Intolerance of uncertainty and psychological flexibility as predictors of mental health from adolescence to old age.

PURPOSE: The COVID-19 pandemic brought with it significant social, economic and health uncertainties. These were proposed to impact young people more compared to adults, leading adolescents to report more mental health problems during the pandemic. The current study examined whether differences in cognitive risk (tolerance of uncertainty) and protective (psychological flexibility) factors accounted for age-related differences in depression and anxiety. METHODS: These associations were investigated in the COVID-19 Risks Across the Lifespan (CORAL) cohort (N = 2280, 11-89 years). RESULTS: The results showed that adolescents experienced greater intolerance of uncertainty and lower psychological flexibility compared to adults and older adults. Tolerance of uncertainty did not account for age-related differences in depression or anxiety. However, psychological flexibility conferred more protective advantage for anxiety in adults compared to adolescents. CONCLUSION: The observed age-related differences in risk and protective factors advance our understanding of developmental vulnerabilities to depression and anxiety. Implications for mental health interventions in the context of future pandemics are discussed.

Occipital-temporal cortical tuning to semantic and affective features of natural images predicts associated behavioral responses.

In everyday life, people need to respond appropriately to many types of emotional stimuli. Here, we investigate whether human occipital-temporal cortex (OTC) shows co-representation of the semantic category and affective content of visual stimuli. We also explore whether OTC transformation of semantic and affective features extracts information of value for guiding behavior. Participants viewed 1620 emotional natural images while functional magnetic resonance imaging data were acquired. Using voxel-wise modeling we show widespread tuning to semantic and affective image features across OTC. The top three principal components underlying OTC voxel-wise responses to image features encoded stimulus animacy, stimulus arousal and interactions of animacy with stimulus valence and arousal. At low to moderate dimensionality, OTC tuning patterns predicted behavioral responses linked to each image better than regressors directly based on image features. This is consistent with OTC representing stimulus semantic category and affective content in a manner suited to guiding behavior.

Assessing the correlation between fecal calprotectin, blood markers and disease activity in pediatric inflammatory bowel disease.

Background: Crohn's disease (CD) and ulcerative colitis (UC) are the 2 main types of inflammatory bowel disease (IBD), a chronic inflammatory condition of the gastrointestinal tract. Management of IBD necessitates frequent clinical monitoring, including blood tests and occasionally endoscopy. Fecal calprotectin (FC) is a non-invasive measurement of luminal inflammatory activity, and can therefore be used as a useful monitoring tool. This study aimed to assess the relationship between FC, IBD activity indices and the commonly used blood markers in pediatric IBD. Methods: Electronic patient records were accessed to retrospectively collect patient data from a tertiary pediatric hospital from 2015-2021. CD and UC disease activity was quantified using the Pediatric CD Activity Index (PCDAI) and Pediatric UC Activity Index (PUCAI), respectively. The Paris classification was used for phenotype identification. Results: A total of 208 patients were included in the study, 115 with CD (18% <10 years and 82% 10-17 years) and 93 with UC (32% <10 years and 68% 10-17 years). There was a positive correlation between FC and PCDAI (rs=0.546, P<0.001) and between FC and PUCAI (rs=0.485, P<0.001). FC and activity indices were correlated positively with inflammatory markers/platelets and negatively with albumin and hemoglobin. FC correlated positively with PCDAI in all CD phenotypes, including isolated ileal disease. Conclusion: In pediatric IBD, FC shows a positive correlation with the clinical picture and blood markers in all disease phenotypes, and can provide an accurate non-invasive measure of disease activity.

Highly controlled multiplex electrospinning.

Applications of electrospinning (ES) range from fabrication of biomedical devices and tissue regeneration scaffolds to light manipulation and energy conversion, and even to deposition of materials that act as growth platforms for nanoscale catalysis. One major limitation to wide adoption of ES is stochastic fiber deposition resulting from the chaotic motion of the polymer stream as is approaches the deposition surface. In the past, fabrication of structures or materials with precisely determined mesoscale morphology has been accomplished through modification of electrode shape, use of multi-dimensional electrodes or pins, deposition onto weaving looms, hand-held electrospinning devices that allow the user to guide deposition, or electric field manipulation by lensing elements or apertures. In this work, we demonstrate an ES system that contains multiple high voltage power supplies that are independently controlled through a control algorithm implemented in LabVIEW. The end result is what we term "multiplex ES" where multiple independently controlled high-voltage signals are combined by the ES fiber to result in unique deposition control. COMSOL Multiphysics® software was used to model the electric field produced in this novel ES system. Using the multi-power supply system, we demonstrate fabrication of woven fiber materials that do not require complex deposition surfaces. Time-varied sinusoidal wave inputs were used to create electrospun torus shapes. The outer diameter of the tori was found, through parametric analysis, to be rather insensitive to frequency used during deposition, while inner diameter was inversely related to frequency, resulting in overall width of the tori increasing with frequency. Multiplex ES has a high-frequency cutoff based on the time response of the high voltage electrical circuit. These time constants were measured and minimized through the addition of parallel resistors that decreased impedance of the system and improved the high-frequency cutoff by up to 63%.

Sigh generation in preBötzinger Complex.

We explored neural mechanisms underlying sighing. Photostimulation of parafacial (pF) neuromedin B (NMB) or gastrin releasing peptide (GRP), or preBötzinger Complex (preBötC) NMBR or GRPR neurons elicited ectopic sighs with latency inversely related to time from preceding endogenous sigh. Of particular note, ectopic sighs could be produced without involvement of these peptides or their receptors in preBötC. Moreover, chemogenetic or optogenetic activation of preBötC SST neurons induced sighing, even in the presence of NMBR and/or GRPR antagonists. We propose that an increase in the excitability of preBötC NMBR or GRPR neurons not requiring activation of their peptide receptors activates partially overlapping pathways to generate sighs, and that preBötC SST neurons are a downstream element in the sigh generation circuit that converts normal breaths into sighs.

Protein Interaction Kinetics Delimit the Performance of Phosphorylation-Driven Protein Switches.

Post-translational modifications (PTMs) such as phosphorylation and dephosphorylation can rapidly alter protein surface chemistry and structural conformation, which can switch protein-protein interactions (PPIs) within signaling networks. Recently, de novo-designed phosphorylation-responsive protein switches have been created that harness kinase- and phosphatase-mediated phosphorylation to modulate PPIs. PTM-driven protein switches are promising tools for investigating PTM dynamics in living cells, developing biocompatible nanodevices, and engineering signaling pathways to program cell behavior. However, little is known about the physical and kinetic constraints of PTM-driven protein switches, which limits their practical application. In this study, we present a framework to evaluate two-component PTM-driven protein switches based on four performance metrics: effective concentration, dynamic range, response time, and reversibility. Our computational models reveal an intricate relationship between the binding kinetics, phosphorylation kinetics, and switch concentration that governs the sensitivity and reversibility of PTM-driven protein switches. Building upon the insights of the interaction modeling, we built and evaluated novel phosphorylation-driven protein switches consisting of phosphorylation-sensitive coiled coils as sensor domains fused to fluorescent proteins as actuator domains. By modulating the phosphorylation state of the switches with a specific protein kinase and phosphatase, we demonstrate fast, reversible transitions between "on" and "off" states. The response of the switches linearly correlated to the kinase concentration, demonstrating its potential as a biosensor for kinase measurements in real time. As intended, the switches responded to specific kinase activity with an increase in the fluorescence signal and our model could be used to distinguish between two mechanisms of switch activation: dimerization or a structural rearrangement. The protein switch kinetics model developed here should enable PTM-driven switches to be designed with ideal performance for specific applications.

Reanalysis of Immunopeptidomics Datasets Provides Mechanistic Insight into TAPBPR-Mediated Peptide Editing on HLA-A, -B and -C Molecules.

Background: Major histocompatibility class I (MHC-I, human leukocyte antigen [HLA]-I in humans) molecules present small fragments of the proteome on the cell surface for immunosurveillance, which is pivotal to control infected and malignant cells. Immunogenic peptides are generated and selected in the MHC-I antigen processing and presentation pathway. In this pathway, two homologous molecules, tapasin and TAPBPR, optimise the MHC-I peptide repertoire that is ultimately presented at the plasma membrane. Peptide exchange on HLA-I by human TAPBPR involves the flexible loop region K22-D35, with the leucine at position 30 (L30) involved in mediating peptide dissociation. However, our understanding of the exact molecular mechanisms governing TAPBPR-mediated peptide exchange on HLA-I allotypes remains incomplete. Methods: Here, in-depth re-analyses of published immunopeptidomics datasets was used to further examine TAPBPR peptide editing activity and mechanism of action on HLA-I. The role of the TAPBPR editing loop in opening the HLA-I peptide binding groove was assessed using a molecular dynamics simulation. Results: We show that TAPBPR shapes the peptide repertoire on HLA-A, -B and -C allotypes. The TAPBPR editing loop was not essential to allow HLA-I to adopt an open state. L30 in the TAPBPR editing loop was typically sufficient to mediate peptide repertoire restriction on the three HLA-I allotypes expressed by HeLa cells. TAPBPR was also able to load peptides onto HLA-I in a loop-dependent manner. Conclusions: These results unify the previously hypothesised scoop loop and peptide trap mechanisms of TAPBPR-mediated peptide exchange, with the former involved in peptide filtering and the latter in peptide loading.

Major histocompatibility complex (MHC) class I molecules play an essential role in alerting the immune system to infection and cellular changes. They do this by displaying small fragments of proteins (peptides) from pathogen-infected cells and tumours on the cell surface to immune cells. When activated, immune cells can then destroy the target cell. In 2015, we discovered that a novel accessory protein, called TAPBPR, assists in the selection of peptides displayed on MHC class I molecules for immune surveillance. A specific region in the TAPBPR protein ­ the editing loop ­ is known to be involved in removing peptides from MHC class I. However, our understanding of the process of peptide selection on MHC class I molecules remains incomplete. Here, we show that TAPBPR is not only involved in removing peptides from MHC class I molecules but also assists in peptide loading. Additionally, we demonstrate that the TAPBPR editing loop is involved in both removing and loading of peptides. Our results suggest that TAPBPR fine-tunes the peptide repertoire displayed on three different types of MHC class I molecules. Developing our understanding of the mechanisms of peptide selection on MHC class I molecules has important implications in disease and the development of new therapies.

Anthropogenic noise disrupts early-life development in a fish with paternal care.

Anthropogenic noise is a global pollutant but its potential impacts on early life-stages in fishes are largely unknown. Here, using controlled laboratory experiments, we tested for impacts of continuous or intermittent exposure to low-frequency broadband noise on early life-stages of the common goby (Pomatoschistus microps), a marine fish with exclusive paternal care. Neither continuous nor intermittent noise exposure had an effect on filial cannibalism, showing that males were capable and willing to care for their broods. However, broods reared in continuous noise covered a smaller area and contained fewer eggs than control broods. Moreover, although developmental rate was the same in all treatments, larvae reared by males in continuous noise had, on average, a smaller yolk sac at hatching than those reared in the intermittent noise and control treatments, while larvae body length did not differ. Thus, it appears that the increased consumption of the yolk sac reserve was not utilised for increased growth. This suggests that exposure to noise in early life-stages affects fitness-related traits of surviving offspring, given the crucial importance of the yolk sac reserve during the early life of pelagic larvae. More broadly, our findings highlight the wide-ranging impacts of anthropogenic noise on aquatic wildlife living in an increasingly noisy world.

Biopsychosocial Assessments for Pubertal Suppression to Treat Adolescent Gender Dysphoria.

Transgender and gender diverse (TGD) adolescents are those whose gender identity is incongruent with societal expectations based on their sex assigned at birth.1 Some TGD adolescents may meet criteria for gender dysphoria, which requires at least 6 months of psychological distress related to gender incongruence.1 Such adolescents may seek pubertal suppression with a gonadotropin-releasing hormone agonist (GnRHa).1 GnRHa temporarily and reversibly suppress endogenous puberty, allowing adolescents to engage with care without puberty-related psychological distress.2 Most patients treated with GnRHa proceed to treatment with gender-affirming hormones (eg, estrogen or testosterone to induce puberty that aligns with their gender identity), whereas a small number choose to stop GnRHa treatment, allowing endogenous puberty to proceed.2 Longitudinal cohort and cross-sectional studies link GnRHa treatment to improved mental health outcomes, when provided in accordance with clinical guidelines.2.

Removal, Kill, and Transfer of Bacteria from Hands by Antibacterial or Nonantibacterial Soaps After Handling Raw Poultry.

Hand hygiene is broadly recognized as a critical intervention in reducing the spread of disease-causing pathogens in both professional and personal uses. In this study, the impact of antibacterial (AB) or nonantibacterial soaps on the removal and postwash transfer of E. coli following the handling of raw poultry was assessed. Baseline bacterial contamination ranged between 107 and 109 CFU per hand. Hands were washed for 30 s in 40°C ± 2°C tap water using 2 mL of AB soap (0.5% and 1.0% Chloroxylenol, 0.5% Benzalkonium Chloride, or 4.0% Chlorhexidine Gluconate), non-AB soap (cosmetic/plain soap), or water. Postwash, water, and non-AB soap had a mean 3.63 and 3.65 Log10 reduction of E. coli on hands. AB treatments had a mean 4.19-4.35 Log10 reduction. Rinse water had mean bacterial counts of 8.62 and 8.88 Log10 CFU/mL for non-AB soap and water and 5.37-6.90 Log10 CFU/mL for AB treatments. Bacterial transfer was assessed by following the test subject's handling of a sterile polymer knife handle for 30 s postwash. E. coli transfer ranged from 263 to 903 CFU/handle for AB soaps and 1572 or 1709 CFU/handle for water and non-AB soap. Differences between AB and non-AB treatments were statistically significant (p < 0.0001) for hands and rinse water. Differences in transfer from hands to knife handle were not statistically significant (p = 0.139). Combined, these data highlight significant differences in the performance of AB soaps relative to non-AB soaps in a food handling environment-specific usage example and provide an unexplored assessment of the bactericidal vs. removal effects of AB vs. non-AB soaps on bacteria removed from the hands. These data reinforce the importance of hand hygiene, provide new details on the differences between AB vs. non-AB soaps, and highlight potential differences to inform food handling environment operators and public health personnel on how these products may impact food safety.

Binding of Per- and Polyfluoroalkyl Substances to ß-Lactoglobulin from Bovine Milk.

Per- and polyfluoroalkyl substances (PFAS) are known for their high environmental persistence and potential toxicity. The presence of PFAS has been reported in many dairy products. However, the mechanisms underlying the accumulation of PFAS in these products remain unclear. Here, we used native mass spectrometry and molecular dynamics simulations to probe the interactions between 19 PFAS of environmental concern and two isoforms of the major bovine whey protein ß-lactoglobulin (ß-LG). We observed that six of these PFAS bound to both protein isoforms with low- to mid-micromolar dissociation constants. Based on quantitative, competitive binding experiments with endogenous ligands, PFAS can bind orthosterically and preferentially to ß-LG's hydrophobic ligand-binding calyx. ß-Cyclodextrin can also suppress binding of PFAS to ß-LG owing to the ability of ß-cyclodextrin to directly sequester PFAS from solution. This research sheds light on PFAS-ß-LG binding, suggesting that such interactions could impact lipid-fatty acid transport in bovine mammary glands at high PFAS concentrations. Furthermore, our results highlight the potential use of ß-cyclodextrin in mitigating PFAS binding, providing insights toward the development of strategies to reduce PFAS accumulation in dairy products and other biological systems.

Medicaid Acceptance Varies by Physician Seniority and Specialty in California.

Given varied insurance acceptances and differing pay between insurances, our objective was to examine the number of California physicians enrolled in Medicare and Medicaid (Medi-Cal), stratified by specialty and graduation year. Medi-Cal and Medicare providers were extracted from publicly available databases (Centers for Medicare & Medicaid Services and California Health and Human Services) and were subsequently merged into one dataset using National Provider Identifier. From there, we stratified physicians by specialty and graduation year. We found that emergency medicine, radiology, pathology, anesthesiology, general surgery, and internal medicine had the highest percent of Medi-Cal-accepting physicians, whereas dermatology, psychiatry, physical medicine & rehabilitation, and plastic & reconstructive surgery physicians had the lowest. There also appears to be an inverse relationship between acceptance of Medi-Cal and earlier year of graduation (P < 0.05). This study demonstrated striking variability in Medi-Cal acceptance based upon physician years in practice and specialty. Older, experienced physicians, as well as physicians of certain specialties, are less likely to accept Medi-Cal.

Phospholipids Differentially Regulate Ca2+ Binding to Synaptotagmin-1.

Synaptotagmin-1 (Syt-1) is a calcium sensing protein that is resident in synaptic vesicles. It is well established that Syt-1 is essential for fast and synchronous neurotransmitter release. However, the role of Ca2+ and phospholipid binding in the function of Syt-1, and ultimately in neurotransmitter release, is unclear. Here, we investigate the binding of Ca2+ to Syt-1, first in the absence of lipids, using native mass spectrometry to evaluate individual binding affinities. Syt-1 binds to one Ca2+ with a KD ∼ 45 µM. Each subsequent binding affinity (n ≥ 2) is successively unfavorable. Given that Syt-1 has been reported to bind anionic phospholipids to modulate the Ca2+ binding affinity, we explored the extent that Ca2+ binding was mediated by selected anionic phospholipid binding. We found that phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) and dioleoylphosphatidylserine (DOPS) positively modulated Ca2+ binding. However, the extent of Syt-1 binding to phosphatidylinositol 3,5-bisphosphate (PI(3,5)P2) was reduced with increasing [Ca2+]. Overall, we find that specific lipids differentially modulate Ca2+ binding. Given that these lipids are enriched in different subcellular compartments and therefore may interact with Syt-1 at different stages of the synaptic vesicle cycle, we propose a regulatory mechanism involving Syt-1, Ca2+, and anionic phospholipids that may also control some aspects of vesicular exocytosis.

Innovations in pharmacovigilance studies of medicines in older people.

Pharmacovigilance is defined by the World Health Organization as "the science and activities relating to the detection, assessment, understanding and prevention of adverse effects or any other medicine/vaccine related problem". Pharmacovigilance studies are critical for detecting and assessing adverse events of medicines that may not have been observed in clinical trials. This activity is especially important in older people who are often excluded from clinical trials as they have multiple chronic conditions and use multiple medicines for longer durations than the clinical trials. In this narrative review we describe innovative methods in pharmacovigilance studies of medicines in older people that leverage the increasing availability of digital health technologies, electronic health records and real-world health data to identify and quantify medication related harms in older people.

On the reliable estimation of sequential Monod kinetic parameters.

While dozens of studies have attempted to estimate the Monod kinetic parameters of microbial reductive dechlorination, published values in the literature vary by 2-6 orders of magnitude. This lack of consensus can be attributed in part to limitations of both experimental design and parameter estimation techniques. To address these issues, Hamiltonian Monte Carlo was used to produce more than one million sets of realistic simulated microcosm data under a variety of experimental conditions. These data were then employed in model fitting experiments using a number of parameter estimation algorithms for determining Monod kinetic parameters. Analysis of data from conventional triplicate microcosms yielded parameter estimates characterized by high collinearity, resulting in poor estimation accuracy and precision. Additionally, confidence intervals computed by commonly used classical regression analysis techniques contained true parameter values much less frequently than their nominal confidence levels. Use of an alternative experimental design, requiring the same number of analyses as conventional experiments but comprised of microcosms with varying initial chlorinated ethene concentrations, is shown to result in order-of-magnitude decreases in parameter uncertainty. A Metropolis algorithm which can be run on a typical personal computer is demonstrated to return more reliable parameter interval estimates.

Kir4.1 channels contribute to astrocyte CO2/H+-sensitivity and the drive to breathe.

Astrocytes in the retrotrapezoid nucleus (RTN) stimulate breathing in response to CO2/H+, however, it is not clear how these cells detect changes in CO2/H+. Considering Kir4.1/5.1 channels are CO2/H+-sensitive and important for several astrocyte-dependent processes, we consider Kir4.1/5.1 a leading candidate CO2/H+ sensor in RTN astrocytes. To address this, we show that RTN astrocytes express Kir4.1 and Kir5.1 transcripts. We also characterized respiratory function in astrocyte-specific inducible Kir4.1 knockout mice (Kir4.1 cKO); these mice breathe normally under room air conditions but show a blunted ventilatory response to high levels of CO2, which could be partly rescued by viral mediated re-expression of Kir4.1 in RTN astrocytes. At the cellular level, astrocytes in slices from astrocyte-specific inducible Kir4.1 knockout mice are less responsive to CO2/H+ and show a diminished capacity for paracrine modulation of respiratory neurons. These results suggest Kir4.1/5.1 channels in RTN astrocytes contribute to respiratory behavior.

Applying Proteomics and Computational Approaches to Identify Novel Targets in Blast-Associated Post-Traumatic Epilepsy.

Traumatic brain injury (TBI) can lead to post-traumatic epilepsy (PTE). Blast TBI (bTBI) found in Veterans presents with several complications, including cognitive and behavioral disturbances and PTE; however, the underlying mechanisms that drive the long-term sequelae are not well understood. Using an unbiased proteomics approach in a mouse model of repeated bTBI (rbTBI), this study addresses this gap in the knowledge. After rbTBI, mice were monitored using continuous, uninterrupted video-EEG for up to four months. Following this period, we collected cortex and hippocampus tissues from three groups of mice: those with post-traumatic epilepsy (PTE+), those without epilepsy (PTE-), and the control group (sham). Hundreds of differentially expressed proteins were identified in the cortex and hippocampus of PTE+ and PTE- relative to sham. Focusing on protein pathways unique to PTE+, pathways related to mitochondrial function, post-translational modifications, and transport were disrupted. Computational metabolic modeling using dysregulated protein expression predicted mitochondrial proton pump dysregulation, suggesting electron transport chain dysregulation in the epileptic tissue relative to PTE-. Finally, data mining enabled the identification of several novel and previously validated TBI and epilepsy biomarkers in our data set, many of which were found to already be targeted by drugs in various phases of clinical testing. These findings highlight novel proteins and protein pathways that may drive the chronic PTE sequelae following rbTBI.