Search Results (5,175)

BRCA1 and BRCA2 are associated with advanced prostate cancer progression and poor prognosis. Copy number variants (CNVs) of these genes play a crucial role in guiding targeted treatments, particularly for patients receiving PARP inhibitors. However, CNV detection using multiplex ligation-dependent probe amplification (MLPA) is often limited by tumor heterogeneity, leading to ambiguous results. This study therefore aimed to evaluate BRCA1/2 CNVs in advanced prostate cancer patients using droplet digital PCR (ddPCR) and compare the results with MLPA. DNA from 11 advanced prostate cancer tissues was analyzed using both methods, in parallel with four cell lines and seven healthy volunteers. Our findings revealed that ddPCR effectively classified normal CNV groups—including normal control cell lines, healthy volunteers, and samples with normal MLPA final ratios—from deletion groups, which included deletion control cell lines, samples with deletion final ratios from MLPA, and cases with previously ambiguous results. Interestingly, two cases involving BRCA1 and one case involving BRCA2 exhibited ambiguous results using MLPA; however, ddPCR enabled more precise classification by applying the Youden Index from ROC analysis and identifying optimal cutoff values of 1.35 for BRCA1 and 1.55 for BRCA2. These optimal thresholds allowed ddPCR to effectively reclassify the ambiguous MLPA cases into the deletion group. Overall, ddPCR could offer a more sensitive and reliable approach for CNV detection in heterogeneous tissue samples and demonstrates strong potential as a biomarker tool for guiding targeted therapy in advanced prostate cancer patients. However, further validation in larger cohorts is necessary to optimize cutoff precision, confirm diagnostic performance, and evaluate the full clinical utility of ddPCR. Full article

Tissue hypoxia is commonly observed in head cancers and contributes to both molecular and functional changes in tumour cells. It is known to stimulate erythropoiesis, angiogenesis, and metabolic alterations within tumour cells. Glioblastoma, a type of brain tumour, is characterized by rapid proliferation and aggressive growth. Recent studies have indicated that natural products may hold potential as components of cancer therapy. Among these, Polish propolis and its active compound, quercetin, have demonstrated promising anti-cancer properties. The aim of this study was to evaluate the concentrations of selected cytokines—specifically IL-6, IL-9, vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF-BB), interferon gamma-induced protein 10 (IP-10), and monocyte chemoattractant protein-1 (MCP-1)—produced by astrocytes of the CCF-STTG1 cell line. The cytotoxic effects of ethanolic extract of propolis (EEP) and quercetin were assessed using the MTT assay. Astrocytes were stimulated with lipopolysaccharide (LPS, 200 ng/mL) and/or IFN-α (100 U/mL), followed by treatment with EEP or quercetin (25–50 µg/mL) under hypoxic conditions for two hours. Cytokine concentrations were measured using the xMAP Luminex Multiplex Immunoassay and the Multiplex Bead-Based Cytokine Kit. Our study demonstrated that Polish propolis and its component quercetin modulate the tumour microenvironment in vitro, primarily by altering the levels of specific cytokines. The HCA analysis revealed that IL-6 and MCP-1 formed a distinct cluster at the highest linkage distance (approximately 100% of Dmax), suggesting that their expression patterns are significantly different from those of the other cytokines and that they are more similar to each other than to the rest. PCA analysis showed that EEP-PL (50 μg/mL) with IFN-α and EEP-PL (50 μg/mL) with LPS exert similar activities on cytokine secretion by astrocytes. Similar effects were demonstrated for EEP-PL 50 μg/mL + LPS + IFN-α, EEP-PL 25 μg/mL + IFN-α and EEP-PL 25 μg/mL + LPS + IFN-α. Our findings suggest that Polish propolis and quercetin may serve as promising natural agents to support the treatment of stage IV malignant astrocytoma. Nonetheless, further research is needed to confirm these results. Full article

We propose and experimentally demonstrate an Integrated Multiband-Mode Multiplexing Photonic Lantern (IM3PL) that enables the selective excitation of high-order modes and stable modal preservation across multiple wavelength bands. As a proof-of-concept configuration, the IM3PL integrates a custom-designed input fiber array composed of three 980 nm single-mode fibers (SMFs) and two few-mode fibers (FMFs) operating at 1310 nm and 1550 nm, respectively. Simulations verify that 980 nm input signals can selectively excite $L{P}_{01}$, $L{P}_{11a}$, and $L{P}_{11b}$ modes at the FMF output, while the modal integrity of high-order linear polarized modes is preserved at 1310 nm and 1550 nm. The fabricated IM3PL device is experimentally validated via near-field pattern measurements, confirming the selective excitation at 980 nm and low-loss, mode-preserving transmission at the signal bands. This work offers a scalable and reconfigurable solution for multiband high-order-mode multiplexing, with promising applications in mode-division multiplexed fiber communication systems and multiband high-mode fiber lasers. Full article

Background/Objectives: The role of tertiary lymphoid structures (TLSs) in cancer prognosis is well established, yet their significance in early-stage EGFR-mutant lung adenocarcinoma remains unclear. While outcomes for early-stage lung cancer are generally better than those of late-stage disease, recurrence remains a significant challenge. This study investigates the prognostic value of TLSs and their molecular characteristics in early-stage EGFR-mutant lung adenocarcinoma. Methods: TLSs were identified in tumor samples using multiplex immunohistochemistry (IHC), and their density was quantified. The PD-L1 tumor proportion score (TPS) and TLS density were analyzed for associations with disease-free survival (DFS). Gene expression profiling was performed to compare tumor microenvironment signatures between high- and low-TLS-density groups. Results: High TLS density correlated with significantly longer DFS (43 vs. 20.5 months, p = 0.0082). No relationship was found between TLS density and PD-L1 TPS or EGFR mutation subtype. Transcriptomic analysis revealed upregulated immune response genes in the high-TLS-density group, including those involved in T and B cell activation. Low-TLS-density tumors exhibited gene signatures promoting tumor growth, such as cell cycle and WNT pathway activation. Conclusions: In summary, TLS density is a potential prognostic biomarker for DFS in early-stage EGFR-mutant lung adenocarcinoma, independent of PD-L1 TPS or EGFR mutation subtype. Enhanced immune activation in high-TLS-density tumors highlights TLSs as a potential target for improving outcomes in these patients. Full article

The rapid and accurate detection of pathogenic bacteria and viruses is critical for infectious disease control and public health protection. While conventional methods (e.g., culture, microscopy, serology, and PCR) are widely used, they are often limited by lengthy processing times, high costs, and specialized equipment requirements. In recent years, metal nanocluster (MNC)-based biosensors have emerged as powerful diagnostic platforms due to their unique optical, catalytic, and electrochemical properties. This systematic review comprehensively surveys advancements in MNC-based biosensors for bacterial and viral pathogen detection, focusing on optical (colorimetric and fluorescence) and electrochemical platforms. Three key aspects are emphasized: (1) detection mechanisms, (2) nanocluster types and properties, and (3) applications in clinical diagnostics, environmental monitoring, and food safety. The literature demonstrates that MNC-based biosensors provide high sensitivity, specificity, portability, and cost-efficiency. Moreover, the integration of nanotechnology with biosensing platforms enables real-time and point-of-care diagnostics. This review also discusses the limitations and future directions of the technology, emphasizing the need for enhanced stability, multiplex detection capability, and clinical validation. The findings offer valuable insights for developing next-generation biosensors with improved functionality and broader applicability in microbial diagnostics. Full article

We developed the Global–Local Integrated Topology inverse design algorithm (denoted as the GLINT algorithm), which employs a trajectory-based optimization strategy with waveguide–substrate material-flipping structural modifications, enabling the direct optimization of discrete waveguide–substrate binary structures. Compared to the conventional Direct Binary Search (DBS), the GLINT algorithm not only significantly enhances computational efficiency through its global search–local refinement framework but also achieves a superior 20 nm × 20 nm optimization resolution while maintaining its optimization speed—substantially advancing the design capability. Utilizing this algorithm, we designed and experimentally demonstrated a 3.5 µm × 3.5 µm dual-port wavelength division multiplexer (WDM), achieving a minimum crosstalk of −11.3 dB and a 2 µm × 2 µm 90-degree bending waveguide exhibiting a 0.31–0.52 dB insertion loss over the 1528–1600 nm wavelength range, both fabricated on silicon-on-insulator (SOI) wafers. Additionally, a 4.5 µm × 4.5 µm three-port WDM structure was also designed and simulated, demonstrating crosstalk as low as −36.5 dB. Full article

Innate immune cells appear to have an important implication in the resolution and/or the aggravation of the COVID-19 pathogenesis after infection with SARS-CoV-2. To better appreciate the role of these cells during COVID-19, changes in blood eosinophil, the neutrophil and monocyte count, and levels of surface protein markers have been reported. However, analyses at several timepoints of multiple surface markers on granulocytes and monocytes over a period of one month after a SARS-CoV-2 infection are missing. Therefore, in this study, we performed blood eosinophil, neutrophil, and monocyte phenotyping using a list of surface proteins and flow cytometry during a period of 30 days after the hospitalization of patients with severe SARS-CoV-2 infections. Blood cell counts were reported at seven different timepoints over the 30-day period as well as measures of multiple mediators in serum using a targeted multiplex assay approach. Our results indicate a 95% drop in the blood eosinophil count by D1, with eosinophils displaying a phenotype defined as CD69/CD63/CD125^high and CCR3/CD44^low during the early phases of hospitalization. Conversely, by D7 the neutrophil count increased significantly and displayed an immature, activated, and immunosuppressive phenotype (i.e., 3% of CD10/CD16^low and CD10^lowCD177^high, 6.7% of CD11b^highCD62L^low, and 1.6% of CD16^highCD62L^low), corroborated by enhanced serum proteins that are markers of neutrophil activation. Finally, our results suggest a rapid recruitment of non-classical monocytes leaving CD163/CD64^high and CD32^low monocytes in circulation during the very early phase. In conclusion, our study reveals potential very early roles for eosinophils and monocytes in the pathogenesis of COVID-19 with a likely reprogramming of eosinophils in the bone marrow. The exact roles of the pro-inflammatory neutrophils and the functions of the eosinophils and the monocytes, as well as these innate immune cell types, interplays need to be further investigated. Full article

CRISPR technology, which is derived from the bacterial adaptive immune system, has transformed traditional genetic engineering techniques, made strain engineering significantly easier, and become a very versatile genome editing system that allows for precise, programmable modifications to a wide range of microbial genomes. The economies of fermentation-based manufacturing are changing because of its quick acceptance in both academic and industry labs. CRISPR processes have been used to modify industrially significant bacteria, including the lactic acid producers, Clostridium spp., Escherichia coli, and Corynebacterium glutamicum, in order to increase the yields of bioethanol, butanol, succinic acid, acetone, and polyhydroxyalkanoate precursors. CRISPR-mediated promoter engineering and single-step multiplex editing have improved inhibitor tolerance, raised ethanol titers, and allowed for the de novo synthesis of terpenoids, flavonoids, and recombinant vaccines in yeasts, especially Saccharomyces cerevisiae and emerging non-conventional species. While enzyme and biopharmaceutical manufacturing use CRISPR for quick strain optimization and glyco-engineering, food and beverage fermentations benefit from starter-culture customization for aroma, texture, and probiotic functionality. Off-target effects, cytotoxicity linked to Cas9, inefficient delivery in specific microorganisms, and regulatory ambiguities in commercial fermentation settings are some of the main challenges. This review provides an industry-specific summary of CRISPR–Cas9 applications in microbial fermentation and highlights technical developments, persisting challenges, and industrial advancements. Full article

Background/Objectives: Concurrent outbreaks of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza A and B viruses (IAV/IBV), and respiratory syncytial virus (RSV) necessitate rapid and precise differential laboratory diagnostic methods. This study aimed to evaluate the multiplex molecular diagnostic performance of the geneLEAD VIII system (Precision System Science Co., Ltd., Matsudo, Japan), a fully automated sample-to-result precision instrument, in conjunction with the VIASURE SARS-CoV-2, Flu & RSV Real Time PCR Detection Kit (CerTest Biotec, S.L., Zaragoza, Spain). Methods: The specific detection capabilities of SARS-CoV-2, IAV/IBV, and RSV genes were evaluated using virus-spiked saliva and nasal swab samples. Using saliva samples, the viral titer detection limits of geneLEAD/VIASURE and manual referent singleplex RT-qPCR assays were compared. The performance of geneLEAD/VIASURE in analyzing single- and multiple-infection models was scrutinized. The concordance between the geneLEAD/VIASURE and the manual assays was assessed. Results: The geneLEAD/VIASURE successfully detected all the virus genes in the saliva and nasal swab samples despite some differences in the Ct values. The viral titer detection limits in the saliva samples for SARS-CoV-2, IAV, IBV, and RSV using geneLEAD/VIASURE were 10⁰, ≤10⁻², 10⁰, and 10² TCID₅₀/mL, respectively, compared to ≤10⁻¹, ≤10⁰, ≤10⁰, and ≤10⁴ TCID₅₀/mL, respectively, in the manual assays. geneLEAD/VIASURE yielded similar Ct values in the single- and multiple-infection models, with some exceptions noted in the triple-infection models when low titers of RSV were spiked with high titers of the other viruses. The concordance between geneLEAD/VIASURE and the manual assays was high, with Pearson’s R² values of 0.90, 0.85, 0.92, and 0.95 for SARS-CoV-2, IAV, IBV, and RSV, respectively. Conclusions: geneLEAD/VIASURE is a reliable diagnostic tool for detecting SARS-CoV-2, IAV/IBV, and RSV in single- and multiple-infection scenarios. Full article

Background: Mucosal immunity plays a pivotal role in preventing infections with SARS-CoV-2. While COVID-19 mRNA vaccines induce robust systemic immune responses in patients with inflammatory bowel disease (IBD), little is known about their efficacy in the mucosal immune compartment. In this sub-investigation of the ongoing STAR-SIGN study, we present the first analysis of mucosal immunity elicited by XBB.1.5 mRNA vaccines in immunocompromised patients with IBD. Methods: IgG and IgA antibodies targeting the receptor-binding domain of the SARS-CoV-2 JN.1 variant were quantified longitudinally in the saliva of IBD patients using the multiplex immunoassay MultiCoV-Ab. Antibody levels were quantified before and 2–4 weeks after vaccination with XBB.1.5 mRNA vaccines. All patients previously received three doses with original COVID-19 vaccines. Results: Mucosal IgG antibodies were readily induced by XBB.1.5 mRNA vaccines (p = 0.0013 comparing pre- and post-vaccination levels). However, mucosal IgA levels were comparable before and after vaccination (p = 0.8233). Consequently, mucosal IgG and IgA antibody levels correlated only moderately before and after immunization (pre-vaccination: r = 0.5294; p = 0.0239; post-vaccination: r = 0.4863; p = 0.0407). Contrary to a previous report in healthy individuals, vaccination did not induce serum IgA in patients with IBD (p = 0.5841 comparing pre- and post-vaccination levels). These data suggest that COVID-19 mRNA vaccines fail to elicit mucosal IgA in patients with IBD. Conclusions: Since mucosal IgA plays a pivotal role in infection control, the lack of IgA induction indicates that patients lack sufficient protection against SARS-CoV-2 infections which warrants the development of mucosal COVID-19 vaccines. Full article

This paper studied the general mechanism of spatial-frequency-shift (SFS) super-resolution imaging based on multiplex illumination modulation. The theory of SFS joint intensity was first proposed. Experiments on parallel slots with discrete spatial frequency (SF) distribution and V-shape slots with continuous SF distribution were carried out, and their real-space images and k-space images were obtained. The influence of single illumination with different SFS and mixed illumination with various combinations on SFS super-resolution imaging was analyzed. The phenomena of sample SF coverage were discussed. The SFS super-resolution imaging characteristics based on low-coherence illumination and highly localized light fields were discovered. The phenomenon of image magnification during SFS super-resolution imaging process was discussed. The differences and connections between the SF spectrum of objects and the k-space images obtained in SFS super-resolution imaging process were explained. This provides certain support for optimization of high-throughput SFS super-resolution imaging. Full article

Enteroviruses (EVs) cause broad clinical manifestations, particularly in children. Certain serotypes have been implicated in respiratory infections; however, epidemiological studies analyzing EV circulation based on clinical respiratory specimens are limited in Korea. This retrospective study evaluates EV detection patterns in respiratory specimens to demonstrate their clinical and epidemiological significance as respiratory pathogens in Korea. Respiratory samples collected from outpatient and hospitalized patients with respiratory symptoms at Dankook University Hospital between 2018 and 2024 were analyzed. EV detection patterns were analyzed by year, season, sex, and age. EVs were detected in 303/6292 respiratory specimens. The highest and lowest positivity rates were in 2018 (8.2%) and 2020 (1.6%), likely due to non-pharmaceutical interventions during the COVID-19 pandemic. The highest positivity rates were in summer and autumn, and in children aged 2–11 years and infants aged 0–1 years. EV positivity did not differ significantly between sexes. Significant differences were identified across years, seasons, and age groups. EVs can be detected in respiratory specimens from symptomatic patients and exhibit a marked seasonal distribution and elevated positivity rates in pediatric populations. Hence, EVs may act as atypical respiratory pathogens, underscoring the need for integrated public health surveillance and seasonal prevention strategies. Full article

The diagnosis of allergic diseases and anaphylaxis is complex and encompasses a broad spectrum of in vitro and in vivo diagnostic tests. The choice of diagnostic tests is related to the presumed pathophysiological mechanism of the allergic reaction. In the past decade the implementation of component-resolved diagnostics (CRD) into clinical practice has significantly improved the depicting of sensitization profiles, which has aided in the assessment of clinically relevant allergen components that are associated with true allergy, as well as the levels of risk of severe anaphylactic reactions. Recently, multiplex-specific immunoglobulin E (IgE) platforms have emerged for better selection of patients at risk for anaphylaxis and have improved the selection criteria for patients undergoing allergen immunotherapy, including novel regimes such as oral immunotherapy. This review describes the advantages of the utilization of component-resolved diagnostics and multiplex assays in clinical settings, especially in cases of anaphylaxis when no clear trigger is recognized or where multiple culprits are suspected. As multiplex component-resolved diagnostics becomes more readily available globally and with the use of novel approaches, CRD will certainly be a crucial tool in personalized and individually tailored management plans and reduce the financial burden of anaphylaxis. Full article

The increasing demand for high-speed internet and advanced digital services necessitates the deployment of robust and scalable broadband infrastructure, particularly in smaller urban and rural areas. This paper presents the design and implementation of a passive optical network (PON) based on a gigabit-capable passive optical network (GPON) standard to deliver fiber-to-the-home (FTTH) services in a small-town setting. The proposed solution prioritizes cost-effectiveness, scalability, and minimal energy consumption by leveraging passive splitters and unpowered network elements. We detail the topology planning, splitter architecture, installation practices, and technical specifications that ensure efficient signal distribution and future network expansion. The results demonstrate the successful implementation of an optical access infrastructure that supports high-speed internet, Internet Protocol television (IPTV), and voice services while maintaining flexibility for diverse urban layouts and housing types. Full article