Autofluorescence-based tissue characterization enhances clinical prospects of light-sheet-microscopy.

Light sheet fluorescence microscopy (LSFM) is a transformative imaging method that enables the visualization of non-dissected specimen in real-time 3D. Optical clearing of tissues is essential for LSFM, typically employing toxic solvents. Here, we test the applicability of a non-hazardous alternative, ethyl cinnamate (ECi). We comprehensively characterized autofluorescence (AF) spectra in diverse murine tissues-ocular globe, knee, and liver-employing LSFM under various excitation wavelengths (405-785 nm) to test the feasibility of unstained samples for diagnostic purposes, in particular regarding percutaneous biopsies, as they constitute to most harvested type of tissue sample in clinical routine. Ocular globe structures were best discerned with 640 nm excitation. Knee tissue showed complex variation in AF spectra variation influenced by tissue depth and structure. Liver exhibited a unique AF pattern, likely linked to vasculature. Hepatic tissue samples were used to demonstrate the compatibility of our protocol for antibody staining. Furthermore, we employed machine learning to augment raw images and segment liver structures based on AF spectra. Radiologists rated representative samples transferred to the clinical assessment software. Learning-generated images scored highest in quality. Additionally, we investigated an actual murine biopsy. Our study pioneers the application of AF spectra for tissue characterization and diagnostic potential of optically cleared unstained percutaneous biopsies, contributing to the clinical translation of LSFM.

Inclusion of Dental Services in Medicare to Improve Oral and General Health for Older Americans.

Poor oral health negatively impacts overall health, quality of life and well-being. Increasing evidence suggests that provision of basic dental care for elderly Americans would improve outcomes for a variety of systemic diseases and reduce the overall cost of health care. As a result, recent changes have been implemented to include some dental benefits in the Medicare program. This article outlines evidence, rationale and approaches required for inclusion of dental benefits for more Americans through the Medicare program. Improving access to dental services through Medicare to help prevent and manage common chronic diseases is an important step towards integration of dental care with general healthcare to improve the overall health, quality of life, and well-being for many older Americans.

Economically optimized forage utilization choices in drylands for adapting to economic, ecological, and climate stress.

Improving the economic performance of range forage in drylands internationally faces challenges from economic, ecological, and climate stress. Stakeholders in these drylands wish to protect range forage ecosystems while assuring economic viability of ranching. Despite several recent research achievements, little work to date has integrated relationships among precipitation, grazing pressure, animal performance, and forage production to protect ranching incomes faced with economic, ecological, and climate stress in dryland areas. This work addresses that gap by developing an empirical mathematical programming model for optimizing economic performance of livestock grazing on range forage ecosystems that adapt to several stressors. Its unique contribution is to formulate and apply a ranch income optimization model calibrated using positive mathematical programming. The model replicates observed economic, forage, and climate conditions while accounting for interacting relations among stocking rates, forage conditions, grazing pressure, animal performance, and ranch economic productivity. Results show ranch incomes ranging from about $5 to $88 per acre and marginal values of forage ranging from $0.01 to $0.12 per pound of forage, depending on economic, ecological, and climate conditions. Results reveal how all these stressors affect economically optimized choices of grazing levels, ranch income, and economic values of forage for a range of six biomes seen in the US west. Results help livestock ranchers to adjust stocking and forage choices as well as farm policymakers who seek flexible government programs to adapt to changes in economic, ecological, and climate conditions. The work's importance comes from applicability to forage management problems in dry regions internationally.

Addressing critical gaps in protected area coverage for bird habitats in China.

Currently, protected areas cover approximately 14% of the Earth's land surface, yet 12.2% of the world's bird species remain unprotected by any designated areas and face significant threats. This study investigates the current status of bird conservation in China, aiming to evaluate the effectiveness of existing protected areas, analyze why certain bird species are not adequately protected, and propose strategies for optimizing protected area configurations. Utilizing citizen science data and the zonation optimization algorithm, we comprehensively assessed the conservation value of birds in China. We then employed anthropogenic stressor data to evaluate the conservation of threatened bird habitats through a binary conflict intensity model. Finally, we conducted a spatial overlap analysis to determine the coverage and effectiveness of Chinese nature reserves in regions with high conservation value and high conflict risk. Our findings indicate that only 10.0% of the highest conservation value bird habitats are covered by protected areas, and just 7.3% of these protected areas effectively safeguard these critical habitats. Additionally, only 5.9% of bird habitats impacted by human activity conflicts are within protected areas, and merely 22.0% of the total protected areas can effectively conserve high conflict risk habitats. Overall, China's current protected area system has substantial shortcomings in safeguarding bird habitats and requires further optimization and expansion to maximize conservation benefits.

A polyphosphazene elastomer containing 2,2,2-trifluoroethoxy groups as a dielectric in electrically responsive soft actuators.

The adaptive structure and excellent actuation of dielectric elastomer actuators (DEAs) make them promising candidates for soft robotics, haptic interfaces and artificial muscles. A wide variety of elastomers have been synthesised and investigated as dielectrics. Inorganic polymers such as polysiloxanes and polyphosphazenes have a low glass transition temperature. While polydimethylsiloxane (PDMS) has made its way into DEAs, the latter has received little attention in this field. Here, we present a dielectric elastomer based on polyphosphazene modified with 2,2,2,-trifluoroethoxy groups as the dielectric, which exhibits a dielectric permittivity two times higher than polydimethylsiloxanes (PDMS), excellent elasticity and a high dielectric breakdown field. These properties enable fast, reliable actuation and higher electrostatic forces than conventional PDMS. The actuators can withstand repeated actuation cycles and are suitable for long-term reliability applications.

Non-binary patients in ART: new challenges and considerations.

OBJECTIVES: To discuss the increasing visibility of non-binary individuals and inclusive policies and practices in Assisted Reproductive Technologies (ART). METHODS: Comparison between traditional ART approach designed for binary genders and propose approach in non-binary individuals. RESULTS: Traditional ART services, designed for binary genders, must adapt to address the unique fertility needs of non-binary patients. This includes using gender-neutral language, providing comprehensive fertility assessments, and offering hormone therapy and fertility preservation options. Children of non-binary parents benefit from open communication about gender diversity, enhancing psychological well-being. Positive societal attitudes and inclusive environments in schools are crucial for preventing discrimination and promoting mental health. A systemic approach is required to make ART inclusive. This involves training staff, adjusting facilities, updating documentation, and advocating for supportive legislation. Addressing the reproductive needs of non-binary individuals ensures equitable access to care and supports their family-building goals. CONCLUSIONS: Ensuring the well-being of children in non-binary families requires psychological support, inclusive healthcare, legal recognition, social support, and comprehensive education.

Addressing Teenage Pregnancy and Reproductive Health Among Latinx Adolescents and Young Adults.

Despite a national decline in teenage pregnancy rates, Latinxs and Black individuals continue to have higher teenage birth rates compared to white teens. In the United States, Latinx females (ages 15-19) are more than twice as likely to have a teenage birth compared to non-Latinx white teens. With an increasingly diverse nation, a shift towards culturally inclusive approaches to care is critical to achieving equitable patient outcomes. Improving access to preventative care, workforce diversity, and insurance coverage will lead to cost-savings and also help restore trust in a system that has failed past generations. In this commentary, targeted recommendations tailored specifically towards Latinx women will be provided including culturally competent birth control counseling, and a focus on reproductive justice. Not speaking English, low socioeconomic status, acculturation, legal status, and being uninsured are all teenage pregnancy risk factors unique to Latinx women. Cultural preferences and attitudes towards contraception must also be considered. Further expansion of Medicaid insurance coverage is also necessary to increase reproductive access among vulnerable populations. A cultural humility framework to reproductive health counseling and services is recommended. Within this framework family engagement is encouraged and women are free from reproductive coercion; they have the freedom to access over-the-counter contraception and receive guidance and support from providers and community health workers who are cognizant of culture and heritage. An integrated community-based approach that is culturally sensitive and in tune with Latinx heterogenicity is necessary to lower teenage pregnancy rates and achieve reproductive justice.

Impact of the Novel MRI Contrast Agent Gadopiclenol on Radiotherapy Decision Making in Patients With Brain Metastases.

PURPOSE: The aim of this study was to assess the effect of gadopiclenol versus gadobenate dimeglumine contrast-enhanced magnetic resonance imaging (MRI) on decision-making between whole-brain radiotherapy (WBRT) and stereotactic radiosurgery (SRS) for treatment of brain metastases (BMs). METHODS: Patients with BMs underwent 2 separate MRI examinations in a double-blind crossover phase IIb comparative study between the MRI contrast agents gadopiclenol and gadobenate dimeglumine, both administered at 0.1 mmol/kg. The imaging data of a single site using identical MRI scanners and protocols were included in this post hoc analysis. Patients with 1 or more BMs in any of both MRIs were subjected to target volume delineation for treatment planning. Two radiation oncologists contoured all visible lesions and decided upon SRS or WBRT, according to the number of metastases. For each patient, SRS or WBRT treatment plans were calculated for both MRIs, considering the gross target volume (GTV) as the contrast-enhancing aspects of the tumor. Mean GTVs and volume of healthy brain exposed to 12 Gy (V12), as well as Dice similarity coefficient scores, were obtained. The Spearman rank (ρ) correlation was additionally calculated for assessing linear differences. Three different expert radiation oncologists blindly rated the contrast enhancement for contouring purposes. RESULTS: Thirteen adult patients were included. Gadopiclenol depicted additional BM as compared with gadobenate dimeglumine in 7 patients (54%). Of a total of 63 identified metastatic lesions in both MRI sets, 3 subgroups could be defined: A, 48 (24 pairs) detected equal GTVs visible in both modalities; B, 13 GTVs only visible in the gadopiclenol set (mean ± SD, 0.16 ± 0.37 cm3); and C, 2 GTVs only visible in the gadobenate dimeglumine set (mean ± SD, 0.01 ± 0.01). Treatment indication was changed for 2 (15%) patients, 1 from no treatment to SRS and for 1 from SRS to WBRT. The mean GTVs and brain V12 were comparable between both agents (P = 0.694, P = 0.974). The mean Dice similarity coefficient was 0.70 ± 0.14 (ρ = 0.82). According to the readers, target volume definition was improved in 63.9% of cases (23 of 36 evaluations) with gadopiclenol and 22.2% with gadobenate dimeglumine (8 of 36), whereas equivalence was obtained in 13.9% (5 of 36). CONCLUSIONS: Gadopiclenol-enhanced MRI improved BM detection and characterization, with a direct impact on radiotherapy treatment decision between WBRT and SRS. Additionally, a more exact target delineation and planning could be performed with gadopiclenol. A prospective evaluation in a larger cohort of patients is required to confirm these findings.

Penetration Thresholds of Porcine Limbs for Low Sectional Density Projectiles in High-Rate Impact.

INTRODUCTION: With similar prevalence to injuries from fires, stings, and natural disasters, soft tissue injuries may occur from fireworks, industrial accidents, or other explosives. Surgeons are less familiar with treating high-velocity penetration from small debris, which may increase the chance of infection and subsequent fatality. Penetration risk curves have been developed to predict V50, the velocity with 50% probability of penetration, for various sized projectiles. However, there has been limited research using nonmetallic materials to achieve lower density projectiles less than 1 g cm-2, such as sand or rocks. MATERIAL AND METHODS: To emulate the size and density of these energized particles, 14 ball bearings of stainless steel, silicon nitride, or Delrin acetal plastic ranging from 1.59 mm (1/16") to 9.53 mm (3/8") with sectional densities between 0.3 g cm-2 and 5 g cm-2 were launched toward porcine legs at a range of velocities to determine the penetration thresholds. High-speed videography was captured laterally at 40 kHz and impact velocity was captured using a physics-based tracking software. A generalized linear model with repeated measures and a logit link function was used to predict probability of penetration for each projectile. A total of 600 impacts were conducted to achieve at least 15 penetrating impacts for each projectile over a range of velocities. RESULTS: Higher impact velocities were required to penetrate the skin as sectional density of the projectile decreased, and the relationship between velocity and sectional density exhibited an exponential relationship (V50, $ = 184.6*S{D^{ - 0.385}}$, R2 = 0.95) with substantial change for nonlinearity in sectional densities ranging from 0.3 g cm-2 to 1 g cm-2. Compared to previous studies, the empirical relationship was consistent in the linear region (2-5 g cm-2), and novel experimentation filled in the gaps for sectional densities less than 1 g cm-2, which expressed more nonlinearity than previously estimated. For low-density projectiles with diameters of 1.59 (1/16") or 3.18 (1/8"), 32 impacts were lodged into the epidermis but did not penetrate through the dermis; however, penetration was defined as displacement into or through the dermis. CONCLUSIONS: These experimental results may be used to develop and validate finite element simulations of low-density projectile impacts to address complex, multivariate loading conditions for the development of protective clothing to reduce wounding and subsequent infection rates.

The unfolded protein response sensor PERK mediates mechanical stress-induced maturation of focal adhesion complexes in glioblastoma cells.

Stiffening of the brain extracellular matrix (ECM) in glioblastoma promotes tumor progression. Previously, we discovered that protein kinase R (PKR)-like endoplasmic reticulum kinase (PERK) plays a role in glioblastoma stem cell (GSC) adaptation to matrix stiffness through PERK/FLNA-dependent F-actin remodeling. Here, we examined the involvement of PERK in detecting stiffness changes via focal adhesion complex (FAC) formation. Compared to control GSCs, PERK-deficient GSCs show decreased vinculin and tensin expression, while talin and integrin-ß1 remain constant. Furthermore, vimentin was also reduced while tubulin increased, and a stiffness-dependent increase of the differentiation marker GFAP expression was absent in PERK-deficient GSCs. In conclusion, our study reveals a novel role for PERK in FAC formation during matrix stiffening, which is likely linked to its regulation of F-actin remodeling.

Upright versus inverted catching and crating end-of-lay hens: a trade-off between animal welfare, ergonomic and financial concerns.

This study explores upright versus inverted catching and crating of spent laying hens. Both catching methods were compared using a cost-benefit analysis that focused on animal welfare, ergonomic, and financial considerations. Data were collected on seven commercial farms (one floor system and six aviary systems) during depopulation of approximately 3,000 hens per method per flock. Parameters such as wing flapping frequency, catcher bird interaction, incidence of catching damage and hens dead on arrival (DOA) were measured and compared between catching methods. Ergonomic evaluations were performed via catcher surveys and expert assessment of video recordings. The wing flapping frequency was lower (3.1 ± 0.6 vs. 4.0 ± 0.5, P < 0.001) and handling was gentler (1.9 ± 0.5 vs. 4.4 ± 0.5, P < 0.001), both on a 7-point Likert scale, for upright versus inverted catching. However, more person-hours per 1000 hens were required for upright than inverted catching (8.2 ± 3.2 h vs. 4.8 ± 2.0 h, P = 0.011), with only wing bruises being significantly less common for upright than inverted catching (1.1 ± 0.6 % vs. 1.7 ± 0.7%, P = 0.04). Upright catching was 1.8 times more expensive than inverted catching; compensation for this cost would require a premium price of approximately €0.0005 extra per egg. Ergonomically, both catching methods were considered demanding, although catchers (n = 29) preferred inverted catching. In conclusion, this study showed animal welfare benefits of upright vs. inverted catching. Industry adoption of upright catching will depend on compensation of the additional labor costs, adjustments to labor conditions and shorter loading times.

The Benefit of Hearing Aids as Measured by Listening Accuracy, Subjective Listening Effort, and Functional Near Infrared Spectroscopy.

There is broad consensus that listening effort is an important outcome for measuring hearing performance. However, there remains debate on the best ways to measure listening effort. This study sought to measure neural correlates of listening effort using functional near-infrared spectroscopy (fNIRS) in experienced adult hearing aid users. The study evaluated impacts of amplification and signal-to-noise ratio (SNR) on cerebral blood oxygenation, with the expectation that easier listening conditions would be associated with less oxygenation in the prefrontal cortex. Thirty experienced adult hearing aid users repeated sentence-final words from low-context Revised Speech Perception in Noise Test sentences. Participants repeated words at a hard SNR (individual SNR-50) or easy SNR (individual SNR-50 + 10 dB), while wearing hearing aids fit to prescriptive targets or without wearing hearing aids. In addition to assessing listening accuracy and subjective listening effort, prefrontal blood oxygenation was measured using fNIRS. As expected, easier listening conditions (i.e., easy SNR, with hearing aids) led to better listening accuracy, lower subjective listening effort, and lower oxygenation across the entire prefrontal cortex compared to harder listening conditions. Listening accuracy and subjective listening effort were also significant predictors of oxygenation.

Sulfotransferase 1C2 Increases Mitochondrial Respiration by Converting Mitochondrial Membrane Cholesterol to Cholesterol Sulfate.

HYPOTHESIS: In this communication, we test the hypothesis that sulfotransferase 1C2 (SULT1C2, UniProt accession no. Q9WUW8) can modulate mitochondrial respiration by increasing state-III respiration. METHODS AND RESULTS: Using freshly isolated mitochondria, the addition of SULT1C2 and 3-phosphoadenosine 5 phosphosulfate (PAPS) results in an increased maximal respiratory capacity in response to the addition of succinate, ADP, and rotenone. Lipidomics and thin-layer chromatography of mitochondria treated with SULT1C2 and PAPS showed an increase in the level of cholesterol sulfate. Notably, adding cholesterol sulfate at nanomolar concentration to freshly isolated mitochondria also increases maximal respiratory capacity. In vivo studies utilizing gene delivery of SULT1C2 expression plasmids to kidneys result in increased mitochondrial membrane potential and confer resistance to ischemia/reperfusion injury. Mitochondria isolated from gene-transduced kidneys have elevated state-III respiration as compared with controls, thereby recapitulating results obtained with mitochondrial fractions treated with SULT1C2 and PAPS. CONCLUSION: SULT1C2 increases mitochondrial respiratory capacity by modifying cholesterol, resulting in increased membrane potential and maximal respiratory capacity. This finding uncovers a unique role of SULT1C2 in cellular physiology and extends the role of sulfotransferases in modulating cellular metabolism.

In vitro skin puncture methodology for material characterization.

Quantifying the mechanical behavior of skin has been foundational in applications of cosmetics, surgical techniques, forensic science, and protective clothing development. However, previous puncture studies have lacked consistent and physiological boundary conditions of skin. To determine natural skin tension, excision of in situ porcine skin resulted in significantly different diameter reduction (shrinkage) in leg (19.5 %) and abdominal skin (38.4 %) compared to flank skin (28.5 %) (p = 0.047). To examine effects of initial tension and pre-conditioning, five conditions of initial tension (as percentage of diameter increase) and pre-conditioning were tested in quasistatic puncture with a 5 mm spherical impactor using an electrohydraulic load frame and custom clamping apparatus. Samples with less than 5 % initial tension resulted in significantly greater (p = 0.011) force at failure (279.2 N) compared to samples with greater than 25 % initial tension (195.1 N). Eight pre-conditioning cycles of 15 mm displacement reduced hysteresis by 45 %. The coefficient of variance was substantially reduced for force, force normalized by cutis thickness, displacement, stiffness, and strain energy up to 46 %. Pre-conditioned samples at physiological initial tension (14-25 %) resulted in significantly greater (p = 0.03) normalized forces at failure (278.3 N/mm) compared to non-conditioned samples of the same initial tension (234.4 N/mm). Pre-conditioned samples with 14-25 % initial tension, representing physiological boundary conditions, resulted in the most appropriate failure thresholds with the least variation. For in vitro puncture studies, the magnitude of applied initial tension should be defined based on anatomical location, through a shrinkage experimentation, to match natural tension of skin. Characterizing the biological behavior and tolerances of skin may be utilized in finite element models to aid in protective clothing development and forensic science analyses.

A Protocol to Determine Circadian Phase by At-Home Salivary Dim Light Melatonin Onset Assessment.

Internal circadian phase assessment is increasingly acknowledged as a critical clinical tool for the diagnosis, monitoring, and treatment of circadian rhythm sleep-wake disorders and for investigating circadian timing in other medical disorders. The widespread use of in-laboratory circadian phase assessments in routine practice has been limited, most likely because circadian phase assessment is not required by formal diagnostic nosologies, and is not generally covered by insurance. At-home assessment of salivary dim light melatonin onset (DLMO, a validated circadian phase marker) is an increasingly accepted approach to assess circadian phase. This approach may help meet the increased demand for assessments and has the advantages of lower cost and greater patient convenience. We reviewed the literature describing at-home salivary DLMO assessment methods and identified factors deemed to be important to successful implementation. Here, we provide specific protocol recommendations for conducting at-home salivary DLMO assessments to facilitate a standardized approach for clinical and research purposes. Key factors include control of lighting, sampling rate, and timing, and measures of patient compliance. We include findings from implementation of an optimization algorithm to determine the most efficient number and timing of samples in patients with Delayed Sleep-Wake Phase Disorder. We also provide recommendations for assay methods and interpretation. Providing definitive criteria for each factor, along with detailed instructions for protocol implementation, will enable more widespread adoption of at-home circadian phase assessments as a standardized clinical diagnostic, monitoring, and treatment tool.

Distinct epigenomic landscapes underlie tissue-specific memory T cell differentiation.

The memory CD8+ T cell pool contains phenotypically and transcriptionally heterogeneous subsets with specialized functions and recirculation patterns. Here, we examined the epigenetic landscape of CD8+ T cells isolated from seven non-lymphoid organs across four distinct infection models, alongside their circulating T cell counterparts. Using single-cell transposase-accessible chromatin sequencing (scATAC-seq), we found that tissue-resident memory T (TRM) cells and circulating memory T (TCIRC) cells develop along distinct epigenetic trajectories. We identified organ-specific transcriptional regulators of TRM cell development, including FOSB, FOS, FOSL1, and BACH2, and defined an epigenetic signature common to TRM cells across organs. Finally, we found that although terminal TEX cells share accessible regulatory elements with TRM cells, they are defined by TEX-specific epigenetic features absent from TRM cells. Together, this comprehensive data resource shows that TRM cell development is accompanied by dynamic transcriptome alterations and chromatin accessibility changes that direct tissue-adapted and functionally distinct T cell states.