Role of dipicolinic acid in heat resistance of spores of Clostridium botulinum and Clostridium sporogenes PA3679 by thermal and pressure-assisted thermal processing.

Dipicolinic acid (DPA) is a major constituent of spores and reportedly provides protection against inactivation by various thermal processes, however the relationship between DPA and resistance towards pressure-assisted thermal processing is not well understood. Thermal and pressure-assisted thermal inactivation studies of Clostridium botulinum nonproteolytic strains QC-B and 610-F, proteolytic strain Giorgio-A, and thermal surrogate Clostridium sporogenes PA3679 spores suspended in ACES buffer (0.05 M, pH 7.0) were performed to determine if a relationship exists between DPA release and log reduction of spores. Thermal inactivation at 80, 83, and 87°C for nonproteolytic strains and 101, 105, and 108°C for the proteolytic strain and thermal surrogate were conducted. Pressure-assisted thermal inactivation for nonproteolytic strains at 83°C/600 MPa and for the proteolytic strain and thermal surrogate at 105°C/600 MPa were performed. Surviving spores were enumerated by 5-tube MPN method for log reductions and analyzed for released DPA by liquid chromatography-tandem mass spectrometry. The correlation between MPN log reductions, released DPA, and D-values were calculated. A positive correlation between released DPA and log reduction of spores was observed for QC-B and 610-F at 80 and 83°C (r = 0.6073 - 0.7755; P <0.01). At 87°C, a positive correlation was detected for 610-F (r = 0.4242, P < 0.05) and no correlation observed for QC-B (r = 0.1641; P > 0.05). A strong, positive correlation (r = 0.8359 - 0.9284; P < 0.05) between released DPA and log reduction of spores was observed for Giorgio-A at 101, 105, and 108°C and a strong, positive correlation (r = 0.8402; P < 0.05) was observed for PA3679 at 101°C. A positive correlation (r = 0.5646 - 0.6724; P <0.01) was observed for QC-B, 610-F, and Giorgio-A after pressure-assisted thermal treatment. No correlation (r = 02494; P > 0.05) was found for PA3679 after pressure-assisted thermal treatment. These results suggest a correlation exists between DPA release and heat resistance, however, the level of correlation varied between strains and temperatures. The findings from this research may aid in the development of spore inactivation strategies targeting the thermal resistance profiles of various strains of C. botulinum spores.

Searching for the Scale-Able Weight Loss Intervention: Can One Size Fit All? Lessons to Be Learned from the EQUAL Trial.

Overweight and obesity affect 71.2% of adults in the United States, with cancer survivors not far behind at 70.3%. Subgroups such as those diagnosed with acute lymphoblastic leukemia (ALL) face even greater challenges. The Exercise and Quality Diet after Leukemia (EQUAL) trial sought to address weight management issues among ALL survivors by implementing a remotely delivered weight loss intervention, leveraging the previously proven Practice-based Opportunities for Weight Reduction (POWER) program. Despite a strong foundation and design, the EQUAL trial yielded null results. Key differences in study populations and intervention contexts between the EQUAL and POWER trials, such as the lack of primary care physician involvement in EQUAL, contributed to these outcomes. EQUAL's failure to meet its accrual target and poor adherence among participants highlighted challenges in engaging this unique population. Contrary to EQUAL's conclusions, evidence from other studies supports the efficacy of remote interventions for weight loss among cancer survivors. The lack of qualitative assessment among ALL survivors and key integration to inform intervention adaptations undermined EQUAL's impact. However, EQUAL's impressive retention rate offers valuable insights. Lessons from EQUAL underscore the need for well-fitted, remotely delivered interventions and the importance of thoughtfully adapted and tailored approaches to specific survivor populations. See related article by Fiedmann et al., p. 1158.

Soft tissue laxity is highly variable in patients undergoing total knee arthroplasty.

BACKGROUND: One major goal of total knee arthroplasty (TKA) is to achieve balanced medial and lateral gaps in flexion and extension. While bone resections are planned by the surgeon, soft tissue laxity is largely intrinsic and patient-specific in the absence of additional soft tissue releases. We sought to determine the variability in soft tissue laxity in patients undergoing TKA. METHODS: We retrospectively reviewed 113 patients undergoing TKA. Data on preoperative knee deformity were collected. Data from a dynamic intraoperative stress examination were collected by a robotic tracking system to quantify maximal medial and lateral opening in flexion (85-95 degrees) and extension (-5-20 degrees). T-tests were used to assess the differences between continuous variables. RESULTS: A valgus stress opened the medial compartment a mean of 4.3 ± 2.3 mm (0.0-12.4 mm) in extension and 4.6 ± 2.3 mm (0.0-12.9 mm) in flexion. A varus stress opened the lateral compartment a mean of 5.4 ± 2.4 mm (0.3-12.6 mm) in extension and 6.2 ± 2.5 mm (0.0-13.4 mm) in flexion. The medial compartment of varus knees opened significantly more in response to valgus stress than valgus knees in both extension (5.2 mm vs. 2.6 mm; P < 0.0001) and flexion (5.4 mm vs 3.3 mm; P < 0.0001). The lateral compartment of valgus knees opened significantly more in response to varus stress than varus knees in both extension (6.7 mm vs. 4.8 mm; P < 0.0001) and flexion (7.4 mm vs. 5.8 mm; P = 0.0003). CONCLUSIONS: Soft tissue laxity is highly variable in patients undergoing TKA, contributing anywhere from 0-13 mm to the post-resection gap. Only a small part of this variability is predictable by preoperative deformity. These findings have implications for either measured-resection or gap-balancing techniques. LEVEL OF EVIDENCE: Level III.

Computational investigation of the role of ventricular remodelling in HFpEF: The key to phenotype dissection.

Recent clinical studies have reported that heart failure with preserved ejection fraction (HFpEF) can be divided into two phenotypes based on the range of ejection fraction (EF), namely HFpEF with higher EF and HFpEF with lower EF. These phenotypes exhibit distinct left ventricle (LV) remodelling patterns and dynamics. However, the influence of LV remodelling on various LV functional indices and the underlying mechanics for these two phenotypes are not well understood. To address these issues, this study employs a coupled finite element analysis (FEA) framework to analyse the impact of various ventricular remodelling patterns, specifically concentric remodelling (CR), concentric hypertrophy (CH), and eccentric hypertrophy (EH), with and without LV wall thickening on LV functional indices. Further, the geometries with a moderate level of remodelling from each pattern are subjected to fibre stiffening and contractile impairment to examine their effect in replicating the different features of HFpEF. The results show that with severe CR, LV could exhibit the characteristics of HFpEF with higher EF, as observed in recent clinical studies. Controlled fibre stiffening can simultaneously increase the end-diastolic pressure (EDP) and reduce the peak longitudinal strain (ell) without significant reduction in EF, facilitating the moderate CR geometries to fit into this phenotype. Similarly, fibre stiffening can assist the CH and 'EH with wall thickening' cases to replicate HFpEF with lower EF. These findings suggest that potential treatment for these two phenotypes should target the bio-origins of their distinct ventricular remodelling patterns and the extent of myocardial stiffening.

Motivation Matters: Understanding the Antidepressant Mechanism of Physical Activity among Young Adults.

A negative association between physical activity and depressive symptoms is consistently reported within scientific literature and physical self-concept has been suggested to mediate this pathway. However, for whom these associations are strongest remains poorly understood, and little is known about how other psychosocial factors might be implicated. Consequently, we examined how various exercise motivations, specifically appearance, physical health, and mental health, might moderate the indirect effect of physical activity on depressive symptoms through physical self-concept. Canadian young adults (N = 496, Mage = 20.36, SD = 1.87) completed an online questionnaire. Mediation and moderated-mediation models were tested using PROCESS macro in RStudio. A significant indirect effect (ß = -0.18, CI [-0.005, -0.003]) of physical activity on depressive symptoms through physical self-concept was found. Exercise motivations moderated the association between physical activity and physical self-concept, such that the association was stronger when individuals were motivated by physical health. Thus, the effect of physical activity on depressive symptoms varied according to physical self-concept and physical health-exercise motivations. We conclude that motivation should be considered when developing and delivering physical activity prevention efforts for depressive symptoms.

Surgeon-Applied Stress and a Ligament Tensor Instrument Provide a Similar Assessment of Preresection Flexion Laxity During Robotic Total Knee Arthroplasty.

Background: Robotic-assisted total knee arthroplasty (RA-TKA) allows surgeons to perform intraoperative soft tissue laxity assessments prior to bone resections and is used to alter resections to achieve gap balance. This study compared 2 techniques for flexion gap laxity assessment during RA-TKA. Methods: A prospective study of 50 primary RA-TKAs performed by a single surgeon was conducted between February and October 2023. Following full exposure, anterior tibial dislocation, and osteophyte removal, maximal medial and lateral compartment flexion laxity was quantified to the nearest 0.5 mm by the robotic system using a dynamic, surgeon-applied stress (SURGEON). This data was used to plan a balanced flexion gap by adjusting the femoral component size, rotation, and anterior-posterior translation. Flexion laxity was quantified again after distal femoral and proximal tibial resections using a ligament tensor instrument (TENSOR). These new data were used to plan for the same desired flexion gap using the same variables. Paired-samples t-tests and a simple linear regression were used for analysis. Results: Both methods produced near-identical recommendations for femoral component sizing (mean deviation 0.06 sizes, range -1 to +1 size; P = .569), rotation (deviation mean 1.0°, range -3.0° to +3.0°; P = .741), and anterior-posterior translation (deviation mean 0.13 mm, range -0.5 to +0.5 mm, P = .785). SURGEON femoral component rotation predicted TENSOR rotation (R2 = 0.157; 95% confidence interval = 0.124, 0.633; P = .004). Conclusions: Assessing flexion laxity with a surgeon-applied stress vs a ligament tensor produced near-identical laxity data in RA-TKA, suggesting surgeons may comfortably choose either technique as a reliable method. Level of Evidence: Level III.

Recruiting and Engaging Heterosexual-Identified Men Who have Sex with Men: A Brief Report of Considerations for Sex Researchers.

Heterosexual-identified men who have sex with men (H-MSM) are a unique population difficult to identify and recruit for research and practice. Yet, engaging H-MSM remains a top research priority to learn more about this population's health needs. A scoping review was conducted to develop a stronger understanding of recruitment patterns involving H-MSM in research. The search and screening procedures yielded 160 total articles included in the present study. Most studies relied on venue-based and internet-based recruitment strategies. Thematic analysis was then used to identify three themes. Locations of H-MSM's sexual encounters related to where sex researchers may recruit participants; sociocultural backgrounds of H-MSM related to important characteristics researchers should acknowledge and consider when working with H-MSM; and engagement with health services related to how H-MSM interact with or avoid HIV/STI testing and treatment and other public health services. Findings suggest H-MSM have sex with other men in a variety of venues (e.g. bathhouses, saunas) but tend to avoid gay-centric venues. H-MSM also are diverse, and these unique identities should be accounted for when engaging them. Finally, H-MSM are less likely to access healthcare services than other MSM, highlighting the need for targeted advertisements and interventions specific for H-MSM.

Sensitivity of Meloidogyne incognita and Rotylenchulus reniformis to cyclobutrifluram.

Cyclobutrifluram, a succinate dehydrogenase inhibitor fungicide, is being evaluated as a seed-applied nematicide in cotton and soybean to manage plant-parasitic nematodes. Currently, there is no information on the toxicity, ovicidal activity, nematode recovery, or effects on nematode infection for Meloidogyne incognita or Rotylenchulus reniformis after exposure to low concentrations of cyclobutrifluram. Nematode toxicity assays were performed in aqueous solutions of cyclobutrifluram, while root infection assays were conducted on tomato. Nematode paralysis was observed after 2 h of exposure to 0.5 µg/ml cyclobutrifluram for both nematode species. Based on an assay of nematode motility, the 2-hr EC50 value for M. incognita and R. reniformis was 0.48 and 1.07 µg/ml, respectively. In a comparable assay with a similar nematicide, continuous exposure to 0.5 µg/ml cyclobutrifluram for 24 h resulted in at least 45% more immotile nematodes for both species compared to those treated with 0.5 µg/ml fluopyram. Continuous exposure to concentrations >1.0 µg/ml suppressed hatching for both species compared to the water control. Nematode recovery from paralysis was greater than 80% for M. incognita and R. reniformis 24 h after nematodes were rinsed and removed from a 1-h treatment to their respective 2-hr EC50 concentrations. Nematode infection of tomato roots was reduced following a 1-h treatment with aqueous solutions of cyclobutrifluram, ranging from 0.12 to 0.48 µg/ml for M. incognita and 0.27 to 1.07 µg/ml for R. reniformis. Overall, the toxicity of cyclobutrifluram to these nematode species was greater than that of fluopyram and although the effects of cyclobutrifluram were reversible, low concentrations were effective at reducing the ability of both nematodes to infect tomato roots.

Biochemical characterisation and production kinetics of high molecular-weight (HMW) putative antibacterial proteins of insect pathogenic Brevibacillus laterosporus isolates.

BACKGROUND: Bacterial genomes often encode structures similar to phage capsids (encapsulins) and phage tails which can be induced spontaneously or using genotoxic compounds such as mitomycin C. These high molecular-weight (HMW) putative antibacterial proteins (ABPs) are used against the competitive strains under natural environment. Previously, it was unknown whether these HMW putative ABPs originating from the insect pathogenic Gram-positive, spore-forming bacterium Brevibacillus laterosporus (Bl) isolates (1821L, 1951) are spontaneously induced during the growth and pose a detrimental effect on their own survival. Furthermore, no prior work has been undertaken to determine their biochemical characteristics. RESULTS: Using a soft agar overlay method with polyethylene glycol precipitation, a narrow spectrum of bioactivity was found from the precipitated lysate of Bl 1951. Electron micrographs of mitomycin C- induced filtrates showed structures similar to phage capsids and contractile tails. Bioactivity assays of cell free supernatants (CFS) extracted during the growth of Bl 1821L and Bl 1951 suggested spontaneous induction of these HMW putative ABPs with an autocidal activity. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis of spontaneously induced putative ABPs showed appearance of ~ 30 kDa and ~ 48 kDa bands of varying intensity across all the time intervals during the bacterial growth except in the initial hours. Statistically, spontaneously induced HMW putative ABPs of Bl 1951 exhibited a significant decrease in the number of viable cells of its producer strain after 18 h of growth in liquid. In addition, a significant change in pH and prominent bioactivity of the CFS of this particular time period was noted. Biochemically, the filtered supernatant derived from either Bl 1821L or Bl 1951 maintained bioactivity over a wide range of pH and temperature. CONCLUSION: This study reports the spontaneous induction of HMW putative ABPs (bacteriocins) of Bl 1821L and Bl 1951 isolates during the course of growth with potential autocidal activity which is critically important during production as a potential biopesticide. A narrow spectrum of putative antibacterial activity of Bl 1951 precipitate was found. The stability of HMW putative ABPs of Bl 1821L and Bl 1951 over a wide range of pH and temperature can be useful in expanding the potential of this useful bacterium beyond the insecticidal value.

Does Lack of Initial Collar-Calcar Contact Influence Performance of Collared Cementless Femoral Stems?

Background: Initial stability of cementless stems is important to minimize the risk of subsidence, pain, and periprosthetic fracture after total hip arthroplasty (THA). Collared stems improve initial component stability when contacting the femoral calcar. Direct contact is not always achieved, and collared stem performance has not been studied in this context. We hypothesized that collared stems achieving direct contact would demonstrate reduced subsidence. Methods: A single-surgeon retrospective study of 482 consecutive primary THAs implanted between February 2020 and May 2023 using collared cementless stems was performed. The 2 cohorts included stems with initial collar-calcar contact vs stems without. Subsidence was evaluated by comparing intraoperative fluoroscopy to postoperative 8-week radiographs. Binary logistic regression identified independent risk factors for subsidence. Chi-square tests were used for categorical variables and t-tests for continuous variables. Results: Of stems, 63.9% achieved initial collar-calcar contact, while 36.1% did not. The rate (1.3% vs 19.0%; P < .001) and magnitude (0.02 mm, range 0-3 mm vs 0.35 mm, range 0-3 mm; P < .001) of subsidence were significantly higher among stems without initial contact. Stems without initial collar-calcar contact (P < .001) and male gender (P = .007) were independent risk factors for subsidence. Two patients with initial contact had nondisplaced calcar cracks and <3 mm of subsidence at 4 weeks, which healed with protected weight-bearing. Stem survivorship was 100% in both groups, with all achieving osteointegration and none needing revision. Conclusions: Excellent performance of collared cementless stems was observed at 8 weeks after primary THA. Initial collar-calcar contact lowered the risk and magnitude of minor subsidence but did not affect survivorship or fracture risk. Level of Evidence: Level III.

High-Throughput, Label-Free Detection of DNA Origami in Single-Cell Suspensions Using origamiFISH-Flow.

Structural DNA nanotechnology enables custom fabrication of nanoscale devices and promises diverse biological applications. However, the effects of design on DNA nanostructure (DN)-cell interactions in vitro and in vivo are not yet well-characterized. origamiFISH is a recently developed technique for imaging DNs in cells and tissues. Compared to the use of fluorescent tags, origamiFISH offers label-free and structure-agnostic detection of DNs with significantly improved sensitivity. Here, the origamiFISH technique is extended to quantify DNs in single-cell suspensions, including in nonadherent cells such as subsets of immune cells, via readout by flow cytometry. This method, referred to as origamiFISH-Flow, is high-throughput (e.g., 10 000 cells per second) and compatible with immunostaining for concurrent cell-type and cell-state characterization. It is shown that origamiFISH-Flow provides 20-fold higher signal-to-noise ratio for DN detection compared to dye labeling approaches, leading to the capture of >25-fold more DN+ cells under single-picomolar DN uptake concentrations. Additionally, the use of origamiFISH-Flow is validated to profile the uptake of various DN shapes across multiple cell lines and splenocytes, as well as to quantify in vivo DN accumulation in lymphoid organs. Together, origamiFISH-Flow offers a new tool to interrogate DN interactions with cells and tissues, while providing insights for tailoring their designs in bio-applications.

ERMA (TMEM94) is a P-type ATPase transporter for Mg2+ uptake in the endoplasmic reticulum.

Intracellular Mg2+ (iMg2+) is bound with phosphometabolites, nucleic acids, and proteins in eukaryotes. Little is known about the intracellular compartmentalization and molecular details of Mg2+ transport into/from cellular organelles such as the endoplasmic reticulum (ER). We found that the ER is a major iMg2+ compartment refilled by a largely uncharacterized ER-localized protein, TMEM94. Conventional and AlphaFold2 predictions suggest that ERMA (TMEM94) is a multi-pass transmembrane protein with large cytosolic headpiece actuator, nucleotide, and phosphorylation domains, analogous to P-type ATPases. However, ERMA uniquely combines a P-type ATPase domain and a GMN motif for ERMg2+ uptake. Experiments reveal that a tyrosine residue is crucial for Mg2+ binding and activity in a mechanism conserved in both prokaryotic (mgtB and mgtA) and eukaryotic Mg2+ ATPases. Cardiac dysfunction by haploinsufficiency, abnormal Ca2+ cycling in mouse Erma+/- cardiomyocytes, and ERMA mRNA silencing in human iPSC-cardiomyocytes collectively define ERMA as an essential component of ERMg2+ uptake in eukaryotes.

Functional outcomes in MRI-guided laser interstitial therapy for temporal lobe epilepsy: a systematic review and meta-analysis.

OBJECTIVE: The use of MRI-guided laser interstitial thermal therapy (MRgLITT) has emerged as a promising treatment option for patients with drug-resistant temporal lobe epilepsy (TLE). Although the minimally invasive approach holds promise as an effective treatment for achieving seizure freedom, a comprehensive review of its impact on functional outcomes is still warranted. To address this need, this review aims to summarize data pertaining to the functional and neurocognitive outcomes following MRgLITT for TLE. METHODS: Four primary electronic databases were screened following PRISMA guidelines by two independent reviewers. All functional data related to cognitive, behavioral, and emotional outcomes were gathered and analyzed as well as the neuropsychological tests issued to assess pre- and postoperative outcomes. The functional outcomes assessed were grouped into the 5 most common categories: verbal cognition, visual cognition, cognitive emotion, visual deficits, and other higher-order cognitive functioning. RESULTS: A total of 4184 studies were screened and ultimately 408 patients from 14 studies were included for analysis. Changes in functional areas were assessed by comparing pre- and postoperative scores across a comprehensive set of 31 different functional and cognitive assessments, and were tabulated as the percentage of patients whose status improved, declined, or was maintained, where possible. In verbal (n = 112) and visual (n = 42) cognition, the rates of patients experiencing a decline were 20.4% and 13.5%, respectively, and the rates of improvement were 24.9% and 16.7%, respectively. Other functional outcomes assessed, including cognitive emotion (n = 150), visual deficits (n = 325), and higher-order cognitive processes like attention/processing (n = 19), motor cognition (n = 18), and general executive function (n = 4), exhibited varying rates of decline, ranging from 10.5% to 25%. CONCLUSIONS: MRgLITT is an effective and minimally invasive surgical alternative treatment for TLE, but there is an observable impact on patient functioning and cognitive status. This review demonstrates the need for standardized methods that can accurately capture and quantify the associated risk of MRgLITT to optimize its effect on patient quality of life moving forward.

How are Oral Antibiotics Being Used in Total Joint Arthroplasty? A Review of the Literature.

While the role and benefit of perioperative intravenous (IV) antibiotics in patients undergoing total joint arthroplasty (TJA) is well-established, oral antibiotic use in TJA remains a controversial topic with wide variations in practice patterns. With this review, we aimed to better educate the orthopedic surgeon on when and how oral antibiotics may be used most effectively in TJA patients, and to identify gaps in the literature that could be clarified with targeted research. Extended oral antibiotic prophylaxis (EOAP) use in high-risk primary, aseptic revision, and exchange TJA for infection may be useful in decreasing periprosthetic joint infection (PJI) rates. When prescribing oral antibiotics either as EOAP or for draining wounds, patient factors, type of surgery, and type of infectious organisms should be considered in order to optimally prevent and treat PJI. It is important to maintain antibiotic stewardship by administering the proper duration, dose, and type of antibiotics and by consulting infectious disease when necessary.

Management of Soybean Cyst Nematode and Sudden Death Syndrome with Nematode-Protectant Seed Treatments Across Multiple Environments in Soybean.

As soybean (Glycine max) production continues to expand in the United States and Canada, so do pathogens and pests that directly threaten soybean yield potential and economic returns for farmers. One such pathogen is the soybean cyst nematode (SCN; Heterodera glycines). SCN has traditionally been managed using SCN-resistant cultivars and rotation with nonhost crops, but the interaction of SCN with sudden death syndrome (SDS; caused by Fusarium virguliforme) in the field makes management more difficult. Nematode-protectant seed treatments have become options for SCN and SDS management. The objectives of this study were to evaluate nematode-protectant seed treatments for their effects on (i) early and full season SCN reproduction, (ii) foliar symptoms and root-rot caused by SDS, and (iii) soybean yield across environments accounting for the above factors. Using a standard protocol, field trials were implemented in 13 states and one Canadian province from 2019 to 2021 constituting 51 site-years. Six nematode-protectant seed treatment products were compared with a fungicide + insecticide base treatment and a nontreated check. Initial (at soybean planting) and final (at soybean harvest) SCN egg populations were enumerated, and SCN females were extracted from roots and counted at 30 to 35 days postplanting. Foliar disease index (FDX) and root rot caused by the SDS pathogen were evaluated, and yield data were collected for each plot. No seed treatment offered significant nematode control versus the nontreated check for in-season and full-season nematode response, no matter the initial SCN population or FDX level. Of all treatments, ILEVO (fluopyram) and Saltro (pydiflumetofen) provided more consistent increases in yield over the nontreated check in a broader range of SCN environments, even when FDX level was high.

Setting up EEG Source Imaging in Practice.

SUMMARY: Adding EEG source imaging to a clinical practice has clear advantages over visual inspection of EEG. This article offers insight on incorporating EEG source imaging into an EEG laboratory and the best practices for producing optimal source analysis results.

The application of synthetic antibacterial minerals to combat topical infections: exploring a mouse model of MRSA infection.

The development of new antibiotics has stalled, and novel strategies are needed as we enter the age of antibiotic resistance. Certain naturally occurring clays have been shown to be effective in killing antibiotic resistant bacteria. However, these natural clays are too variable to be used in clinical settings. Our study shows that synthetic antibacterial minerals exhibit potent antibacterial activity against topical MRSA infections and increase the rate of wound closure relative to controls. The antibacterial minerals maintain a redox cycle between Fe2+/Fe3+ and the surfaces of pyrite minerals, which act as a semiconductor and produce reactive oxygen species (ROS), while smectite minerals act as a cation exchange reservoir. Acidic conditions are maintained throughout the application of the hydrated minerals and can mitigate the alkaline pH conditions observed in chronic non-healing wounds. These results provide evidence for the strategy of 'iron overload' to combat antibiotic resistant infections through the maintained release of Fe2+ and generation of ROS via distinct geochemical reactions that can break the chronic wound damage cycle.

Universal, label-free, single-molecule visualization of DNA origami nanodevices across biological samples using origamiFISH.

Structural DNA nanotechnology enables the fabrication of user-defined DNA origami nanostructures (DNs) for biological applications. However, the role of DN design during cellular interactions and subsequent biodistribution remain poorly understood. Current methods for tracking DN fates in situ, including fluorescent-dye labelling, suffer from low sensitivity and dye-induced artifacts. Here we present origamiFISH, a label-free and universal method for the single-molecule fluorescence detection of DNA origami nanostructures in cells and tissues. origamiFISH targets pan-DN scaffold sequences with hybridization chain reaction probes to achieve 1,000-fold signal amplification. We identify cell-type- and DN shape-specific spatiotemporal distribution patterns within a minute of uptake and at picomolar DN concentrations, 10,000× lower than field standards. We additionally optimize compatibility with immunofluorescence and tissue clearing to visualize DN distribution within tissue cryo-/vibratome sections, slice cultures and whole-mount organoids. Together, origamiFISH enables the accurate mapping of DN distribution across subcellular and tissue barriers for guiding the development of DN-based therapeutics.

Differences in Tic Severity Among Adolescent Girls and Boys with Tourette Syndrome During the Pandemic.

Limited data are available regarding the impact of the coronavirus disease 2019 (COVID-19) pandemic on adolescents with Tourette syndrome (TS). We sought to compare sex differences in tic severity experienced by adolescents before and during the COVID-19 pandemic. We extracted from the electronic health record and retrospectively reviewed Yale Global Tic Severity Scores (YGTSS) from adolescents (ages 13 through 17) with TS presenting to our clinic before (36 months) and during (24 months) the pandemic. A total of 373 unique adolescent patient encounters (prepandemic: 199; pandemic: 173) were identified. Compared with prepandemic, girls accounted for a significantly greater proportion of visits during the pandemic (p < 0.001). Prepandemic, tic severity did not differ between girls and boys. During the pandemic, compared with girls, boys had less clinically severe tics (p = 0.003). During the pandemic, older girls, but not boys, had less clinically severe tics (ρ =- 0.32, p = 0.003). These findings provide evidence that, regarding tic severity assessed with YGTSS, the experiences of adolescent girls and boys with TS have differed during the pandemic.