Search

Abstract. Introduction:. Chronic primary pain conditions (CPPCs) are linked to catecholamine activation of peripheral adrenergic receptors. Yet, catecholamine-dependent epigenetic mechanisms, such as microRNA (miRNA) regulation of mRNA transcripts, remain largely unknown. Objectives:. We sought to identify RNA species correlated with case status in 3 pain cohorts, to validate RNAs found to be dysregulated in a mouse model of CPPC onset, and to directly test the role of adrenergic receptors in miRNA regulation. Furthermore, we tested antinociceptive effects of miR-374 overexpression. Methods:. We used RNA-seq and quantitative reverse transcription polymerase chain reaction to measure RNA expression in 3 pain cohorts. Next, we validated identified RNAs with quantitative reverse transcription polymerase chain reaction in a mouse model of CPPC onset, measuring expression in plasma, peripheral (adipose, muscle, dorsal root ganglia [DRG]), and central (spinal cord) tissues. Then, we stimulated adrenergic receptors in primary adipocyte and DRG cultures to directly test regulation of microRNAs by adrenergic signaling. Furthermore, we used in vitro calcium imaging to measure the antinociceptive effects of miR-374 overexpression. Results:. We found that one miRNA family, miR-374, was downregulated in the plasma of individuals with temporomandibular disorder, fibromyalgia syndrome, or widespread pain following a motor vehicle collision. miR-374 was also downregulated in plasma, white adipose tissue, and spinal cord from mice with multisite mechanical sensitivity. miR-374 downregulation in plasma and spinal cord was female specific. Norepinephrine stimulation of primary adipocytes, but not DRG, led to decreased miR-374 expression. Furthermore, we identified tissue-specific and sex-specific changes in the expression of predicted miR-374 mRNA targets, including known (HIF1A, NUMB, TGFBR2) and new (ATXN7, CRK-II) pain targets. Finally, we demonstrated that miR-374 overexpression in DRG neurons reduced capsaicin-induced nociceptor activity. Conclusions:. Downregulation of miR-374 occurs between adrenergic receptor activation and mechanical hypersensitivity, and its adipocyte source implicates adipose signaling in nociception. Further study of miR-374 may inform therapeutic strategies for the millions worldwide who experience CPPCs.

Abstract Raman spectroscopy has tremendous potential for material analysis with its molecular fingerprinting capability in many branches of science and technology. It is also an emerging omics technique for metabolic profiling to shape precision medicine. However, precisely attributing vibration peaks coupled with specific environmental, instrumental, and specimen noise is problematic. Intelligent Raman spectral preprocessing to remove statistical bias noise and sample-related errors should provide a powerful tool for valuable information extraction. Here, we propose a novel Raman spectral preprocessing scheme based on self-supervised learning (RSPSSL) with high capacity and spectral fidelity. It can preprocess arbitrary Raman spectra without further training at a speed of ~1 900 spectra per second without human interference. The experimental data preprocessing trial demonstrated its excellent capacity and signal fidelity with an 88% reduction in root mean square error and a 60% reduction in infinite norm ( $${L}_{{\infty }}$$ L ∞ ) compared to established techniques. With this advantage, it remarkably enhanced various biomedical applications with a 400% accuracy elevation (ΔAUC) in cancer diagnosis, an average 38% (few-shot) and 242% accuracy improvement in paraquat concentration prediction, and unsealed the chemical resolution of biomedical hyperspectral images, especially in the spectral fingerprint region. It precisely preprocessed various Raman spectra from different spectroscopy devices, laboratories, and diverse applications. This scheme will enable biomedical mechanism screening with the label-free volumetric molecular imaging tool on organism and disease metabolomics profiling with a scenario of high throughput, cross-device, various analyte complexity, and diverse applications.

Summary: The severe acute respiratory syndrome coronavirus 2 pandemic is characterized by the emergence of novel variants of concern (VOCs) that replace ancestral strains. Here, we dissect the complex selective pressures by evaluating variant fitness and adaptation in human respiratory tissues. We evaluate viral properties and host responses to reconstruct forces behind D614G through Omicron (BA.1) emergence. We observe differential replication in airway epithelia, differences in cellular tropism, and virus-induced cytotoxicity. D614G accumulates the most mutations after infection, supporting zoonosis and adaptation to the human airway. We perform head-to-head competitions and observe the highest fitness for Gamma and Delta. Under these conditions, RNA recombination favors variants encoding the B.1.617.1 lineage 3′ end. Based on viral growth kinetics, Alpha, Gamma, and Delta exhibit increased fitness compared to D614G. In contrast, the global success of Omicron likely derives from increased transmission and antigenic variation. Our data provide molecular evidence to support epidemiological observations of VOC emergence.

Soliton molecules in optical resonators have attracted remarkable attention in nonlinear dynamics, driven by their compelling analogies with matter molecules. So far, while extensive research has been conducted on their generation, pulsations, and dissociation behaviors, the investigation of their quasi-periodic dynamics has been relatively limited. Here, we present a systematic exploration of the quasi-periodic dynamics of soliton molecules using advanced balanced optical cross-correlation techniques. The incommensurable quasi-period bifurcations constituted of cascaded Hopf bifurcations are found, providing an unambiguous pathway toward chaotic soliton molecules. The chaotic intramolecular dynamics are analyzed by time series, radio frequency spectra, phase portraits, and Lyapunov exponent analysis. In addition, we reveal an intrinsic frequency entrainment phenomenon experimentally. Such frequency entrainment provides a novel perspective on synchronization in optical resonators, encompassing the competition and interaction of oscillations across multiple temporal scales. Our experimental findings offer clear proof that the gain dynamics serve as the origin of the binding forces between solitons within the molecule, which are well supported by the numerical simulations. By advancing the understanding of sub-femtosecond resolved quasi-period dynamics of optical soliton molecules, this study contributes to the broader field of complex nonlinear dynamics, paving the way for future explorations into the intricate behaviors of solitons within optical resonators and relevant fields.

Hyper-secretion and/or hyper-concentration of mucus is a defining feature of multiple obstructive lung diseases, including chronic obstructive pulmonary disease (COPD). Mucus itself is composed of a mixture of water, ions, salt and proteins, of which the gel-forming mucins, MUC5AC and MUC5B, are the most abundant. Recent studies have linked the concentrations of these proteins in sputum to COPD phenotypes, including chronic bronchitis (CB) and acute exacerbations (AE). We sought to determine whether common genetic variants influence sputum mucin concentrations and whether these variants are also associated with COPD phenotypes, specifically CB and AE. We performed a GWAS to identify quantitative trait loci for sputum mucin protein concentration (pQTL) in the Sub-Populations and InteRmediate Outcome Measures in COPD Study (SPIROMICS, n = 708 for total mucin, n = 215 for MUC5AC, MUC5B). Subsequently, we tested for associations of mucin pQTL with CB and AE using regression modeling (n = 822-1300). Replication analysis was conducted using data from COPDGene (n = 5740) and by examining results from the UK Biobank. We identified one genome-wide significant pQTL for MUC5AC (rs75401036) and two for MUC5B (rs140324259, rs10001928). The strongest association for MUC5B, with rs140324259 on chromosome 11, explained 14% of variation in sputum MUC5B. Despite being associated with lower MUC5B, the C allele of rs140324259 conferred increased risk of CB (odds ratio (OR) = 1.42; 95% confidence interval (CI): 1.10-1.80) as well as AE ascertained over three years of follow up (OR = 1.41; 95% CI: 1.02-1.94). Associations between rs140324259 and CB or AE did not replicate in COPDGene. However, in the UK Biobank, rs140324259 was associated with phenotypes that define CB, namely chronic mucus production and cough, again with the C allele conferring increased risk. We conclude that sputum MUC5AC and MUC5B concentrations are associated with common genetic variants, and the top locus for MUC5B may influence COPD phenotypes, in particular CB.

A fiber speckle sensor (FSS) based on a tapered multimode fiber (TMMF) has been developed to measure liquid analyte refractive index (RI) in this work. By the lateral and axial offset of input light into TMMF, several high-order modes are excited in TMMF, and the speckle pattern is spatially modulated, which affects an asymmetrical speckle pattern with a random intensity distribution at the output of TMMF. When the TMMF is immersed in the liquid analyte with RI variation, it influences the guided modes, as well as the mode interference, in TMMF. A digital image correlations method with zero-mean normalized cross-correlation coefficient is explored to digitize the speckle image differences, analyzing the RI variation. It is found that the lateral- and axial-offsets-induced speckle sensor can enhance the RI sensitivity from 6.41 to 19.52 RIU−1 compared to the one without offset. The developed TMMF speckle sensor shows an RI resolution of 5.84 × 10−5 over a linear response range of 1.3164 to 1.3588 at 1550 nm. The experimental results indicate the FSS provides a simple, efficient, and economic approach to RI sensing, which exhibits an enormous potential in the image-based ocean-sensing application.

The vast majority of people with cystic fibrosis (CF) are now eligible for CF transmembrane regulator (CFTR) modulator therapy. The remaining individuals with CF harbor premature termination codons (PTCs) or rare CFTR variants with limited treatment options. Although the clinical modulator response can be reliably predicted using primary airway epithelial cells, primary cells carrying rare CFTR variants are scarce. To overcome this obstacle, cell lines can be created by overexpression of mouse Bmi-1 and human TERT (hTERT). Using this approach, we developed 2 non-CF and 6 CF airway epithelial cell lines, 3 of which were homozygous for the W1282X PTC variant. The Bmi-1/hTERT cell lines recapitulated primary cell morphology and ion transport function. The 2 F508del-CFTR cell lines responded robustly to CFTR modulators, which was mirrored in the parent primary cells and in the cell donors’ clinical response. Cereblon E3 ligase modulators targeting eukaryotic release factor 3a (eRF3a) rescued W1282X-CFTR function to approximately 20% of WT levels and, when paired with G418, rescued G542X-CFTR function to approximately 50% of WT levels. Intriguingly, eRF3a degraders also diminished epithelial sodium channel (ENaC) function. These studies demonstrate that Bmi-1/hTERT cell lines faithfully mirrored primary cell responses to CFTR modulators and illustrate a therapeutic approach to rescue CFTR nonsense mutations.

Abstract Background Foxp3+ regulatory T cells (Tregs) play essential roles in immune homeostasis and repair of damaged lung tissue. We hypothesized that patients whose lung injury resolves quickly, as measured by time to liberation from mechanical ventilation, have a higher percentage of Tregs amongst CD4+ T cells in either airway, bronchoalveolar lavage (BAL) or peripheral blood samples. Methods We prospectively enrolled patients with ARDS requiring mechanical ventilation and collected serial samples, the first within 72 h of ARDS diagnosis (day 0) and the second 48–96 h later (day 3). We analyzed immune cell populations and cytokines in BAL, tracheal aspirates and peripheral blood, as well as cytokines in plasma, obtained at the time of bronchoscopy. The study cohort was divided into fast resolvers (FR; n = 8) and slow resolvers (SR; n = 5), based on the median number of days until first extubation for all participants (n = 13). The primary measure was the percentage of CD4+ T cells that were Tregs. Results The BAL of FR contained more Tregs than SR. This finding did not extend to Tregs in tracheal aspirates or blood. BAL Tregs expressed more of the full-length FOXP3 than a splice variant missing exon 2 compared to Tregs in simultaneously obtained peripheral blood. Conclusion Tregs are present in the bronchoalveolar space during ARDS. A greater percentage of CD4+ cells were Tregs in the BAL of FR than SR. Tregs may play a role in the resolution of ARDS, and enhancing their numbers or functions may be a therapeutic target.

Cystic fibrosis (CF) is a severe genetic disorder that can cause multiple comorbidities affecting the lungs, the pancreas, the luminal digestive system and beyond. In our previous genome-wide association studies (GWAS), we genotyped approximately 8,000 CF samples using a mixture of different genotyping platforms. More recently, the Cystic Fibrosis Genome Project (CFGP) performed deep (approximately 30×) whole genome sequencing (WGS) of 5,095 samples to better understand the genetic mechanisms underlying clinical heterogeneity among patients with CF. For mixtures of GWAS array and WGS data, genotype imputation has proven effective in increasing effective sample size. Therefore, we first performed imputation for the approximately 8,000 CF samples with GWAS array genotype using the Trans-Omics for Precision Medicine (TOPMed) freeze 8 reference panel. Our results demonstrate that TOPMed can provide high-quality imputation for patients with CF, boosting genomic coverage from approximately 0.3–4.2 million genotyped markers to approximately 11–43 million well-imputed markers, and significantly improving polygenic risk score (PRS) prediction accuracy. Furthermore, we built a CF-specific CFGP reference panel based on WGS data of patients with CF. We demonstrate that despite having approximately 3% the sample size of TOPMed, our CFGP reference panel can still outperform TOPMed when imputing some CF disease-causing variants, likely owing to allele and haplotype differences between patients with CF and general populations. We anticipate our imputed data for 4,656 samples without WGS data will benefit our subsequent genetic association studies, and the CFGP reference panel built from CF WGS samples will benefit other investigators studying CF.

Primary human bronchial epithelial cultures (HBECs) are used to study airway physiology, disease, and drug development. HBECs often replicate human airway physiology/pathophysiology. Indeed, in the search for cystic fibrosis (CF) transmembrane conductance regulator (CFTR) therapies, HBECs were seen as the “gold standard” in preclinical studies. However, HBECs are not without their limitations: they are non-immortalized and the requirement for human donors, especially those with rare genetic mutations, can make HBECs expensive and/or difficult to source. For these reasons, researchers may opt to expand HBECs by passaging. This practice is common, but to date, there has not been a robust analysis of the impact of expanding HBECs on their phenotype. Here, we used functional studies of airway surface liquid (ASL) homeostasis, epithelial barrier properties, and RNA-seq and Western blotting to investigate HBEC changes over two passage cycles. We found that passaging impaired CFTR-mediated ASL secretion and led to a reduction in the plasma membrane expression of the epithelial sodium channel (ENaC) and CFTR. Passaging also resulted in an increase in transepithelial resistance and a decrease in epithelial water permeability. We then looked for changes at the mRNA level and found that passaging significantly affected 323 genes, including genes involved in inflammation, cell growth, and extracellular matrix remodeling. Collectively, these data highlight the potential for HBEC expansion to impact research findings.

A significant promotion of growth and accumulation of metabolites of freshwater microalga Euglena gracilis was obtained by adding phytic acid to the culture. Phytic acid concentration of 0.05% (v/v) showed a maximum biomass increase of 1.6-fold compared to the control group. Synchronous variation in the content and proportion of photosynthetic pigments was also observed. The total chlorophyll content increased with phytic acid concentration, suggesting the enhanced capacity of absorbing light. Cell length, an important biomarker for E. gracilis, was prolonged to a certain extent under light condition, indicating the state of the cells was more active. Since phytic acid is derived from agro waste of rice bran, it is promising as a low-cost but high-efficient growth promoter of E. gracilis.

A stimulated thermal Rayleigh scattering (STRS)-based self-injection locking technique is proposed and utilized in fiber laser to improve its spectral purity. Both theoretical and experimental analyses of the STRS-based filter and the interaction mechanism between the STRS-based filter and self-injection locking are demonstrated in detail. These analyses verify effects of the proposed method on single-longitudinal-mode (SLM) selection and linewidth compression. A fiber laser employing STRS-based self-injection locking configuration is designed, and a significant improvement of spectral purity comparing to commercially available lasers is achieved. Experimental results indicate that this fiber laser operates at SLM regime and has a linewidth of 45 Hz, and can be widely applied in distributed sensing, optical communication, and high-resolution optical spectrum analysis.

Genome wide association studies (GWAS) have identified several genomic loci with candidate modifiers of cystic fibrosis (CF) lung disease, but only a small proportion of the expected genetic contribution is accounted for at these loci. We leveraged expression data from CF cohorts, and Genotype-Tissue Expression (GTEx) reference data sets from multiple human tissues to generate predictive models, which were used to impute transcriptional regulation from genetic variance in our GWAS population. The imputed gene expression was tested for association with CF lung disease severity. By comparing and combining results from alternative approaches, we identified 379 candidate modifier genes. We delved into 52 modifier candidates that showed consensus between approaches, and 28 of them were near known GWAS loci. A number of these genes are implicated in the pathophysiology of CF lung disease (e.g., immunity, infection, inflammation, HLA pathways, glycosylation, and mucociliary clearance) and the CFTR protein biology (e.g., cytoskeleton, microtubule, mitochondrial function, lipid metabolism, endoplasmic reticulum/Golgi, and ubiquitination). Gene set enrichment results are consistent with current knowledge of CF lung disease pathogenesis. HLA Class II genes on chr6, and CEP72, EXOC3, and TPPP near the GWAS peak on chr5 are most consistently associated with CF lung disease severity across the tissues tested. The results help to prioritize genes in the GWAS regions, predict direction of gene expression regulation, and identify new candidate modifiers throughout the genome for potential therapeutic development.

Polarized low-coherence interferometry (PLCI) is widely used for the demodulation of Fabry–Perot (F-P) sensors. To avoid the influence of noise and dispersion on interference fringes, this paper proposes a data processing method in which the wavelet tools are applied to extract useful information from the extremum locations and envelope center of the fringes. Firstly, the wavelet threshold denoising (WTD) algorithm is used to remove electrical noise, and the complex Morlet wavelet is used to extract the fringe envelope. Based on this, the envelope center is used to predict the extremum locations of the specified order in its adjacent interval, the predicted locations are used as references to track the exact extremum locations, and the middle location of the peak and valley values is obtained to demodulate the F-P cavity accurately. The validity of this demodulation theory is verified by an air F-P cavity whose cavity length varies from 17 to 20 μm. With a sampling interval of 30 nm, the experimental results indicate that the repeatability accuracy is higher than 6.04 nm, and the resolution is better than 4.0 nm.

Cigarette smoke is well recognized to cause injury to the airways and the alveolar walls over time. This injury usually requires many years of exposure, suggesting that the lungs may rapidly develop responses that initially protect it from this repetitive injury. Our studies tested the hypotheses that smoke induces an inflammatory response and changes in mRNA profiles that are dependent on sex and the health status of the lung, and that the response of the lungs to smoke differs after 1 day compared to 5 days of exposure. Male and female wildtype (WT) and Scnn1b-transgenic (βENaC) mice, which have chronic bronchitis and emphysematous changes due to dehydrated mucus, were exposed to cigarette smoke or sham air conditions for 1 or 5 days. The inflammatory response and gene expression profiles were analyzed in lung tissue. Overall, the inflammatory response to cigarette smoke was mild, and changes in mediators were more numerous after 1 than 5 days. βENaC mice had more airspace leukocytes than WT mice, and smoke exposure resulted in additional significant alterations. Many genes and gene sets responded similarly at 1 and 5 days: genes involved in oxidative stress responses were upregulated while immune response genes were downregulated. However, certain genes and biological processes were regulated differently after 1 compared to 5 days. Extracellular matrix biology genes and gene sets were upregulated after 1 day but downregulated by 5 days of smoke compared to sham exposure. There was no difference in the transcriptional response to smoke between WT and βENaC mice or between male and female mice at either 1 or 5 days. Taken together, these studies suggest that the lungs rapidly alter gene expression after only one exposure to cigarette smoke, with few additional changes after four additional days of repeated exposure. These changes may contribute to preventing lung damage.

Micro absolute distance measurement (MADM) is widely used in industrial and military fields. To achieve high accuracy and frequency response, a polarized low-coherence interferometry (PLCI)-based method for MADM is proposed. The nearly linear relationship between the envelope center and m-order PLCI fringe (PLCIF) peak center is found and verified. Dispersion compensation is achieved by fringe peak position estimation and polynomial fitting to get rid of the dependence on an a priori model and birefringence parameters, and make this method very robust. Meanwhile, the zero-order PLCIF center is estimated and located to demodulate the measured displacement. Then, the measurement accuracy is raised by polynomial fittings. In comparison to conventional methods, the proposed method can effectively avoid jump errors and has a higher accuracy. Experimental results indicate that the measurement accuracy is higher than 19.51 nm, the resolution is better than 2 nm, and its processing data rate can reach 35 kHz.

Optical fiber distributed sensing that is based on optical frequency domain reflectometer (OFDR) is a promising technology for achieving a highest spatial resolution downwards to several millimeters. An OFDR signal demodulation method that is based on Morlet wavelet transformation (WT) is demonstrated in detail to improve the resolution of distributed sensing physical quantity under a high spatial resolution, aiming at the trade-off between spatial and spectrum resolution. The spectrum resolution, spatial interval of the measured gauges, and spatial resolution can be manually controlled by adjusting the wavelet parameters. The experimental results that were achieved by the wavelet transformation (WT) method are compared with these by short time Fourier transformation (STFT) method and they indicate that significant improvements, such as strain resolution of 1 με, spatial resolution of 5 mm, average repeatability of 4.3 με, and stability of 7.3 με within one hour, have been achieved. The advantages of this method are high spatial and spectral resolution, robust, and applicability with current OFDR systems.

The principal resonance of a delayed piecewise-smooth (DPWS) system with negative stiffness under narrow-band random excitation is investigated in aspects of multiscale analysis, design methodology of the controller, and response properties. The amplitude-frequency response and steady-state moments together with the corresponding stability conditions of the controlled stochastic system are derived, in which the degradation case is also under consideration. Then, from the perspective of the equivalent damping, the comparisons of the response characteristics of the controlled system to the uncontrolled system, such as the phenomenon of frequency island, are fulfilled. Furthermore, sensitivity of the system response to feedback gain and time delay is studied and interesting dynamic properties are found. Meanwhile, the classification of the steady-state solution is also discussed. To control the maximum amplitude, the feedback parameters are determined by the frequency response together with stability boundaries which must be utilized to exclude the combinations of the unstable parameters. For the case with small noise intensity, mean-square responses present the similar characteristics to what is discussed in the deterministic case.

A novel optical frequency domain reflectometer (OFDR) processing algorithm is proposed to enhance the measurable range and data processing rate using a narrow swept spectrum range and reducing the time consuming of the process distributed sensing results. To reduce the swept wavelength range and simultaneously enhance strain measurable range, the local similarity characteristics of Rayleigh scattering fingerprint spectrum is discovered and a new similarity evaluation function based on least-square method is built to improve the data processing rate and sensing performance. By this method, the strain measurable range is raised to 3000 µε under a highest spatial resolution of 3 mm when the swept spectrum range is only 10 nm and the data processing rate is improved by at least 10 times. Experimental results indicate that a nonlinearity of less than 0.5%, a strain resolution of better than 10 µε, a repeatability at zero strain of below ±0.4 GHz and a full-scale accuracy is lower than 0.85 GHz under a highest spatial resolution of 3 mm can be achieved. Advantages of this method are fast processing rate, large strain measurable range, high SNR, and applicability with current OFDR systems.

Three-dimensional micro-scale sensors are in high demand in the fields of metrology, precision manufacturing and industry inspection. To extend the minimum measurable dimension and enhance the accuracy, a tapered four-cores fiber Bragg grating (FBG) probe is proposed. The sensing model is built to investigate the micro-scale sensing characteristics of this method and the design of the tapered stylus is found to influence the accuracy. Therefore, a π/2 phase-shift point is introduced into the FBGs comprised in the probe to suppress spectrum distortion and improve accuracy. Then, the manufacturing method based on capillary self-assembly is proposed to form the probe and the critical length to form a square array for four cylindrical fibers is verified to be effective for the tapered fibers. Experimental results indicate that the design of the tapered stylus can extend the minimum measurable dimension by twofold and has nearly no influence on its sensitivity. The three-dimensional measurement repeatability is better than 31.1 nm and the stability is better than 200 nm within once measuring process. Furthermore, the measurement precision of the three-dimensional micro-scale measurement results is less than 150 nm. It would be widely used in measuring micro-scale features for industry inspection or metrology.

To achieve a narrow bandwidth optical filter with a wide swept range for new generation optical spectrum analysis (OSA) of high performance optical sensors, an optoelectronic equivalent narrowband filter (OENF) was investigated and a swept optical filter with bandwidth of several MHz and sweep range of several tens of nanometers was built using electric filters and a sweep laser as local oscillator (LO). The principle of OENF is introduced and analysis of the OENF system is presented. Two electric filters are optimized to be RBW filters for high and medium spectral resolution applications. Both simulations and experiments are conducted to verify the OENF principle and the results show that the power uncertainty is less than 1.2% and the spectral resolution can reach 6 MHz. Then, a real-time wavelength calibration system consisting of a HCN gas cell and Fabry–Pérot etalon is proposed to guarantee a wavelength accuracy of ±0.4 pm in the C-band and to reduce the influence of phase noise and nonlinear velocity of the LO sweep. Finally, OSA experiments on actual spectra of various optical sensors are conducted using the OENF system. These experimental results indicate that OENF system has an excellent capacity for the analysis of fine spectrum structures.

Background: Non-invasive multi-cancer early detection (MCED) tests have shown promise in enhancing early cancer detection. However, their clinical utility across diverse populations remains underexplored, limiting their routine implementation. This study aims to validate the clinical utility of a multimodal non-invasive circulating tumor DNA (ctDNA)-based MCED test, SPOT-MAS (Screening for the Presence Of Tumor by DNA Methylation And Size). Methods: We conducted a multicenter prospective study, K-DETEK (ClinicalTrials.gov identifier: NCT05227261), involving 9057 asymptomatic individuals aged 40 years or older across 75 major hospitals and one research institute in Vietnam. Participants were followed for 12 months. Results: Of the 9024 eligible participants, 43 (0.48%) tested positive for ctDNA. Among these, 17 were confirmed with malignant lesions in various primary organs through standard-of-care (SOC) imaging and biopsy, with 9 cases matching our tissue of origin (TOO) predictions. This resulted in a positive predictive value of 39.53% (95%CI 26.37–54.42) and a TOO accuracy of 52.94% (95%CI 30.96–73.83). Among the 8981 participants (99.52%) who tested negative, 8974 were confirmed cancer-free during a 12-month period after testing, yielding a negative predictive value of 99.92% (95% CI 99.84–99.96). The test demonstrated an overall sensitivity of 70.83% (95%CI 50.83–85.09) and a specificity of 99.71% (95% CI 99.58–99.80) for detecting various cancer types, including those without SOC screening options. Conclusions: This study presents a prospective validation of a multi-cancer early detection (MCED) test conducted in a lower middle-income country, demonstrating the potential of SPOT-MAS for early cancer detection. Our findings indicate that MCED tests could be valuable additions to national cancer screening programs, particularly in regions where such initiatives are currently limited. Trial registration: ClinicalTrials.gov ID: NCT05227261. Date of registration: 07/02/2022. [ABSTRACT FROM AUTHOR]

Aim: Cancers lacking standard screening (LSS) options account for approximately 70% of cancer-related deaths due to late-stage diagnosis. Circulating tumor DNA (ctDNA) is a promising biomarker for multi-cancer early detection. We previously developed SPOT-MAS, a multimodal ctDNA-based assay analyzing methylation and fragmentomic profiles, effective in detecting common cancers (breast, colorectal, liver, lung and gastric). This study extends the analysis to five LSS cancers: endometrial, esophageal, head and neck, ovarian and pancreatic. Methods: SPOT-MAS was applied to profile cfDNA methylation and fragmentomic patterns in 739 healthy individuals and 135 LSS cancer patients. Results: We identified 347 differentially methylated regions and observed genome-wide hypomethylation across all five LSS cancers. Esophageal and head and neck cancers showed an enrichment of short cfDNA fragments (<150 bp). Eleven 4-mer end motifs were consistently altered in cfDNA fragments across all LSS cancers. Many significant signatures were consistent with previous observations in common cancers. Notably, SPOT-MAS achieved 96.2% specificity and 74.8% overall sensitivity, with a lower sensitivity of 60.7% in early-stage cancers. Conclusion: This proof-of-concept study demonstrates that SPOT-MAS a non-invasive test trained on five common cancer types, could detect a number of LSS cancer cases, potentially complementing existing screening programs. Plain Language Summary Many cancers do not have standard tests, so they are often found too late, which leads to about 70% of cancer deaths. We've created a blood test that can help find cancer early. This test has already worked well for common cancers like breast and lung cancer, and now we're testing it on five harder-to-detect cancers: endometrial, esophageal, head and neck, ovarian and pancreatic cancers. In our study, we tested our blood test on 739 healthy people and 135 patients with these difficult cancers. Our method correctly identified healthy people 96.2% of the time and found cancer cases 74.8% of the time. This new test could help with screening for types of cancer that do not have good tests right now. Article highlights Approximately 70% of cancer deaths result from late-stage diagnoses in cancers lacking standard-of-care screening (LSS). The SPOT-MAS test is a ctDNA-based assay designed to detect five common cancers (breast, lung, colorectal, gastric and liver) and has been extended to five additional LSS cancers: endometrial, esophageal, head and neck, ovarian and pancreatic. The assay analyzed cfDNA methylation and fragmentomic patterns in 135 LSS cancer patients and 739 healthy individuals. It identified 347 differentially methylated regions and specific fragmentomic alterations. SPOT-MAS achieved 96.2% specificity and 74.8% overall sensitivity, with 60.7% sensitivity for early-stage detection. This study suggests that SPOT-MAS could serve as a non-invasive method for detecting LSS cancers. [ABSTRACT FROM AUTHOR]

Background: Haematococcus pluvialis, a green microalga, is a rich source of natural astaxanthin and a potent antioxidant with high commercial value. This study investigates the biological characteristics and potential of H. pluvialis HB isolated from Hoa Binh, Vietnam, for growth and astaxanthin accumulation using a two-phase culture method. Methods: H. pluvialis HB was cultured in a C/RM medium at 25 °C, and morphological characteristics were examined. NMR spectroscopy was used to determine the structure of the astaxanthin, which was extracted using the Soxhlet method. Results: After 22 days, the highest cell density (4.96 × 106 cells mL−1) was achieved under optimized light and ultraviolet conditions. Nutrient deprivation followed by bicarbonate supplementation resulted in a maximal astaxanthin accumulation of 48.8 mg g−1 dry cell weight within two days. The extracted astaxanthin demonstrated potent antioxidant activity (IC50: 3.74 mg mL−1) compared to ascorbic acid (IC50: 18.53 µg mL−1) and exhibited strong acetylcholinesterase inhibition (IC50: 297.99 µg mL−1). It also showed neuroprotective effects against H2O2 and amyloid beta-induced neurotoxicity in C6 cells. Conclusions: This study highlights H. pluvialis HB as a promising source for large-scale astaxanthin production with potential applications in neuroprotective health products. [ABSTRACT FROM AUTHOR]