Search Results (347)

Accurate detection and semantic labeling of intervertebral discs (IVDs) in magnetic resonance imaging (MRI) are crucial for evaluating and treating spinal-related disorders. Conventional approaches typically utilize convolutional neural networks (CNNs) to extract contextual features from MRI images, but they often overlook the inherent geometric structure of the vertebral column, leading to inaccuracies in IVD localization and segmentation. Addressing this limitation, we propose a novel prompt encoder method that incorporates geometric information to enhance the semantic labeling of IVDs in MRI images. Our approach effectively learns the skeleton structure of the spinal column and adaptively adjusts its predictions to conform to this anatomical framework. Extensive evaluations on multi-center spine datasets demonstrate that our method outperforms existing state-of-the-art techniques, consistently achieving superior performance in both T1w and T2w MRI modalities. The incorporation of geometric information significantly improves the accuracy and robustness of IVD semantic labeling, paving the way for more precise and reliable assessments of spine health and disease progression. Full article

Background. Routine lymphocyte counting requires quality assurance validation using quality controls (QCs) that closely resemble fresh human blood leukocytes. This study aimed to evaluate a novel artificial product designed to mimic leukocyte light scatters and marker expression. Methods. FlowCytes and TruCytes, “artificial cell mimics” (Slingshot Biosciences, Emeryville, CA, USA), were tested on CE-IVD-certified systems, namely FACSCanto, FACSLyric (BD Biosciences), Navios, and DxFlex (Beckman Coulter), using routine staining, lysing, fixation, and no-wash procedures for T, B, and NK counting. Results. FlowCytes and TruCytes provided forward and side scatter profiles comparable to human leukocytes on the FACSCanto, FACSLyric, and DxFlex systems but not on Navios despite adapting the process to be slightly different from the manufacturer’s recommendations. TruCytes demonstrated robust immunolabeling of CD3, CD4, CD8, and CD19 on the FACSCanto, FACSLyric, and DxFlex systems, with fluorescence intensities and subset distributions being similar to those usually observed in fresh human blood. However, CD16 and CD56 labeling was inconsistent and depended on the antibody clones used. Regrettably, monocyte and granulocyte mimics lacked expression of CD4, CD16, and CD14. TruCytes also displayed significantly lower concentrations of TBNK lymphocyte subsets compared to healthy human blood. Conclusions. FlowCytes and TruCytes show promises as internal quality controls for T cell and B cell immunophenotyping, but not NK cells. They are compatible with most CE-IVD cytometers, even when using lysis/fixation/no-wash routine diagnosis procedures. Further multicentric studies are warranted to assess their performance relative to existing products, such as stabilized human blood. Full article

The growing demand for precision diagnostics in cystic fibrosis and other genetic disorders, such as cancers, is driving the need for sequencing platforms that combine analytical robustness, scalability, and cost-efficiency. In this study, we performed a direct comparison between two leading Next-Generation Sequencing (NGS) platforms, MiSeq (Illumina, CA, USA) and DNBSEQ-G99RS (MGI Tech Co., Shenzhen, China), using a CE-IVD-certified CFTR panel (Devyser AB), selected for its complexity and variant spectrum, including SNVs, CNVs, and intronic polymorphisms. A total of 47 genomic DNA samples from routine clinical activity were analyzed on both platforms. Illumina sequencing covered all CFTR variants using standard workflows, while MGI data were generated from residual diagnostic DNA, with informed consent. Sequencing data were processed using Amplicon Suite v3.7.0 for variant calling, annotation, and ACMG classification. Quality control metrics and platform-specific parameters were also evaluated. Both platforms demonstrated complete concordance in variant detection, including SNVs, CNVs, and complex alleles (e.g., Poly-T/TG). Illumina exhibited slightly superior basecalling quality and allelic frequency uniformity, while MGI achieved higher sequencing depth (mean ~2793×) and demultiplexing efficiency. No false positives, false negatives, or discordant HGVS annotations were observed. The use of full-gene CFTR sequencing enabled granular and technically rigorous cross-platform validation. These findings confirm the analytical equivalence of Illumina and MGI for diagnostic genotyping. Moreover, MGI’s greater data output and flow cell capacity may offer tangible advantages in high-throughput settings, including somatic applications such as liquid biopsy and molecular oncology workflows. Full article

Accurate labeling of intervertebral discs (IVDs) in MRI scans is crucial for diagnosing spinal-related diseases such as osteoporosis, vertebral fractures, and IVD herniation. However, automatic IVD labeling remains challenging. The main issues include visual similarity to surrounding bone, anatomical variation across individuals, and inconsistencies between MRI scans. Traditional post-detection disc labeling methods often struggle when localization algorithms miss discs or generate false positives. To address these challenges, we propose MSA-Net, a novel multi-scale attention network designed for semantic IVD labeling, emphasizing the use of prior geometric data. MSA-Net efficiently extracts multi-scale features and models intricate spatial dependencies throughout the spinal structure. We also integrate contrastive learning to enforce feature consistency. This helps the network distinguish IVDs from surrounding tissues. Extensive experiments on multi-center spine datasets demonstrate that MSA-Net consistently outperforms previous methods across MRI T1w and T2w modalities. These improvements demonstrate MSA-Net’s ability to handle variability in disc geometry, tissue contrast, and missed detections that challenge prior methods. Full article

Background/Objectives: The timely diagnosis of schistosomiasis and strongyloidiasis is important because of their potentially severe, even lethal, consequences. European diagnostic laboratories must comply with the European In Vitro Diagnostic (IVD) Regulation, which requires justifying the use of in-house assays when CE-IVD-marked kits are available. We aimed to compare the performance of the Biosynex Helminths AMPLIQUICK^® RT-PCR and the multiplex in-house RT-PCR for the diagnosis of Schistosoma mansoni and Strongyloides stercoralis currently used in our department, an Italian reference centre for tropical diseases. Methods: We conducted a performance comparison study on biobanked frozen stool samples classified as cases or controls according to PCR and/or copromicroscopy at diagnosis. Both RT-PCRs were performed on DNA re-extracted from the same stool aliquot. Sensitivity and specificity were compared using McNemar’s Chi-squared test, while agreement was assessed using Gwet’s AC1 and Cohen’s K coefficients, and Bland–Altman analysis. Results: A total of 45 S. mansoni cases with 52 controls and 29 S. stercoralis cases with 54 controls were analyzed. For both S. mansoni and S. stercoralis, sensitivity and specificity were not significantly different between RT-PCRs (p = 1). Concordance was perfect for controls (AC1 = 1) in both cohorts, but was poor for S. mansoni cases (AC1 = 0.38) and good for S. stercoralis cases (AC1 = 0.78). Conclusions: Performance was not significantly different between in-house and Biosynex RT-PCRs. Nevertheless, careful assessment of the specific molecular targets included in the panels and prospective evaluation of any newly introduced tests should be implemented to minimize the impact of clinically significant discrepancies. Full article

Pentadecanoic acid (C15:0) is an odd-chain fatty acid (OCFA) with significant health benefits, mainly produced by microbial fermentation. To improve C15:0 production, this study compared the effects of different alcohols on C15:0 production in Yarrowia lipolytica CICC1778, identified 1-propanol as the most effective precursor, assessed its optimal concentration, and employed transcriptomic and metabolomic analyses to elucidate the regulatory mechanisms. The results showed that supplementation with 0.5% 1-propanol resulted in a total lipid production of 1.54 g/L in Y. lipolytica CICC1778, showing no differences compared with the negative control (NC) group, while C15:0 production increased to 76.68 mg/L, representing a 794.7% increase compared with the NC group. Integrated omics analysis showed that propionylcarnitine was positively correlated with ADH2, ADH1, ACADSB, ALDH6A1, and CAT2; O-methylmalonylcarnitine was positively correlated with IVD, MCCC2, ACADSB, and ALDH6A1; and (R)-leucic acid and 2-hydroxy-3-methylbutyric acid were positively correlated with IVD, BAT2, MCCC2, and ACADSB and ALDH6A1 and BAT2, respectively. All of these DEGs and DEMs were upregulated in the alcohol-treated (ALC; supplementation with 0.5% 1-propanol) group. Taken together, supplementation with 0.5% 1-propanol was an effective strategy for enhancing C15:0 production in Y. lipolytica CICC1778; 1-propanol underwent dehydrogenation-oxidation and promoted branched-chain amino acid degradation to expand the propionyl-CoA pool, thereby facilitating C15:0 synthesis. Full article

This comprehensive narrative review of bioactive plant compounds, pro-resolving anti-inflammatory lipids, and statins shows their potential in the inhibition of intervertebral disc degeneration (IVDD), pain resolution, tissue repair, and disc regeneration. IVDD is a multifactorial disease involving a multitude of signaling pathways, leading to the loss of normal disc function. An influx of nociceptive mechanoreceptors generate low back pain (LBP). IL6 and IL8 levels are elevated in patients undergoing spinal fusion to alleviate LBP, indicating these pro-inflammatory mediators may be major contributors to the generation of LBP. Apoptosis of disc cells leads to the depletion of key extracellular matrix components that equip the disc with its weight-bearing properties. A biomechanically incompetent degenerated IVD stimulates nociceptor mechanoreceptor activity, generating pain. Myo-tendinous, vertebral body, muscle, and facet joint tissues also contain pain receptors. Disturbance of the normal architecture of the IVD also generates pain in these tissues. Plant compounds have been used in folkloric medicine for centuries. This review attempts to provide a scientific basis for their purported health benefits; however, further studies are still required to substantiate this. Until this evidence is available, it would be prudent to be cautious in the use of such compounds. A diverse range of plant compounds (flavonoids, terpenoids, glycosides, alkaloids, and polyphenolics) inhibit inflammation and apoptosis, reduce spinal pain, and stimulate tissue repair by targeting cell signaling pathways in IVDD. Pro-resolving lipid mediators (lipoxin A4, resolvin D1, protectins, and maresins) also reduce inflammation, maintaining disc health and function. Cholesterol lowering statins disrupt phosphorylation in cell signaling pathways inhibiting IVDD, promoting tissue repair and regeneration. Full article

Background/Objectives: Short-read-sequencing (SRS) is currently the standard for genetic testing in inherited human diseases. Intrinsic limitations include PCR dependency, restricted read length, and challenges in identifying structural variants (SVs), copy number variations (CNVs), and intronic small variants (SNVs/indels). Long-read-sequencing (LRS) enables the sequencing of long DNA molecules, detection of deep intronic variants, rapid testing of few samples, and improved resolution of SVs, CNVs, and SNVs/indels. We therefore aimed to validate Oxford Nanopore Technologies (ONT) LRS for potential clinical application. Methods: We evaluated the ONT’s ability to detect pathogenic/likely pathogenic (P/LP) variants previously identified by SRS and confirmed via Sanger sequencing, Multiplex-Ligation-dependent-Probe-Amplification (MLPA), or quantitative-PCR (qPCR). In total, 509 samples were analyzed, including 393 with P/LP variants and 116 negative controls. We used CE-IVD panels HEVA pro, CARDIO pro, BRaCA panel, and ClinEX pro (4Bases-CH). Sequencing was performed on MinION, GridION, and PromethION-2 platforms. Data were analyzed using the 4eVAR pipeline. Results: ONT successfully identified all P/LP variants across the panels (sensitivity 100%); identified a previously missed CNV in ENG gene; precisely defined the breakpoints of a del(13q) (unsuspected and diagnosed as BRCA2 del ex2–14); improved the coverage profiles in difficult-to-map regions (e.g., ex1 TGFBR1, PSM2CL); expanded the coverage of out-of-target deep intronic regions; and allowed for the set-up of fast-track tests (<24 h) for urgent clinical needs. Conclusions: Our findings demonstrate that ONT LRS provides diagnostic performance comparable to SRS, with significant advantages in resolving complex and previously undetectable variants. Ongoing developments are further increasing read length, expanding detectable targets, and potential clinical applications. Full article

In 40 unique pediatric biopsy samples, this study aimed to compare the obtained results of Helicobacter pylori DNA detection using next-generation sequencing (NGS), a real-time PCR-based IVD-certified kit and an established high resolution melting real-time PCR-based method for H. pylori-specific ureA gene. From the same group, the H. pylori DNA was identified in 16 (40.0%) samples in both real-time PCR-based methods, with quantification cycle (Cq) values ranging from 17.51 to 32.21 for the IVD kit. NGS was able to detect H. pylori DNA in 14 (35.0%) samples, with read counts between 7768 and 42,924. While all three methods showed similar detection rates, both PCR variants were slightly more sensitive, identifying H. pylori in two additional samples not detected by NGS. The study highlights the strengths and limitations of each method. NGS, though promising due to its high sensitivity and ability to detect low bacterial load, is still limited by its cost and complexity. Despite these challenges, NGS could complement PCR in diagnosing difficult or ambiguous cases, enabling the detection of multiple pathogens simultaneously. Especially when other infectious etiologies are suspected, NGS could be considered, though PCR variants remain a more attractive and cost-effective option for routine H. pylori detection. Full article

Isovaleric acidemia (IVA, OMIM 243500) is an inherited disorder of leucine metabolism caused by a deficiency of isovaleryl-CoA dehydrogenase (IVD), leading to an accumulation of isovaleric acid and its derivates 3-hydroxyisovaleric acid, isovaleryl (C5)-carnitine and isovalerylglycine in body fluids. The clinical presentation is highly variable, ranging from life-threatening metabolic crises with metabolic acidosis and hyperammonemia to a clinically asymptomatic only biochemical phenotype. Newborn screening for IVA has been established in many countries. Treatment consists of a protein-restricted diet combined with supplementation of carnitine and/or glycine and emergency treatment in catabolic episodes. Still, evidence-based recommendations for the diagnosis and management of IVA patients with various phenotypes are lacking. Therefore, a systematic search and review of the literature was conducted to make suggestions for the care of patients with IVA based on both the available scientific evidence and consensus-derived expert conclusions. Based on a comprehensive set of literature data published between 1966 and 2024, 15 statements were phrased on the presentation, diagnosis, management, and outcome of IVA involving clinical, biochemical, and nutrition expertise. These statements can serve as a basis for more standardized care for IVA. Full article

Obesity is a recognized risk factor for intervertebral disc (IVD) degeneration, a condition characterized by the progressive loss of extracellular matrix components in the nucleus pulposus. Elevated circulating leptin levels in obese individuals contribute to this degeneration by upregulating matrix metalloproteinase-1 (MMP-1) expression. Targeting MMP-1 expression with low-toxicity natural compounds may provide a promising strategy to prevent or mitigate IVD degeneration. In this study, we examined the effects of cinnamaldehyde (CA), a natural compound derived from Cinnamomum osmophloeum Kaneh, on leptin-induced MMP-1 expression in human IVD cartilage endplate-derived stem cells (SV40 cell line). Our results showed that leptin induced MMP-1 expression via activation of leptin receptor-mediated JAK2/STAT3, JAK2/RhoA/STAT3, and RhoA/ERK1/2/NF-κB signaling pathways. CA significantly reduced MMP-1 expression by inhibiting phosphorylation of the leptin receptor and STAT3 and blocking RhoA and NF-κB activation, without affecting JAK2 and ERK1/2 phosphorylation. These findings suggest that CA suppresses leptin-induced MMP-1 expression by modulating specific signaling pathways, highlighting its potential as a therapeutic agent for IVD degeneration associated with obesity. Full article

The growing demand for traceable, high-precision attenuation measurements in electromagnetic compatibility (EMC) testing and low-frequency wireless communication systems has driven the development of a primary attenuation standard covering 1 kHz to 10 MHz. The system employs a dual channel null-detection method using an inductive voltage divider (IVD) as a reference, ensuring the highest accuracy and traceability while eliminating sensitivity to detector nonlinearity. Attenuation at 1 kHz, 9 kHz, and 10 kHz is measured directly against the IVD ratio, while higher-frequency measurements (100 kHz–10 MHz) are performed via heterodyne detection, down-converting signals to 1 kHz for comparison. To ensure comparable accuracy at higher attenuation levels, a double-step method is applied at 9 kHz and 10 kHz to mitigate the increased IVD uncertainty above 1 kHz. Linearity is ensured by suppressing common-mode currents with toroidal ferrite chokes and minimizing inter-channel coupling. Type B (non-statistical) measurement uncertainties are evaluated, with major contributions from the IVD reference, system errors, and mismatch. The expanded uncertainties are 2.2 × 10⁻³ dB at 20 dB, 3.0 × 10⁻³ dB at 40 dB, and 4.0 × 10⁻³ dB at 60 dB attenuation. To facilitate wider dissemination and extend the calibration range, a resistive step attenuator with 10 dB pads is evaluated as a practical transfer standard, providing a simple and robust solution for traceable attenuation calibration in this frequency range. Full article

Background: Intervertebral disc degeneration (IVDD) is closely linked to low back pain (LBP), a leading cause of disability worldwide. IVDD is characterized by the loss of proteoglycans (PGs), extracellular matrix (ECM) degradation, and reduced hydration of the nucleus pulposus (NP). Extracellular vesicles (EVs) derived from human umbilical cord mesenchymal stem cells (hUC-MSCs) exhibit tissue repair and immunomodulatory effects and are emerging as promising cell-free therapeutics. Methods: We established a rat IVDD model via fluoroscopy-guided needle puncture of three consecutive coccygeal discs and confirmed degeneration through Alcian Blue and hematoxylin & eosin (H&E) staining. The gene expression of inflammatory and pain markers (ADRβ2, COMP, CXCL1, COX2, PPTA, MMP13, YKL40) was measured by qPCR. Subsequently, we implanted hUC-MSCs or EVs to evaluate their reparative potential. Results: Upregulation of inflammatory and pain genes in IVDD was associated with an immunomodulatory response. Tracking DiI-labelled hUC-MSCs and EVs revealed enhanced survival of hUC-MSCs, retention of EVs, and dispersion within rat tail discs; EVs showed greater retention than hUC-MSCs. Implanted EVs were internalized by NP cells and remained within degenerative IVDs. EVs passively diffused, accumulated at the injury site, interacted with host cells, and enhanced function, as shown by increased expression of human chondrocyte-related markers (SOX9, TGFβ1, TGFβ2, COL2) compared to hUC-MSC treatment. Histological analysis of two weeks post-transplantation showed NP cellular patterns resembling chondromas in treated discs. EVs integrated into and distributed within degenerated NP regions, with greater glycosaminoglycan (GAG) content. Conclusions: Overall, hUC-MSC EVs demonstrated superior regenerative capacity, supporting a safe, cell-free strategy for disc repair. Full article

The intervertebral disc (IVD) is a biphasic tissue in which the extracellular matrix (ECM) acts as a structural scaffold and regulates hydration and solute transport. The influence of hydration on the swelling and mechanical properties of the IVD, particularly the annulus fibrosus (AF), is not fully described in the literature. Hydration is assumed to affect inter- and intramolecular hydrogen bonding and hydrophilic interactions, thereby modulating tissue mechanics. This study aimed to assess the effect of hydration time on free swelling of AF and its impact on mechanical performance. AF specimens were divided into five groups, hydrated for 0, 10, 20, 30, or 40 min and subjected to uniaxial tensile testing until failure. Swelling-related geometric changes were correlated with tensile properties. Results demonstrated that hydration duration significantly influenced AF’s structural and mechanical characteristics in anterior and posterior IVD regions. Hydration increases rapidly within 10–20 min, causing cross-sections to swell, stress capacity to decrease, and stiffness to remain unchanged. However, after 40 min, the tissue becomes swollen beyond physiological balance. These findings identify hydration duration as a critical factor regulating AF function and provide important insights for experimental standardization, numerical modeling, and hydrogels designed for intervertebral disc regeneration. Full article

The aim of this study was to treat sheep oocytes with resveratrol during in vitro maturation (IVM) and use them as cytoplasts in ISCNT via handmade cloning (HMC), evaluating the effect of resveratrol on the in vitro development of cloned Mexican bighorn sheep embryos. Post mortem skin fibroblasts from an adult male specimen were frozen for 8 years, thawed, and reseeded for eight cell passages. For IVM, Ovis aries oocytes were treated with 0, 0.5, or 1.0 µM resveratrol. Matured oocytes were manually enucleated, and triplets (O. aries cytoplast–Ovis canadensis mexicana karyoplast–O. aries cytoplast) were formed and electrically fused. The reconstructed embryos were chemically activated and cultured for in vitro development (IVD). The IVM rate was 81.8 ± 10.4% for CG, 81.9 ± 6.7% for EG1, and 76.3 ± 7.7% for EG2, with no significant differences between groups. For IVD, EG1 showed a statistically significant increase (p < 0.05) in the blastocyst rate (31 ± 12.0%) and a statistically significant decrease in the fragmented embryo rate (25 ± 10.4) when compared with the other groups. It was concluded that better rates of cloned bighorn sheep blastocysts could be obtained in ISCNT via HMC when fusing O. aries oocytes supplemented with resveratrol during IVM with post mortem adult male O. c. mexicana fibroblasts that had been cryopreserved for 8 years. Full article