The obesity-related mutation gene on nonalcoholic fatty liver disease.

The prevalence of overweight and obesity is increasing, leading to metabolic-associated fatty liver disease (MAFLD) characterized by excessive accumulation of liver fat and a risk of developing hepatocellular carcinoma (HCC). The driver gene mutations may play the roles of passengers that occur in single 'hotspots' and can promote tumorigenesis from benign to malignant lesions. We investigated the impact of high body weight and BMI on HCC survival using The Cancer Genome Atlas Liver Hepatocellular Carcinoma (TCGA-LIHC) dataset. To explore the effects of obesity-related gene mutations on HCC, we collected driver mutation genes in 34 TCGA patients with BMI ≥ 27 and 23 TCGA patients with BMI < 27. The digital PCR performing the PBMC samples for the variant rate by clinical cohort of 96 NAFLD patients. Our analysis showed that obesity leads to significantly worse survival outcomes in HCC. Using cbioportal, we identified 414 driver mutation genes in patients with obesity and 127 driver mutation genes in non-obese patients. Functional analysis showed that obese-related genes significantly enriched the regulated lipid and insulin pathways in HCC. The insulin secretion pathway in patients with obesity HCC-specific survival identified ABCC8 and PRKCB as significant genes (p < 0.001). It revealed significant differences in gene mutation and gene expression profiles compared to non-obese patients. The digital PCR test ABCC8 variants were detected in PBMC samples and caused a 14.5% variant rate, significantly higher than that of non-obese NAFLD patients. The study findings showed that the gene ABCC8 was a patient with the obesity-related gene in NAFLD, which provides the probability that ABCC8 mutation contributes to the pre-cancer lesion biomarker for HCC.

Accelerating pandemic response with the emergency Omicron RT-PCR test: A comprehensive solution for COVID-19 diagnosis and tracking.

The Omicron variant of concern (VOC) has surged in many countries and replaced the previously reported VOC. To identify different Omicron strains/sublineages on a rapid, convenient, and precise platform, we report a novel multiplex real-time reverse transcriptase polymerase chain reaction (RT-PCR) method in one tube based on the Omicron lineage sequence variants' information. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) subvariants were used in a PCR-based assay for rapid identification of Omicron sublineage genotyping in 1000 clinical samples. Several characteristic mutations were analyzed using specific primers and probes for the spike gene, del69-70, and F486V. To distinguish Omicron sublineages (BA.2, BA.4, and BA.5), the NSP1:141-143del in the ORF1a region and D3N mutation in membrane protein occurring outside the spike protein region were analyzed. Results from the real-time PCR assay for one-tube accuracy were compared to those of whole genome sequencing. The developed PCR assay was used to analyze 400 SARS-CoV-2 positive samples. Ten samples determined as BA.4 were positive for NSP1:141-143del, del69-70, and F486V mutations; 160 BA.5 samples were positive for D3N, del69-70, and F486V mutations, and 230 BA.2 samples were without del69-70. Screening these samples allowed the identification of epidemic trends at different time intervals. Our novel one-tube multiplex PCR assay was effective in identifying Omicron sublineages.

Host-Pathogen Interactions in K. pneumoniae Urinary Tract Infections: Investigating Genetic Risk Factors in the Taiwanese Population.

BACKGROUND: Klebsiella pneumoniae (K. pneumoniae) urinary tract infections pose a significant challenge in Taiwan. The significance of this issue arises because of the growing concerns about the antibiotic resistance of K. pneumoniae. Therefore, this study aimed to uncover potential genomic risk factors in Taiwanese patients with K. pneumoniae urinary tract infections through genome-wide association studies (GWAS). METHODS: Genotyping data are obtained from participants with a history of urinary tract infections enrolled at the Tri-Service General Hospital as part of the Taiwan Precision Medicine Initiative (TPMI). A case-control study employing GWAS is designed to detect potential susceptibility single-nucleotide polymorphisms (SNPs) in patients with K. pneumoniae-related urinary tract infections. The associated genes are determined using a genome browser, and their expression profiles are validated via the GTEx database. The GO, Reactome, DisGeNET, and MalaCards databases are also consulted to determine further connections between biological functions, molecular pathways, and associated diseases between these genes. RESULTS: The results identified 11 genetic variants with higher odds ratios compared to controls. These variants are implicated in processes such as adhesion, protein depolymerization, Ca2+-activated potassium channels, SUMOylation, and protein ubiquitination, which could potentially influence the host immune response. CONCLUSIONS: This study implies that certain risk variants may be linked to K. pneumoniae infections by affecting diverse molecular functions that can potentially impact host immunity. Additional research and follow-up studies are necessary to elucidate the influence of these risk variants on infectious diseases and develop targeted interventions for mitigating the spread of K. pneumoniae urinary tract infections.

Monitoring coronavirus disease progression and clinical impact through quantitative viral load testing.

BACKGROUND: The viral load (VL) in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-infected individuals is critical for improving clinical treatment strategies, care, and decisions. Several studies have reported that the initial SARS-CoV-2 VL is associated with disease severity and mortality. Cycle threshold (Ct) values and/or copies/mL are often used to quantify VL. However, a multitude of platforms, primer/probe sets of different SARS-CoV-2 target genes, and reference material manufacturers may cause inconsistent interlaboratory interpretations. The first International Standard for SARS-CoV-2 RNA quantitative assays has allowed diagnostic laboratories to transition SARS-CoV-2 VL results into international units per milliliter (IU/mL). The Cobas SARS-CoV-2 Duo quantitative assay provides VL results expressed in IU/mL. MATERIALS AND METHODS: We enrolled 145 and 50 SARS-CoV-2-positive, hospitalized and 50-negative individuals at the Tri-Service General Hospital, Taiwan from January to May 2022. Each participant's electronic medical record was reviewed to determine asymptomatic, mild, moderate, and severe cases. Nasopharyngeal swabs were collected using universal transport medium. We investigated the association of SARS-CoV-2 VL with disease severity using the Cobas SARS-CoV-2 Duo quantitative assay and its functionality in clinical assessment and decision making to further improve clinical treatment strategies. Limit of detection (LOD) was assessed. RESULTS: All 50 SARS-CoV-2-negative samples confirmed negative for SARS-CoV-2, demonstrating 100 % specificity of the Cobas SARS-CoV-2 Duo assay. Patients with severe symptoms had longer hospital stays, and the length of hospital stay (30.56 days on average) positively correlated with the VL (8.22 ± 1.21 log10 IU/mL). Asymptomatic patients had the lowest VL (5.54 ± 2.06 log10 IU/mL) at admission and the shortest hospital stay (14.1 days on average). CONCLUSIONS: VL is associated with disease severity and duration of hospitalization; therefore, its quantification should be considered when making clinical care decisions and treatment strategies. The Cobas SARS-CoV-2 Duo assay provides a commutable unitage IU/mL for interlaboratory interpretations.

Intraoperative Detection of Surgical Gauze Using Deep Convolutional Neural Network.

During laparoscopic surgery, surgical gauze is usually inserted into the body cavity to help achieve hemostasis. Retention of surgical gauze in the body cavity may necessitate reoperation and increase surgical risk. Using deep learning technology, this study aimed to propose a neural network model for gauze detection from the surgical video to record the presence of the gauze. The model was trained by the training group using YOLO (You Only Look Once)v5x6, then applied to the testing group. Positive predicted value (PPV), sensitivity, and mean average precision (mAP) were calculated. Furthermore, a timeline of gauze presence in the video was drawn by the model as well as human annotation to evaluate the accuracy. After the model was well-trained, the PPV, sensitivity, and mAP in the testing group were 0.920, 0.828, and 0.881, respectively. The inference time was 11.3 ms per image. The average accuracy of the model adding a marking and filtering process was 0.899. In conclusion, surgical gauze can be successfully detected using deep learning in the surgical video. Our model provided a fast detection of surgical gauze, allowing further real-time gauze tracing in laparoscopic surgery that may help surgeons recall the location of the missing gauze.

Prediction of antidepressant responses to non-invasive brain stimulation using frontal electroencephalogram signals: Cross-dataset comparisons and validation.

BACKGROUND: 10-Hz repetitive transcranial magnetic stimulation(rTMS) and intermittent theta-burst stimulation(iTBS) over left prefrontal cortex are FDA-approved, effective options for treatment-resistant depression (TRD). Optimal prediction models for iTBS and rTMS remain elusive. Therefore, our primary objective was to compare prediction accuracy between classification by frontal theta activity alone and machine learning(ML) models by linear and non-linear frontal signals. The second objective was to study an optimal ML model for predicting responses to rTMS and iTBS. METHODS: Two rTMS and iTBS datasets (n = 163) were used: one randomized controlled trial dataset (RCTD; n = 96) and one outpatient dataset (OPD; n = 67). Frontal theta and non-linear EEG features that reflect trend, stability, and complexity were extracted. Pretreatment frontal EEG and ML algorithms, including classical support vector machine(SVM), random forest(RF), XGBoost, and CatBoost, were analyzed. Responses were defined as ≥50 % depression improvement after treatment. Response rates between those with and without pretreatment prediction in another independent outpatient cohort (n = 208) were compared. RESULTS: Prediction accuracy using combined EEG features by SVM was better than frontal theta by logistic regression. The accuracy for OPD patients significantly dropped using the RCTD-trained SVM model. Modern ML models, especially RF (rTMS = 83.3 %, iTBS = 88.9 %, p-value(ACC > NIR) < 0.05 for iTBS), performed significantly above chance and had higher accuracy than SVM using both selected features (p < 0.05, FDR corrected for multiple comparisons) or all EEG features. Response rates among those receiving prediction before treatment were significantly higher than those without prediction (p = 0.035). CONCLUSION: The first study combining linear and non-linear EEG features could accurately predict responses to left PFC iTBS. The bootstraps-based ML model (i.e., RF) had the best predictive accuracy for rTMS and iTBS.

Clinical Evaluation of Direct Reverse Transcription PCR for Detection of SARS-CoV-2 Compared to Conventional RT-PCR in Patients with Positive Rapid Antigen Test Results during Circulation of Emerging Viral Variants.

The emergence of the Omicron (B.1.1.529) variant of SARS-CoV-2 has precipitated a new global wave of the COVID-19 pandemic. The rapid identification of SARS-CoV-2 infection is imperative for the effective mitigation of transmission. Diagnostic modalities such as rapid antigen testing and real-time reverse transcription polymerase chain reaction (RT-PCR) offer expedient turnaround times of 10-15 min and straightforward implementation. This preliminary study assessed the correlation between outcomes of commercially available rapid antigen tests for home use and conventional reverse transcription polymerase chain reaction (RT-PCR) assays using a limited set of clinical specimens. Patients aged 5-99 years presenting to the emergency department for SARS-CoV-2 testing were eligible for enrollment (n = 5652). Direct PCR and conventional RT-PCR were utilized for the detection of SARS-CoV-2. The entire cohort of 5652 clinical specimens was assessed by both modalities to determine the clinical utility of the direct RT-PCR assay. Timely confirmation of SARS-CoV-2 infection may attenuate viral propagation and guide therapeutic interventions. Additionally, direct RT-PCR as a secondary confirmatory test for at-home rapid antigen test results demonstrated sensitivity comparable to conventional RT-PCR, indicating utility for implementation in laboratories globally, especially in resource-limited settings with constraints on reagents, equipment, and skilled personnel. In summary, direct RT-PCR enables the detection of SARS-CoV-2 with a sensitivity approaching that of conventional RT-PCR while offering expedient throughput and shorter turnaround times. Moreover, direct RT-PCR provides an open-source option for diagnostic laboratories worldwide, particularly in low- and middle-income countries.

Multi-omics profiling of chemotactic characteristics of brain microglia and astrocytoma.

Brain cancer is a deadly disease with low survival rates for over 70 % of patients. Therefore, there is a critical need to develop better treatment methods and strategies to improve patient outcomes. In this study, we explored the tumor microenvironment and discovered unique characteristics of microglia to interact with astrocytoma cells and promote proliferation and migration of collisions. The conditioned medium from the collisions expressed cell chemoattraction and anti-inflammatory responses. To further understand the interactions between microglia and astrocytoma cells, we used flow sorting and protein analysis found that the protein alterations were related to biogenesis in the astrocytoma cells and metabolic processes in the microglia. Both types of cells were involved in binding and activity in cell-cell interactions. Using STRING to demonstrate the protein cross-interaction between the cells. Furthermore, PHB and RDX interact with oncogenic proteins, which were significantly expressed in patients with Glioblastoma Multiforme (GBM) and low-grade glioma (LGG) according to GEPIA. To study the role of RDX in chemoattraction, the inhibitor-NSC668394 suppressed collision formation and migration in BV2 cells in vitro by down-regulating F-actin. Additionally, it suppressed macrophage infiltration in infiltrating islands in vivo of intracranial tumor-bearing mice. These findings provide evidence for the role of resident cells in mediating tumor development and invasiveness and suggest that potential interacting molecules may be a strategy for controlling tumor growth by regulating the infiltration of tumor-associated microglia in the brain tumor microenvironment.

Monitoring algorithm of hospitalized patients in a medical center with SARS-CoV-2 (Omicron variant) infection: clinical epidemiological surveillance and immunological assessment.

Purpose: Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is a major healthcare threat worldwide. Since it was first identified in November 2021, the Omicron (B.1.1.529) variant of SARS-CoV-2 has evolved into several lineages, including BA.1, BA.2-BA.4, and BA.5. SARS-CoV-2 variants might increase transmissibility, pathogenicity, and resistance to vaccine-induced immunity. Thus, the epidemiological surveillance of circulating lineages using variant phenotyping is essential. The aim of the current study was to characterize the clinical outcome of Omicron BA.2 infections among hospitalized COVID-19 patients and to perform an immunological assessment of such cases against SARS-CoV-2. Patients and Methods: We evaluated the analytical and clinical performance of the BioIC SARS-CoV-2 immunoglobulin (Ig)M/IgG detection kit, which was used for detecting antibodies against SARS-CoV-2 in 257 patients infected with the Omicron variant. Results: Poor prognosis was noted in 38 patients, including eight deaths in patients characterized by comorbidities predisposing them to severe COVID-19. The variant-of-concern (VOC) typing and serological analysis identified time-dependent epidemic trends of BA.2 variants emerging in the outbreak of the fourth wave in Taiwan. Of the 257 specimens analyzed, 108 (42%) and 24 (9.3%) were positive for anti-N IgM and IgG respectively. Conclusion: The VOC typing of these samples allowed for the identification of epidemic trends by time intervals, including the B.1.1.529 variant replacing the B.1.617.2 variant. Moreover, antibody testing might serve as a complementary method for COVID-19 diagnosis. The combination of serological testing results with the reverse transcription-polymerase chain reaction cycle threshold value has potential value in disease prognosis, thereby aiding in epidemic investigations conducted by clinicians or the healthcare department.

The application of a novel 5-in-1 multiplex reverse transcriptase-polymerase chain reaction assay for rapid detection of SARS-CoV-2 and differentiation between variants of concern.

OBJECTIVES: We have established a novel 5-in-1 VOC assay to rapidly detect SARS-CoV-2 and immediately distinguish whether positive samples represent variants of concern (VOCs). METHODS: This assay could distinguish among five VOCs: Alpha, Beta, Gamma, Delta, and Omicron, in a single reaction tube. The five variants exhibit different single nucleotide polymorphisms (SNPs) in their viral genome, which can be used to distinguish them. We selected target SNPs in the spike gene, including N501Y, P681R, K417N, and deletion H69/V70 for the assay. RESULTS: The limit of detection of each gene locus was 80 copies per polymerase chain reaction. We observed a high consistency among the results when comparing the performance of our 5-in-1 VOC assay, whole gene sequencing, and the Roche VirSNiP SARS-CoV-2 test in retrospectively analyzing 150 clinical SARS-CoV-2 variant positive samples. The 5-in-1 VOC assay offers an alternative and rapid high-throughput test for most diagnostic laboratories in a flexible sample-to-result platform. CONCLUSION: The assay can also be applied in a commercial platform with the completion of the SARS-CoV-2 confirmation test and identification of its variant within 2.5 hours.

Electromembrane Extraction of Posaconazole for Matrix-Assisted Laser Desorption/Ionization Mass Spectrometric Detection.

A new mode of electromembrane extraction (EME) has been developed for detection via matrix-assisted laser desorption/ionization mass spectrometry (MALDI/MS). Posaconazole, extracted from 8 mL of a 10 mM trifluoroacetic acid solution onto a thin polyvinylidene difluoride (PVDF) membrane, was used as a model analyte. The transport was forced by an electrical potential difference between two electrodes inside the lumen of a hollow fiber and glass tube. Under an application of 80 V, cationic posaconazole in the sample solution moved toward the negative electrode inside the glass tube and was trapped by the PVDF membrane on the side. After 15 min of extraction, 3 µL of α-cyano-4-hydroxycinnamic acid (CHCA) solution was applied on top of the membrane, which was then analyzed by MALDI/MS. Under optimal extraction conditions, the calibration curve of posaconazole was linear over a concentration range of 0.10-100.00 nM. The limit of detection (LOD) at a signal-to-noise ratio of 3 was 0.03 nM with an enhancement factor of 138 for posaconazole. The application of this method to the determination of posaconazole in human serum samples was also successfully demonstrated.

Critical pediatric neurological illness associated with COVID-19 (Omicron BA.2.3.7 variant) infection in Taiwan: immunological assessment and viral genome analysis in tertiary medical center.

OBJECTIVES: Since April 2022, another wave of the Omicron epidemic has struck Taiwanese society, and children with severe neurological complications have been reported frequently. A few cases even developed acute fulminant encephalitis. To investigate the possible causes of the increased incidence of such complications in Taiwan, we reviewed several cases of pediatric patients with severe neurological symptoms. METHODS: We collected the medical records of pediatric patients with COVID-19 infection who presented with severe neurological symptoms. The COVID-19 infection was diagnosed by nasal swab reverse transcriptase-polymerase chain reaction. The remaining samples were sent for whole genome sequencing and spike (S) protein amino acid variation mapping. RESULTS: The increase of several inflammatory markers was observed in all patients included in this study. However, none of the cerebrospinal fluid samples tested positive for SARS-CoV-2. The result of whole genome sequencing showed that all the sequences belonged to the lineage BA.2.3.7. However, the sequences had a K97E mutation in the S protein that differed from other BA.2.3.7 lineage strains, which was located at the S protein N-terminal domain. CONCLUSION: The new mutation in the S protein, which had not previously been observed but was discovered in this study, potentially explains the sudden increase in incidence of extremely adverse neurological symptoms in pediatric patients.

Cellular senescence as a driver of cognitive decline triggered by chronic unpredictable stress.

When an individual is under stress, the undesired effect on the brain often exceeds expectations. Additionally, when stress persists for a long time, it can trigger serious health problems, particularly depression. Recent studies have revealed that depressed patients have a higher rate of brain aging than healthy subjects and that depression increases dementia risk later in life. However, it remains unknown which factors are involved in brain aging triggered by chronic stress. The most critical change during brain aging is the decline in cognitive function. In addition, cellular senescence is a stable state of cell cycle arrest that occurs because of damage and/or stress and is considered a sign of aging. We used the chronic unpredictable stress (CUS) model to mimic stressful life situations and found that, compared with nonstressed control mice, CUS-treated C57BL/6 mice exhibited depression-like behaviors and cognitive decline. Additionally, the protein expression of the senescence marker p16INK4a was increased in the hippocampus, and senescence-associated ß-galactosidase (SA-ß-gal)-positive cells were found in the hippocampal dentate gyrus (DG) in CUS-treated mice. Furthermore, the levels of SA-ß-gal or p16INK4a were strongly correlated with the severity of memory impairment in CUS-treated mice, whereas clearing senescent cells using the pharmacological senolytic cocktail dasatinib plus quercetin (D + Q) alleviated CUS-induced cognitive deficits, suggesting that targeting senescent cells may be a promising candidate approach to study chronic stress-induced cognitive decline. Our findings open new avenues for stress-related research and provide new insight into the association of chronic stress-induced cellular senescence with cognitive deficits.

Influence of background music on work attention in clients with chronic schizophrenia.

BACKGROUND: Work attention in persons with chronic schizophrenia is an important issue in vocational rehabilitation. Some of the research literature indicates that background music may influence visual attention performance. OBJECTIVES: Based on the theory of occupational therapy, environmental sounds, colors and decorations may affect individual performance, this study thus examined the influence of music on work attention in persons with schizophrenia. PARTICIPANTS: Participants were recruited from a halfway house in Taipei. Forty-nine (49) patients with chronic schizophrenia volunteered. They had been accepted into vocational rehabilitation and a work-seeking program. The sample included 20 females and 29 males. The participant ages ranged between 29 and 63 years old, and their average age was 47 years old. METHODS: Using a randomized controlled trial (RCT) study, the participants were assigned to one of three conditions: quiet environment as the control group (n= 16), classical light music as background music (n= 16), and popular music as background music (n= 17). RESULTS: For Group 1 (control group/quiet environment), there was no significant variance (sig = 0.172). For Group 2 (Classical light music), the intervention revealed significant variance (sig = 0.071*). For Group 3 (popular music), the intervention had significant variance (sig = 0.048**). CONCLUSIONS: The introduction of background music tended to increase attention test scores of persons with schizophrenia. Moreover, the increase in test attention scores was statistically significant when popular music was played in the background. This result suggested that background music may improve attention performance of persons with chronic schizophrenia. Future research is required with a larger sample size to support the study results.

1,2-Bridged Tricyclic Cyclopropenes: Tricyclo[3.2.1.0(2,4)]octa-2(4),6-diene and Tricyclo[3.2.1.0(2,4)]oct-2(4)-ene.

Two 1,2-bridged tricyclic cyclopropenes, tricyclo[3.2.1.0(2,4)]oct-2(4)-ene (6) and tricyclo[3.2.1.0(2,4)]octa-2(4),6-diene (7), have been synthesized by elimination of 2-bromo-4-chlorotricyclo[3.2.1.0(2,4)]octane (17) and 2-bromo-4-chlorotricyclo[3.2.1.0(2,4)]oct-6-ene (8), respectively. Both 6 and 7 were trapped with diphenylisobenzofuran to form two isomers: exo-addition of cyclopropenes and exo-addition of bicyclo[2.2.1]heptenes (exo-exo adducts) and endo-addition of cyclopropenes and exo-addition of bicyclo[2.2.1]heptenes (endo-exo adducts). The stereoselectivity of the Diels-Alder reactions of 6 and 7 with DPIBF is different. The exo-exo/endo-exo ratios of 6 and 7 with DPIBF are 2/1 and 1/2, respectively. Both exo-exo adducts 12 and 18 are stable at refluxing chloroform temperature either with or without DPIBF, but endo-exo adducts 15 and 22 are unstable at room temperature and either isomerize to styrenes 13 and 19 or react with oxygen in the absence of catalyst to generate epoxides 14 and 20. Both styrenes 13 and 19 can be photooxidized by oxygen to give epoxides 14 and 20.