Identification of compounds that cause axonal dieback without cytotoxicity in dorsal root ganglia explants and intervertebral disc cells with potential to treat pain via denervation.

Low back pain, knee osteoarthritis, and cancer patients suffer from chronic pain. Aberrant nerve growth into intervertebral disc, knee, and tumors, are common pathologies that lead to these chronic pain conditions. Axonal dieback induced by capsaicin (Caps) denervation has been FDA-approved to treat painful neuropathies and knee osteoarthritis but with short-term efficacy and discomfort. Herein, we propose to evaluate pyridoxine (Pyr), vincristine sulfate (Vcr) and ionomycin (Imy) as axonal dieback compounds for denervation with potential to alleviate pain. Previous literature suggests Pyr, Vcr, and Imy can cause undesired axonal degeneration, but no previous work has evaluated axonal dieback and cytotoxicity on adult rat dorsal root ganglia (DRG) explants. Thus, we performed axonal dieback screening using adult rat DRG explants in vitro with Caps as a positive control and assessed cytotoxicity. Imy inhibited axonal outgrowth and slowed axonal dieback, while Pyr and Vcr at high concentrations produced significant reduction in axon length and robust axonal dieback within three days. DRGs treated with Caps, Vcr, or Imy had increased DRG cytotoxicity compared to matched controls, but overall cytotoxicity was minimal and at least 88% lower compared to lysed DRGs. Pyr did not lead to any DRG cytotoxicity. Further, neither Pyr nor Vcr triggered intervertebral disc cell death or affected cellular metabolic activity after three days of incubation in vitro. Overall, our findings suggest Pyr and Vcr are not toxic to DRGs and intervertebral disc cells, and there is potential for repurposing these compounds for axonal dieback compounds to cause local denervation and alleviate pain.

Evaluation of Pm2.5 Influence on Human Lung Cancer Cells Using a Microfluidic Platform.

In this study, we developed a microfluidic device that is able to monitor cell biology under continuous PM2.5 treatment. The effects of PM2.5 on human alveolar basal epithelial cells, A549 cells, and uncovered several significant findings were investigated. The results showed that PM2.5 exposure did not lead to a notable decrease in cell viability, indicating that PM2.5 did not cause cellular injury or death. However, the study found that PM2.5 exposure increased the invasion and migration abilities of A549 cells, suggesting that PM2.5 might promote cell invasiveness. Results of RNA sequencing revealed 423 genes that displayed significant differential expression in response to PM2.5 exposure, with a particular focus on pathways associated with the generation of reactive oxygen species (ROS) and mitochondrial dysfunction. Real-time detection demonstrated an increase in ROS production in A549 cells after exposure to PM2.5. JC1 assay, which indicated a loss of mitochondrial membrane potential (ΔΨm) in A549 cells exposed to PM2.5. The disruption of mitochondrial membrane potential further supports the detrimental effects of PM2.5 on A549 cells. These findings highlight several adverse effects of PM2.5 on A549 cells, including enhanced invasion and migration capabilities, altered gene expression related to ROS pathways, increased ROS production and disruption of mitochondrial membrane potential. These findings contribute to our understanding of the potential mechanisms through which PM2.5 can impact cellular function and health.

Photodynamic treatment increases the lifespan and oxidative stress resistance of Caenorhabditis elegans.

Photodynamic therapy is a noninvasive treatment in which specific photosensitizers and light are used to produce high amounts of reactive oxygen species (ROS), which can be employed for targeted tissue destruction in cancer treatment or antimicrobial therapy. However, it remains unknown whether lower amounts of ROS produced by mild photodynamic therapy increase lifespan and stress resistance at the organism level. Here, we introduce a novel photodynamic treatment (PDTr) that uses 20 µM hypericin, a photosensitizer that originates from Hypericum perforatum, and orange light (590 nm, 5.4 W/m2, 1 min) to induce intracellular ROS formation (ROS), thereby resulting in lifespan extension and improved stress resistance in C. elegans. The PDTr-induced increase in longevity was abrogated by N-acetyl cysteine, suggesting the hormetic response was driven by prooxidative mechanisms. PDTr activated the translocation of SKN-1/NRF-2 and DAF-16/FOXO, leading to elevated expression of downstream oxidative stress-responsive genes, including ctl-1, gst-4, and sod-3. In summary, our findings suggest a novel PDTr method that extends the lifespan of C. elegans under both normal and oxidative stress conditions through the activation of SKN-1 and DAF-16 via the involvement of many antioxidant genes.

Patient Education Materials for Immobilisation Masks in Radiation Therapy for Adult Head and Neck Cancer Patients: A Scoping Review.

Immobilisation masks (IMs) are used for people with head and neck cancer (HNC) undergoing radiation therapy (RT) treatment to ensure accuracy and reproducibility between treatments. Claustrophobia-related mask anxiety in HNC patients is common and can compromise treatment due to patient distress. This scoping review aimed to describe the content of publicly available Patient Education Materials (PEMs) for people with HNC undergoing RT. Three search engines (Bing, Yahoo, and Google) were systematically searched using standard terms. PEMs in audio-visual or written formats were eligible for inclusion if the target readership was adults with HNC and included content on IMs for RT. Content was appraised using the Patient Education Materials Assessment Tool for Printable and Audio-Visual Materials to assess understandability and actionability. In total, 304 PEMs were identified of which 20 met the inclusion criteria. Sixteen PEMs were webpages, three were PDF format, and one was a standalone video. The understandability and actionability of PEMs ranged between 47 to 100% and 0 to 80%, respectively. PEMs authored by Foundations/Organisations scored higher in understandability (80-100%) and were more likely to discuss mask anxiety coping strategies. In comparison, News sites and IM manufacturers published PEMs with the lowest understandability scores (20-80%). The significant variations in the quality of IM PEMs identified suggest that some sources may be more effective at informing patients about IMs. Although multiple aspects of the PEMs were consistent across the reviewed materials, many PEMs lacked information, and a stronger focus on understandability and actionability is required.

Ultra-High-Frequency ECG in Cardiac Pacing and Cardiac Resynchronization Therapy: From Technical Concept to Clinical Application.

Identifying electrical dyssynchrony is crucial for cardiac pacing and cardiac resynchronization therapy (CRT). The ultra-high-frequency electrocardiography (UHF-ECG) technique allows instantaneous dyssynchrony analyses with real-time visualization. This review explores the physiological background of higher frequencies in ventricular conduction and the translational evolution of UHF-ECG in cardiac pacing and CRT. Although high-frequency components were studied half a century ago, their exploration in the dyssynchrony context is rare. UHF-ECG records ECG signals from eight precordial leads over multiple beats in time. After initial conceptual studies, the implementation of an instant visualization of ventricular activation led to clinical implementation with minimal patient burden. UHF-ECG aids patient selection in biventricular CRT and evaluates ventricular activation during various forms of conduction system pacing (CSP). UHF-ECG ventricular electrical dyssynchrony has been associated with clinical outcomes in a large retrospective CRT cohort and has been used to study the electrophysiological differences between CSP methods, including His bundle pacing, left bundle branch (area) pacing, left ventricular septal pacing and conventional biventricular pacing. UHF-ECG can potentially be used to determine a tailored resynchronization approach (CRT through biventricular pacing or CSP) based on the electrical substrate (true LBBB vs. non-specified intraventricular conduction delay with more distal left ventricular conduction disease), for the optimization of CRT and holds promise beyond CRT for the risk stratification of ventricular arrhythmias.

Metabolites from Streptomyces aureus (VTCC43181) and Their Inhibition of Mycobacterium tuberculosis ClpC1 Protein.

Tuberculosis is one of the most common infectious diseases in the world, caused by Mycobacterium tuberculosis. The outbreak of multiple drug-resistant tuberculosis has become a major challenge to prevent this disease worldwide. ClpC1 is a Clp ATPase protein of Mycobacterium tuberculosis, functioning as a chaperon when combined with the Clp complex. ClpC1 has emerged as a new target to discover anti-tuberculosis drugs. This study aimed to explore the ClpC1 inhibitors from actinomycetes, which have been known to provide abundant sources of antibiotics. Two cyclic peptides, including nocardamin (1), halolitoralin A (3), and a lactone pleurone (2), were isolated from the culture of Streptomyces aureus (VTCC43181). The structures of these compounds were determined based on the detailed analysis of their spectral data and comparison with references. This is the first time these compounds have been isolated from S. aureus. Compounds 1-3 were evaluated for their affection of ATPase activity of the recombinant ClpC1 protein. Of these compounds, halolitoralin A (1), a macrocyclic peptide, was effective for the ATPase hydrolysis of the ClpC1 protein.

Interaction between sleep duration and trouble sleeping on depressive symptoms among U.S. adults, NHANES 2015-2018.

BACKGROUND: This study aims to examine the associations and interaction effects of sleep duration and trouble sleeping on depressive symptoms among U.S. adults. METHODS: National Health and Nutrition Examination Survey (NHANES) data from 2015 to 2018 were analyzed (N = 10,044). Trouble sleeping and sleep duration were self-reported. Sleep duration was defined as short (≤6 h) or long (≥9 h), compared with normal (>6 and < 9 h). Depressive symptoms were determined by the Patient Health Questionnaire-9 score ≥ 10. Both multiplicative interaction and additive interaction were reported. RESULTS: There was a significant positive additive interaction between short sleep duration and trouble sleeping on depressive symptoms in the fully adjusted model (Relative excess risk due to interaction, RERIOR = 4.42, 95 % CI: 1.12, 7.73), with 43 % of the association with depressive symptoms attributed to the interaction (attributable proportion of interaction, AP = 0.43, 95 % CI: 0.22, 0.64). Similarly, a significant positive additive interaction between long sleep duration and trouble sleeping on depressive symptoms was found (RERIOR = 4.17, 95 % CI: 0.96, 7.38), with 41 % of the association with depressive symptoms attributed to the interaction (AP = 0.41, 95 % CI: 0.21, 0.60). No multiplicative interaction was detected between short or long sleep duration and trouble sleeping. LIMITATIONS: The cross-sectional design limits the ability to draw causal inferences. CONCLUSIONS: Findings suggest that different aspects of sleep health interact synergistically, accounting for a substantial portion of the association with depressive symptoms. This underscores the importance of simultaneously considering multiple dimensions of sleep health in relation to depressive symptoms.

Exploring QRS Area beyond Patient Selection in CRT-Can It Guide Left Ventricular Lead Placement?

Vectorcardiographic QRS area is a promising tool for patient selection and implantation guidance in cardiac resynchronization therapy (CRT). Research has mainly focused on the role of QRS area in patient selection for CRT. Recently, QRS area has been proposed as a tool to guide left ventricular lead placement in CRT. Theoretically, vector-based electrical information of ventricular fusion pacing, calculated from the basic 12-lead ECG, can give real-time insight into the extent of resynchronization at any LV lead position, as well as any selected electrode on the LV lead. The objective of this review is to provide an overview of the background of vectorcardiographic QRS area and its potential in optimizing LV lead location in order to optimize the benefits of CRT.

Type I collagen concentration affects neurite outgrowth of adult rat DRG explants by altering mechanical properties of hydrogels.

Type I collagen is a predominant fibrous protein that makes up the extracellular matrix. Collagen enhances cell attachment and is commonly used in three-dimensional culture systems, to mimic the native extracellular environment, for primary sensory neurons such as dorsal root ganglia (DRG). However, the effects of collagen concentration on adult rat DRG neurite growth have not been assessed in a physiologically relevant, three-dimensional culture. This study focuses on the effects of type I collagen used in a methacrylated hyaluronic acid (MAHA)-laminin-collagen gel (triple gel) on primary adult rat DRG explants in vitro. DRGs were cultured in triple gels, and the neurite lengths and number of support cells were quantified. Increased collagen concentration significantly reduced neurite length but did not affect support cell counts. Mechanical properties, fiber diameter, diffusivity, and mesh size of the triple gels with varying collagen concentration were characterized to further understand the effects of type I collagen on hydrogel property that may affect adult rat DRG explants. Gel stiffness significantly increased as collagen concentration increased and is correlated to DRG neurite length. Collagen concentration also significantly impacted fiber diameter but there was no correlation with DRG neurite length. Increasing collagen concentration had no significant effect on mesh size and diffusivity of the hydrogel. These data suggest that increasing type I collagen minimizes adult rat DRG explant growth in vitro while raising gel stiffness. This knowledge can help develop more robust 3D culture platforms to study sensory neuron growth and design biomaterials for nerve regeneration applications.

Left ventricular lead placement in cardiac resynchronization therapy: Current data and potential explanations for the lack of benefit.

The present article reviews the literature on image-guided cardiac resynchronization therapy (CRT) studies. Improved outcome to CRT has been associated with the placement of a left ventricular (LV) lead in the latest activated segment free from scar. The majority of randomized controlled trials investigating guided LV lead implantation did not show superiority over conventional implantation approaches. Several factors may contribute to this paradoxical observation, including inclusion criteria favoring patients with left bundle branch block who already respond well to conventional anatomical LV lead implantation, differences in activation wavefronts during simultaneous right ventricular and LV pacing, incorrect definition of target regions, and limitations in coronary venous anatomy that prevent access to target regions that are detected by imaging. It is imperative that exclusion of patients lacking access to target regions from these studies would lead to larger benefit of image-guided CRT.

Biosynthesis of green CuO@C nanocomposite using Combretum indicum flower extract for organic dye removal: adsorption performance, modeling, and recyclability studies.

Water contamination becomes one of the most high-priority environmental concerns, calling for the efficient treatment techniques. Bionanocomposites can be robust adsorbents, but the synthesis requires toxic chemicals or energy consuming and cause the secondary pollution. Green nanocomposites can be biogenically synthesized using the plant extract to end up with a critically safe strategy. Herein, we used the flower extract of Combretum indicum plant as a bio-based reductant and carbonaceous source for the green CuO@C nanocomposite. This green nanoadsorbent obtained a specific surface area of 17.33 m2/g, good crystallinity, and functional group-containing surface, i.e., -OH and -CONH-. We also conducted the optimization of parameters, i.e., concentration, CuO@C dose, pH, time, and temperature, and reached removal efficiencies towards malachite green (MG, 83.23%), Congo red (CR, 84.60%), brilliant blue (BB, 71.39%), and methylene blue (MB, 23.67%). The maximum adsorption capacities were found as ordered, MG (46.387 mg/g) > MB (23.154 mg/g) > BB (22.8 mg/g) > CR dye (11.063 mg/g). Through the intra-particle diffusion kinetic model, MG and BB adsorption endured a three-step process, while CR and MB adsorption was a two-step process. The recyclability of the green CuO@C nanocomposite was three cycles with 67.54% for the final cycle of BB removal. Moreover, the nanoadsorbent displayed a high stability, checked by X-ray diffraction, FT-IR analysis, EDX spectra, and SEM images. It is recommended that the green CuO@C nanocomposite biosynthesized using the Combretum indicum flower extract can be a good alternative for the dye treatment from wastewater.

A Bayesian spatiotemporal approach to modelling arboviral diseases in Mexico.

BACKGROUND: The objective of this study was to evaluate the spatial and temporal patterns of disease prevalence clusters of dengue (DENV), chikungunya (CHIKV) and Zika (ZIKV) virus and how socio-economic and climatic variables simultaneously influence the risk and rate of occurrence of infection in Mexico. METHODS: To determine the spatiotemporal clustering and the effect of climatic and socio-economic covariates on the rate of occurrence of disease and risk in Mexico, we applied correlation methods, seasonal and trend decomposition using locally estimated scatterplot smoothing, hotspot analysis and conditional autoregressive Bayesian models. RESULTS: We found cases of the disease are decreasing and a significant association between DENV, CHIKV and ZIKV cases and climatic and socio-economic variables. An increment of cases was identified in the northeastern, central west and southeastern regions of Mexico. Climatic and socio-economic covariates were significantly associated with the rate of occurrence and risk of the three arboviral disease cases. CONCLUSION: The association of climatic and socio-economic factors is predominant in the northeastern, central west and southeastern regions of Mexico. DENV, CHIKV and ZIKV cases showed an increased risk in several states in these regions and need urgent attention to allocate public health resources to the most vulnerable regions in Mexico.

Physician-level determinants of HCV screening during pregnancy in a U.S. sample.

PURPOSE: The purpose of this study was to assess the association between select determinants and HCV screening guideline adherence among physicians who provide prenatal care. RESEARCH QUESTION: What factors may act as determinants of guideline adherence to HCV screening among physicians who provide prenatal care? METHODS: We surveyed a national sample of physicians who provided prenatal care in 2021. The survey included questions from the Clinician Guideline Determinant (CGD) questionnaire, demographic characteristics, and medical practice characteristics. We estimated odds ratios and 95% confidence intervals (CIs) using semi-Bayesian logistic regression for the association between determinants and guideline adherence. RESULTS: Participants included 224 physicians in the United States who reported providing prenatal care. Most physicians practiced in private practice (65%) and the majority were members of the American College of Obstetricians and Gynecologists (ACOG; 91%). Less than half (43%; 95% CI: 36%-49%) of physicians reported regular use of the HCV screening guideline. Physicians who reported general knowledge about HCV (OR = 9.0, 95% CI 3.1-30) or endorsed agreement with ease of implementation (OR = 8.0, 95% CI 2.7-25) had higher odds of adherence to the HCV screening guideline. CONCLUSION: Our study suggests that less than half of practicing prenatal care physicians adhere to HCV screening guidelines for pregnant patients. Our results may be useful as a preliminary screening of select determinants of guideline use for further investigation.

A village in a dish model system for population-scale hiPSC studies.

The mechanisms by which DNA alleles contribute to disease risk, drug response, and other human phenotypes are highly context-specific, varying across cell types and different conditions. Human induced pluripotent stem cells are uniquely suited to study these context-dependent effects but cell lines from hundreds or thousands of individuals are required. Village cultures, where multiple induced pluripotent stem lines are cultured and differentiated in a single dish, provide an elegant solution for scaling induced pluripotent stem experiments to the necessary sample sizes required for population-scale studies. Here, we show the utility of village models, demonstrating how cells can be assigned to an induced pluripotent stem line using single-cell sequencing and illustrating that the genetic, epigenetic or induced pluripotent stem line-specific effects explain a large percentage of gene expression variation for many genes. We demonstrate that village methods can effectively detect induced pluripotent stem line-specific effects, including sensitive dynamics of cell states.

University lecturers' conceptions of online teaching in distance education courses in Vietnamese higher education.

This study aimed to characterise academics' conceptions of teaching in fully online undergraduate distance education courses with no on-campus component. The study aimed to fill a gap in the literature, as previous research had examined conceptions of teaching in face-to-face courses, with a few studies of blended teaching via the Internet in on-campus courses. Fourteen academics from five faculties in a Vietnamese regional university were interviewed, with the study taking place shortly after the outbreak of Covid-19. Grounded theory was used for data analysis. The results revealed four categories of conceptions of online teaching, namely online teaching (1) as transmitting structured knowledge and skills, (2) as guiding students to acquire knowledge and skills, (3) as facilitating students' understanding via interaction and (4) as developing students' understanding and capabilities. The four categories of conceptions were defined and distinguished by a set of six dimensions, which included e-Learning/LMS. The set of categories had some similarities to those found for face-to-face teaching, but also some distinctions which could be explained by the nature of online teaching and learning. The study, therefore, makes a major contribution by establishing a category scheme for conceptions of teaching in online distance education, with detailed characterisation of the four categories of conceptions. The descriptors of the conceptions showed cultural influences, particularly from a Confucian heritage, which is of significance as research into face-to-face conceptions had not found cultural variations.

LBD18 and IAA14 antagonistically interact with ARF7 via the invariant Lys and acidic residues of the OPCA motif in the PB1 domain.

MAIN CONCLUSION: LBD18 and IAA14 antagonistically interact with ARF7 through the electrostatic faces in the ARF7PB1 domain, modulating ARF7 transcriptional activity. Auxin Response Factor 7 (ARF7)/ARF19 control lateral root development by directly activating Lateral Organ Boundaries Domain 16 (LBD16)/LBD18 genes in Arabidopsis. LBD18 upregulates ARF19 expression by binding to the ARF19 promoter. It also interacts with ARF7 through the Phox and Bem1 (PB1) domain to enhance the ARF7 transcriptional activity, forming a dual mode of positive feedback loop. LBD18 competes with the repressor indole-3-acetic acid 14 (IAA14) for ARF7 binding through the PB1 domain. In this study, we examined the molecular determinant of the ARF7 PB1 domain for interacting with LBD18 and showed that the electronic faces in the ARF7 PB1 domain are critical for interacting with LBD18 and IAA14/17. We used a luminescence complementation imaging assay to determine protein-protein interactions. The results showed that mutation of the invariant lysine residue and the OPCA motif in the PB1 domain in ARF7 significantly reduces the protein interaction between ARF7 and LBD18. Transient gene expression assays with Arabidopsis protoplasts showed that IAA14 suppressed transcription-enhancing activity of LBD18 on the LUC reporter gene fused to the ARF19 promoter harboring an auxin response element, but mutation of the invariant lysine residue and OPCA motif in the PB1 domain of IAA14 reduced the repression capability of IAA14 for transcription-enhancing activity of LBD18. We further showed that the same mutation in the PB1 domain of IAA14 reduces its repression capability, thereby increasing the LUC activity induced by both ARF7 and LBD18 compared with IAA14. These results suggest that LBD18 competes with IAA14 for ARF7 binding via the electrostatic faces of the ARF7 PB1 domain to modulate ARF7 transcriptional activity.

Moderation effects of serotype on dengue severity across pregnancy status in Mexico.

BACKGROUND: Pregnancy increases a woman's risk of severe dengue. To the best of our knowledge, the moderation effect of the dengue serotype among pregnant women has not been studied in Mexico. This study explores how pregnancy interacted with the dengue serotype from 2012 to 2020 in Mexico. METHOD: Information from 2469 notifying health units in Mexican municipalities was used for this cross-sectional analysis. Multiple logistic regression with interaction effects was chosen as the final model and sensitivity analysis was done to assess potential exposure misclassification of pregnancy status. RESULTS: Pregnant women were found to have higher odds of severe dengue [1.50 (95% CI 1.41, 1.59)]. The odds of dengue severity varied for pregnant women with DENV-1 [1.45, (95% CI 1.21, 1.74)], DENV-2 [1.33, (95% CI 1.18, 1.53)] and DENV-4 [3.78, (95% CI 1.14, 12.59)]. While the odds of severe dengue were generally higher for pregnant women compared with non-pregnant women with DENV-1 and DENV-2, the odds of disease severity were much higher for those infected with the DENV-4 serotype. CONCLUSION: The effect of pregnancy on severe dengue is moderated by the dengue serotype. Future studies on genetic diversification may potentially elucidate this serotype-specific effect among pregnant women in Mexico.

Whole Blood Resuscitation is Safe in Pediatric Trauma Patients: A Multicenter Study.

INTRODUCTION: Whole blood (WB) resuscitation has been associated with a mortality benefit in trauma patients. Several small series report the safe use of WB in the pediatric trauma population. We performed a subgroup analysis of the pediatric patients from a large prospective multicenter trial comparing patients receiving WB or blood component therapy (BCT) during trauma resuscitation. We hypothesized that WB resuscitation would be safe compared to BCT resuscitation in pediatric trauma patients. METHODS: This study included pediatric trauma patients (0-17 y), from ten level-I trauma centers, who received any blood transfusion during initial resuscitation. Patients were included in the WB group if they received at least one unit of WB during their resuscitation, and the BCT group was composed of patients receiving traditional blood product resuscitation. The primary outcome was in-hospital mortality with secondary outcomes being complications. Multivariate logistic regression was performed to assess for mortality and complications in those treated with WB vs BCT. RESULTS: Ninety patients, with both penetrating and blunt mechanisms of injury (MOI), were enrolled in the study (WB: 62 (69%), BCT: 28 (21%)). Whole blood patients were more likely to be male. There were no differences in age, MOI, shock index, or injury severity score between groups. On logistic regression, there was no difference in complications. Mortality was not different between the groups (P = .983). CONCLUSION: Our data suggest WB resuscitation is safe when compared to BCT resuscitation in the care of critically injured pediatric trauma patients.

Label-free optical imaging of cell function and collagen structure for cell-based therapies.

Cell-based therapies harness functional cells or tissues to mediate healing and treat disease. Assessment of cellular therapeutics requires methods that are non-destructive to ensure therapies remain viable and uncontaminated for use in patients. Optical imaging of endogenous collagen, by second-harmonic generation, and the metabolic coenzymes NADH and FAD, by autofluorescence microscopy, provides tissue structure and cellular information. Here, we review applications of label-free nonlinear optical imaging of cellular metabolism and collagen second-harmonic generation for assessing cell-based therapies. Additionally, we discuss the potential of label-free imaging for quality control of cell-based therapies, as well as the current limitations and potential future directions of label-free imaging technologies.