesis of dopamine (DA), mutations had previously been described in the non-neurodegenerative movement disorder dopa-responsive dystonia (DRD). Rather than causing DAergic cell death (as previously hypothesized by others), we now demonstrate that GCH1 impairs tyrosine hydroxylase (Th) homeostasis and activates innate immune mechanisms in the brain and provide evidence of microglial activation and phagocytic activity.

Aim We aimed at identifying potential roles of vesicular monoamine transporter 2, also known as Solute Carrier protein 18 A2 (SLC18A2) (hereafter, Vmat2), in brain monoamine regulation, their turnover, behaviour and brain development using a novel zebrafish model. Methods A zebrafish strain lacking functional Vmat2 was generated with the CRISPR/Cas9 system. Larval behaviour and heart rate were monitored. Monoamines and their metabolites were analysed with high-pressure liquid chromatography. Amine synthesising and degrading enzymes, and genes essential for brain development, were analysed with quantitative PCR, in situ hybridisation and immunocytochemistry. Results The 5-bp deletion in exon 3 caused an early frameshift and was lethal within 2 weeks post-fertilisation. Homozygous mutants (hereafter, mutants) displayed normal low locomotor activity during night-time but aberrant response to illumination changes. In mutants dopamine, noradrenaline, 5-hydroxytryptamine and histamine levels were reduced, whereas levels of dopamine and 5-hydroxytryptamine metabolites were increased, implying elevated monoamine turnover. Consistently, there were fewer histamine, 5-hydroxytryptamine and dopamine immunoreactive cells. Cellular dopamine immunostaining, in wild-type larvae more prominent in tyrosine hydroxylase 1 (Th1)-expressing than in Th2-expressing neurons, was absent in mutants. Despite reduced dopamine levels, mutants presented upregulated dopamine-synthesising enzymes. Further, in mutants the number of histidine decarboxylase-expressing neurons was increased, notch1a and pax2a were downregulated in brain proliferative zones. Conclusion Lack of Vmat2 increases monoamine turnover and upregulates genes encoding amine-synthesising enzymes, including histidine decarboxylase. Notch1a and pax2a, genes implicated in stem cell development, are downregulated in mutants. The zebrafish vmat2 mutant strain may be a useful model to study how monoamine transport affects brain development and function, and for use in drug screening.

Laboratory-based X-ray absorption spectroscopy (XAS) and especially X-ray absorption near-edge structure (XANES) offers new opportunities in catalyst characterization and presents not only an alternative, but also a complementary approach to precious beamtime at synchrotron facilities. We successfully designed a laboratory-based setup for performing operando, quasi-simultaneous XANES analysis at multiple K-edges, more specifically, operando XANES of mono-, bi-, and trimetallic CO2 hydrogenation catalysts containing Ni, Fe, and Cu. Detailed operando XANES studies of the multielement solid catalysts revealed metal-dependent differences in the reducibility and re-oxidation behavior and their influence on the catalytic performance in CO2 hydrogenation. The applicability of operando laboratory-based XANES at multiple K-edges paves the way for advanced multielement catalyst characterization complementing detailed studies at synchrotron facilities., Angewandte Chemie. International Edition, 61 (48), ISSN:1433-7851, ISSN:1521-3773, ISSN:0570-0833

Introduction: Porous silicon (PSi) nanoparticles are capable of delivering therapeutic payloads providing targeted delivery and sustained release of the payloads. In this work we describe the development and proof-of-concept in vivo evaluation of thermally hydrocarbonized porous silicon (PSi) nanoparticles that are implanted with radioactive $^{155}$Tb atoms and coated with red blood cell (RBC) membrane ($^{155}$Tb-THCPSi). The developed nanocomposites can be utilized as an intravenous delivery platform for theranostic radionuclides. Methods: THCPSi thin films were implanted with $^{155}$Dy ions that decay to 155Tb at the ISOLDE radioactive ion-beam (RIB) facility at CERN. The films were processed to nanoparticles by ball-milling and sonication, and subsequently coated with either a solid lipid and RBC membrane or solely with RBC membrane. The nanocomposites were evaluated in vitro for stability and in vivo for circulation half-life and ex vivo for biodistribution in Balb/c mice. Results: Nanoporous THCPSi films were successfully implanted with $^{155}$Tb and processed to coated nanoparticles. The in vitro stability of the particles in plasma and buffer solutions was not significantly different between the particle types, and therefore the RBC membrane coated particles with less laborious processing method were chosen for the biological evaluation. The RBC membrane coating enhanced significantly the blood halflife compared to bare THCPSi particles. In the ex vivo biodistribution study a pronounced accumulation to the spleen was found, with lower uptake in the liver and a minor uptake in the lung, gall bladder and bone marrow. Conclusions: We have demonstrated, using $^{155}$Tb RIB-implanted PSi nanoparticles coated with mouse RBC membranes, the feasibility of using such a theranostic nanosystem for the delivery of RIB based radionuclides with prolonged circulation time. Advances in knowledge and implications for patient care: For the first time, the RIB implantation technique has been utilized to produce PSi nanoparticle with a surface modified for better persistence in circulation. When optimized, these particles could be used in targeted radionuclide therapy with a combination of chemotherapeutic payload within the PSi structure.

The Ang II (Angiotensin II)-Angiotensin-(1-7) axis of the Renin Angiotensin System encompasses 3 enzymes that form Angiotensin-(1-7) [Ang-(1-7)] directly from Ang II: ACE2 (angiotensin-converting enzyme 2), PRCP (prolylcarboxypeptidase), and POP (prolyloligopeptidase). We investigated their relative contribution to Ang-(1-7) formation in vivo and also ex vivo in serum, lungs, and kidneys using models of genetic ablation coupled with pharmacological inhibitors. In wild-type (WT) mice, infusion of Ang II resulted in a rapid increase of plasma Ang-(1-7). In ACE2 −/− / PRCP −/− mice, Ang II infusion resulted in a similar increase in Ang-(1-7) as in WT (563±48 versus 537±70 fmol/mL, respectively), showing that the bulk of Ang-(1-7) formation in circulation is essentially independent of ACE2 and PRCP. By contrast, a POP inhibitor, Z-Pro-Prolinal reduced the rise in plasma Ang-(1-7) after infusing Ang II to control WT mice. In POP −/− mice, the increase in Ang-(1-7) was also blunted as compared with WT mice (309±46 and 472±28 fmol/mL, respectively P =0.01), and moreover, the rate of recovery from acute Ang II-induced hypertension was delayed ( P =0.016). In ex vivo studies, POP inhibition with ZZP reduced Ang-(1-7) formation from Ang II markedly in serum and in lung lysates. By contrast, in kidney lysates, the absence of ACE2, but not POP, obliterated Ang-(1-7) formation from added Ang II. We conclude that POP is the main enzyme responsible for Ang II conversion to Ang-(1-7) in the circulation and in the lungs, whereas Ang-(1-7) formation in the kidney is mainly ACE2-dependent.

Natural biopolymer nanoparticles (NPs), including nanocrystalline cellulose (CNC) and lignin, have shown potential as scaffolds for targeted drug delivery systems due to their wide availability, cost‐efficient preparation, and anticipated biocompatibility. Since both CNC and lignin can potentially cause complications in cell viability assays due to their ability to scatter the emitted light and absorb the assay reagents, we investigated the response of bioluminescent (CellTiter‐Glo®), colorimetric (MTT® and AlamarBlue®) and fluorometric (LIVE/DEAD®) assays for the determination of the biocompatibility of the multimodal CNC and lignin constructs in murine RAW 264.7 macrophages and 4T1 breast adenocarcinoma cell lines. Here, we have developed multimodal CNC and lignin NPs harboring the radiometal chelator DOTA (1,4,7,10‐tetraazacyclododecane‐1,4,7,10‐tetraacetic acid) and the fluorescent dye Cyanine 5 for the investigation of nanomaterial biodistribution in vivo with nuclear and optical imaging, which were then used as the model CNC and lignin nanosystems in the cell viability assay comparison. CellTiter‐Glo® based on the detection of ATP‐dependent luminescence in viable cells revealed to be the best assay for both nanoconstructs for its robust linear response to increasing NP concentration and lack of interference from either of the NP types. Both multimodal CNC and lignin NPs displayed low cytotoxicity and favorable interactions with the cell lines, suggesting that they are good candidates for nanosystem development for targeted drug delivery in breast cancer and for theranostic applications. Our results provide useful guidance for cell viability assay compatibility for CNC and lignin NPs and facilitate the future translation of the materials for in vivo applications.

Pharmacological research in mice and human genetic analyses suggest that the kallikrein-kinin system (KKS) may regulate anxiety. We examined the role of the KKS in anxiety and stress in both species. In human genetic association analysis, variants in genes for the bradykinin precursor (KNG1) and the bradykinin receptors (BDKRB1 and BDKRB2) were associated with anxiety disorders (p Pgc1a and the bradykinin precursor gene Kng2 was upregulated after cold stress. Our data suggests that the bradykinin system modulates a variety of stress responses through B2 receptor-mediated effects, but systemic antagonists of the B2 receptor were not anxiolytic in mice. Genetic variants in the bradykinin receptor genes may predispose to anxiety disorders in humans by affecting their function.

Most non–small cell lung cancers (NSCLC) contain nontargetable mutations, including KRAS, TP53, or STK11/LKB1 alterations. By coupling ex vivo drug sensitivity profiling with in vivo drug response studies, we aimed to identify drug vulnerabilities for these NSCLC subtypes. Primary adenosquamous carcinoma (ASC) or adenocarcinoma (AC) cultures were established from KrasG12D/+;Lkb1fl/fl (KL) tumors or AC cultures from KrasG12D/+;p53fl/fl (KP) tumors. Although p53-null cells readily propagated as conventional cultures, Lkb1-null cells required conditional reprograming for establishment. Drug response profiling revealed short-term response to MEK inhibition, yet long-term clonogenic assays demonstrated resistance, associated with sustained or adaptive activation of receptor tyrosine kinases (RTK): activation of ERBBs in KL cultures, or FGFR in AC cultures. Furthermore, pan-ERBB inhibition reduced the clonogenicity of KL cultures, which was exacerbated by combinatorial MEK inhibition, whereas combinatorial MEK and FGFR inhibition suppressed clonogenicity of AC cultures. Importantly, in vivo studies confirmed KL-selective sensitivity to pan-ERBB inhibition, which correlated with high ERBB ligand expression and activation of ERBB receptors, implying that ERBB network activity may serve as a predictive biomarker of drug response. Interestingly, in human NSCLCs, phosphorylation of EGFR or ERBB3 was frequently detected in ASCs and squamous cell carcinomas. We conclude that analysis of in situ ERBB signaling networks in conjunction with ex vivo drug response profiling and biochemical dissection of adaptive RTK activities may serve as a valid diagnostic approach to identify tumors sensitive to ERBB network inhibition.

Previous studies have implicated several brain cell types in schizophrenia (SCZ), but the genetic impact of astrocytes is unknown. Considering their high complexity in humans, astrocytes are likely key determinants of neurodevelopmental diseases, such as SCZ. Human induced pluripotent stem cell (hiPSC)-derived astrocytes differentiated from five monozygotic twin pairs discordant for SCZ and five healthy subjects were studied for alterations related to high genetic risk and clinical manifestation of SCZ in astrocyte transcriptomics, neuron-astrocyte co-cultures, and in humanized mice. We found gene expression and signaling pathway alterations related to synaptic dysfunction, inflammation, and extracellular matrix components in SCZ astrocytes, and demyelination in SCZ astrocyte transplanted mice. While Ingenuity Pathway Analysis identified SCZ disease and synaptic transmission pathway changes in SCZ astrocytes, the most consistent findings were related to collagen and cell adhesion associated pathways. Neuronal responses to glutamate and GABA differed between astrocytes from control persons, affected twins, and their unaffected co-twins and were normalized by clozapine treatment. SCZ astrocyte cell transplantation to the mouse forebrain caused gene expression changes in synaptic dysfunction and inflammation pathways of mouse brain cells and resulted in behavioral changes in cognitive and olfactory functions. Differentially expressed transcriptomes and signaling pathways related to synaptic functions, inflammation, and especially collagen and glycoprotein 6 pathways indicate abnormal extracellular matrix composition in the brain as one of the key characteristics in the etiology of SCZ.

Background Computed tomography perfusion (CTP) is the mainstay to determine possible eligibility for endovascular thrombectomy (EVT), but there is still a need for alternative methods in patient triage. Purpose To study the ability of a computed tomography angiography (CTA)-based convolutional neural network (CNN) method in predicting final infarct volume in patients with large vessel occlusion successfully treated with endovascular therapy. Materials and Methods The accuracy of the CTA source image-based CNN in final infarct volume prediction was evaluated against follow-up CT or MR imaging in 89 patients with anterior circulation ischemic stroke successfully treated with EVT as defined by Thrombolysis in Cerebral Infarction category 2b or 3 using Pearson correlation coefficients and intraclass correlation coefficients. Convolutional neural network performance was also compared to a commercially available CTP-based software (RAPID, iSchemaView). Results A correlation with final infarct volumes was found for both CNN and CTP-RAPID in patients presenting 6–24 h from symptom onset or last known well, with r = 0.67 ( p < 0.001) and r = 0.82 ( p < 0.001), respectively. Correlations with final infarct volumes in the early time window (0–6 h) were r = 0.43 ( p = 0.002) for the CNN and r = 0.58 ( p < 0.001) for CTP-RAPID. Compared to CTP-RAPID predictions, CNN estimated eligibility for thrombectomy according to ischemic core size in the late time window with a sensitivity of 0.38 and specificity of 0.89. Conclusion A CTA-based CNN method had moderate correlation with final infarct volumes in the late time window in patients successfully treated with EVT.

The diversity and safety of nanofibrillated cellulose (NFC) hydrogels have gained a vast amount of interest at the pharmaceutical site in recent years. Moreover, this biomaterial has a high potential to be utilized as a protective matrix during the freeze-drying of heat-sensitive pharmaceuticals and biologics to increase their properties for long-term storing at room temperature and transportation. Since freeze-drying and subsequent reconstitution have not been optimized for this biomaterial, we must find a wider understanding of the process itself as well as the molecular level interactions between the NFC hydrogel and the most suitable lyoprotectants. Herein we optimized the reconstitution of the freeze-dried NFC hydrogel by considering critical quality attributes required to ensure the success of the process and gained insights of the obtained experimental data by simulating the effects of the used lyoprotectants on water and NFC. We discovered the correlation between the measured characteristics and molecular dynamics simulations and obtained successful freeze-drying and subsequent reconstitution of NFC hydrogel with the presence of 300 mM of sucrose. These findings demonstrated the possibility of using the simulations together with the experimental measurements to obtain a more comprehensive way to design a successful freeze-drying process, which could be utilized in future pharmaceutical applications.

BackgroundChronic pulmonary embolism (CPE) is a life-threatening disease easily misdiagnosed on computed tomography. We investigated a three-dimensional convolutional neural network (CNN) algorithm for detecting hypoperfusion in CPE from computed tomography pulmonary angiography (CTPA).MethodsPreoperative CTPA of 25 patients with CPE and 25 without pulmonary embolism were selected. We applied a 48%–12%–40% training-validation-testing split (12 positive and 12 negative CTPA volumes for training, 3 positives and 3 negatives for validation, 10 positives and 10 negatives for testing). The median number of axial images per CTPA was 335 (min–max, 111–570). Expert manual segmentations were used as training and testing targets. The CNN output was compared to a method in which a Hounsfield unit (HU) threshold was used to detect hypoperfusion. Receiver operating characteristic area under the curve (AUC) and Matthew correlation coefficient (MCC) were calculated with their 95% confidence interval (CI).ResultsThe predicted segmentations of CNN showed AUC 0.87 (95% CI 0.82–0.91), those of HU-threshold method 0.79 (95% CI 0.74–0.84). The optimal global threshold values were CNN output probability ≥ 0.37 and ≤ -850 HU. Using these values, MCC was 0.46 (95% CI 0.29–0.59) for CNN and 0.35 (95% CI 0.18–0.48) for HU-threshold method (average difference in MCC in the bootstrap samples 0.11 (95% CI 0.05–0.16). A high CNN prediction probability was a strong predictor of CPE.ConclusionsWe proposed a deep learning method for detecting hypoperfusion in CPE from CTPA. This model may help evaluating disease extent and supporting treatment planning.

We studied the social hierarchy in zebrafish and assessed differences in neurotransmitters and behavior in the F1 generation offspring of dominant and subordinate zebrafish (Danio rerio). We used behavioral assays to study locomotion, ability to complete cognitive tasks, social interaction and aggression. To study the neurochemical changes, we applied quantitative polymerase chain reaction, high pressure liquid chromatography and immunohistochemistry. Social hierarchies were formed both by males and females when animals were kept in same sex pairs in the dyadic dominant-subordinate hierarchy test. The offspring of dominant animals were the leaders in social interactions, however aggression in the mirror-test was not altered in any group. Serotonin and noradrenaline levels were lower in the F1 generation subordinate animals when compared with dominant animals, but not compared with animals that were naive to social hierarchy. The mRNA level of the rate-limiting enzyme in histamine synthesis, histidine decarboxylase, was significantly lower in dominant and subordinate larval zebrafish when compared with control animals. In the dominant adult zebrafish tyrosine hydroxylase 1 mRNA level was lower compared with control animals, whereas tyrosine hydroxylase 2 mRNA was not different. The result was verified with immunohistochemistry. There were gender specific differences between the dominant and subordinate animals, where the dominant females performed better in cognitive tasks such as the T-maze than subordinate females. This was not observed in males, as the behavior of the dominant and subordinate males did not differ. These results add to the understanding of the plastic nature of the central nervous system and show that neurochemical features in aminergic neurotransmitter systems are associated with social leadership and dominance.

This study aims at assessing the thermal nuclei motion effects on the multipole transition channels involved in two core-level spectroscopies, x-ray absorption spectroscopy (XAS) and x-ray Raman scattering (XRS). Temperature effects on the $1s\ensuremath{\rightarrow}s$ monopole, $1s\ensuremath{\rightarrow}p$ dipole, and $1s\ensuremath{\rightarrow}d$ quadrupole transitions are investigated using two reference systems for which we present original experimental data: $\ensuremath{\alpha}\text{\ensuremath{-}}{\mathrm{Al}}_{2}{\mathrm{O}}_{3}$ at the Al $K$ edge probed by XRS at room temperature and rutile $\mathrm{Ti}{\mathrm{O}}_{2}$ at the Ti $K$ pre-edge probed by XAS at temperatures ranging from 6 to 700 K. Through the latter, this work enlightens the part of the pre-edge peak enhancement due to temperature in the $K$ pre-edge region of $3d$ transition metal, which is known to be routinely used to determine the concentration, valence or symmetry of the probed element in a given sample. Nuclear thermal fluctuations are taken into account using a method based on density functional theory that consists in averaging spectra over atomic configurations, generated within the harmonic approximation and obeying quantum statistics at finite temperature. Since only a finite number of such configurations are used, the numerically averaged spectra generally lose the symmetry of the equilibrium crystal positions. In this paper, we demonstrate that the physical average has to be symmetric and propose a method to restore the physical angular dependence of the spectra. The approach is successfully applied to investigate the angular dependent XAS spectra in rutile as a function of temperature. The two systems under study allow to draw general conclusions regarding the effect of nuclear quantum fluctuations on the different transition channels available to both core-level spectroscopies.

Objectives: To assess radiographic brain abnormalities and investigate volumetric differences in adults born preterm at very low birth weight (

Monoamine oxidase (MAO) deficiency and imbalanced levels of brain monoamines have been associated with developmental delay, neuropsychiatric disorders and aggressive behavior. Animal models are valuable tools to gain mechanistic insight into outcomes associated with MAO deficiency. Here, we report a novel genetic model to study the effects of mao loss of function in zebrafish. Quantitative PCR, in situ hybridization and immunocytochemistry were used to study neurotransmitter systems and expression of relevant genes for brain development in zebrafish mao mutants. Larval and adult fish behavior was evaluated through different tests. Stronger serotonin immunoreactivity was detected in mao+/− and mao−/− larvae compared with their mao+/+ siblings. mao−/− larvae were hypoactive, and presented decreased reactions to visual and acoustic stimuli. They also had impaired histaminergic and dopaminergic systems, abnormal expression of developmental markers and died within 20 days post-fertilization. mao+/− fish were viable, grew until adulthood, and demonstrated anxiety-like behavior and impaired social interactions compared with adult mao+/+ siblings. Our results indicate that mao−/− and mao+/− mutants could be promising tools to study the roles of MAO in brain development and behavior. This article has an associated First Person interview with the first author of the paper.

Funding Information: Financial support from the Finnish Cultural Foundation (grant no. 00190375), the Academy of Finland (decision nos. 318422, 278056, 298481, 302102, and 331659), and the University of Helsinki Research Funds are gratefully acknowledged. Finnish Center for Laboratory Animal Pathology (FCLAP) at HiLIFE, University of Helsinki, is acknowledged for the tissue preparation and staining services. The authors also thank Dr. Jaakko Sarparanta for help with phospho‐MEK western blot, Ms. Karina Moslova for assistance with the elemental analysis, Ms. Alexandra Correia for assistance with cell culture, and Ms. Phetcharada Imlimthan for the assistance with the images. Publisher Copyright: © 2021 The Authors. Small published by Wiley-VCH GmbH Copyright: Copyright 2021 Elsevier B.V., All rights reserved. Metastatic melanoma can be difficult to detect until at the advanced state that decreases the survival rate of patients. Several FDA-approved BRAF inhibitors have been used for treatment of metastatic melanoma, but overall therapeutic efficacy has been limited. Lutetium-177 (177Lu) enables simultaneous tracking of tracer accumulation with single-photon emission computed tomography and radiotherapy. Therefore, the codelivery of 177Lu alongside chemotherapeutic agents using nanoparticles (NPs) might improve the therapeutic outcome in metastatic melanoma. Cellulose nanocrystals (CNC NPs) can particularly deliver payloads to lung capillaries in vivo. Herein, 177Lu-labeled CNC NPs loaded with vemurafenib ([177Lu]Lu-CNC-V NPs) is developed and the therapeutic effect in BRAF V600E mutation-harboring YUMM1.G1 murine model of lung metastatic melanoma is investigated. The [177Lu]Lu-CNC-V NPs demonstrate favorable radiolabel stability, drug release profile, cellular uptake, and cell growth inhibition in vitro. In vivo biodistribution reveals significant retention of the [177Lu]Lu-CNC-V NPs in the lung, liver, and spleen. Ultimately, the median survival time of animals is doubly increased after treatment with [177Lu]Lu-CNC-V NPs compared to control groups. The enhanced therapeutic efficacy of [177Lu]Lu-CNC-V NPs in the lung metastatic melanoma animal model provides convincing evidence for the potential of clinical translation for theranostic CNC NP-based drug delivery systems after intravenous administration.

Compton scattering imaging using high-energy synchrotron x-rays allows the visualization of the spatio-temporal lithiation state in lithium-ion batteries probed in-operando. Here, we apply this imaging technique to the commercial 18650-type cylindrical lithium-ion battery. Our analysis of the lineshapes of the Compton scattering spectra taken from different electrode layers reveals the emergence of inhomogeneous lithiation patterns during the charge-discharge cycles. Moreover, these patterns exhibit oscillations in time where the dominant period corresponds to the time scale of the charging curve., 11 pages, 5 figures

Clinical problems related to intestinal sand accumulation in horses are common in certain geographic areas, but the clinical signs appear nonspecific and the course of the accumulation remains somewhat obscure. This study examined the association between the presence and size of intestinal sand accumulations and owner-reported clinical signs, management, and feeding practices, as well as behavioral patterns in horses with radiographic diagnosis of sand accumulation. Owners of the horses filled in an online questionnaire. A total of 447 responses met the inclusion criteria. The size of the sand accumulation detected in the radiographs was not significantly associated with the age, body condition score, sex, or use of the horses. Horses reported to have expressed colic had significantly larger sand accumulations than those without this sign, and a similar association was detected in horses with poor performance. The highest odds ratio for sand accumulation was for the combination of colic and poor performance, followed by colic combined with diarrhea/loose feces or hyperesthesia to touch of the abdominal wall. Larger sand accumulations were detected in greedy horses that eat all their roughage, whereas dominant position in group hierarchy was associated with less sand. The possibility of abdominal sand accumulation should be considered as one of the differentials in horses with multiple owner-reported clinical signs such as colic, poor performance, diarrhea, and hyperesthesia to touch of the abdomen. (C) 2019 The Authors. Published by Elsevier Inc.

Efficient, abundant and low-cost catalysts for the oxygen evolution reaction (OER) are required for energy conversion and storage. In this study, a doping-etching route has been developed to access defect rich Fe-Co-Al (Fe-Co-Al-AE) ternary hydroxide nanosheets for superior electrochemical oxygen evolution. After partial etching of Al, ultrathin Fe3Co2Al2-AE electrocatalysts with a rich pore structure are obtained with a shift of the cobalt valence state towards higher valence (Co2+-> Co3+), along with a substantial improvement in the catalytic performance. Fe3Co2Al2-AE shows a notably lower overpotential of only 284 mV at a current density of 10 mA cm(-2) and double the OER mass activity of the etching-free Fe3Co2Al2 with an overpotential of 350 mV. Density functional theory shows the leaching of Al changes the rate-determining step of the OER from conversion of *OOH into O-2 on Fe3Co2Al2 to formation of OOH from *O on the Al-defective catalysts. This work demonstrates an effective route to design and synthesize transition metal electrocatalysts and provides a promising alternative for the further development of oxygen evolution catalysts.

Mesoporous silicon (PSi) is biocompatible and tailorable material with high potential in drug delivery applications. Here, we report of an evaluation of PSi as a carrier platform for theranostics by delivering a radioactive ion beam- (RIB-) based radioactive lanthanoid into tumors in a mouse model of prostate carcinoma. Thermally hydrocarbonized porous silicon (THCPSi) wafers were implanted with 159Dy at the facility for radioactive ion beams ISOLDE located at CERN, and the resulting [159Dy]THCPSi was postprocessed into particles. The particles were intratumorally injected into mice bearing prostate cancer xenografts. The stability of the particles was studied in vivo, followed by ex vivo biodistribution and autoradiographic studies. We showed that the process of producing radionuclide-implanted PSi particles is feasible and that the [159Dy]THCPSi particles stay stable and local inside the tumor over seven days. Upon release of 159Dy from the particles, the main site of accumulation is in the skeleton, which is in agreement with previous studies on the biodistribution of dysprosium. We conclude that THCPSi particles are a suitable platform together with RIB-based radiolanthanoids for theranostic purposes as they are retained after administration inside the tumor and the radiolanthanoid remains embedded in the THCPSi.

Concerns have been expressed that the engagement shown by committed individuals is not fully utilized by their organizations while there is insufficient knowledge of which conditions facilitate teaching collaboration and lead to improvements in university education. Portfolios of 43 life science academics applying to enter to the University of Helsinki Teachers' Academy were analyzed through content analysis. Five categories of interactive or collaborative practices emerged from the data: (1) Interacting with peers for personal development, (2) Sharing good teaching practices, (3) Teaching together, (4) Producing educational artefacts, (5) Developing education systematically. The practices occurred in both formal and informal settings, and both settings were present in all categories. In contrast with the formal practices, the informal practices were described in an enthusiastic way. The engagement shown by the scholarly teachers was mostly realized in informal settings. There is probably unrealized potential in the scholarly teachers' teaching-related practices through which they could contribute to the development of teaching in academia. Formal communities related to teaching should be developed to promote deeper collaboration and to foster the participants' feeling of personal commitment and ownership.

Recent genome-wide association studies and mouse models have identified LIN28B as a gene affecting several pubertal timing-related traits and vertebrate growth. However, the exact biological mechanisms underlying the associations remain unknown. We have explored the mechanisms linking LIN28B with growth regulation by combining human gene expression data with functional models. Specifically, we show that 1) pubertal timing-associated genetic variation correlates with LIN28B expression in the pituitary and hypothalamus, 2) downregulating lin28b in zebrafish embryos associates with aberrant development of kiss2-neurons, and 3) increasing lin28b expression transiently by synthetic mRNA injections during embryogenesis results in sustained enhancement of zebrafish growth. Unexpectedly, the mRNA injections resulted in advanced sexual maturation of female fish, suggesting that lin28b may influence pubertal timing through multiple developmental mechanisms. Overall, these results provide novel insight into LIN28B function in vertebrate growth regulation, emphasizing the importance of the gene and related genetic pathways for embryonic and juvenile development.

Hypolimnetic withdrawal provides a way to remove phosphorus (P) from eutrophic lakes, but the method is still rarely combined with water treatment for capturing this P. Thus, little is known about the chemical interactions of P and other elements upon the treatment of hypolimnetic lake water. We investigated these chemical processes in a hypolimnetic withdrawal and treatment system (HWTS) in which hypolimnetic water is first led into a treatment unit for dissolved P (dP) precipitation and subsequently filtered before being circulated back into the lake. We studied three different field-scale treatment unit setups and water treatments (aeration only, aeration + calcium hydroxide (Ca(OH)(2)), aeration + biopolymer) to compare their effectiveness for dP removal and the geochemical properties of the resulting precipitate. In the aeration only treatment, most of the dissolved iron (dFe) (91-95%) and dP (71-91%) were removed when sand filters were used. The addition of Ca (OH)(2) and biopolymer enhanced Fe flocculation, leading to more effective removal of dFe (d99-100%) and dP (88-95%) from the water. Regardless of the water treatment method, dP was always precipitated by amorphous Fe oxides formed in the hypolimnetic water upon aeration. The P content of the resulting precipitates was somewhat lower than expected (2 439-4 145 mg kg(-1)), which may be linked to chemical interactions between Fe and other components in the hypolimnetic water, such as organic matter. The precipitates also contained some heavy metals such as copper and zinc. We conclude that all the tested water treatments were effective in removing dP from hypolimnetic water, but the enhanced precipitation by the addition of treatment chemicals is beneficial when a mesh or other rapid filtration method is used, or when there is only negligible accumulation of dFe in the hypolimnion of the treated lake. Depending on the water treatment method and the water chemistry of the treated lake, the precipitate may have potential for nutrient recycling, although it may sometimes require preliminary processing to enhance bioavailability for plants and to reduce the concentration of heavy metals.

Faba bean is a potential ingredient due to its high protein yield and its possible cultivation in colder climate regions. In this study, meat analogues made from faba bean protein isolate (FPI) and concentrate (FPC) blends were produced using high moisture extrusion. The aim of this study was to investigate the effect of the FPI content (FPIc), feed water content (FWC), and temperature of the long cooling die (LT) during extrusion on the mechanical and physicochemical properties as well as on the structure of the meat analogues. Increased FPIc resulted in higher values in hardness, gumminess, chewiness, and cutting strengths as well as in darker colour and decreased water absorption capacity. The effect of increased FWC on these properties was weaker and the opposite. Images from microtomography revealed that higher FPIc led to a less organised fibrous structure. In conclusion, fibrous structures can be achieved by utilising a mixture of faba bean protein ingredients, and a higher FPC content seemed to promote fibre formation in the meat analogue.

Mephedrone (4-MMC), despite its illegal status, is still a widely used psychoactive substance. Its effects closely mimic those of the classical stimulant drug methamphetamine (METH). Recent research suggests that unlike METH, 4-MMC is not neurotoxic on its own. However, the neurotoxic effects of 4-MMC may be precipitated under certain circumstances, such as administration at high ambient temperatures. Common use of 4-MMC in conjunction with alcohol raises the question whether this co-consumption could also precipitate neurotoxicity. A total of six groups of adolescent rats were treated twice daily for four consecutive days with vehicle, METH (5 mg/kg) or 4-MMC (30 mg/kg), with or without ethanol (1.5 g/kg). To investigate persistent delayed effects of the administrations at two weeks after the final treatments, manganese-enhanced magnetic resonance imaging brain scans were performed. Following the scans, brains were collected for Golgi staining and spine analysis. 4-MMC alone had only subtle effects on neuronal activity. When administered with ethanol, it produced a widespread pattern of deactivation, similar to what was seen with METH-treated rats. These effects were most profound in brain regions which are known to have high dopamine and serotonin activities including hippocampus, nucleus accumbens and caudate-putamen. In the regions showing the strongest activation changes, no morphological changes were observed in spine analysis. By itself 4-MMC showed few long-term effects. However, when co-administered with ethanol, the apparent functional adaptations were profound and comparable to those of neurotoxic METH.

Parvalbumin-positive interneurons (PV+) are a key component of inhibitory networks in the brain and are known to modulate memory and learning by shaping network activity. The mechanisms of PV+ neuron generation and maintenance are not fully understood, yet current evidence suggests that signalling via the glial cell line-derived neurotrophic factor (GDNF) receptor GFR alpha 1 positively modulates the migration and differentiation of PV+ interneurons in the cortex. Whether GDNF also regulates PV+ cells in the hippocampus is currently unknown. In this study, we utilized a Gdnf "hypermorph" mouse model where GDNF is overexpressed from the native gene locus, providing greatly increased spatial and temporal specificity of protein expression over established models of ectopic expression. Gdnf(wt/hyper) mice demonstrated impairments in long-term memory performance in the Morris water maze test and an increase in inhibitory tone in the hippocampus measured electrophysiologically in acute brain slice preparations. Increased PV+ cell number was confirmed immunohistochemically in the hippocampus and in discrete cortical areas and an increase in epileptic seizure threshold was observed in vivo. The data consolidate prior evidence for the actions of GDNF as a regulator of PV+ cell development in the cortex and demonstrate functional effects upon network excitability via modulation of functional GABAergic signalling and under epileptic challenge.

Boron neutron capture therapy (BNCT) is a noninvasive binary therapeutic modality applicable to the treatment of cancers. While BNCT offers a tumor-targeting selectivity that is difficult to match by other means, the last obstacles preventing the full harness of this potential come in the form of the suboptimal boron delivery strategies presently used in the clinics. To address these challenges, we have developed delivery agents that target the glucose transporter GLUT1. Here, we present the chemical synthesis of a number of ortho-carboranylmethyl-substituted glucoconjugates and the biological assessment of all positional isomers. Altogether, the study provides protocols for the synthesis and structural characterization of such glucoconjugates and insights into their essential properties, for example, cytotoxicity, GLUT1-affinity, metabolism, and boron delivery capacity. In addition to solidifying the biochemical foundations of a successful GLUT1-targeting approach to BNCT, we identify the most promising modification sites in d-glucose, which are critical in order to further develop this strategy toward clinical use.

Human imaging data suggest that the motivational processes associated with alcohol reward are reflected in the patterns of neural activation after alcohol or alcohol-related cues. In animal models of alcohol drinking, however, the changes in brain activation during voluntary alcohol ingestion are poorly known. In order to improve the translational utility of animal models, we examined alcohol-induced functional brain activation in Alko Alcohol (AA) and Marchigian-Sardinian alcohol-preferring (msP) rats that drink voluntarily high levels of alcohol, but exhibit widely different neurochemical and behavioral traits cosegregated with alcohol preference. Brain imaging was performed using manganese-enhanced MRI (MEMRI), which is based on accumulation of Mn2+ ions in activated neurons, allowing the identification of functional neuronal networks recruited during specific behaviors in awake animals during a subsequent imaging session under anesthesia. MEMRI was performed following 4 weeks of voluntary alcohol drinking, using water drinking as the control. Despite similar levels of alcohol drinking, strikingly different alcohol-induced neuronal activity patterns were observed in AA and msP rats. Overall, functional activation in the AA rats was more widespread, involving large cortical areas and subcortical structures, such as the bed nucleus of the stria terminalis, preoptic area, hypothalamus, periaqueductal grey, and substantia nigra. In the msP rats, however, alcohol-related activation was largely confined to prefrontal cortical regions and insular cortex, and olfactory areas. Overlapping areas of activation found in both rat lines included the nucleus accumbens, prelimbic, orbital, and insular cortex. In conclusion, our data reveal strikingly different brain circuits associated with alcohol drinking in two genetically different rat lines and suggest innately different motivational and behavioral processes driving alcohol drinking. These findings have important implications for the use of these lines in translational alcohol research., The work was supported under the ERA-Net NEURON framework with grants given to the project “Translational Neuroimaging in Alcoholism” (TRANSALC) by the Academy of Finland (TRANSALC 01EW1112), and the Spanish Ministry of Science and Innovation (PIM2010ERN-00679). Further funding was obtained from the Spanish State Research Agency through the Severo Ochoa Program for Centres of Excellence in R&D (SEV- 2017-0723), the Ministerio de Economía y Competitividad (MINECO) and FEDER funds (BFU2015-64380-C2-1-R, BFU2015-64380-C2-2-R), the Generalitat Valencia through the Prometeo Program (PROMETEO/2019/015), the Spanish Ministerio de Sanidad, Servicios Sociales e Igualdad (#2017I065), the Bundesministerium für Bildung und Forschung (FKZ01EW1112), the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) – Project-ID 402170461 – TRR 265 B02 (Heinz et al., 2020), and the European Union’s Horizon 2020 program (668863-SyBil-AA).

In the recent years, progress in nanotechnology has significantly contributed to the development of novel pharmaceutical formulations to overcome the drawbacks of conventional treatments and improve the therapeutic outcome in many diseases, especially cancer. Nanoparticle vectors have demonstrated the potential to concomitantly deliver diagnostic and therapeutic payloads to diseased tissue. Due to their special physical and chemical properties, the characteristics and function of nanoparticles are tunable based on biological molecular targets and specific desired features (e.g., surface chemistry and diagnostic radioisotope labeling). Within the past decade, several theranostic nanoparticles have been developed as a multifunctional nanosystems which combine the diagnostic and therapeutic functionalities into a single drug delivery platform. Theranostic nanosystems can provide useful information on a real-time systemic distribution of the developed nanosystem and simultaneously transport the therapeutic payload. In general, the diagnostic functionality of theranostic nanoparticles can be achieved through labeling gamma-emitted radioactive isotopes on the surface of nanoparticles which facilitates noninvasive detection using nuclear molecular imaging techniques, such as positron emission tomography (PET) and single-photon emission computed tomography (SPECT), meanwhile, the therapeutic effect arises from the potent drug released from the nanoparticle. Moreover, some radioisotopes can concurrently emit both gamma radiation and high-energy particles (e.g., alpha, beta, and Auger electrons), prompting the use either alone for radiotheranostics or synergistically with chemotherapy. This chapter provides an overview of the fundamentals of radiochemistry and relevant radiolabeling strategies for theranostic nanosystem development as well as the methods for the preclinical evaluation of radiolabeled nanoparticles. Furthermore, preclinical case studies of recently developed theranostic nanosystems will be highlighted.

LRRK2 gain-of-function is considered a major cause of Parkinson’s disease (PD) in humans. However, pathogenicity of LRRK2 loss-of-function in animal models is controversial. Here we show that deletion of the entire zebrafish lrrk2 locus elicits a pleomorphic transient brain phenotype in maternal-zygotic mutant embryos (mzLrrk2). In contrast to lrrk2, the paralog gene lrrk1 is virtually not expressed in the brain of both wild-type and mzLrrk2 fish at different developmental stages. Notably, we found reduced catecholaminergic neurons, the main target of PD, in specific cell populations in the brains of mzLrrk2 larvae, but not adult fish. Strikingly, age-dependent accumulation of monoamine oxidase (MAO)-dependent catabolic signatures within mzLrrk2 brains revealed a previously undescribed interaction between LRRK2 and MAO biological activities. Our results highlight mzLrrk2 zebrafish as a tractable tool to study LRRK2 loss-of-function in vivo, and suggest a link between LRRK2 and MAO, potentially of relevance in the prodromic stages of PD., Author summary Parkinson’s disease is the second most common degenerative disorder of the brain. Mutations of the LRRK2 gene are the most recurrent genetic cause of pathology, and are thought to result in a more active LRRK2 protein, a large enzyme whose biological function is unclear. Therefore, LRRK2 inhibitors are regarded as promising therapeutics. However, mouse models do not reproduce human pathology unless they also lack LRRK1, and there is evidence of dominant negative effects of LRRK2 mutations. Therefore, the characterization of reliable LRRK2 knockout models might provide insights. In our study, we used the zebrafish as a tractable model to study both early developmental and adult phenotypes resulting from the loss of the entire endogenous lrrk2 gene. We found that mutant embryos displayed subtle brain phenotypes, including reduction of catecholaminergic neurons, the main target of human disease, that spontaneously resolved with development, and a late-onset and progressive increase of dopamine and serotonin degradation consistent with increased MAO enzyme activity. Our results suggest that similar defects might occur in the pre-symptomatic stage of the disease in humans, and warrant further evaluation of LRRK2 inhibition in a therapeutic perspective.

While interest in psychedelic drugs in the fields of psychiatry and neuroscience has re-emerged in recent last decades, the general understanding of the effects of these drugs remains deficient. In particular, there are gaps in knowledge on executive functions and goal-directed behaviors both in humans and in commonly used animal models. The effects of acute doses of psychedelic lysergic acid diethylamide (LSD) on reward-driven decision making were explored using the mouse version of the Iowa Gambling Task. A total of 15 mice were trained to perform in a touch-screen adaptation of the rodent version of the Iowa Gambling Task, after which single acute doses of LSD (0.025, 0.1, 0.2, 0.4 mg/kg), serotonin 2A receptor-selective agonist 25CN-NBOH (1.5 mg/kg), d-amphetamine (2.0 mg/kg), and saline were administered before the trial. 25CN-NBOH and the three lowest doses of LSD showed no statistically significant changes in option selection or in general functioning during the gambling task trials. The highest dose of LSD (0.4 mg/kg) significantly decreased premature responding and increased the omission rate, but had no effect on option selection in comparison with the saline control. Amphetamine significantly decreased the correct responses and premature responding while increasing the omission rate. In conclusion, mice can perform previously learned, reward-driven decision-making tasks while under the acute influence of LSD at a commonly used dose range. While interest in psychedelic drugs in the fields of psychiatry and neuroscience has re-emerged in recent last decades, the general understanding of the effects of these drugs remains deficient. In particular, there are gaps in knowledge on executive functions and goal-directed behaviors both in humans and in commonly used animal models. The effects of acute doses of psychedelic lysergic acid diethylamide (LSD) on reward-driven decision making were explored using the mouse version of the Iowa Gambling Task. A total of 15 mice were trained to perform in a touch-screen adaptation of the rodent version of the Iowa Gambling Task, after which single acute doses of LSD (0.025, 0.1, 0.2, 0.4 mg/kg), serotonin 2A receptor-selective agonist 25CN-NBOH (1.5 mg/kg), d-amphetamine (2.0 mg/kg), and saline were administered before the trial. 25CN-NBOH and the three lowest doses of LSD showed no statistically significant changes in option selection or in general functioning during the gambling task trials. The highest dose of LSD (0.4 mg/kg) significantly decreased premature responding and increased the omission rate, but had no effect on option selection in comparison with the saline control. Amphetamine significantly decreased the correct responses and premature responding while increasing the omission rate. In conclusion, mice can perform previously learned, reward-driven decision-making tasks while under the acute influence of LSD at a commonly used dose range.

The anterior insular cortex is hypothesized to represent interoceptive effects of drug reward in the service of goal-directed behavior. The insula is richly connected, but the insula circuitry in addiction remains poorly characterized. We examined the involvement of the anterior insula, amygdala, and nucleus accumbens, as well as the projections of the anterior insula to the central amygdala, basolateral amygdala (BLA), and nucleus accumbens core in voluntary alcohol drinking. We trained alcohol-preferring Alko Alcohol (AA) rats to drink alcohol during intermittent 2-h sessions. We then expressed excitatory or inhibitory designer receptors [designer receptors exclusively activated by designer drugs (DREADDs)] in the anterior insula, nucleus accumbens, or amygdala by means of adenovirus-mediated gene transfer and activated the DREADDs with clozapine-N-oxide (CNO) prior to the drinking sessions. Next, to examine the role of specific insula projections, we expressed FLEX-DREADDs in the efferent insula → nucleus accumbens core, insula → central amygdala, and insula → BLA projections by means of a retrograde AAV-Cre vector injected into the insula projection areas. In the anterior insula and amygdala, excitatory Gq-DREADDs significantly attenuated alcohol consumption. In contrast, in the nucleus accumbens, the Gq-DREADD stimulation increased alcohol drinking, and the inhibitory Gi-DREADDs suppressed it. The Gq-DREADDs expressed in the insula → nucleus accumbens core and insula → central amygdala projections increased alcohol intake, whereas inhibition of these projections had no effect. These data demonstrate that the anterior insula, along with the amygdala and nucleus accumbens, has a key role in controlling alcohol drinking by providing excitatory input to the central amygdala and nucleus accumbens to enhance alcohol reward.

Lymphedema can occur when tissue fluid cannot enter or leaks from the lymphatic system into surrounding tissues. Some genetic causes of primary lymphedema are known, but these currently explain a minority of cases. Previous studies have shown that dominant-negative mutations in angiopoietin 2 (ANGPT2), which is involved in lymphatic vessel formation and maturation, promote lymphangiogenesis in mice. Leppänen et al. now show that inactivating mutations in angiopoietin 2 associate with primary lymphedema in humans.Primary lymphedema is caused by developmental and functional defects of the lymphatic vascular system that result in accumulation of protein-rich fluid in tissues, resulting in edema. The 28 currently known genes causing primary lymphedema can explain