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Objective: GRIN1 -related neurodevelopmental disorder ( GRIN1 -NDD) is characterized by clinically significant variation in the GRIN1 gene, which encodes the obligatory GluN1 subunit of N-methyl-D-aspartate receptors (NMDARs). The identified p.Tyr647Ser (Y647S) variant - carried by a 33-year-old female with seizures and intellectual disability - is located in the M3 helix in the GluN1 transmembrane domain. This study builds upon initial in vitro investigations of the functional impacts of the GRIN1 Y647S variant and examines its in vivo consequences in a mouse model. Methods: To investigate in vitro functional impacts of NMDARs containing GluN1-Y647S variant subunits, GluN1-Y647S was co-expressed with wildtype GluN2A or GluN2B subunits in Xenopus laevis oocytes and HEK cells. Grin1 Y647S/+ mice were created by CRISPR-Cas9 endonuclease-mediated transgenesis and the molecular, electrophysiological, and behavioural consequences of the variant were examined. Results: In vitro , NMDARs containing GluN1-Y647S show altered sensitivity to endogenous agonists and negative allosteric modulators, and reduced cell surface trafficking. Grin1 Y647S/+ mice displayed a reduction in whole brain GluN1 levels and deficiency in NMDAR-mediated synaptic transmission in the hippocampus. Behaviourally, Grin1 Y647S/+ mice exhibited spontaneous seizures, altered vocalizations, muscle strength, sociability, and problem-solving. Interpretation: The Y647S variant confers a complex in vivo phenotype, which reflects largely diminished properties of NMDAR function. As a result, Grin1 Y647S/+ mice display atypical behaviour in domains relevant to the clinical characteristics of GRIN1 -NDD and the individual carrying the variant. Ultimately, the characterization of Grin1 Y647S/+ mice accomplished in the present work expands our understanding of the mechanisms underlying GRIN1 -NDD and provides a foundation for the development of novel therapeutics.

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Importance: Sudden infant death syndrome (SIDS) is a major cause of infant death in the US. Previous research suggests that inborn errors of metabolism may contribute to SIDS, yet the relationship between SIDS and biomarkers of metabolism remains unclear. Objective: To evaluate and model the association between routinely measured newborn metabolic markers and SIDS in combination with established risk factors for SIDS. Design, Setting, and Participants: This was a case-control study nested within a retrospective cohort using data from the California Office of Statewide Health Planning and Development and the California Department of Public Health. The study population included infants born in California between 2005 and 2011 with full metabolic data collected as part of routine newborn screening (NBS). SIDS cases were matched to controls at a ratio of 1:4 by gestational age and birth weight z score. Matched data were split into training (2/3) and testing (1/3) subsets. Data were analyzed from January 2005 to December 2011. Exposures: Metabolites measured by NBS and established risk factors for SIDS. Main Outcomes and Measures: The primary outcome was SIDS. Logistic regression was used to evaluate the association between metabolic markers combined with known risk factors and SIDS. Results: Of 2 276 578 eligible infants, 354 SIDS (0.016%) cases (mean [SD] gestational age, 38.3 [2.3] weeks; 220 male [62.1%]) and 1416 controls (mean [SD] gestational age, 38.3 [2.3] weeks; 723 male [51.1%]) were identified. In multivariable analysis, 14 NBS metabolites were significantly associated with SIDS in a univariate analysis: 17-hydroxyprogesterone, alanine, methionine, proline, tyrosine, valine, free carnitine, acetyl-L-carnitine, malonyl carnitine, glutarylcarnitine, lauroyl-L-carnitine, dodecenoylcarnitine, 3-hydroxytetradecanoylcarnitine, and linoleoylcarnitine. The area under the receiver operating characteristic curve for a 14-marker SIDS model, which included 8 metabolites, was 0.75 (95% CI, 0.72-0.79) in the training set and was 0.70 (95% CI, 0.65-0.76) in the test set. Of 32 infants in the test set with model-predicted probability greater than 0.5, a total of 20 (62.5%) had SIDS. These infants had 14.4 times the odds (95% CI, 6.0-34.5) of having SIDS compared with those with a model-predicted probability less than 0.1. Conclusions and Relevance: Results from this case-control study showed an association between aberrant metabolic analytes at birth and SIDS. These findings suggest that we may be able to identify infants at increased risk for SIDS soon after birth, which could inform further mechanistic research and clinical efforts focused on monitoring and prevention.

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Agrobacterium-mediated transformation is a preferred method for genetic engineering and genome editing of plants due to its numerous advantages, although not all species exhibit transformability. Genetic engineering and plant genome editing methods are technically challenging in recalcitrant crop plants. Factors affecting the poor rate of transformation in such species include host genotype, Agrobacterium genotype, type of explant, physiological condition of the explant, vector, selectable marker, inoculation method, chemical additives, antioxidative compounds, transformation-enhancing compounds, medium formulation, optimization of culture conditions, and pre-treatments. This review provides novel insights into the key factors involved in gene transfer facilitated by Agrobacterium and proposes potential solutions to overcome existing barriers to transformation in recalcitrant species, thereby contributing to improvement programs for these species. This review introduces the key factors that impact the effectiveness of a molecular breeding program using Agrobacterium-mediated transformation, specifically focusing on recalcitrant plant species.

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This cohort study evaluates whether in-office biopsies significantly improve time to biopsy and time to treatment compared with operating room biopsies.

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There are two major subtypes of adipose tissue, i.e., white adipose tissue (WAT) and brown adipose tissue (BAT). It has been known for a long time that WAT mediates obesity and impairs healthful longevity. More recently, interest has focused on BAT, which, unlike WAT, actually augments healthful aging. The goal of this review is to examine the role of BAT in mediating healthful longevity. A major role for BAT and its related beige adipose tissue is thermogenesis, as a mechanism to maintain body temperature by producing heat through uncoupling protein 1 (UCP1) or through UCP1-independent thermogenic pathways. Our hypothesis is that healthful longevity is, in part, mediated by BAT. BAT protects against the major causes of impaired healthful longevity, i.e., obesity, diabetes, cardiovascular disorders, cancer, Alzheimer's disease, reduced exercise tolerance, and impaired blood flow. Several genetically engineered mouse models have shown that BAT enhances healthful aging and that their BAT is more potent than wild-type (WT) BAT. For example, when BAT, which increases longevity and exercise performance in mice with disruption of the regulator of G protein signaling 14 (RGS14), is transplanted to WT mice, their exercise capacity is enhanced at 3 days after BAT transplantation, whereas BAT transplantation from WT to WT mice also resulted in increased exercise performance, but only at 8 weeks after transplantation. In view of the ability of BAT to mediate healthful longevity, it is likely that a pharmaceutical analog of BAT will become a novel therapeutic modality.

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The Rhynie Chert (Lower Devonian, Scotland) hosts a remarkably well-preserved early terrestrial ecosystem. Organisms including plants, fungi, arthropods, and bacteria were rapidly silicified due to inundation by silica-rich hot spring fluids. Exceptional molecular preservation has been noted by many authors, including some of the oldest evidence of lignin in the fossil record. The evolution of lignin was a critical factor in the diversification of land plants, providing structural support and defense against herbivores and microbes. However, the timing of the evolution of lignin decay processes remains unclear. Studies placing this event near the end of the Carboniferous are contradicted by evidence for fungal pathogenesis in Devonian plant fossils, including from the Rhynie Chert. We conducted organic geochemical analyses on a Rhynie Chert sample, including hydropyrolysis (HyPy) of kerogen and high-resolution mass spectrometric mapping of a thin section, to elucidate the relationship between lignin and the potential fungal marker perylene. HyPy of kerogen showed an increase in relative abundance of perylene supporting its entrapment within the silicate matrix of the chert. Lignin monomers were isolated through an alkaline oxidation process, showing a distribution dominated by H-type monomers. G- and S-type monomers were also detected, preserved by rapid silicification. Polycyclic aromatic hydrocarbons including perylene, a known marker for lignin-degrading fungi, were also concentrated in the kerogen and found to be localized within silicified plant fragments. Our results strongly link perylene in the Rhynie Chert to the activity of phytopathogenic fungi, demonstrating the importance of fungal degradation processes as far back as the Early Devonian.

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Widespread fragmentation and degradation of habitats make organisms increasingly vulnerable to declines in population size. Immigration is a key process potentially affecting the rescue and persistence of populations in the face of such pressures. Field research addressing severe demographic declines in the context of immigration among interconnected local populations is limited owing to difficulties in detecting such demographic events and the need for long-term monitoring of populations. In a 17-subpopulation metapopulation of the butterfly, Parnassius smintheus, all adults observed in two adjacent patches were removed over eight consecutive generations. Despite this severe and long-term reduction in survival and reproduction, the targeted populations did not go extinct. Here, we use genetic data to assess the role of immigration versus in situ reproduction in allowing the persistence of these populations. We genotyped 471 samples collected from the targeted populations throughout the removal experiment at 152 single nucleotide polymorphisms. We found no reduction in the genetic diversity of the targeted populations over time, but a decrease in the number of loci in Hardy-Weinberg equilibrium, consistent with a high level of immigration from multiple surrounding populations. Our results highlight the role of connectivity and movement in making metapopulations resilient to even severe and protracted localized population reductions.

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Staphylococcus aureus, a common human pathogen, has long been the focus of scientific investigation due to its association with various infections. However, recent research has unveiled a tantalizing enigma surrounding this bacterium and its potential involvement in carcinogenesis. Chronic S. aureus infections have been linked to an elevated risk of certain cancers, including skin cancer and oral cancer. This review explores the current state of knowledge regarding this connection, examining epidemiological evidence, pathogenic mechanisms, and biological interactions that suggest a correlation. Although initial studies point to a possible link, the precise mechanisms through which S. aureus may contribute to cancer development remain elusive. Emerging evidence suggests that the chronic inflammation induced by persistent S. aureus infections may create a tumor-promoting environment. This inflammation can lead to DNA damage, disrupt cellular signaling pathways, and generate an immunosuppressive microenvironment conducive to cancer progression. Additionally, S. aureus produces a variety of toxins and metabolites that can directly interact with host cells, potentially inducing oncogenic transformations. Despite these insights, significant gaps remain in our understanding of the exact biological processes involved. This review emphasizes the urgent need for more comprehensive research to clarify these microbiological mysteries. Understanding the role of S. aureus in cancer development could lead to novel strategies for cancer prevention and treatment, potentially transforming therapeutic approaches.

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In an effort to expedite the publication of articles, AJHP is posting manuscripts online as soon as possible after acceptance. Accepted manuscripts have been peer-reviewed and copyedited, but are posted online before technical formatting and author proofing. These manuscripts are not the final version of record and will be replaced with the final article (formatted per AJHP style and proofed by the authors) at a later time.

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This study explores the impact of positive selection on the genetic composition of a Drosophila serrata population in eastern Australia through a comprehensive analysis of 110 whole genome sequences. Utilizing an advanced deep learning algorithm (partialS/HIC) and a range of inferred demographic histories, we identified that approximately 14% of the genome is directly affected by sweeps, with soft sweeps being more prevalent (10.6%) than hard sweeps (2.1%), and partial sweeps being uncommon (1.3%). The algorithm demonstrated robustness to demographic assumptions in classifying complete sweeps but faced challenges in distinguishing neutral regions from partial sweeps and linked regions under demographic misspecification. The findings reveal the indirect influence of sweeps on nearly two-thirds of the genome through linkage, with an over-representation of putatively deleterious variants suggesting that positive selection drags deleterious variants to higher frequency due to hitchhiking with beneficial loci. Gene ontology enrichment analysis further supported our confidence in the accuracy of sweep detection as several traits expected to be under positive selection due to evolutionary arms races (e.g. immunity) were detected in hard sweeps. This study provides valuable insights into the direct and indirect contributions of positive selection in shaping genomic variation in natural populations.

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Importance: Traumatic brain injuries (TBI) represent an important, potentially modifiable risk factor for dementia. Despite frequently observed vascular imaging changes in individuals with TBI, the relationships between TBI-associated changes in brain imaging and clinical outcomes have largely been overlooked in community cases of TBI. Objective: To assess whether TBI are associated with and interact with midlife changes in neuroimaging and clinical features in otherwise healthy individuals. Design, Setting, and Participants: This cross-sectional analysis used baseline data from the PREVENT Dementia program collected across 5 sites in the UK and Ireland between 2014 and 2020. Eligible participants were cognitively healthy midlife adults aged between 40 and 59 years. Data were analyzed between January 2023 and April 2024. Exposure: Lifetime TBI history was assessed using the Brain Injury Screening Questionnaire. Main Outcomes and Measures: Cerebral microbleeds and other markers of cerebral small vessel disease (white matter hyperintensities [WMH], lacunes, perivascular spaces) were assessed on 3T magnetic resonance imaging. Clinical measures were cognition, sleep, depression, gait, and cardiovascular disease (CVD) risk, assessed using Computerized Assessment of Information Processing (COGNITO), Pittsburgh Sleep Quality Index, Center for Epidemiologic Studies Depression Scale, clinical interviews, and the Framingham Risk Score, respectively. Results: Of 617 participants (median [IQR] age, 52 [47-56] years; 380 female [61.6%]), 223 (36.1%) had a history of TBI. TBI was associated with higher microbleed count (ß = 0.10; 95% CI, 0.01-0.18; P = .03), with a dose-response association observed with increasing number of TBI events (ß = 0.05; 95% CI, 0.01-0.09; P = .03). Conversely, TBI was not associated with other measures of small vessel disease, including WMH. Furthermore, TBI moderated microbleed associations with vascular risk factors and clinical outcomes, such that associations were present only in the absence of TBI. Importantly, observations held when analyses were restricted to individuals reporting only mild TBI. Conclusions and Relevance: In this cross-sectional study of healthy middle-aged adults, detectable changes in brain imaging and clinical features were associated with remote, even mild, TBI in the general population. The potential contribution of vascular injury to TBI-related neurodegeneration presents promising avenues to identify potential targets, with findings highlighting the need to reduce TBI through early intervention and prevention in both clinical care and policymaking.

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BACKGROUND: Over the last decade, there has been an increase in the evidence base supporting the efficacy of video consultations (VCs) in mental health services. Furthermore, the potential of VC treatment was also demonstrated during the COVID-19 pandemic. Despite these promising results and conducive conditions for VCs, several studies have highlighted that the uptake and implementation of VCs continues to be slow, even after the pandemic. To facilitate and strengthen the implementation of VCs and exploit their potential as a useful tool for mental health disorder treatment, there is a need for a deeper understanding of the issues and experiences of implementing and using VCs as a treatment modality in clinical practice. OBJECTIVE: The aim of this study was to investigate patients' and clinicians' experiences and attitudes toward using VCs in clinical practice. METHODS: Treatment was conducted through the VC modality. Semistructured interviews were conducted individually with patients (n=10) and focus group interview were conducted with clinicians (n=4). Patients had participated in weekly VC treatment over 2 months as part of mental health outpatient services in Denmark. Data from these interviews were analyzed using thematic analysis. RESULTS: Thematic analysis of the patient interviews yielded two main themes: (1) adjusting to the practicalities of the VC format and (2) the practice of therapy using VCs. Patients experienced that using VCs was easy and convenient, and it was possible to establish and maintain a therapeutic alliance. They also described the contact as different to in-person therapy. The thematic analysis conducted on clinicians' experiences of using VCs yielded three themes: (1) a shift in mindset from resistance to acceptance, (2) the contact is different when using the VC modality, and (3) adapting to a new way of working. Clinicians experienced that their initial concerns and resistance toward VC implementation gradually diminished over time as they gained clinical experience of using the modality. They expressed that contact with patients can be different when using the VC modality and that it took time to adjust to a new way of working therapeutically. CONCLUSIONS: Both patients and clinicians experienced that VCs could enhance access to treatment and be meaningfully integrated into clinical practice. In addition, both groups described the contact when using the VC modality as being different to in-person therapy. Future research could examine patients' and clinicians' perceived differences regarding contact when using the VC modality and the implications for therapeutic interventions.

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Ras and Rap small GTPases of the Ras superfamily act as molecular switches to control diverse cellular processes as part of different signaling pathways. Dictyostelium expresses several Ras and Rap proteins, and their study has and continues to greatly contribute to our understanding of their role in eukaryote biology. To study the activity of Ras and Rap proteins in Dictyostelium, several assays based on their interaction with the Ras binding domain of known eukaryotic Ras/Rap effectors have been developed and proved extremely useful to study their regulation and cellular roles. Here, we describe methods to assess Ras/Rap activity biochemically using a pull-down assay and through live-cell imaging using fluorescent reporters.

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Planetary boundaries represent thresholds in major Earth system processes that are sensitive to human activity and control global-scale habitability and stability. These processes are interconnected such that movement of one planetary boundary process can alter the likelihood of crossing other boundaries. Here we argue that the observed deoxygenation of the Earth's freshwater and marine ecosystems represents an additional planetary boundary process that is critical to the integrity of Earth's ecological and social systems, and both regulates and responds to ongoing changes in other planetary boundary processes. Research on the rapid and ongoing deoxygenation of Earth's aquatic habitats indicates that relevant, critical oxygen thresholds are being approached at rates comparable to other planetary boundary processes. Concerted global monitoring, research and policy efforts are needed to address the challenges brought on by rapid deoxygenation, and the expansion of the planetary boundaries framework to include deoxygenation as a boundary helps to focus those efforts.

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The optimal efficacy of xenogeneically generated proteins intended for application in humans requires that their own antigenicity be minimized. This necessary adaptation of antibodies to a humanized version poses challenges since modifications even distant from the binding sites can greatly influence antigen recognition and this is the primary feature that must be maintained during all modifications. Current strategies often rely on grafting and/or randomization/selection to arrive at a humanized variant retaining the binding properties of the original molecule. However, in terms of speed and efficiency, rationally directed approaches can be superior, provided the requisite structural information is available. We present here a humanization procedure based on the high-resolution X-ray structure of a chimaeric IgG against a marker for multiple myeloma. Based on in silico modelling of humanizing amino acid substitutions identified from sequence alignments, we devised a straightforward cloning procedure to rapidly evaluate the proposed sequence changes. Careful inspection of the structure allowed the identification of a potentially problematic amino acid change that indeed disrupted antigen binding. Subsequent optimization of the antigen binding loop sequences resulted in substantial recovery of binding affinity lost in the completely humanized antibody. X-ray structures of the humanized and optimized variants demonstrate that the antigen binding mode is preserved, with surprisingly few direct contacts to antibody atoms. These results underline the importance of structural information for the efficient optimization of protein therapeutics. KEY MESSAGES: Structure-based humanization of an IgG against BCMA, a marker for Multiple Myeloma. Identification of problematic mutations and unexpected modification sites. Structures of the modified IgG-antigen complexes verified predictions. Provision of humanized high-affinity IgGs against BCMA for therapeutic applications.

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BACKGROUND: Drug resistance in Plasmodium falciparum is a major threat to malaria control efforts. Pathogen genomic surveillance could be invaluable for monitoring current and emerging parasite drug resistance. METHODS: Data from two decades (2000-2020) of continuous molecular surveillance of P. falciparum parasites from Senegal were retrospectively examined to assess historical changes in malaria drug resistance mutations. Several known drug resistance markers and their surrounding haplotypes were profiled using a combination of single nucleotide polymorphism (SNP) molecular surveillance and whole genome sequence based population genomics. RESULTS: This dataset was used to track temporal changes in drug resistance markers whose timing correspond to historically significant events such as the withdrawal of chloroquine (CQ) and the introduction of sulfadoxine-pyrimethamine (SP) in 2003. Changes in the mutation frequency at Pfcrt K76T and Pfdhps A437G coinciding with the 2014 introduction of seasonal malaria chemoprevention (SMC) in Senegal were observed. In 2014, the frequency of Pfcrt K76T increased while the frequency of Pfdhps A437G declined. Haplotype-based analyses of Pfcrt K76T showed that this rapid increase was due to a recent selective sweep that started after 2014. DISCUSSION (CONCLUSION): The rapid increase in Pfcrt K76T is troubling and could be a sign of emerging amodiaquine (AQ) resistance in Senegal. Emerging AQ resistance may threaten the future clinical efficacy of artesunate-amodiaquine (ASAQ) and AQ-dependent SMC chemoprevention. These results highlight the potential of molecular surveillance for detecting rapid changes in parasite populations and stress the need to monitor the effectiveness of AQ as a partner drug for artemisinin-based combination therapy (ACT) and for chemoprevention.

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OBJECTIVE: To describe and illustrate septal perforation shape through the documentation and analysis of length and height measurements. A secondary objective is to correlate perforation size to surgical and nonsurgical etiologies. STUY DESIGN: Retrospective case series. SETTING: Tertiary academic medical center. METHODS: Length and height of consecutively treated perforations over a 3-year period were measured directly or through computed tomography. Mean differences in length and height measurements were compared and regression analysis used to determine perforation shape and the effect of etiology on perforation size. Perforations were classified by length into small (1-5 mm), medium (6-15 mm), and large (>15 mm) and correlated to shape and etiology. RESULTS: One hundred twenty-four patients (mean age 50.4 years, 60.5% female) met study inclusion criteria. Height was less than length in 93% of perforations 5 mm or greater in length. Mean perforation height was significantly less than length for medium and large perforations (P < .001). Mean length and height measurements of nonsurgical perforations were greater than those for surgical perforations (P < .001). CONCLUSION: Height is less than length in over 90% of septal perforations. Most perforations assume an elliptical shape as they enlarge. Accurate measurement and presentation of length and height is relevant information to perforation management decisions and for the evaluation of treatment outcomes.

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OBJECTIVE: This study explores the experiences of young adults with psychosis using a smartphone application to promote patient activation and support shared decision making in their outpatient treatment. METHOD: Semistructured interviews were conducted with eight participants who had access to the app while receiving mental health treatment. Qualitative data from the interviews were analyzed using thematic analysis aimed at experiences of interacting with the app. RESULTS: Four themes were extracted from the interviews: supporting users with memory difficulties, giving symptoms substance, a new source of information to guide conversations, and the challenge of capturing complex experiences digitally. While the majority of the themes highlight the benefits of using the app in ways that may facilitate communication between patient and provider, the participants also described some negative experiences when interacting with the app concerning failure to communicate nuances and emotional states satisfyingly. CONCLUSION AND IMPLICATIONS FOR PRACTICE: Experiences with the app were double sided. On the one hand, the use of the app supports communication, and conversely, interaction with the app can create limitations and new challenges for communication. There is a need for more research to understand the use of mental health smartphone apps and their role in supporting interactive processes such as shared decision making in mental health. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

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We retrospectively reviewed the clinical and radiologic findings in 17 children with an aberrant cisternal CN7 and found that these patients had additional anomalies involving other pontine cranial nerves. The hallmark imaging feature identified in all patients was aberrant cisternal segment of an enlarged appearing CN7. The abnormal nerve coursed anteriorly towards the Gasserian ganglion where it fanned out towards the internal auditory canal, Meckel's cave or both. This finding was accompanied by a small cisternal CN5 which often had a lateral bowed appearance. CN5 and CN7 were abnormally close to each other. Meckel's cave appeared widened posteriorly and often was close to or merged with the internal auditory canal. Other abnormalities in the pontine cranial nerves included CN8 deficiency in the majority of children and variable CN6 deficiency. This constellation of findings was most often discovered in children having MR evaluation for sensorineural hearing loss and the majority of patients had preserved facial nerve function. In patients with available genetic testing, no pathogenic variants were observed. Interestingly, in 13 children with available birth history, 9 were notable for maternal or gestational diabetes (69%), suggesting a possible early intrauterine insult to the developing nerves.ABBREVIATIONS: CN= cranial nerve; OAVS= Oculo-Auriculo-Vertebral Spectrum; IAC= Internal Auditory Canal; PTCD= Pontine Tegmental Cap Dysplasia; EMR= Electronic Medical Record; SNHL= sensorineural hearing loss.