Genetic Factors Associated with Clinical Response in Melanoma Patients Treated with Talimogene Laherparapvec: A Single-Institution Retrospective Analysis.

BACKGROUND: Talimogene laherparapvec (T-VEC) is a modified herpes simplex virus type 1 (HSV-1) and the first oncolytic virus to be approved for the treatment of unresectable melanoma. We assessed whether there are tumor-intrinsic genetic factors that are associated with tumor control. METHODS: A single-institution, retrospective analysis of melanoma patients treated with T-VEC was performed. Demographics, histopathologic reports, treatment history, clinical outcomes, and tumor genomic analysis of approximately 100 genes were collected. RESULTS: Ninety-three patients who had received T-VEC were identified, of whom 84 (91%) were diagnosed with cutaneous melanoma. Sixty-nine (69) patients received more than one dose of T-VEC and had sufficient data available for clinical analysis. Of these patients 30.0% (n = 21) had evidence of a complete response, defined as complete regression of all lesions without the need for additional treatment or procedures. Stage III disease (p < 0.001), absence of macroscopic nodal disease (p < 0.001), and absence of visceral/central nervous system metastases (p = 0.004) were all associated with evidence of any clinical response or local control by univariate analysis. At the time of analysis, 54 patients had tumor genetic data available. Sixty genes were mutated in at least one patient, and all but one patient had at least one gene mutation identified. Presence of TERT promotor mutation was associated with evidence of any clinical response (p = 0.043) or local control (p = 0.039) by multivariate analysis. CONCLUSIONS: This work describes the experience using T-VEC in melanoma at a single institution and highlights the presence of TERT promotor mutations as a possible driver of clinical response.

Hybrid Sparse Transformer and Wavelet Fusion-Based Deep Unfolding Network for Hyperspectral Snapshot Compressive Imaging.

Recently, deep unfolding network methods have significantly progressed in hyperspectral snapshot compressive imaging. Many approaches directly employ Transformer models to boost the feature representation capabilities of algorithms. However, they often fall short of leveraging the full potential of self-attention mechanisms. Additionally, current methods lack adequate consideration of both intra-stage and inter-stage feature fusion, which hampers their overall performance. To tackle these challenges, we introduce a novel approach that hybridizes the sparse Transformer and wavelet fusion-based deep unfolding network for hyperspectral image (HSI) reconstruction. Our method includes the development of a spatial sparse Transformer and a spectral sparse Transformer, designed to capture spatial and spectral attention of HSI data, respectively, thus enhancing the Transformer's feature representation capabilities. Furthermore, we incorporate wavelet-based methods for both intra-stage and inter-stage feature fusion, which significantly boosts the algorithm's reconstruction performance. Extensive experiments across various datasets confirm the superiority of our proposed approach.

Novel rapid molecular diagnosis methods for comprehensive genetic analysis of 21-hydroxylase deficiency.

BACKGROUND: Molecular analysis of the CYP21A2 gene is highly important for understanding the aetiology of 21-hydroxylase deficiency (21-OHD). The aim of this study was to use a novel approach named CNVplex, together with the SNaPshot assay and direct sequencing, to identify CYP21A2 mutations efficiently and comprehensively. Targeted CYP21A2 mutation analysis was performed in 113 patients and 226 parents. Large rearrangements of CYP21A2 were characterized by CNVplex; twenty prevalent mutations, including nine common micro-conversions and eleven high-frequency mutations reported in the literature, were detected by SNaPshot; and rare mutations were investigated by direct sequencing. RESULTS: Among the 113 21-OHD patients, 95.6% of the affected alleles were detected accurately by SNaPshot and CNVplex. Prevalent mutations were detected in 69.5% of the alleles; 62.4% of alleles contained pseudogene-derived micro-conversions, 1.8% contained nonpseudogene-derived mutations, and 5.3% contained complex variations resulting from multiple recombinations between CYP21A2 and CYP21A1P. Large rearrangements were identified in 27.0% of the alleles, including five types (CH-1, CH-3, CH-4, CH-5 and CH-8) of chimeric CYP21A1P/CYP21A2 genes. Two novel CYP21A2 haplotypes and four de novo CYP21A2 mutations were characterized. A rare haplotype with a c.955 C > T mutation in the duplicated CYP21A2 gene was found in 0.9% of the probands and 33.3% of the parents. In addition, four parents were also diagnosed with 21-OHD. CONCLUSION: CNVplex and SNaPshot appear to be highly efficient and reliable techniques for use in a molecular diagnosis laboratory, and combined with direct sequencing based on locus-specific PCR, they might constitute a definitive way to detect almost all common and rare 21-OHD-related alleles.

Spectral analysis comparison of pushbroom and snapshot hyperspectral cameras for in vivo brain tissues and chromophore identification.

Significance: Hyperspectral imaging sensors have rapidly advanced, aiding in tumor diagnostics for in vivo brain tumors. Linescan cameras effectively distinguish between pathological and healthy tissue, whereas snapshot cameras offer a potential alternative to reduce acquisition time. Aim: Our research compares linescan and snapshot hyperspectral cameras for in vivo brain tissues and chromophore identification. Approach: We compared a linescan pushbroom camera and a snapshot camera using images from 10 patients with various pathologies. Objective comparisons were made using unnormalized and normalized data for healthy and pathological tissues. We utilized the interquartile range (IQR) for the spectral angle mapping (SAM), the goodness-of-fit coefficient (GFC), and the root mean square error (RMSE) within the 659.95 to 951.42 nm range. In addition, we assessed the ability of both cameras to capture tissue chromophores by analyzing absorbance from reflectance information. Results: The SAM metric indicates reduced dispersion and high similarity between cameras for pathological samples, with a 9.68% IQR for normalized data compared with 2.38% for unnormalized data. This pattern is consistent across GFC and RMSE metrics, regardless of tissue type. Moreover, both cameras could identify absorption peaks of certain chromophores. For instance, using the absorbance measurements of the linescan camera, we obtained SAM values below 0.235 for four peaks, regardless of the tissue and type of data under inspection. These peaks are one for cytochrome b in its oxidized form at λ = 422 nm , two for HbO 2 at λ = 542 nm and λ = 576 nm , and one for water at λ = 976 nm . Conclusion: The spectral signatures of the cameras show more similarity with unnormalized data, likely due to snapshot sensor noise, resulting in noisier signatures post-normalization. Comparisons in this study suggest that snapshot cameras might be viable alternatives to linescan cameras for real-time brain tissue identification.

Design and implementation of a portable snapshot multispectral imaging crop-growth sensor.

The timely and accurate acquisition of crop-growth information is a prerequisite for implementing intelligent crop-growth management, and portable multispectral imaging devices offer reliable tools for monitoring field-scale crop growth. To meet the demand for obtaining crop spectra information over a wide band range and to achieve the real-time interpretation of multiple growth characteristics, we developed a novel portable snapshot multispectral imaging crop-growth sensor (PSMICGS) based on the spectral sensing of crop growth. A wide-band co-optical path imaging system utilizing mosaic filter spectroscopy combined with dichroic mirror beam separation is designed to acquire crop spectra information over a wide band range and enhance the device's portability and integration. Additionally, a sensor information and crop growth monitoring model, coupled with a processor system based on an embedded control module, is developed to enable the real-time interpretation of the aboveground biomass (AGB) and leaf area index (LAI) of rice and wheat. Field experiments showed that the prediction models for rice AGB and LAI, constructed using the PSMICGS, had determination coefficients (R²) of 0.7 and root mean square error (RMSE) values of 1.611 t/ha and 1.051, respectively. For wheat, the AGB and LAI prediction models had R² values of 0.72 and 0.76, respectively, and RMSE values of 1.711 t/ha and 0.773, respectively. In summary, this research provides a foundational tool for monitoring field-scale crop growth, which is important for promoting high-quality and high-yield crops.

Snapshot computed tomographic microscopic imaging spectrometer and its video-level tracking of poisonous Microcystis aeruginosa cells in mixed algae.

Conventional microscopic spectral imaging suffers from extended scanning times across wavelength or spatial dimension. To improve capabilities of dynamic microscopic spectral imaging, we developed a snapshot computed tomographic microscopic imaging spectrometer (CTMIS) based on the zeroth and first orders dispersive diffraction of a two-dimensional grating. Utilizing the CTMIS-UNET reconstruction algorithm, we can reconstruct a spectral cube (541x541x26) for each frame of micro spectral imaging video. Experimental results demonstrate a sub-4 µm spatial resolution achievable through a 20x objective lens and a spectral resolution better than 10 nm among 450-700 nm, while maintaining spectral cosine similarities exceeding 0.9989 when comparing reconstructed spectra with ground truth data. Spectral imaging videos of four species of algae and mixed algae were captured under 10 ms exposure time using the CTMIS system. Leveraging the self-developed UNET-SI26 algae recognition network, precise identification and tracking of four types of algae and poisonous microcysts aeruginosa in mixed algae were conducted. The pixel-level recognition accuracy exceeds 95 %, while the accuracy for counting the numbers of different types of cells surpasses 85 %, offering an efficient and accurate spectral imaging method for real-time monitoring and early warning of harmful algae at the cellular level.

Classification of melanocytic lesions using direct illumination multispectral imaging.

With rising melanoma incidence and mortality, early detection and surgical removal of primary lesions is essential. Multispectral imaging is a new, non-invasive technique that can facilitate skin cancer detection by measuring the reflectance spectra of biological tissues. Currently, incident illumination allows little light to be reflected from deeper skin layers due to high surface reflectance. A pilot study was conducted at the University Hospital Basel to evaluate, whether multispectral imaging with direct light coupling could extract more information from deeper skin layers for more accurate dignity classification of melanocytic lesions. 27 suspicious pigmented lesions from 23 patients were included (6 melanomas, 6 dysplastic nevi, 12 melanocytic nevi, 3 other). Lesions were imaged before excision using a prototype snapshot mosaic multispectral camera with incident and direct illumination with subsequent dignity classification by a pre-trained multispectral image analysis model. Using incident light, a sensitivity of 83.3% and a specificity of 58.8% were achieved compared to dignity as determined by histopathological examination. Direct light coupling resulted in a superior sensitivity of 100% and specificity of 82.4%. Convolutional neural network classification of corresponding red, green, and blue lesion images resulted in 16.7% lower sensitivity (83.3%, 5/6 malignant lesions detected) and 20.9% lower specificity (61.5%) compared to direct light coupling with multispectral image classification. Our results show that incorporating direct light multispectral imaging into the melanoma detection process could potentially increase the accuracy of dignity classification. This newly evaluated illumination method could improve multispectral applications in skin cancer detection. Further larger studies are needed to validate the camera prototype.

Improving image quality and in-stent restenosis diagnosis with high-resolution "double-low" coronary CT angiography in patients after percutaneous coronary intervention.

Objective: This study aims to investigate the image quality of a high-resolution, low-dose coronary CT angiography (CCTA) with deep learning image reconstruction (DLIR) and second-generation motion correction algorithms, namely, SnapShot Freeze 2 (SSF2) algorithm, and its diagnostic accuracy for in-stent restenosis (ISR) in patients after percutaneous coronary intervention (PCI), in comparison with standard-dose CCTA with high-definition mode reconstructed by adaptive statistical iterative reconstruction Veo algorithm (ASIR-V) and the first-generation motion correction algorithm, namely, SnapShot Freeze 1 (SSF1). Methods: Patients after PCI and suspected of having ISR scheduled for high-resolution CCTA (randomly for 100 kVp low-dose CCTA or 120 kVp standard-dose) and invasive coronary angiography (ICA) were prospectively enrolled in this study. After the basic information pairing, a total of 105 patients were divided into the LD group (60 patients underwent 100 kVp low-dose CCTA reconstructed with DLIR and SSF2) and the SD group (45 patients underwent 120 kVp standard-dose CCTA reconstructed with ASIR-V and SSF1). Radiation and contrast medium doses, objective image quality including CT value, image noise (standard deviation), signal-to-noise ratio (SNR), and contrast-to-noise ratio (CNR) for the aorta, left main artery (LMA), left ascending artery (LAD), left circumflex artery (LCX), and right coronary artery (RCA) of the two groups were compared. A five-point scoring system was used for the overall image quality and stent appearance evaluation. Binary ISR was defined as an in-stent neointimal proliferation with diameter stenosis ≥50% to assess the diagnostic performance between the LD group and SD group with ICA as the standard reference. Results: The LD group achieved better objective and subjective image quality than that of the SD group even with 39.1% radiation dose reduction and 28.0% contrast media reduction. The LD group improved the diagnostic accuracy for coronary ISR to 94.2% from the 83.8% of the SD group on the stent level and decreased the ratio of false-positive cases by 19.2%. Conclusion: Compared with standard-dose CCTA with ASIR-V and SSF1, the high-resolution, low-dose CCTA with DLIR and SSF2 reconstruction algorithms further improves the image quality and diagnostic performance for coronary ISR at 39.1% radiation dose reduction and 28.0% contrast dose reduction.

Snapshot Multi-Wavelength Birefringence Imaging.

A snapshot multi-wavelength birefringence imaging measurement method was proposed in this study. The RGB-LEDs at wavelengths 463 nm, 533 nm, and 629 nm were illuminated with circularly polarized light after passing through a circular polarizer. The transmitted light through the birefringent sample was captured by a color polarization camera. A single imaging process captured light intensity in four polarization directions (0°, 45°, 90°, and 135°) for each of the three RGB spectral wavelength channels, and subsequently measured the first three elements of Stokes vectors (S0, S1, and S2) after the sample. The birefringence retardance and fast-axis azimuthal angle were determined simultaneously. An experimental setup was constructed, and polarization response matrices were calibrated for each spectral wavelength channel to ensure the accurate detection of Stokes vectors. A polymer true zero-order quarter-wave plate was employed to validate measurement accuracy and repeatability. Additionally, stress-induced birefringence in a PMMA arch-shaped workpiece was measured both before and after the application of force. Experimental results revealed that the repeatability of birefringence retardance and fast-axis azimuthal angle was better than 0.67 nm and 0.08°, respectively. This approach enables multispectral wavelength, high-speed, high-precision, and high-repeatability birefringence imaging measurements through a single imaging session.

Comparative analysis of SNaPshot and massively parallel sequencing for body fluid-specific DNA methylation markers.

The identification of tissue-specific differentially methylated regions has significantly contributed to the field of forensic genetics, particularly in body fluid identification crucial for linking evidence to crimes. Among the various approaches to analyzing DNA methylation, the SNaPshot assay has been popularly studied in numerous researches. However, there is a growing interest in exploring alternative methods such as the use of massively parallel sequencing (MPS), which can process a large number of samples simultaneously. This study compares SNaPshot and MPS multiplex assays using nine cytosine-phosphate-guanine markers for body fluid identification. As a result of analyzing 112 samples, including blood, saliva, vaginal fluid, menstrual blood, and semen, both methods demonstrated high sensitivity and specificity, indicating their reliability in forensic investigations. A total of 92.0% samples were correctly identified by both methods. Although both methods accurately identified all blood, saliva, and semen samples, some vaginal fluid samples showed unexpected methylation signals at nontarget loci in addition to the target loci. In the case of menstrual blood samples, due to their complexity, independent typing criteria were applied, and successful menstrual blood typing was possible, whereas a few samples showed profiles similar to vaginal fluid. The MPS method worked better in vaginal fluid samples, and the SNaPshot method performed better in menstrual blood samples. This study offers valuable insights into body fluid identification based on the characteristics of the SNaPshot and MPS methods, which may help in more efficient forensic applications.

An Internet Snapshot Survey Assessing the sale of Synthetic Cannabinoid Receptor Agonists for use with Electronic Vaping Devices.

INTRODUCTION: Synthetic cannabinoid receptor agonists (SCRAs) are associated with significant toxicity and are increasingly used in electronic vaping devices. We assessed the availability of SCRA vaping products to UK purchasers on the surface web. METHODS: An internet snapshot survey was performed between October 2022 and January 2023 on 'google.com' using the search terms "buy c-liquid vape", "buy herbal incense vape liquid", "buy cannabis vape liquid", "buy hashish vape liquid", "buy K2 vape liquid". RESULTS: 62 websites selling 128 SCRA vaping brands were identified. Most were purportedly based in the USA (41 websites, 66%) and most sold other controlled substances. Purchase incentives offered included discreet packaging (38, 61%), discounts for bulk purchase (34, 55%) and tracked delivery (30, 48%). Many websites stated SCRA products were: not for human consumption (41, 66%), for research purposes only (15, 24%), or legal (28, 45%). Websites sold a median (IQR) of 16 (7-25) SCRA vaping brands. Almost all were bottles of vaping liquid (1220/1225, 99.6%). The most common bottle size was 5mL (60%), the median (IQR) total volume of SCRA liquid per sale was 50mL (10-200mL). Median (IQR) price was £3.39/mL (£2.01/mL- £5.29/mL). Price decreased with increasing volume purchased (£6.58/mL for ≤ 5mL, £1.60/mL for > 200mL). CONCLUSION: SCRA vaping products are easily obtainable online, in both small and bulk quantities. Information provided to purchasers on safety and legality is lacking or misleading. Further studies are needed to confirm the chemistry of these products. Policymakers should consider steps to limit the potential harm caused by the purchase and use of these products.

A method of identifying the high-risk mutations of sudden cardiac death at KCNQ1 and KCNH2 genes.

Sudden Cardiac Death (SCD) often shows negative anatomy results after a systemic autopsy and the gene mutations of potassium channel play a key role in the etiology of SCD. We established a feasible system to detect SCD-related mutations and investigated the mutations at KCNQ1 and KCNH2 genes in the Chinese population. We established a mutation detection system combined with multiplex PCR, SNaPshot technique, and capillary electrophoresis. We genotyped 101 putative mutations at KCNQ1 and KCNH2 genes in 60 SCD of negative anatomy and 50 controls using the established assay and compared Odd Ratio (OR). Four coding variants were identified in the KCNQ1 gene: S546S, I145I, P448R, and G643S. The mutations of I145I and S546S did not differ significantly in the SCD compared with controls. 21 SCD individuals (35 %) and 1 control individual (2 %) showed a genotype of C/G at P448R (OR = 17.5, 95 % CI [2.40-127.82]). 24 SCD individuals (40 %) and 1 control individual (2 %) showed a genotype of C/G at G643S (OR = 20.0, 95 % CI [2.75-145.25]). We established a robust assay for rapid screening the putative SCD-related mutations in KCNQ1 and KCNH2 genes. The new assay in our study is easily amenable to the majority of laboratories without the need for new specialized equipment. Our method will meet the increasing requirement of mutation screening for SCD in regular DNA laboratories and will help screen mutations in those dead of SCD and their relatives.

A multiplex SNaPshot assay for the detection of single nucleotide polymorphism associated with clinical severity of enterovirus infections.

Non-polio enterovirus infections are known to cause a variety of diseases and neurological complications. It is also known that the severity of these diseases largely differs among individuals with different genotypes and alleles. The Single Nucleotide Polymorphisms (SNPs) within specific genes have a considerable effect on the immune response to enteroviruses and on the outcome of disease, leading to variations in complications and infection susceptibility. Knowing the distribution of such SNPs can be valuable for individual case management and studying epidemiological parameters of enterovirus infections. In this feasibility study, a multiplex version of the primer extension-based technique called the SNaPshot Assay has been developed to examine SNPs in various relevant genes for predicting the clinical severity of enterovirus infections. It is already established that this technique is precise, consistent, scalable, and likely to exhibit high throughput. The multiplex SNaPshot can investigate multiple genetic susceptibility markers simultaneously, and the assay can be used to identify vulnerable populations, understand the epidemiology of infections, and manage the outbreaks of enteroviruses. Based on the literature, 15 SNPs were identified which are suspected for higher susceptibility to the worst outcomes after enterovirus infection and the assay was developed. Blood samples of 100 healthy volunteers were collected and tested for assay feasibility as well as to know the proportions of 15 selected SNPs. After the analysis, seven SNPs have been identified and suggested to be considered for future assays. Based on the pilot test results, it appears that positivity for any three out of the identified seven SNPs might indicate a higher risk, and future studies correlated with clinical studies among patients with and without severe diseases utilizing this assay will provide robust parameters to determine at-risk individuals more accurately.

Development and validation of a new multiplex panel using SNaPshot-based DIP-TriSNP markers for forensic DNA mixtures.

Short tandem repeat analysis is challenging when dealing with unbalanced mixtures in forensic cases due to the presence of stutter peaks and large amplicons. In this research, we propose a novel genetic marker called DIP-TriSNP, which combines deletion/insertion polymorphism (DIP) with tri-allelic single nucleotide polymorphism in less than 230 bp length of human genome. Based on multiplex PCR and SNaPShot, a panel, including 14 autosomal DIP-TriSNPs and one Y chromosomal DIP-SNP, had been developed and applied to genotyping 102 unrelated Han Chinese individuals in Sichuan of China and simulated a mixture study. The panel sensitivity can reach as low as 0.1 ng DNA template, and the minor contributor of DNA can be detected with the highest ratio of 19:1, as indicated by the obtained results. In the Sichuan Han population, the cumulative probability of informative genotypes reached 0.997092, with a combined power of discrimination of 0.999999998801. The panel was estimated to detect more than two alleles in at least one locus in 99.69% of mixtures of the Sichuan Han population. In conclusion, DIP-TriSNPs have shown promising as an innovative DNA marker for identifying the minor contributor in unbalanced DNA mixtures, offering advantages such as short amplifications, increased polymorphism, and heightened sensitivity.

Meta-Attention Network Based Spectral Reconstruction with Snapshot Near-Infrared Metasurface.

Near-infrared (NIR) spectral information is important for detecting and analyzing material compositions. However, snapshot NIR spectral imaging systems still pose significant challenges owing to the lack of high-performance NIR filters and bulky setups, preventing effective encoding and integration with mobile devices. This study introduces a snapshot spectral imaging system that employs a compact NIR metasurface featuring 25 distinct C4 symmetry structures. Benefitting from the sufficient spectral variety and low correlation coefficient among these structures, center-wavelength accuracy of 0.05 nm and full width at half maximum accuracy of 0.13 nm are realized. The system maintains good performance within an incident angle of 1°. A novel meta-attention network prior iterative denoising reconstruction (MAN-IDR) algorithm is developed to achieve high-quality NIR spectral imaging. By leveraging the designed metasurface and MAN-IDR, the NIR spectral images, exhibiting precise textures, minimal artifacts in the spatial dimension, and little crosstalk between spectral channels, are reconstructed from a single grayscale recording image. The proposed NIR metasurface and MAN-IDR hold great promise for further integration with smartphones and drones, guaranteeing the adoption of NIR spectral imaging in real-world scenarios such as aerospace, health diagnostics, and machine vision.

DNA methylation-based organ tissue identification: Marker identification, SNaPshot multiplex assay development, and interlaboratory comparison.

Identifying body fluids and organ tissues is highly significant as they can offer crucial evidence in criminal investigations and aid the court in making informed decisions, primarily through evaluating the biological source and possibly at the activity level up to death or fatal damage. In this study, organ tissue-specific CpG markers were identified from Illumina's methylation EPIC array data of nine organ tissues, including epidermis, dermis, heart, skeletal muscle, blood, kidney, brain, lung, and liver, from autopsies of 10 Koreans. Through the validation test using 43 samples, 18 hypomethylation markers, with two markers for each organ tissue type, were selected to construct a SNaPshot assay. Two multiplex assays involving forward and reverse SBE primers were designed to help investigators accurately determine the organ origin of the analyzed tissue samples through repeated analysis of the same PCR products for markers. The developed multiplex demonstrated high accuracy, achieving 100.0 % correct detection of the presence of nine organ tissue types in 88 samples from autopsies of 10 Asians. However, two lung samples showed additional positive indications of the presence of blood. An interlaboratory comparison using 80 autopsy samples (heart, skeletal muscle, blood, kidney cortex, kidney medulla, brain, lung, and liver) from 10 individuals in Germany revealed overall comparable results with correct detection of the presence of eight organ tissue types in 92.5 % samples (74 of 80 samples). In the case of six samples, it was impossible to determine the correct tissue successfully due to drop-outs of unmethylation signals at target tissue marker loci. One of these lung samples revealed only non-intended off-target signals for blood. The observed differences might be due to differences in sample collection during routine autopsy, technical differences due to the PCR cycler, and the threshold used for signal calling. Indicating the presence of additional tissue type and off-target unmethylation signals seems alleviated by applying more stringent hypomethylation thresholds. Therefore, the developed SNaPshot multiplex assays will be valuable for forensic investigators dealing with organ tissue identification, as well as for prosecutors and defense aiming to establish the circumstances that occurred at the crime scene.

Protocol for kinetic mode potassium channel assays on common plate readers and microscopes.

Fluorescence-based potassium channel assays are typically run on expensive, hard to obtain, fluorescence imaging kinetic plate readers that are uncommon in most laboratories. Here we describe the use of the Brilliant Thallium Snapshot assay to conduct an endpoint potassium channel assay, so that it can be used across multiple plate reader platforms that are more common in many labs. These methods will allow users to identify modulators of potassium channels. For this work, we have taken a kinetic mode Molecular Devices FLIPR based protocol and adapted it to be utilized on endpoint plate readers, such as the BMG Labtech PHERAstar, to identify activators of GIRK channels in CHO cells. We demonstrate that both plate readers are functionally competent at generating excellent Z' values which makes them ideally suited to finding corollary hits from the Sigma LOPAC 1,280 screening collection. Importantly, this assay has also been validated using a high content reader, demonstrating the possibility of spatially resolving signals from individual cells within a mixed cell population. The compendium of these results shows the flexibility, accessibility and functionality of endpoint-compatible potassium channel assay readouts on more common plate readers.

Impaired cerebral autoregulation detected in early prevasospasm period is associated with unfavorable outcome after spontaneous subarachnoid hemorrhage: an observational prospective pilot study.

BACKGROUND: Subarachnoid hemorrhage (SAH) patients with cerebral autoregulation (CA) impairment at an early post-SAH period are at high risk of unfavorable outcomes due to delayed cerebral ischemia (DCI) or other complications. Limited evidence exists for an association between early-stage CA impairments and SAH patient outcomes. The objective of this prospective study was to explore associations between CA impairments detected in early post-SAH snapshot examinations and patient outcomes. METHODS: The pilot observational study included 29 SAH patients whose CA status was estimated 2-3 days after spontaneous aneurysm rupture and a control group of 15 healthy volunteers for comparison. Inflatable leg recovery boots (reboots.com, Germany) were used for the safe controlled generation of arterial blood pressure (ABP) changes necessary for reliable CA examination. At least 5 inflation‒deflation cycles of leg recovery boots with a 2-3 min period were used during examinations. CA status was assessed according to the delay time (∆TCBFV) measured between ABP(t) and cerebral blood flow velocity (CBFV(t)) signals during artificially induced ABP changes at boot deflation cycle. CBFV was measured in middle cerebral artery by using transcranial Doppler device. RESULTS: Statistically significant differences in ∆TCBFV were found between SAH patients with unfavorable outcomes (∆TCBFV = 1.37 ± 1.23 s) and those with favorable outcomes (∆TCBFV = 2.86 ± 0.99 s) (p < 0.001). Early assessment of baroreflex sensitivity (BRS) during the deflation cycle showed statistically significant differences between the DCI and non-DCI patient groups (p = 0.039). CONCLUSIONS: A relatively small delay of ∆TCBFV <1.6 s between CBFV(t) and ABP(t) waves could be an early warning sign associated with unfavorable outcomes in SAH patients. The BRS during boot deflation can be used as a biomarker for the prediction of DCI. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT06028906. Registered 31 August 2023 - Retrospectively registered, https://www. CLINICALTRIALS: gov/study/NCT06028906 .

Enhanced deep unrolling networks for snapshot compressive hyperspectral imaging.

Snapshot compressive hyperspectral imaging necessitates the reconstruction of a complete hyperspectral image from its compressive snapshot measurement, presenting a challenging inverse problem. This paper proposes an enhanced deep unrolling neural network, called EDUNet, to tackle this problem. The EDUNet is constructed via the deep unrolling of a proximal gradient descent algorithm and introduces two innovative modules for gradient-driven update and proximal mapping reflectivity. The gradient-driven update module leverages a memory-assistant descent approach inspired by momentum-based acceleration techniques, for enhancing the unrolled reconstruction process and improving convergence. The proximal mapping is modeled by a sub-network with a cross-stage spectral self-attention, which effectively exploits the inherent self-similarities present in hyperspectral images along the spectral axis. It also enhances feature flow throughout the network, contributing to reconstruction performance gain. Furthermore, we introduce a spectral geometry consistency loss, encouraging EDUNet to prioritize the geometric layouts of spectral curves, leading to a more precise capture of spectral information in hyperspectral images. Experiments are conducted using three benchmark datasets including KAIST, ICVL, and Harvard, along with some real data, comprising a total of 73 samples. The experimental results demonstrate that EDUNet outperforms 15 competing models across four metrics including PSNR, SSIM, SAM, and ERGAS.

A preliminary report on the exploration of salivary bacterial diversity by the multiplex SNaPshot assay.

Salivary bacterial community composition is associated with the host's internal and environmental factors, which have potential applications in forensic practice. The 16S rRNA gene sequencing is the most commonly used strategy for detecting salivary bacterial diversity; however, its platforms are not compatible with capillary electrophoresis (CE) platforms commonly used for forensic applications. Therefore, we attempted to detect the salivary bacterial diversity using a single nucleotide polymorphism (SNP) assay. Salivary bacterial diversity varies among diverse geographic locations, making it a potential supplementary biomarker for forensic geographic sourcing. To evaluate the performance of the multiplex SNaPshot assay, saliva samples from three geographic locations in China were analyzed using the multiplex SNaPshot assay and 16S rRNA gene sequencing. We screened SNPs from two high-relative-abundance salivary genera (Streptococcus and Veillonella) to construct a multiplex SNaPshot system that can be used on the CE platform. The stability and sensitivity of the multiplex SNaPshot system were also tested. A random forest classification model was used to classify samples from different regions to explore the ability of salivary bacteria to discriminate between geographic sources. Six bacterial SNPs were screened and a multiplex SNaPshot system was constructed. The stability results showed that the typing of salivary stains that were placed indoors for different days was not affected in this study. Two-thirds of mocked salivary stain samples showed more than 90% of typing results obtained for salivary stain samples with an input of 0.1 µl saliva. The results of principal coordinate analysis based on salivary bacterial diversity showed significant differences between samples from the three different geographic locations. The accuracy of the random forest classification was 66.67% based on the multiplex SNaPshot assay and 83.33% based on the 16S rRNA gene sequencing. In conclusion, this is the first attempt to detect salivary bacterial diversity using a multiplex SNaPshot bacterial SNP assay. The geographic difference in human salivary bacterial community composition was significant, as revealed by the multiplex SNaPshot assay; however, its performance in discriminating geographic sources was lower than that of 16S rRNA gene sequencing. This strategy based on bacterial SNP loci may favor the detection of human bacterial diversity in common forensic laboratories but requires further exploration in larger sample sizes and more bacterial SNP loci.