Search Results (329)

Doxorubicin (DOX) is widely used for the treatment of several tumors, but considerable dose-dependent side effects on many normal tissues, including bones, have been reported. The aim of the present study was to follow for the first time the kinetics of DOX accumulation/clearance in the non-vascularized intervertebral disc (IVD), as well as to assess the drug’s biological action in the annulus fibrosus (AF) and nucleus pulposus (NP) IVD cells and tissues. DOX was administered intravenously to rabbits before the isolation of IVDs, in which DOX quantification was performed using a highly sensitive LC-HRMS/MS analytical method. The effect of the drug on IVD cells’ physiology was assessed in vitro, while IVD tissue quality post-DOX administration was studied in vivo through histological analysis. DOX delivery was found significantly lower in the IVD compared to the highly vascularized skin, declining from the outer AF to the inner NP. The low DOX concentrations reaching the IVDs had marginal effects on cells’ viability, intracellular redox status, and p38 MAPK activation, while they did not evoke cellular senescence. Most importantly, the drug did not negatively affect ECM integrity, as collagen and proteoglycan content remained stable in vitro and in vivo. Full article

In this study, an accurate attenuation measurement system with high attenuation capability (≥100 dB) is presented, covering a broad radio frequency range from 1 GHz to 25 GHz. The system employs a dual-channel intermediate frequency (IF) substitution method, utilizing a programmable inductive voltage divider (IVD) that provides precise voltage ratios at a 1 kHz operating IF, serving as the primary attenuation standard. To ensure optimal inter-channel isolation, essential for accurate high-attenuation measurements, an optical fiber assembly, consisting of a laser diode, a wideband external electro-optic modulator, and a photodetector, is integrated between the channels. A comprehensive performance evaluation is presented, with particular emphasis on the programmable IVD calibration technique, which achieves an accuracy better than 0.001 dB across all attenuation levels, and on the role of the optical fiber assembly in enhancing isolation, demonstrating levels exceeding 120 dB across the entire frequency range. The system demonstrates measurement capabilities with expanded uncertainties (k = 2) of 0.004 dB, 0.008 dB, and 0.010 dB at attenuation levels of 20 dB, 60 dB, and 100 dB, respectively. Full article

In dogs with intervertebral disc extrusion (IVDE), the Glasgow Composite Measure Pain Scale—Short Form (GCMPS) and the Sharp and Wheeler Grading Scale (SWGS) are routinely used in the evaluation of pain (GCMPS) and neurological function (SWGS). Additionally, quantitative sensory tests (QSTs) are increasingly being incorporated into veterinary clinical practice for pain characterisation. The aim was to investigate a possible relationship between the GCMPS, the SWGS, and mechanical thresholds (MTs) in 31 client-owned dogs with thoracolumbar IVDEs. Dogs were always assessed in the same order, starting with pain rating using the GCMPS, followed by classifying neurological severity using the SWGS, before determining MTs using a handheld pressure algometer. Dogs were evaluated over a five-day testing period (before surgery and on days one, two, three, and ten after surgery). The GCMPS and the SWGS data remained consistent across all days of testing. No statistically significant correlation or difference was observed between the scores. MTs showed a significant negative correlation with the GCMPS (r = −0.311; p < 0.001) and a positive one with the SWGS (r = 0.282; p = 0.002). The GCMPS and MTs showed a slight divergence in their progression. MTs might be more sensitive than GCMPS in reflecting clinical improvement and should be considered for clinical practice. Full article

Accurate and sensitive detection of protein biomarkers is critical for advancing in vitro diagnostics (IVD), yet conventional enzyme-linked immunosorbent assays (ELISA) often fall short in terms of sensitivity compared to nucleic acid-based tests. Bridging this sensitivity gap is essential for improving diagnostic accuracy, particularly in diseases where protein levels better reflect disease progression than nucleic acid biomarkers. In this review, we present strategies developed to enhance the sensitivity of ELISA, structured according to the sequential steps of the assay workflow. Beginning with surface modifications, we then discuss the methodologies to improve mixing and washing efficiency, followed by a summary of recent advances in signal generation and amplification techniques. In particular, we highlight the emerging role of cell-free synthetic biology in augmenting ELISA sensitivity. Recent developments such as expression immunoassays, CRISPR-linked immunoassays (CLISA), and T7 RNA polymerase–linked immunosensing assays (TLISA) demonstrate how programmable nucleic acid and protein synthesis systems can be integrated into ELISA workflows to surpass the present sensitivity, affordability, and accessibility. By combining synthetic biology-driven amplification and signal generation mechanisms with traditional immunoassay formats, ELISA is poised to evolve into a highly modular and adaptable diagnostic platform, representing a significant step toward the next generation of highly sensitive and programmable immunoassays. Full article

6-PPD quinone (6-PPDQ) is a derivative from 6-PPD, an antioxidant added in tires. Leucine is an important amino acid that needs to be obtained from the diet. In Caenorhabditis elegans, we examined the effect of 6-PPDQ exposure at environmentally relevant concentrations (ERCs) on the content of leucine and underlying mechanisms. In nematodes, 0.1–10 μg/L of 6-PPDQ decreased leucine content. The expression of the aat-1-encoding amino acid transmembrane transporter was decreased by 0.1–10 μg/L of 6-PPDQ, and leucine content was reduced by aat-1 RNAi. Meanwhile, the expression of bcat-1-encoding branched-chain amino acid transferase was increased by 0.1–10 μg/L of 6-PPDQ, and leucine content was increased by bcat-1 RNAi. Additionally, the expressions of dbt-1 and ivd-1 encoding two enzyme genes governing NADH and FADH₂ generations were decreased by 0.1–10 μg/L of 6-PPDQ, and their expressions in 6-PPDQ exposed nematodes were increased by bcat-1 RNAi. After 6-PPDQ exposure, NADH content was reduced by dbt-1 RNAi, and FADH₂ content was reduced by ivd-1 RNAi. Moreover, 6-PPDQ-induced mitochondrial dysfunction and other aspects of toxicity (such as intestinal ROS generation and lipofuscin accumulation, inhibited locomotion, and reduced brood size) were suppressed by bcat-1 RNAi and strengthened by dbt-1 and ivd-1 RNAi. The 6-PPDQ-induced toxicity and the decrease in dbt-1 and ivd-1 expressions could be inhibited by following leucine (5 mM) treatment. Our results demonstrate the important association of leucine adsorption and catabolism with 6-PPDQ toxicity induction. Full article

Background/Objectives: Lumbar disc herniation (LDH) is the most common condition associated with low back pain, and it adversely impacts individuals’ health. The interplay between energy metabolism and apoptosis is critical, as the loss of viable cells in the intervertebral disc (IVD) can lead to a cascade of degenerative changes. Efferocytosis is a key biological process that maintains homeostasis by removing apoptotic cells, resolving inflammation, and promoting tissue repair. Therefore, enhancing mitochondrial energy metabolism and efferocytosis function in IVD cells holds great promise as a potential therapeutic approach for LDH. Methods: In this study, energy metabolism and efferocytosis-related differentially expressed genes (EMERDEGs) were identified from the transcriptomic datasets of LDH. Machine learning approaches were used to identify key genes. Functional enrichment analyses were performed to elucidate the biological roles of these genes. The functions of the hub genes were validated by RT-qPCR. The CIBERSORT algorithm was used to compare immune infiltration between LDH and Control groups. Additionally, we used single-cell RNA sequencing dataset to analyze cell-specific expression of the hub genes. Results: By using bioinformatics methods, we identified six EMERDEGs hub genes (IL6R, TNF, MAPK13, ELANE, PLAUR, ABCA1) and verified them using RT-qPCR. Functional enrichment analysis revealed that these genes were primarily associated with inflammatory response, chemokine production, and cellular energy metabolism. Further, we identified candidate drugs as potential treatments for LDH. Additionally, in immune infiltration analysis, the abundance of activated dendritic cells, neutrophils, and gamma delta T cells varied significantly between the LDH group and Control group. The scRNA-seq analysis showed that these hub genes were mainly expressed in chondrocyte-like cells. Conclusions: The identified EMERDEG hub genes and pathways offer novel insights into the molecular mechanisms underlying LDH and suggest potential therapeutic targets. Full article

Following the application of epigenetic and epigenomics research into tumor diseases, cardiovascular disease, diabetes, hereditary diseases, and rare diseases, in vitro diagnostics (IVD) epigenetic and epigenomics are increasingly employed for those patients. Here, we review a clinical sampling of epigenetics and epigenomics from patients. We then present procedures, including the detection procedure of clinical epigenetic approaches from clinical samples, clinical epigenomic methods applied to those samples, the small cell number of epigenetics, and epigenomics. Finally, we present the current IVD of epigenetics and epigenomics used for clinical analysis and diagnosis, along with the development of approaches. To improve clinical study and diagnosis, we also introduce clinical research and clinical applications to develop a more comprehensive strategy to maximize the sensitivity and specificity of the epigenetics and epigenomics analysis and diagnosis. Full article

Lumbar spine Magnetic Resonance Imaging (MRI) is commonly used for intervertebral disc (IVD) and vertebral body (VB) evaluation during low back pain. Segmentation of these tissues can provide useful quantitative information such as shape and volume. The objective of the study was to determine the performances of Segment Anything Model (SAM) and medical SAM (MedSAM), two “zero-shot” deep learning models, in segmenting lumbar IVD and VB from MRI images and compare against the nnU-Net model. This cadaveric study used 82 donor spines. Manual segmentation was performed to serve as ground truth. Two readers processed the spine MRI using SAM and MedSAM by placing points or drawing bounding boxes around regions of interest (ROI). The outputs were compared against ground truths to determine Dice score, sensitivity, and specificity. Qualitatively, results varied but overall, MedSAM produced more consistent results than SAM, but neither matched the performance of nnU-Net. Mean Dice scores for MedSAM were 0.79 for IVDs and 0.88 for VBs, and significantly higher (each p < 0.001) than those for SAM (0.64 for IVDs, 0.83 for VBs). Both were lower compared to nnU-Net (0.99 for IVD and VB). Sensitivity values also favored MedSAM. These results demonstrated the feasibility of “zero-shot” DL models to segment lumbar spine MRI. While performance falls short of recent models, these zero-shot models offer key advantages in not needing training data and faster adaptation to other anatomies and tasks. Validation of a generalizable segmentation model for lumbar spine MRI can lead to more precise diagnostics, follow-up, and enhanced back pain research, with potential cost savings from automated analyses while supporting the broader use of AI and machine learning in healthcare. Full article

Objectives: To investigate whether preservation of the posterior elements protects the spine from degeneration and improves postoperative symptoms in lumbar spine laminoplasty. Methods: Eighty-five consecutive patients who underwent lumbar spine laminoplasty were retrospectively reviewed. They were non-randomly stratified into two groups, the posterior elements resection (R) group and the preservation (P) group, and they were followed for two years after surgery. We radiographically analyzed the conditions of the spine and intervertebral disc (IVD) two years after surgery. The Japanese Orthopaedic Association Back Pain Evaluation Questionnaire (JOABPEQ) was used for symptom assessments. Logistic regression analysis was performed to determine whether the kissing spine was a significant factor for the outcomes in group R. Results: The 2-year D score increment and 2-year IVD height decrement was lower in group P. No difference was found in the flexion–extension angles or incidence of instability between groups. The JOABPEQ revealed higher scores in walking ability, social life function, and mental health in group P one year after surgery. Walking ability was the only score that remained higher two years after surgery. The visual analog scale of pain in the buttocks and lower limbs was lower in group P only one year after surgery. Finally, the kissing spine was not a significant factor in any outcome. Conclusions: The preserved posterior elements were considered to protect the IVD in lumbar spine laminoplasty. In addition, they positively affected postoperative health status from multiple aspects. Full article

The human neck is highly vulnerable in motor vehicle crashes, and cervical spine response data are essential to improve injury prediction tools (e.g., crash test dummies, human body models). This feasibility study aimed to implement the use of pressure sensors in whole-body post-mortem human subject (PMHS) cervical spine intervertebral discs (IVDs) to confirm the feasibility and repeatability of cervical IVD pressure response to biomechanic research. Two fresh frozen whole-body PMHSs were instrumented with miniature pressure sensors (Model 060S, Precision Measurement Company, Ann Arbor, MI, USA) at three cervical IVD levels (C3/C4, C5/C6, and C7/T1) using minimally invasive surgical insertion techniques. Each PMHS underwent three quasistatic motion test trials, and each trial included multiple head/neck motions (i.e., gentle traction, flexion/extension, lateral bending, axial rotation, and forced tension/compression). Results showed marked pressure differences between both the cervical level assessed and the motion undertaken as well as successful intra-subject repeatability between the three motion trials. This study demonstrates that changes in cervical IVD pressure are associated with motion events of the cervical spine. Cervical IVD response data could be utilized to assess and supplement the characterization of the head/neck complex motion, and data could facilitate the continued improvement of injury prediction tools. Full article

Low back pain is a widespread condition that significantly impacts quality of life, with intervertebral disc degeneration (IDD) being a major contributing factor. However, the underlying mechanisms of IDD remain poorly understood, necessitating further investigation. Environmental risk factors, such as mechanical stress and cigarette smoke, elevate reactive oxygen species levels from both endogenous and exogenous sources, leading to redox imbalance and oxidative stress. The endoplasmic reticulum (ER) and mitochondria, two key organelles responsible for protein folding and energy production, respectively, are particularly vulnerable to oxidative stress. Under oxidative stress conditions, ER stress and mitochondrial dysfunction occur, resulting in unfolded protein response activation, impaired biosynthetic processes, and disruptions in the tricarboxylic acid cycle and electron transport chain, ultimately compromising energy metabolism. Prolonged and excessive ER stress can further trigger apoptosis through ER–mitochondrial crosstalk. Given the unique microenvironment of the intervertebral disc (IVD)—characterized by hypoxia, glucose starvation, and region-specific cellular heterogeneity—the differential effects of environmental stressors on distinct IVD cell populations require further investigation. This review explores the potential mechanisms through which environmental risk factors alter IVD cell activities, contributing to IDD progression, and discusses future therapeutic strategies aimed at mitigating disc degeneration. Full article

Intervertebral disc (IVD) disease is typically characterized by the degradation of IVD tissue, secretion of inflammatory and painful factors, and hyperinnervation of the disc. The pro-inflammatory cytokine interleukin-1β (IL-1β) has been regarded as a principal factor in orchestrating disc degeneration. Link N (LN) is a peptide derived from the link protein that has been shown to promote extracellular disc regeneration even in an inflammatory milieu; however, no mechanism(s) has been described for their behaviour to date. Building on prior studies on LN, we hypothesize that LN directly inhibits IL-1β. IVD degeneration was experimentally induced in New Zealand white rabbits, followed by the injection of either sLN or saline as the vehicle control. To determine the expression of markers of pain, histology was performed. Cultured human Nucleus Pulposus disc cells (hNP) were used to determine the effects of LN on IL-1β-induced changes in gene expression, including the effects on IL-1β, TNFα, and IL6 signalling. Isolated murine dorsal root ganglia (DRG) neurons were used to assess the effect of LN on IL-1β-induced neuronal hyperactivity. LN significantly reduced IL-1β-induced NF-κB activation in a dose-dependent manner in disc cells and was further able to modulate IL-1β-induced gene expression, inflammatory mediators, and neurotrophic factors. Peptide docking simulations revealed that LN could interact with IL-1β. A direct interaction of LN and IL-1β was revealed through co-immunoprecipitation experiments. Although IL-1β was able to hypersensitize DRG neurons following a seven-day exposure, as demonstrated by Ca²⁺ imaging, this effect was significantly blunted when co-treated with LN. LN demonstrates a novel mechanism of action by directly inhibiting IL-1β, in addition to mitigating IL-1β-induced hypersensitivity in DRG neurons. These data suggest a potential role for LN in reducing discogenic pain. Full article

This study presents a novel approach for predicting the location and fatigue life of degenerative intervertebral discs (IVDs) under cyclic loading conditions, aiming to improve the understanding of disc degeneration mechanisms. Based on mechanical theories linking IVD degeneration to stress imbalance and water loss, a finite element (FE) model of the L4–L5 lumbar spine was developed, combining probability-weighted anatomical structures, inverse dynamics, and cumulative fatigue mechanics. By quantifying stress variations and calculating cumulative damage across disc regions, stress-concentration areas prone to degeneration were identified, and validation via a case study of a retired weightlifter diagnosed with intervertebral disc disease (IVDD) demonstrated that the predicted degeneration location correlated well with affected areas observed in CT scan images. These findings suggest that prolonged, abnormal stress imbalances within the disc may contribute significantly to degeneration, offering potential clinical applications in preventive assessment and targeted treatment for spine health. Full article

Electrical and mechanical dyssynchrony (MD) underlies left ventricular (LV) contractile dysfunction in patients with heart failure (HF) and left bundle branch block (LBBB). In some cases, cardiac resynchronization therapy (CRT) almost completely reverses LV contractile dysfunction. The LBBB electrocardiographic Strauss criteria and MD assessment were proposed to improve CRT response. However, using these techniques separately does not improve LV contraction in 20–40% of patients after CRT device implantation. We aimed to evaluate whether the combined use of electrocardiography (ECG), speckle-tracking echocardiography (STE) and cardiac scintigraphy could improve the prognosis of CRT super-response in patients with HF and Strauss LBBB criteria during a 6-month follow-up period. The study prospectively included patients with HF, classified as New York Heart Association (NYHA) functional class (FC) II–III in sinus rhythm with Strauss LBBB criteria and reduced left ventricular ejection fraction (LVEF). Before and 6 months after CRT device implantation, ECG, STE and cardiac scintigraphy were performed. The study’s primary endpoint was the NYHA class improvement ≥ 1 and left ventricle end systolic volume decrease > 30% or LVEF improvement > 15% after 6 months of CRT. Based on collected data, we developed a prognostic model regarding the CRT super-response. Out of 54 (100.0%) patients, 39 (72.2%) had a CRT super-response. Patients with CRT super-response were likelier to have a greater S wave amplitude in V₂ lead (p = 0.004), higher rates of global longitudinal strain (GLS) (p = 0.001) and interventricular delay (IVD) (p = 0.005). Only three indicators (S wave amplitude in V₂ lead, GLS and IVD) were independently associated with CRT super-response in univariable and multivariable logistic regression. We created a prognostic model based on the logistic equation and calculated a cut-off value (>0.73). The resulting ROC curve revealed a discriminative ability with an AUC of 0.957 (sensitivity 87.2%; specificity 100.0%). The electrical and mechanical dyssynchrony assessment using ECG, STE and cardiac scintigraphy is useful in the prediction of CRT super-response in patients with HF and Strauss LBBB criteria during a 6-month follow-up period. Our prognostic model can identify patients who are super-responders to CRT. Full article

Lateral flow tests (LFTs) had a pivotal role in combating the spread of the SARS-CoV-2 virus throughout the COVID-19 pandemic thanks to their affordability and ease of use. Most of LFT devices were based on nitrocellulose membrane strips whose industrial upscaling to billions of devices has already been extensively demonstrated. Nevertheless, the assay option in an LFT format is largely restricted to qualitative detection of the target antigens. In this research, we surveyed the potential of UV nanoimprint lithography (UV-NIL) and extrusion coating (EC) for the high-throughput production of disposable capillary-driven, foil-based tests that allow multistep assays to be implemented for quantitative readout to address the inherent lack of on-demand fluid control and sensitivity of paper-based devices. Both manufacturing technologies operate on the principle of imprinting that enables high-volume, continuous structuring of microfluidic patterns in a roll-to-roll (R2R) production scheme. To demonstrate the feasibility of R2R-fabricated foil chips in a point-of-care biosensing application, we adapted a commercial chemiluminescence multiplex test for COVID-19 antibody detection originally developed for a capillary-driven microfluidic chip manufactured with injection molding (IM). In an effort to build a complete ecosystem for the R2R manufacturing of foil chips, we also recruited additional processes to streamline chip production: R2R biofunctionalization and R2R lamination. Compared to conventional fabrication techniques for microfluidic devices, the R2R techniques highlighted in this work offer unparalleled advantages concerning improved scalability, dexterity of seamless handling, and significant cost reduction. Our preliminary evaluation indicated that the foil chips exhibited comparable performance characteristics to the original IM-fabricated devices. This early success in assay translation highlights the promise of implementing biochemical assays on R2R-manufactured foil chips. Most importantly, it underscores the potential utilization of UV-NIL and EC as an alternative to conventional technologies for the future development in vitro diagnostics (IVD) in response to emerging point-of-care testing demands. Full article