Regulation of Absorption and Emission in a Protein/Fluorophore Complex.

Human cellular retinol binding protein II (hCRBPII) was used as a protein engineering platform to rationally regulate absorptive and emissive properties of a covalently bound fluorogenic dye. We demonstrate the binding of a thio-dapoxyl analog via formation of a protonated imine between an active site lysine residue and the chromophore's aldehyde. Rational manipulation of the electrostatics of the binding pocket results in a 204 nm shift in absorption and a 131 nm shift in emission. The protein is readily expressed in mammalian systems and binds with exogenously delivered fluorophore as demonstrated by live-cell imaging experiments.

From concept to reality: A comprehensive exploration into the development and evolution of a virtual emergency department.

Emergency department (ED) overcrowding remains a persistent challenge in global public health, leading to detrimental outcomes for patients and healthcare professionals. Traditional approaches to improve this issue have been insufficient, prompting exploration of novel strategies such as virtual care interventions. Our team developed the first comprehensive statewide virtual ED in Australia, the Victorian Virtual Emergency Department, offering an alternative to in-person care for non-life-threatening emergencies. Here, we present the development and ongoing refinement of this pioneering virtual care service, aiming to provide insights for hospital administrators and policymakers seeking to implement patient-centric care solutions worldwide. By sharing our model of care, we hope to guide further work toward addressing the global problem of over crowded EDs.

A perspective on brain-age estimation and its clinical promise.

Brain-age estimation has gained increased attention in the neuroscientific community owing to its potential use as a biomarker of brain health. The difference between estimated and chronological age based on neuroimaging data enables a unique perspective on brain development and aging, with multiple open questions still remaining in the brain-age research field. This Perspective presents an overview of current advancements in the field and envisions the future evolution of the brain-age framework before its potential deployment in hospital settings.

Perceptions of advice for acute low back pain: a content analysis of qualitative data collected in a randomised experiment.

OBJECTIVES: To explore how people perceive three different forms of advice for acute low back pain (LBP). DESIGN: Content analysis of qualitative data collected in a three-arm randomised experiment. PARTICIPANTS: 2200 participants with acute LBP (ie, pain duration for ≤6 weeks) were randomly assigned to receive three types of advice: guideline advice and guideline advice with the addition of either brief pain science or ergonomics messages. PRIMARY AND SECONDARY OUTCOMES: After receiving the advice, participants answered two questions: 'If your health professional gave you this advice, how would it make you feel?' and 'If your health professional gave you this advice, what treatments (if any) do you think you would need?' Two researchers coded responses using deductive content analysis. RESULTS: We analysed 4400 free-text responses from 2200 participants. There were little to no differences in participants' feelings, thoughts and expectations after receiving three types of advice for acute LBP. Participants most commonly expressed feeling positive about the advice (38%-35%), reassured (23%-22%) and empowered (10%-8%). Some expressed being unhappy or being frustrated with the advice (4%-3%). Participants most commonly thought they needed no treatment apart from staying active, followed by exercise and medication. CONCLUSIONS: Guideline advice with or without the addition of brief pain science or ergonomics messages generated positive feelings, reassurance or a sense of empowerment in many people with acute LBP, with no difference between types of advice. TRIAL REGISTRATION NUMBER: ACTRN12623000364673.

Disentangling autonomy-supportive and psychologically controlling parenting: A meta-analysis of self-determination theory's dual process model across cultures.

Self-determination theory's (SDT) dual process model claims that parental autonomy support relates positively to child well-being, while psychologically controlling parenting is linked positively to child ill-being. We tested these claims using a combination of one-stage and univariate meta-analytic structural equation modeling with moderation (k = 238; n = 1,040, N = 126,423). In the univariate models, parental autonomy support was linked positively with child well-being, r = 0.30, 95% CI [0.26, 0.33], whereas parental psychological control was positively linked with child ill-being, r = 0.26, 95% CI [0.23, 0.28]. Consistent with SDT's dual process model, the one-stage model that controlled for the intercorrelations between predictors showed that parental autonomy support and psychological control had distinct links to child wellness outcomes. Parental autonomy support was linked positively with child well-being, even when accounting for psychological control, r = 0.26, 95% CI [0.20, 0.31], and parental psychological control was positively linked to child ill-being, controlling for autonomy support, r = 0.20, 95% CI [0.17, 0.23]. Crucially, the beneficial effects of parental autonomy support and the costs of psychological control applied across regions, degrees of national individualism and cultural hierarchy, as well as child developmental periods and sexes. These results help move the field beyond debates about whether autonomy is beneficial toward questions about manifestations of autonomy across groups and variations in its optimal support. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

Upregulation of TET2 and Resistance to DNA Methyltransferase (DNMT) Inhibitors in DNMT1-Deleted Cancer Cells.

BACKGROUND: Ten-eleven-translocation (TET) 2 is a member of the TET family of proteins (TET1-3). DNMT1 gene deletion confers resistance to DNA methyltransferase (DNMT) inhibitors in colorectal, breast, and ovarian cancer cells. Currently, the effect of DNMT1 gene status on TET2 phenotype following DNMT inhibitor treatment is unclear in human malignancies. METHODS: Human colorectal carcinoma HCT116 cells (DNMT+/+) and their isogenic DNMT1 knockout (DNMT1-/-) counterpart were treated with DNMT inhibitors. Expression of TET2 and tumor suppressor (p16ink4A and p15ink4B) proteins were examined by Western blot. Apoptosis and CDKN2A promoter demethylation following drug treatment were detected by Annexin-V apoptosis assay and methylation-specific PCR. RESULTS: TET2 expression was robustly increased in DNMT1-/- cells by 0.5 µM and 5 µM decitabine and azacitidine treatment. Augmentation of TET2 expression was accompanied by re-expression of p16ink4A and p15ink4B proteins and CDKN2A promoter demethylation. TET2 upregulation and tumor suppressor re-expression were associated with resistance conferred by DNMT1 deletion. Treatment with 5-aza-4'-thio-2'-deoxycytidine at a low 0.5 µM dose only upregulated TET2 and reduced CDKN2A promoter methylation, and re-expression of p16ink4A in DNMT1-/- cells. DNMT inhibitors showed minimal effects on TET2 upregulation and re-expression of tumor suppressor proteins in cells with intact DNMT1. CONCLUSIONS: DNMT1 gene deletion made cancer cells prone to TET2 upregulation and activation of tumor suppressor expression upon DNMT inhibitor challenge. TET2 augmentation is concomitant with resistance to DNMT inhibitors in a DNMT1-deleted state.

Arm Injuries and Prevention Proficiency in High School Fastpitch Softball Players: A Survey of Players, Parents, and Coaches.

Arm injuries are common in youth fastpitch softball players. To characterize arm injuries and assess injury prevention knowledge, we administered a cross-sectional online survey and injury prevention quiz to high school fastpitch softball players, parents, and coaches. Associations between throwing habits and adverse arm outcomes were computed. Surveys were collected from 123 high school players, 59 parents, and 28 coaches, totaling 210 responses. Nearly half (48.8%) of players saw a doctor for arm pain. Riseball pitchers were less likely to undergo surgery than non-riseball pitchers (P = .008). Outfield/utility players trended toward more frequent arm pain compared to alternative position groups (P = .086). On injury prevention quizzes, players, parents, and coaches averaged scores of 56-65% correct, with players scoring below parents (P = .011) and coaches (P = .006). In conclusion, high school softball players frequently seek medical attention for arm pain, and injury prevention knowledge is consistently lacking.

Transgenerational phenotypic responses to herbicide stress are more rapid than to shade and simulated herbivory in Arabidopsis.

Weeds in agricultural settings continually adapt to stresses from ecological and anthropogenic sources, in some cases leading to resistant populations. However, consequences of repeated sub-lethal exposure of these stressors on fitness and stress "memory" over generations remain poorly understood. We measured plant performance over a transgenerational experiment with Arabidopsis thaliana where plants were exposed to sub-lethal stress induced by the herbicides glyphosate or trifloxysulfuron, stresses from clipping or shading in either one (G1) or four successive generations (G1-G4), and control plants that never received stress. We found that fourth-generation (G4) plants that had been subjected to three generations of glyphosate or trifloxysulfuron stress produced higher post-stress biomass, seed weight, and rosette area as compared to that produced by plants that experienced stress only in the first generation (G1). By the same measure, clipping and shade were more influential on floral development time (shade) and seed weight (clipping) but did not show responsive phenotypes for vegetative metrics after multiple generations. Overall, we found that plants exhibited more rapid transgenerational vegetative "stress memory" to herbicides while reproductive plasticity was stressor dependent and similar between clipping/shade and anthropogenic stressors. Our study suggests that maternal plant stress memory aids next-generation plants to respond and survive better under the same stressors.

Insect Flight: State of the Field and Future Directions.

The evolution of flight in an early winged insect ancestral lineage is recognized as a key adaptation explaining the unparalleled success and diversification of insects. Subsequent transitions and modifications to flight machinery, including secondary reductions and losses, also play a central role in shaping the impacts of insects on broadscale geographic and ecological processes and patterns in the present and future. Given the importance of insect flight, there has been a centuries-long history of research and debate on the evolutionary origins and biological mechanisms of flight. Here, we revisit this history from an interdisciplinary perspective, discussing recent discoveries regarding the developmental origins, physiology, biomechanics, and neurobiology and sensory control of flight in a diverse set of insect models. We also identify major outstanding questions yet to be addressed and provide recommendations for overcoming current methodological challenges faced when studying insect flight, which will allow the field to continue to move forward in new and exciting directions. By integrating mechanistic work into ecological and evolutionary contexts, we hope that this synthesis promotes and stimulates new interdisciplinary research efforts necessary to close the many existing gaps about the causes and consequences of insect flight evolution.

Consistencies in the dietary and isotopic niche of spotted seatrout, Cynoscion nebulosus, across a salinity gradient within a coastal Louisiana estuary.

Estuaries are essential habitats for recreational and commercial fish that are shaped by both natural and anthropogenic processes. In Louisiana a combination of climate change and planned coastal restoration actions is predicted to increase freshwater introduction to coastal estuaries. As such there is a need to quantify the relationships between estuarine fish ecology and salinity to aid in predicting how species will respond to shifts in salinity. We investigated the relative abundance and dietary niches of adult (24.5 ± 5.4 cm standard length) spotted seatrout Cynoscion nebulosus across varying salinity regimes (oligohaline, mesohaline, and polyhaline) within Barataria Bay, Louisiana, using a combination of net sampling and gut content and stable isotopes analysis. We found that the relative abundance of C. nebulosus was lowest at the oligohaline site, translating to approximately five fewer fish captured for every single psu decrease in a site's average annual salinity. In contrast, we found that diets and, to a lesser extent, isotopic niches had a high degree of overlap across sites with differing salinity regimes. Fish and penaeid shrimp were the most common and important prey taxa recovered from guts at all sites. The small isotopic differences found among sites were likely due to spatial variation in hydrogeochemical baselines, and the observed isotopic overlap provides support for the idea that C. nebulosus move between adjacent salinity regimes and forage throughout Barataria Bay. Our results contribute to a greater understanding of the salinity preference and trophic ecology of C. nebulosus that can aid in predicting their responses to future salinity and habitat changes within Barataria Bay associated with predicted climate change and planned coastal restoration actions.

A Glycemic Status Classification Model Using a Radiofrequency Noninvasive Blood Glucose Monitor.

Despite significant efforts in the development of noninvasive blood glucose (BG) monitoring solutions, delivering an accurate, real-time BG measurement remains challenging. We sought to address this by using a novel radiofrequency (RF) glucose sensor to noninvasively classify glycemic status. The study included 31 participants aged 18-65 with prediabetes or type 2 diabetes and no other significant medical history. During control sessions and oral glucose tolerance test sessions, data were collected from both a RF sensor that rapidly scans thousands of frequencies and concurrently from a venous blood draw measured with an US Food and Drug Administration (FDA)-cleared glucose hospital meter system to create paired observations. We trained a time series forest machine learning model on 80% of the paired observations and reported results from applying the model to the remaining 20%. Our findings show that the model correctly classified glycemic status 93.37% of the time as high, normal, or low.

Reversible two-way tuning of thermal conductivity in an end-linked star-shaped thermoset.

Polymeric thermal switches that can reversibly tune and significantly enhance their thermal conductivities are desirable for diverse applications in electronics, aerospace, automotives, and medicine; however, they are rarely achieved. Here, we report a polymer-based thermal switch consisting of an end-linked star-shaped thermoset with two independent thermal conductivity tuning mechanisms-strain and temperature modulation-that rapidly, reversibly, and cyclically modulate thermal conductivity. The end-linked star-shaped thermoset exhibits a strain-modulated thermal conductivity enhancement up to 11.5 at a fixed temperature of 60 °C (increasing from 0.15 to 2.1 W m-1 K-1). Additionally, it demonstrates a temperature-modulated thermal conductivity tuning ratio up to 2.3 at a fixed stretch of 2.5 (increasing from 0.17 to 0.39 W m-1 K-1). When combined, these two effects collectively enable the end-linked star-shaped thermoset to achieve a thermal conductivity tuning ratio up to 14.2. Moreover, the end-linked star-shaped thermoset demonstrates reversible tuning for over 1000 cycles. The reversible two-way tuning of thermal conductivity is attributed to the synergy of aligned amorphous chains, oriented crystalline domains, and increased crystallinity by elastically deforming the end-linked star-shaped thermoset.

SigmaR1 shapes rough endoplasmic reticulum membrane sheets.

Rough endoplasmic reticulum (ER) sheets are a fundamental domain of the ER and the gateway into the secretory pathway. Although reticulon proteins stabilize high-curvature ER tubules, it is unclear whether other proteins scaffold the flat membranes of rough ER sheets. Through a proteomics screen using ER sheet-localized RNA-binding proteins as bait, we identify the sigma-1 receptor (SigmaR1) as an ER sheet-shaping factor. High-resolution live cell imaging and electron tomography assign SigmaR1 as an ER sheet-localized factor whose levels determine the amount of rough ER sheets in cells. Structure-guided mutagenesis and in vitro reconstitution on giant unilamellar vesicles further support a mechanism whereby SigmaR1 oligomers use their extended arrays of amphipathic helices to bind and flatten the lumenal leaflet of ER membranes to oppose membrane curvature and stabilize rough ER sheets.

Patient Acceptable Symptomatic State and Substantial Clinical Benefit Matter Most to Patients and Must Be Reported Correctly.

Despite a push for a focus on clinical rather than "statistical" significance, and an emphasis on reporting of outcome thresholds such as Patient Acceptable Symptomatic State (PASS) and Substantial Clinical Benefit (SCB), PASS and SCB are rarely reported, and when reported, are often reported incorrectly. Yet, patients require satisfaction (PASS) as a result of our treatments, and patients desire to improve substantially (SCB). Determining whether patients are satisfied and/or substantially improved is simple…just ask them. The questions are known as anchor questions. Obviously, different patients have different thresholds of PASS and SCB, and reliance on previously published literature for values of these thresholds can result in error-thus, the anchor questions. And, each patient must be assessed individually. Outcome thresholds are not group-level metrics, and must be reported as the percentage of individuals who achieve the clinically significant outcome. Certain patients, such as athletes, have high baseline function and may demand Maximum Outcome Improvement or (MOI). In contrast, Minimally Clinically Important Difference (MCID) is a less-than-ideal measure; patients do not desire "minimal" improvement. Journals must do a better job of publishing Patient Reported Outcome Measures (PROM) that matter most to patients: satisfaction and substantial benefit.

Bringing optimised COVID-19 vaccine schedules to immunocompromised populations (BOOST-IC): study protocol for an adaptive randomised controlled clinical trial.

BACKGROUND: Immunocompromised hosts (ICH) experience more breakthrough infections and worse clinical outcomes following infection with COVID-19 than immunocompetent people. Prophylactic monoclonal antibody therapies can be challenging to access, and escape variants emerge rapidly. Immunity conferred through vaccination remains a central prevention strategy for COVID-19. COVID-19 vaccines do not elicit optimal immunity in ICH but boosting, through additional doses of vaccine improves humoral and cellular immune responses. This trial aims to assess the immunogenicity and safety of different COVID-19 vaccine booster strategies against SARS-CoV-2 for ICH in Australia. METHODS: Bringing optimised COVID-19 vaccine schedules to immunocompromised populations (BOOST-IC) is an adaptive randomised trial of one or two additional doses of COVID-19 vaccines 3 months apart in people living with HIV, solid organ transplant (SOT) recipients, or those who have haematological malignancies (chronic lymphocytic leukaemia, non-Hodgkin lymphoma or multiple myeloma). Key eligibility criteria include having received 3 to 7 doses of Australian Therapeutic Goods Administration (TGA)-approved COVID-19 vaccines at least 3 months earlier, and having not received SARS-CoV-2-specific monoclonal antibodies in the 3 months prior to receiving the study vaccine. The primary outcome is the geometric mean concentration of anti-spike SARS-CoV-2 immunoglobulin G (IgG) 28 days after the final dose of the study vaccine. Key secondary outcomes include anti-spike SARS-CoV-2 IgG titres and the proportion of people seroconverting 6 and 12 months after study vaccines, local and systemic reactions in the 7 days after vaccination, adverse events of special interest, COVID-19 infection, mortality and quality of life. DISCUSSION: This study will enhance the understanding of COVID-19 vaccine responses in ICH, and enable the development of safe, and optimised vaccine schedules in people with HIV, SOT, or haematological malignancy. TRIAL REGISTRATION: ClinicalTrials.gov NCT05556720. Registered on 23rd August 2022.

Phosphonodiamidate prodrugs of phosphoantigens (ProPAgens) exhibit potent Vγ9/Vδ2 T cell activation and eradication of cancer cells.

The phosphoantigen (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (HMBPP) is an established activator of Vγ9/Vδ2 T cells and stimulates downstream effector functions including cytotoxicity and cytokine production. In order to improve its drug-like properties, we herein report the design, synthesis, serum stability, in vitro metabolism, and biological evaluation of a new class of symmetrical phosphonodiamidate prodrugs of methylene and difluoromethylene monophosphonate derivatives of HMBPP. These prodrugs, termed phosphonodiamidate ProPAgens, were synthesized in good yields, exhibited excellent serum stability (>7 h), and their in vitro metabolism was shown to be initiated by carboxypeptidase Y. These phosphonodiamidate ProPAgens triggered potent activation of Vγ9/Vδ2 T cells, which translated into efficient Vγ9/Vδ2 T cell-mediated eradication of bladder cancer cells in vitro. Together, these findings showcase the potential of these phosphonodiamidate ProPAgens as Vγ9/Vδ2 T cell modulators that could be further developed as novel cancer immunotherapeutic agents.

Design of Digital Mental Health Platforms for Family Member Cocompletion: Scoping Review.

BACKGROUND: The COVID-19 pandemic placed an additional mental health burden on individuals and families, resulting in widespread service access problems. Digital mental health interventions suggest promise for improved accessibility. Recent reviews have shown emerging evidence for individual use and early evidence for multiusers. However, attrition rates remain high for digital mental health interventions, and additional complexities exist when engaging multiple family members together. OBJECTIVE: As such, this scoping review aims to detail the reported evidence for digital mental health interventions designed for family use with a focus on the build and design characteristics that promote accessibility and engagement and enable cocompletion by families. METHODS: A systematic literature search of MEDLINE, Embase, PsycINFO, Web of Science, and CINAHL databases was conducted for articles published in the English language from January 2002 to March 2024. Eligible records included empirical studies of digital platforms containing some elements designed for cocompletion by related people as well as some components intended to be completed without therapist engagement. Platforms were included in cases in which clinical evidence had been documented. RESULTS: Of the 9527 papers reviewed, 85 (0.89%) met the eligibility criteria. A total of 24 unique platforms designed for co-use by related parties were identified. Relationships between participants included couples, parent-child dyads, family caregiver-care recipient dyads, and families. Common platform features included the delivery of content via structured interventions with no to minimal tailoring or personalization offered. Some interventions provided live contact with therapists. User engagement indicators and findings varied and included user experience, satisfaction, completion rates, and feasibility. Our findings are more remarkable for what was absent in the literature than what was present. Contrary to expectations, few studies reported any design and build characteristics that enabled coparticipation. No studies reported on platform features for enabling cocompletion or considerations for ensuring individual privacy and safety. None examined platform build or design characteristics as moderators of intervention effect, and none offered a formative evaluation of the platform itself. CONCLUSIONS: In this early era of digital mental health platform design, this novel review demonstrates a striking absence of information about design elements associated with the successful engagement of multiple related users in any aspect of a therapeutic process. There remains a large gap in the literature detailing and evaluating platform design, highlighting a significant opportunity for future cross-disciplinary research. This review details the incentive for undertaking such research; suggests design considerations when building digital mental health platforms for use by families; and offers recommendations for future development, including platform co-design and formative evaluation.

Overfeeding copper during rearing affects the liver function and fertility of replacement dairy heifers.

BACKGROUND: Oversupply of dietary copper (Cu) is common among UK dairy herds, but studies on the long-term outcomes of this oversupply are scarce. METHODS: A longitudinal study was undertaken to determine the long-term implications when 80 Holstein‒Friesian heifers with a mean (±standard error) age of 4.1 ± 0.1 months and a mean liveweight of 137 ± 2.4 kg were fed a recommended (R; 16 mg/kg dry matter [DM]) or high (H; 32 mg/kg DM) dietary Cu concentration until 6 weeks prior to calving. RESULTS: Hepatic Cu concentrations in both treatment groups were elevated into the ranges used to diagnose chronic Cu toxicity in cattle at 6.9 months of age (798 ± 46.4 mg/kg DM for H vs. 643 ± 35.4 mg/kg DM for R), with associated evidence of liver damage. Hepatic Cu concentrations then returned to normality but remained higher (p < 0.001) for heifers fed H than for those fed R and were associated with a reduced (p = 0.044) conception rate to first and second services (73.7 ± 8.05% for H vs. 91.2 ± 7.68% for R). LIMITATION: This retrospective analysis identified pre-study liver damage, which may have affected results. CONCLUSIONS: Supplying Cu in excess of requirements resulted in liver damage and reduced conception rates.