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By age 40 years, over 90% of adults with Down syndrome have Alzheimer's disease pathology and most progress to dementia. Despite having few systemic vascular risk factors, individuals with Down syndrome have elevated cerebrovascular disease markers that track with the clinical progression of Alzheimer's disease, suggesting a role of cerebrovascular disease that is hypothesized to be mediated by inflammatory factors. This study examined the pathways through which small vessel cerebrovascular disease contributes to Alzheimer's disease-related pathophysiology and neurodegeneration in adults with Down syndrome. One hundred eighty-five participants from the Alzheimer's Biomarkers Consortium-Down Syndrome [mean (SD) age = 45.2 (9.3) years] with available MRI and plasma biomarker data were included in this study. White matter hyperintensity (WMH) volumes were derived from T2-weighted fluid-attenuated inversion recovery MRI scans, and plasma biomarker concentrations of amyloid beta 42/40, phosphorylated tau 217, astrocytosis (glial fibrillary acidic protein) and neurodegeneration (neurofilament light chain) were measured with ultrasensitive immunoassays. We examined the bivariate relationships of WMH, amyloid beta 42/40, phosphorylated tau 217 and glial fibrillary acidic protein with age-residualized neurofilament light chain across Alzheimer's disease diagnostic groups. A series of mediation and path analyses examined statistical pathways linking WMH and Alzheimer's disease pathophysiology to promote neurodegeneration in the total sample and groups stratified by clinical diagnosis. There was a direct and indirect bidirectional effect through the glial fibrillary acidic protein of WMH on phosphorylated tau 217 concentration, which was associated with neurofilament light chain concentration in the entire sample. Amongst cognitively stable participants, WMH was directly and indirectly, through glial fibrillary acidic protein, associated with phosphorylated tau 217 concentration, and in those with mild cognitive impairment, there was a direct effect of WMH on phosphorylated tau 217 and neurofilament light chain concentrations. There were no associations of WMH with biomarker concentrations among those diagnosed with dementia. The findings from this cross-sectional study suggest that among individuals with Down syndrome, cerebrovascular disease promotes neurodegeneration by increasing astrocytosis and tau pathophysiology in the presymptomatic phases of Alzheimer's disease, but future studies will need to confirm these associations with longitudinal data. This work joins an emerging literature that implicates cerebrovascular disease and its interface with neuroinflammation as a core pathological feature of Alzheimer's disease in adults with Down syndrome.

UNLABELLED: Combinations of immune checkpoint inhibitors (ICI, including anti-PD-1/PD-L1) and chemotherapy have been FDA approved for metastatic and early-stage triple-negative breast cancer (TNBC), but most patients do not benefit. B7-H4 is a B7 family ligand with proposed immunosuppressive functions being explored as a cancer immunotherapy target and may be associated with anti-PD-L1 resistance. However, little is known about its regulation and effect on immune cell function in breast cancers. We assessed murine and human breast cancer cells to identify regulation mechanisms of B7-H4 in vitro. We used an immunocompetent anti-PD-L1-sensitive orthotopic mammary cancer model and induced ectopic expression of B7-H4. We assessed therapy response and transcriptional changes at baseline and under treatment with anti-PD-L1. We observed B7-H4 was highly associated with epithelial cell status and transcription factors and found to be regulated by PI3K activity. EMT6 tumors with cell-surface B7-H4 expression were more resistant to immunotherapy. In addition, tumor-infiltrating immune cells had reduced immune activation signaling based on transcriptomic analysis. Paradoxically, in human breast cancer, B7-H4 expression was associated with survival benefit for patients with metastatic TNBC treated with carboplatin plus anti-PD-L1 and was associated with no change in response or survival for patients with early breast cancer receiving chemotherapy plus anti-PD-1. While B7-H4 induces tumor resistance to anti-PD-L1 in murine models, there are alternative mechanisms of signaling and function in human cancers. In addition, the strong correlation of B7-H4 to epithelial cell markers suggests a potential regulatory mechanism of B7-H4 independent of PD-L1. SIGNIFICANCE: This translational study confirms the association of B7-H4 expression with a cold immune microenvironment in breast cancer and offers preclinical studies demonstrating a potential role for B7-H4 in suppressing response to checkpoint therapy. However, analysis of two clinical trials with checkpoint inhibitors in the early and metastatic settings argue against B7-H4 as being a mechanism of clinical resistance to checkpoints, with clear implications for its candidacy as a therapeutic target.

The gut microbiome (GM) influences multiple processes during host development and maintenance. To study these events, fecal microbiota transfer (FMT) to germ-free (GF) recipients is often performed. Mouse models of disease are also susceptible to GM-dependent effects, and cryo-repositories often store feces from donated mouse strains. Shipping live mice may affect the GM and result in an inaccurate representation of the baseline GM. We hypothesize that the use of such fecal samples for FMT would transfer shipping-induced changes in the donor GM to GF recipients. To test this, donor mice originating from two suppliers were shipped to the University of Missouri. Fecal samples collected pre- and post-shipping were used to inoculate GF mice. Pre- and post-shipping fecal samples from donors and fecal and/or cecal contents were collected from recipients at 1 and 2 weeks post-FMT. 16S rRNA sequencing revealed supplier-dependent effects of shipping on the donor microbiome. FMT efficiency was independent of shipping timepoint or supplier, resulting in the transmission of shipping-induced changes to recipient mice; however, the effect of supplier-origin (SO) microbiome remained evident. While shipping may cause subtle changes in fecal samples collected for FMT, such effects are inconsistent among SO GMs and minor in comparison to other biological variables.

The article examines the specificity of courtroom discourse as a communicative integrity, which is manifested in the comprehensive coherence of its information components. The proposed paper signifi- cantly contributes to the existing studies on the linguistics of courtroom discourse, focusing on the study of its architectonics, a problem that is currently on the periphery of researchers’ scientific interests. The ne- cessity (relevance) of studying courtroom discourse as a comprehensive information cluster, including in- teraction and subordination of its components, is explained by the need to identify the factors ensuring the effectiveness of the communication process. All this determines the novelty of the research. The aim of our study is to define the nature of courtroom discourse and its architectonics from the point of the information it contains, its arrangement, and its impact on the audience. To achieve this aim, both general scientific methods (analysis, synthesis, systematisation, classification, induction, deduction) and strictly linguistic methods (random sampling and observation, lexical-semantic and contextual analysis, interpretation of dictionary definitions) were used. In addition, the method of sociolinguistic analysis of the corpus was employed to study the interdependence between language and society. The aim has led to the following objectives: to establish kinds of information the courtroom discourse is based on; to determine the way of how it is delivered and organised to strongly impact the recipients; to justify the notion of architectonics in using it to courtroom discourse. The corpus material was the texts of the opening and closing speeches of prosecutors and defense lawyers at the two high-profile trials of 1999 – 2000 and 2021 (the Amadou Diallo Trial (1999–2000); the George Floyd Murder (Chauvin) Trial (2021). The findings of the study were as follows: It has been established that communication in court, in terms of its focus on the effective exchange of various kinds of information, affects the following: a) the rational perception (semantic information, impact on the mind); b) the emotional perception (aesthetic information, impact on emotions); c) the irratio- nal perception (synectic information, impact on the unconscious) that form an information cluster. It has been shown for the first time that the effectiveness of the communication process in court is determined by the extent of impact or depth of penetration of resources of each kind of information. The concept of architectonics is clarified as a certain, a coordinated and coherent arrangement of blocks of discourse, not reducible to the sum of these blocks, determined by the overall goal of the author in relation to the delivery and impact of information. It is emphasised that the architectonics of discourse affects and implicitly manages the process of audience’s perception of its content. The research also first establishes that the architectonics of the prosecution discourse is constructed through progressive disclosure. Conversely, the discursive space of the defense is organised through the vi- sualisation of a narrative, with the main theme, idea, or concept at its center. The topics that provide expla- nation are arranged around it. Such construct of architectonics is based on the principle of mindmapping. Both constructs provide legal professionals with almost unlimited scope of the creativity to set up the nec- essary chains of association. It is necessary to mention that personal attitudes should be introduced into the architectonics of courtroom discourse as well as a myth created. This also determines, among other things, the success of courtroom discourse. The classification of types of transgression based on their impact on the recipients has been supple- mented, namely, constructive and disruptive ones. The study shows promise as it would be intriguing to explore this issue from the perspective of the evolution of the architectonics of courtroom discourse.

Abstract Background In prey, patterns of individual habitat selection and movement can be a consequence of an individuals’ anti-predator behavior. Adjustments of anti-predator behavior are important for prey to increase their survival. Hunters may alter the anti-predator behavior of prey. In long-lived animals, experience may cause behavioral changes during individuals’ lifetime, which may result in altered habitat selection and movement. Our knowledge of which specific events related to hunting activity induce behavioral changes in solitary living species is still limited. Methods We used offspring loss in a solitary and long-lived ungulate species, moose (Alces alces), as our model system. We investigated whether offspring loss to hunters induces behavioral changes in a species subjected to heavy human harvest but free from natural predation. To test for behavioral change in relation to two proxies for experience (calf fate and age), we combined movement data from 51 adult female moose with data on their offspring survival and female age. We tested for adjustments in females’ habitat selection and movement following calf harvest using Hidden Markov Models and integrated Step Selection Analysis to obtain behavioral state specific habitat selection coefficients. Results We found that females with a harvested calf modified habitat selection and movement during the following hunting season. Female moose selected for shorter distance to roads during the night, selected for shorter distance to forests and greater distance to human settlements following calf harvest than females who had not lost a calf. The survival of twins in a given hunting season was related to female age. Older females we more likely to have twins survive the hunting season. Conclusions Our findings suggest that losing offspring to human harvest imposes behavioral changes in a long-lived ungulate species, leading to adjustments in females' habitat selection and movement behavior, which may lower the risk of encountering hunters. In our study, female moose that experienced calf loss selected for lower distance to forest and selected for greater distance to human settlements during periods of high hunting pressure compared to females without the experience of calf loss during the previous hunting season. We interpret this as potential learning effects.

Abstract In modern materials discovery, materials are now efficiently screened using machine learning (ML) techniques with target-specific properties for meeting various engineering applications. However, a major challenge that persists with deep generative ML approach is the issue related to lattice reconstruction at the decoding phase, leading to the generation of materials with low symmetry, unfeasible atomic coordination, and triclinic behavioral properties in the crystal lattice. To address this concern, the present research makes a contribution by proposing a Lattice-Constrained Materials Generative Model (LCMGM) for designing new and polymorphic perovskite materials with crystal conformities that are consistent with predefined geometrical and thermodynamic stability constraints at the encoding phase. A comparison with baseline models such as Physics Guided Crystal Generative Model (PGCGM) and Fourier-Transformed Crystal Property (FTCP), confirms the potential of the LCMGM for improved training stability, better chemical learning effect and higher geometrical conformity. The new materials emerging from this research are Density Functional Theory (DFT) validated and openly made available in the Mendeley data repository: https://doi.org/10.17632/m262xxpgn2.1 .

Abstract Background This study introduced an Augmented Reality (AR) navigation system to address limitations in conventional high tibial osteotomy (HTO). The objective was to enhance precision and efficiency in HTO procedures, overcoming challenges such as inconsistent postoperative alignment and potential neurovascular damage. Methods The AR-MR (Mixed Reality) navigation system, comprising HoloLens, Unity Engine, and Vuforia software, was employed for pre-clinical trials using tibial sawbone models. CT images generated 3D anatomical models, projected via HoloLens, allowing surgeons to interact through intuitive hand gestures. The critical procedure of target tracking, essential for aligning virtual and real objects, was facilitated by Vuforia’s feature detection algorithm. Results In trials, the AR-MR system demonstrated significant reductions in both preoperative planning and intraoperative times compared to conventional navigation and metal 3D-printed surgical guides. The AR system, while exhibiting lower accuracy, exhibited efficiency, making it a promising option for HTO procedures. The preoperative planning time for the AR system was notably shorter (4 min) compared to conventional navigation (30.5 min) and metal guides (75.5 min). Intraoperative time for AR lasted 8.5 min, considerably faster than that of conventional navigation (31.5 min) and metal guides (10.5 min). Conclusions The AR navigation system presents a transformative approach to HTO, offering a trade-off between accuracy and efficiency. Ongoing improvements, such as the incorporation of two-stage registration and pointing devices, could further enhance precision. While the system may be less accurate, its efficiency renders it a potential breakthrough in orthopedic surgery, particularly for reducing unnecessary harm and streamlining surgical procedures.

Archaeological research in northern Sweden has customarily proposed models based on assumed migration patterns to portray resource utilization of prehistoric hunter-gatherers. An average hunting household needs about 500km2 for its subsistence. This assumption, as well as the temporal and spatial distribution of animal resources available for hunting households in the interior of Northern Sweden, is investigated using Agent Based Modelling (ABM) with explicitly identified factors and conditions. ABM simulations were run in order to analyse the relationships between hunters, moose (Alces alces), predators, landscapes and how human migration patterns could be adjusted in order to coincide with moose migrations. The results suggest that wolves and human hunters could coexist if the landscape had a moose density of 0.6 moose/km2 or more and if each hunting household possessed territories of 400–500km2. In accordance with the model’s parameters, the simulation identifies those factors that are particularly sensitive to change and those factors that are necessary in order to maintain an ecological balance between hunters and their prey.

In the context of the circular economy, there is an urgent need for transformation into circular material flows by avoiding waste, reducing extraction of virgin raw materials, and extending product life cycles. Within the construction and real estate industry, the reuse of building materials stands out as a critical strategy for value retention. The objective of this paper is to localise the forefront of the practical implementation of reused materials within the Swedish construction and real estate industry. To achieve this, the current state of reuse practices through the perspective of property companies as clients in decision-making positions was conducted through semi-structured interviews to identify key barriers and enablers associated with reuse. The three most significant barriers that emerged were a lack of measurable economic incentives, the absence of a professional reuse market, and obsolete project management. Conversely, the three most significant enablers were new and improved project management methods, enhancing competence and learning within and beyond organisations, and introduce reuse at an early stage. The results emphasise the need for project management to develop and adopt circular economy principles. This is further understood due to responders highlighting the industry's linear approach as a major obstacle to circularity together with uncertainties related to product performance, responsibilities and economics characterising reuse efforts. However, an industry in transition is witnessed, e.g. by the emergence of new roles which suggests a continued need for focused research in organisational matters.

Shigella spp. cause bacillary dysentery (shigellosis) with high morbidity and mortality in low- and middle-income countries. Infection occurs through the fecal-oral route and can be devastating for vulnerable populations, including infants and the elderly. These bacteria invade host cells using a type III secretion system (T3SS). No licensed vaccine yet exists for shigellosis, but we have generated a recombinant fusion protein, L-DBF, combining the T3SS needle tip protein (IpaD), translocator protein (IpaB), and the LTA1 subunit of enterotoxigenic E. coli labile toxin, which offers broad protection in a mouse model of lethal pulmonary infection. The L-DBF vaccine protects high-risk groups, including young and elderly mice. Here, we investigated how the gut microbiota of young and elderly mice responds to intranasal L-DBF vaccination formulated in an oil-in-water emulsion (ME). Samples from lungs, small intestines, and feces were collected on day 14 after 2 or 3 doses of L-DBF in ME. 16S rRNA gene sequencing revealed age-dependent changes in gut microbiota post-vaccination. The vaccine-induced changes were more prominent in the elderly mice and were most significant in the intestinal tract, indicating that vaccination by the intranasal route can have a tremendous impact on the gut environment. These findings provide insight into the communication between the intranasal mucosal surface following subunit vaccination and the microbiota at a distant mucosal site, thereby highlighting the impact of vaccination and the host’s microbiome.

Background: Success in resuscitation depends not only on the timeliness of the maneuvers but also on the quality of chest compressions. Factors such as the rescuer position and arm angle can significantly impact compression quality. Aim: This study explores the influence of rescuer positioning and arm angle on the quality of chest compressions among healthcare professionals experienced in cardiopulmonary resuscitation. Methods: In this international, multicentric, randomized crossover simulation trial with independent groups, healthcare professionals were assigned to one of four positions: kneeling on the floor, standing, standing on a step stool, and kneeling on the bed. Participants performed two 3-minute trials of uninterrupted chest compressions at arm angles of 90° and 105°. Compression quality was assessed, using manikin derived data. Results: A total of 76 participants entered the study. Those using a 90° arm angle exhibited higher compression scores than those at a 105° angle. Rescuers standing on a step stool maintained higher scores over time when compared to other groups. In contrast, rescuers kneeling on the bed consistently scored below 75% throughout the trial, with particularly low scores at the 105° angle. Conclusion: Rescuer position and arm angle significantly influence CPR quality, with a 90° arm angle and elevated positioning optimizing compression depth and effectiveness. The results recommend against kneeling on the bed due to its negative impact on chest compression quality.

Introduction: To minimize the risk of developing foot-ulcers, persons with diabetes are given the advice to daily inspect their feet and to apply skincare formulations. However, commercially available skincare products have rarely been developed and evaluated for diabetes foot care specifically. The primary aim of this randomized controlled trial (RCT) is to evaluate the effects in reducing foot xerosis in persons with diabetes without foot-ulcers using two skincare creams containing different humectants (interventions) against a cream base non-humectant (comparator). Secondary outcomes are to evaluate differences on skin barrier integrity, low-molecular weight biomarkers and skin microbiota, microcirculation including transcutaneous oxygen pressure, degree of neuropathy, and HbA1c between intervention-comparator creams. Methods: Two-armed double-blind RCT, registered in ClinicalTrials.gov Identifier: NCT06427889. With 80 % power, two-tailed significance of 2.5 % in each arm, 39 study persons is needed in each arm, total 78 persons, 98 including dropouts, to be able to prove a reduction of at least one category in the Xerosis Severity Scale with the intervention creams compared to the comparator. In one arm, each participant will treat one foot with one of the intervention creams (Oviderm® or Canoderm®), while the opposite foot will be treated with the comparator cream (Decubal®lipid cream), twice a day. If needed, participants are enrolled after a wash-out period of two weeks. The participants will undergo examinations at baseline, day 14 and day 28. Discussion: This RCT evaluate the potential effects of humectants in skin creams against foot xerosis in persons with diabetes.

Autism spectrum disorders (ASD) are complex human neurodiversities increasing in prevalence within the human population. In search of therapeutics to improve quality-of-life for ASD patients, the gut microbiome (GM) has become a promising target as a growing body of work supports roles for the complex community of microorganisms in influencing host behavior via the gut-brain-axis. However, whether naturally-occurring microbial diversity within the host GM affects these behaviors is often overlooked. Here, we applied a model of population-level differences in the GM to a classic ASD model – the BTBR T+ Itpr3tf/J mouse – to assess how complex GMs affect host behavior. Leveraging the naturally occurring differences between supplier-origin GMs, our data demonstrate that differing, complex GMs selectively effect host ASD-related behavior – especially neonatal ultrasonic communication – and reveal a male-specific effect on behavior not typically observed in this strain. We then identified that the body weight of BTBR mice is influenced by the postnatal GM which was potentially mediated by microbiome-dependent effects on energy harvest in the gut. These data provide insight into how variability within the GM affects host behavior and growth, thereby emphasizing the need to incorporate microbial diversity within the host GM as an experimental factor in biomedical research.

Triticale (× Triticosecale Wittmack), an edible hybrid of rye and wheat, is known for its superior agronomic traits. This study investigates the patterns of 5-n-alk(en)ylresorcinols (AR), phenolic lipids with diverse biological activities, in fresh and dried triticale across developmental stages. Total AR (TAR) content and LC-MS and GC-MS analyses were performed for profiling extracts obtained by solvent-assisted and supercritical fluid extraction methods. All triticale plant parts accumulated ARs, revealing significant distinctions in content and profiles between fresh and dry basis. CO2-extracted dry spikes at late stages and solvent-extracted fresh and dry leaves at the earliest stage exhibited the highest TAR content. Moreover, triticale kernels showcased a broader AR homolog diversity. Semi-quantitative AR levels varied based on plant parts and ontogeny, suggesting constitutive roles during growth. These insights into temporal and spatial distribution offer valuable information for determining optimal harvesting times, quality, and material condition for future breeding studies in triticale.

Abstract In this study, the mass balance, pharmacokinetics (PK) and metabolism of atuliflapon, a novel 5‐lipoxygenase‐activating protein inhibitor, were investigated in healthy male subjects. A single oral dose of 200 mg [14C]atuliflapon suspension was administered to six healthy male subjects. Mass balance, PK and metabolite profiles of atuliflapon were analyzed using radioactivity monitoring and liquid chromatography with mass spectrometry analysis. The safety of atuliflapon was assessed during the study. Atuliflapon was rapidly absorbed with a median tmax of 1.5 h, followed by a biphasic decline in plasma exposure rendering a terminal half‐life of ~20 h. Unchanged atuliflapon was the predominant radioactive component in plasma, accounting for 40.1% of the total drug‐related exposure (DRE), while a direct N‐glucuronide was the only metabolite exceeding 10% of DRE, accounting for 20.9%. Renal excretion of intact atuliflapon accounted for

Abstract Aim To explore Finnish paramedics' perceptions of work‐related performance expectations in relation to work experience, and understand how organizational socialization contributes to understanding paramedics' performance expectations. Design A qualitative design with a deductive‐inductive approach utilizing a social constructivist framework. The organizational socialization framework by Wanberg was used as the theoretical basis. Methods Data were collected between May and August 2023, using group and individual interviews of newly graduated (n = 9) and experienced paramedics (n = 13). Participants were recruited via social media channels. Data were first analyzed deductively, according to constructs of the organizational socialization framework (role clarity, task mastery, and social acceptance), then inductively, using codes not utilized in the deductive phase. Data Sources Interviewed Finnish paramedics (N = 22), both newly graduated paramedics (n = 9) and experienced paramedics (n = 13). The interviews were performed remotely and then transcribed into text. Results Our findings showed comparable performance expectations between newly graduated and experienced paramedics, mismatches in role clarity of paramedic work, challenges in both learning and upholding professional competence, and difficulties of social acceptance into the paramedic community. There were variations in how expectations were perceived between groups, indicating that experience might partly affect how paramedics identify and manage performance expectations. The organizational socialization framework enables the contextualization of these performance expectations. Conclusions and Implications Paramedic work involves challenges to upholding clinical competence, aligning to a professional role, and social integration into the professional community. Our research contributes to understanding how paramedics perceive these challenges as performance expectations in different stages of their careers and how they could be managed utilizing a framework for organizational socialization. The socialization of paramedics into the workforce needs to account for these performance expectations, especially considering the changing paradigm of paramedic work, role, and societal expectations. Patient or Public Contribution No Patient or Public Contribution.

Vitamin D deficiency poses a widespread health challenge, shaped by environmental and genetic determinants. A recent discovery identified a genetic regulator, rs11542462, in the SDR42E1 gene, though its biological implications remain largely unexplored. Our bioinformatic assessments revealed pronounced SDR42E1 expression in skin keratinocytes and the analogous HaCaT human keratinocyte cell lines, prompting us to select the latter as an experimental model. Employing CRISPR/Cas9 gene-editing technology and multi-omics approach, we discovered that depleting SDR42E1 showed a 1.6-fold disruption in steroid biosynthesis pathway (P-value = 0.03), considerably affecting crucial vitamin D biosynthesis regulators. Notably, SERPINB2 (P-value = 2.17 × 10−103), EBP (P-value = 2.46 × 10−13), and DHCR7 (P-value = 8.03 × 10−09) elevated by ∼2–3 fold, while ALPP (P-value

Layered Na3Fe3(PO4)4 can function as a positive electrode for both Li- and Na-ion batteries and may hold advantages from both classical layered and phosphate-based electrode materials. Using a combination of ex-situ and operando synchrotron radiation powder X-ray diffraction, void space analysis, and Mössbauer spectroscopy, we herein investigate the structural evolution of the Na3Fe3(PO4)4 framework during Li- and Na-ion intercalation. We show that during discharge, Li- and Na-intercalation into Na3Fe3(PO4)4 occurs via a solid solution reaction wherein Na-ions appear to be preferentially intercalated into the intralayer sites. The intercalation causes an expansion of the unit cell volume, however at open circuit conditions after ion-intercalation (i.e., after battery discharge), Na3+xFe3(PO4)4 and LixNa3Fe3(PO4)4 undergo a structural relaxation, wherein the unit volume contracts below that of the pristine material. Rietveld refinement suggests that the ions intercalated into the intra-layer sites diffuse to the sites in the inter-layer space during the relaxation. This behavior brings new perspectives to understanding structural relaxation and deviations between structural evolution observed under dynamic and static conditions.

Abstract Mutations in the α-Synuclein (αS) gene promote αS monomer aggregation that causes neurodegeneration in familial Parkinson’s disease (fPD). However, most mouse models expressing single-mutant αS transgenes develop neuronal aggregates very slowly, and few have dopaminergic cell loss, both key characteristics of PD. To accelerate neurotoxic aggregation, we previously generated fPD αS E46K mutant mice with rationally designed triple mutations based on the α-helical repeat motif structure of αS (fPD E46K→3 K). The 3 K variant increased αS membrane association and decreased the physiological tetramer:monomer ratio, causing lipid- and vesicle-rich inclusions and robust tremor-predominant, L-DOPA responsive PD-like phenotypes. Here, we applied an analogous approach to the G51D fPD mutation and its rational amplification (G51D → 3D) to generate mutant mice. In contrast to 3 K mice, G51D and 3D mice accumulate monomers almost exclusively in the cytosol while also showing decreased αS tetramer:monomer ratios. Both 1D and 3D mutant mice gradually accumulate insoluble, higher-molecular weight αS oligomers. Round αS neuronal deposits at 12 mos immunolabel for ubiquitin and pSer129 αS, with limited proteinase K resistance. Both 1D and 3D mice undergo loss of striatal TH+ fibers and midbrain dopaminergic neurons by 12 mos and a bradykinesia responsive to L-DOPA. The 3D αS mice have decreased tetramer:monomer equilibria and recapitulate major features of PD. These fPD G51D and 3D mutant mice should be useful models to study neuronal αS-toxicity associated with bradykinetic motor phenotypes.

Abstract Background Age-associated changes in the gastrointestinal microbiome of young pigs have been robustly described; however, the temporal dynamics of the fecal microbiome of the female pig from early life to first parity are not well understood. Our objective was to describe microbiome and antimicrobial resistance dynamics of the fecal microbiome of breeding sows from early life through estrus, parturition and weaning of the first litter of piglets (i.e., from 3 to 53 weeks of age). Results Our analysis revealed that fecal bacterial populations in developing gilts undergo changes consistent with major maturation milestones. As the pigs progressed towards first estrus, the fecal bacteriome shifted from Rikenellaceae RC9 gut group- and UCG-002-dominated enterotypes to Treponema- and Clostridium sensu stricto 1-dominated enterotypes. After first estrus, the fecal bacteriome stabilized, with minimal changes in enterotype transition and associated microbial diversity from estrus to parturition and subsequent weaning of first litter piglets. Unlike bacterial communities, fecal fungal communities exhibited low diversity with high inter- and intra-pig variability and an increased relative abundance of certain taxa at parturition, including Candida spp. Counts of resistant fecal bacteria also fluctuated over time, and were highest in early life and subsequently abated as the pigs progressed to adulthood. Conclusions This study provides insights into how the fecal microbial community and antimicrobial resistance in female pigs change from three weeks of age throughout their first breeding lifetime. The fecal bacteriome enterotypes and diversity are found to be age-driven and established by the time of first estrus, with minimal changes observed during subsequent physiological stages, such as parturition and lactation, when compared to the earlier age-related shifts. The use of pigs as a model for humans is well-established, however, further studies are needed to understand how our results compare to the human microbiome dynamics. Our findings suggest that the fecal microbiome exhibited consistent changes across individual pigs and became more diverse with age, which is a beneficial characteristic for an animal model system.

Zanthoxylum simulans Hance, commonly known as Sichuan pepper, is a well-known medicinal plant recognized for its potential as a source of bioactive specialized metabolites. As part of our interest in natural antifungal compounds, the present study describes the discovery of an unreported N-alcoxycarbonylbenzo[c]phenanthridinium salt, N-methoxycarbonyl-9,12-dimethoxy-norchelerythrine 1 (a type-III benzo[c]phenanthridine), isolated from Z. simulans seedlings, which were propagated under controlled greenhouse conditions. Six-month seedlings were harvested and subjected to cold acid–base extraction. Chromatographic techniques achieved the isolation of 1 from raw alkaloid extract. The structural elucidation of 1 was accomplished through comprehensive spectroscopic analysis, including nuclear magnetic resonance and high-resolution mass spectrometry. Fusarium oxysporum, a fungal pathogen responsible for substantial agricultural losses, was exposed to different concentrations of the novel compound, exhibiting potent antifungal efficacy (IC50 < 3 µM) and fungicide effects. These findings highlight the potential of benzophenanthridines as antifungal leads and underscore the importance of exploring natural products for agricultural applications.

IntroductionThe concept of a sterile uterus was challenged by recent studies that have described the microbiome of the virgin and pregnant uterus for species including humans and cattle. We designed two studies that tested whether the microbiome is introduced into the uterus when the virgin heifer is first inseminated and whether the origin of the microbiome is the vagina/cervix.MethodsThe uterine microbiome was measured immediately before and after an artificial insemination (AI; Study 1; n = 7 AI and n = 6 control) and 14 d after insemination (Study 2; n = 12 AI and n = 12 control) in AI and non-AI (control) Holstein heifers. A third study (Study 3; n = 5 Holstein heifers) that included additional negative controls was subsequently conducted to support the presence of a unique microbiome within the uterus despite the low microbial biomass and regardless of insemination. Traditional bacteriological culture was performed in addition to 16S rRNA gene sequencing on the same samples to determine whether there were viable organisms in addition to those detected based on DNA sequencing (16S rRNA gene sequence).Results and discussionInseminating a heifer did not lead to a large change in the microbiome when assessed by traditional methods of bacterial culture or metataxonomic (16S rRNA gene) sequencing (results of Studies 1 and 2). Very few bacteria were cultured from the body or horn of the uterus regardless of whether an AI was or was not (negative control) performed. The cultured bacterial genera (e.g., Bacillus, Corynebacterium, Cutibacterium, Micrococcus, Staphylococcus, and Streptococcus) were typical of those found in the soil, environment, skin, mucous membranes, and urogenital tract of animals. Metataxonomic sequencing of 16S rRNA gene generated a large number of amplicon sequence variants (ASV), but these larger datasets that were based on DNA sequencing did not consistently demonstrate an effect of AI on the abundance of ASVs across all uterine locations compared with the external surface of the tract (e.g., perimetrium; positive control samples for environment contamination during slaughter and collection). Major genera identified by 16S rRNA gene sequencing overlapped with those identified with bacterial culture and included Cutibacterium, Staphylococcus, and Streptococcus.