Hyperactive CDK2 Activity in Basal-like Breast Cancer Imposes a Genome Integrity Liability that Can Be Exploited by Targeting DNA Polymerase ε.

Knowledge of fundamental differences between breast cancer subtypes has driven therapeutic advances; however, basal-like breast cancer (BLBC) remains clinically intractable. Because BLBC exhibits alterations in DNA repair enzymes and cell-cycle checkpoints, elucidation of factors enabling the genomic instability present in this subtype has the potential to reveal novel anti-cancer strategies. Here, we demonstrate that BLBC is especially sensitive to suppression of iron-sulfur cluster (ISC) biosynthesis and identify DNA polymerase epsilon (POLE) as an ISC-containing protein that underlies this phenotype. In BLBC cells, POLE suppression leads to replication fork stalling, DNA damage, and a senescence-like state or cell death. In contrast, luminal breast cancer and non-transformed mammary cells maintain viability upon POLE suppression but become dependent upon an ATR/CHK1/CDC25A/CDK2 DNA damage response axis. We find that CDK1/2 targets exhibit hyperphosphorylation selectively in BLBC tumors, indicating that CDK2 hyperactivity is a genome integrity vulnerability exploitable by targeting POLE.

Serum thrombospondin-2 level changes with liver stiffness improvement in patients with type 2 diabetes.

OBJECTIVE: Baseline circulating thrombospondin-2 (TSP2) level was identified as a potential novel hepatic fibrosis biomarker that associates with development and progression of hepatic fibrosis in patients with nonalcoholic fatty liver disease and type 2 diabetes. Here, we investigated whether circulating TSP2 levels changed with improvement in liver stiffness (LS), which reflects liver fibrosis on transient elastography. DESIGN: Serum TSP2 levels were measured in participants from a randomized, open-label intervention study, at baseline and after 24-weeks treatment of either dapagliflozin 10 mg (N = 30) or sitagliptin 100 mg daily (N = 30). Vibration-controlled transient elastography was performed to evaluate the severity of hepatic fibrosis and steatosis using LS and controlled attenuation parameter (CAP), respectively. PATIENTS AND MEASUREMENTS: Among all 60 participants with similar clinical characteristics at baseline (mean HbA1c 8.9%, CAP 289 dB/m and LS 5.8 kPa), despite similar HbA1c lowering, treatment with dapagliflozin, but not sitagliptin, led to significant improvements in body weight (BW) (p = .012), CAP (p = .015) and LS (p = .011) after 24 weeks. RESULTS: Serum TSP2 level decreased significantly from baseline in dapagliflozin-treated participants (p = .035), whereas no significant change was observed with sitagliptin. In correlation analysis, change in serum TSP2 levels only positively correlated with change in LS (r = .487, p = .006), but not with changes in BW, CAP or HbA1c after dapagliflozin treatment. CONCLUSIONS: Serum TSP2 level decreased with LS after dapagliflozin treatment, and was independent of improvements in BW, glycemic control and hepatic steatosis, further supporting the potential of serum TSP2 level as a novel hepatic fibrosis biomarker in type 2 diabetes.

Diagnostic accuracy of non-invasive tests to screen for at-risk MASH-An individual participant data meta-analysis.

BACKGROUND & AIMS: There is a need to reduce the screen failure rate (SFR) in metabolic dysfunction-associated steatohepatitis (MASH) clinical trials (MASH+F2-3; MASH+F4) and identify people with high-risk MASH (MASH+F2-4) in clinical practice. We aimed to evaluate non-invasive tests (NITs) screening approaches for these target conditions. METHODS: This was an individual participant data meta-analysis for the performance of NITs against liver biopsy for MASH+F2-4, MASH+F2-3 and MASH+F4. Index tests were the FibroScan-AST (FAST) score, liver stiffness measured using vibration-controlled transient elastography (LSM-VCTE), the fibrosis-4 score (FIB-4) and the NAFLD fibrosis score (NFS). Area under the receiver operating characteristics curve (AUROC) and thresholds including those that achieved 34% SFR were reported. RESULTS: We included 2281 unique cases. The prevalence of MASH+F2-4, MASH+F2-3 and MASH+F4 was 31%, 24% and 7%, respectively. Area under the receiver operating characteristics curves for MASH+F2-4 were .78, .75, .68 and .57 for FAST, LSM-VCTE, FIB-4 and NFS. Area under the receiver operating characteristics curves for MASH+F2-3 were .73, .67, .60, .58 for FAST, LSM-VCTE, FIB-4 and NFS. Area under the receiver operating characteristics curves for MASH+F4 were .79, .84, .81, .76 for FAST, LSM-VCTE, FIB-4 and NFS. The sequential combination of FIB-4 and LSM-VCTE for the detection of MASH+F2-3 with threshold of .7 and 3.48, and 5.9 and 20 kPa achieved SFR of 67% and sensitivity of 60%, detecting 15 true positive cases from a theoretical group of 100 participants at the prevalence of 24%. CONCLUSIONS: Sequential combinations of NITs do not compromise diagnostic performance and may reduce resource utilisation through the need of fewer LSM-VCTE examinations.

A robust genetic algorithm-based optimal feature predictor model for brain tumour classification from MRI data.

Brain tumour can be cured if it is initially screened and given timely treatment to the patients. This proposed idea suggests a transform- and windowing-based optimization strategy for exposing and segmenting the tumour region in brain pictures. The processes of image processing that are included in the proposed idea include preprocessing, transformation, feature extraction, feature optimization, classification, and segmentation. In order to convert the pixels connected to the spatial domain into a multi-resolution domain, the Gabor transform is first applied to the brain test image. The Gabor converted brain image is then used to extract the parameters of the multi-level features. After that, the Genetic Algorithm (GA) is used to optimize the extracted features, and Neuro Fuzzy System (NFS) is used to classify the optimistic prominent section. Finally, the tumour region in brain images is found and segmented using the normalized segmentation algorithm. The effective detection and classification of brain tumours by the characteristics of sensitivity, specificity, and accuracy are described by the suggested GA-based NFS classification approach. The trial findings are displayed with an average of 99.37% sensitivity, 98.9% specificity, 99.21% accuracy, 97.8% PPV, 91.8% NPV, 96.8% FPR, and 90.4% FNR.

Banana peel extract for CeO2 nanoflower synthesis: Enhancing photocatalytic activity for methyl orange dye removal and bactericidal effects.

The cube architecture associated with the CeO2 nanoflowers (NFs) that generated, which had an average crystallization width of 7 nm, has been confirmed by X-ray crystallographic investigations. The method used is environmentally acceptable since it converts wasted banana peel extracts into CeO2 nanoflower. On the basis of artwork obtained from a High-Resolution Transmission Electron Microscope (HR-TEM), CeO2 nanoparticles have been observed to possess a spherical shape and an average particle diameter of 21 nm. To take the purpose of this study, green-fabricated CeO2-NFs were used to investigate the photocatalytic oxidation of methyl orange (MO) dye when exposed to sunshine. CeO2 nanofibers showed a degradation performance of 98% when compared to methyl orange dye. Evidently is a possibility that this may be caused by the presence of CeO2 nanoflowers, whereby enhance the interaction of electrons, which are holes dissolution, and adherence. Upon a single day of being exposed, the biocidal potential was tested against both gram-positive and gram-negative bacteria, including E. coli, B. cereus, and S. aureus, among others. Due to the fact that its 32 mm minimum inhibitory concentration (MIC) for B. cereus was the highest among conventional medicines. As shown by the extraordinary capabilities of WBP@CeO2 tiny particles, manipulating of flexible tiny particles to feed the purpose of achieving effective and customizable infections and dermatologist advancements is really stunning.

Overall and subgroup prevalence of non-alcoholic fatty liver disease and prevalence of advanced fibrosis in the United States: An updated national estimate in National Health and Nutrition Examination Survey (NHANES) 2011-2018.

INTRODUCTION AND OBJECTIVES: Data on the prevalence of non-alcoholic fatty liver disease (NAFLD) in subgroups of the United States (US) population are limited. This study was conducted to estimate NAFLD prevalence overall and by subgroups, and prevalence of NAFLD with advanced fibrosis. MATERIALS AND METHODS: Using the National Health and Nutrition Examination Survey (NHANES) 2011-2018 data, a cross-sectional study was conducted. NAFLD was defined as having a US Fatty Liver Index (USFLI) ≥ 30 in the absence of other causes of liver disease, including excessive alcohol intake, chronic hepatitis B, and chronic hepatitis C. Likelihood for having advanced fibrosis was determined by the calculated NAFLD fibrosis score (NFS; high ≥ 0.676; low < -1.445) and fibrosis-4 index (FIB-4; high ≥ 2.67; low < 1.30). RESULTS: The weighted national prevalence of NAFLD in US adults was 26.7% (95% confidence interval: 25.3%-28.1%). Prevalence was higher among those aged ≥ 65 years, males, Mexican Americans, with BMI ≥ 35 kg/m2 (class 2 and 3 obesity) and with type 2 diabetes (T2D). Of those meeting the USFLI criterion for NAFLD, 18.1% and 3.7% were determined as having a high probability of advanced fibrosis based on NFS ≥ 0.676 and FIB-4 ≥ 2.67 cut-off values, respectively. CONCLUSIONS: This study supports an increased prevalence of NAFLD in specific subpopulations (aged ≥ 65 years, males, Mexican Americans, obese population, and patients with T2D). The observed difference in the prevalence of advanced fibrosis as estimated by NFS and FIB-4 highlights the challenge of choosing optimal cut-off values.

Synergistic Enhancement of CO2 photoreduction through sulfur defects in (3D/2D) CdS-nanoflowers/CN Binary heterojunction photocatalyst under visible light.

Global warming is the biggest threat to the entire world owing to the continuous release of greenhouse gases such as CO2 from various sources. Herein, we have utilized renewable energy for the conversion of CO2 to valuable feedstocks through a semiconductor-mediated photocatalytic system. The cadmium sulfide nanoflowers (CS-NFs) decorated graphitic carbon nitride (CN) through a solvothermal route to form a Z-scheme CSCN heterojunction. The as-synthesized material has been characterized by various spectroscopic and microscopic tools. The optimal CSCN-0.5 (1:0.5) photocatalyst achieves a CO production rate of 130.9 µmol g-1 under visible light irradiation of 4h (λ > 420 nm), doubling that of pristine CS-NFs and CN. CO, along with CH4 (3.4 µmol g-1) and C2H6 (2.9 µmol g-1), is the sole product detected. Experimental results indicate that the CSCN-0.5 photocatalyst spatially separates electron-hole pairs, suppresses charge carrier recombination, and maintains robust redox ability, enhancing CO2 photoreduction. The CO2 reduction mechanism over CSCN heterojunction was also studied through in-situ DRIFTS and electron spin resonance (ESR) measurements. Therefore, CSCN proves that it could be used as a robust photocatalyst for the CO2 reduction reactions towards C1 and C2 feedstocks.

Pioglitazone treatment increases the cellular acid-labile and protein-bound sulfane sulfur fractions.

BACKGROUND: Iron-sulfur clusters play a central role in cellular function and are regulated by the ATM protein. Iron-sulfur clusters are part of the cellular sulfide pool, which functions to maintain cardiovascular health, and consists of free hydrogen sulfide, iron-sulfur clusters, protein bound sulfides, which constitute the total cellular sulfide fraction. ATM protein signaling and the drug pioglitazone share some cellular effects, which led us to examine the effects of this drug on cellular iron-sulfur cluster formation. Additionally, as ATM functions in the cardiovasculature and its signaling may be diminished in cardiovascular disease, we examined pioglitazone in the same cell type, with and without ATM protein expression. METHODS: We examined the effects of pioglitazone treatment on the total cellular sulfide profile, the glutathione redox state, cystathionine gamma-lyase enzymatic activity, and on double-stranded DNA break formation in cells with and without ATM protein expression. RESULTS: Pioglitazone increased the acid-labile (iron-sulfur cluster) and bound sulfur cellular fractions and reduced cystathionine gamma-lyase enzymatic activity in cells with and without ATM protein expression. Interestingly, pioglitazone also increased reduced glutathione and lowered DNA damage in cells without ATM protein expression, but not in ATM wild-type cells. These results are interesting as the acid-labile (iron-sulfur cluster), bound sulfur cellular fractions, and reduced glutathione are low in cardiovascular disease. CONCLUSION: Here we found that pioglitazone increased the acid-labile (iron-sulfur cluster) and bound sulfur cellular fractions, impinges on hydrogen sulfide synthesis, and exerts beneficial effect on cells with deficient ATM protein signaling. Thus, we show a novel pharmacologic action for pioglitazone.

Prognostic value of non-alcoholic fatty liver disease fibrosis score in patients undergoing cardiac resynchronization therapy.

BACKGROUND: Cardiac resynchronization therapy (CRT) is an effective option in the treatment of patients with heart failure and wide QRS. Non-alcoholic fatty liver disease (NAFLD) fibrosis score (NFS) has been shown to predict cardiac events in several patient populations. However, the relationship between NFS and response to CRT has not been investigated. The aim of the study was to investigate the predictive role of NFS in the assessment of response after CRT. METHODS: Three hundred thirty-six patients with heart failure undergoing CRT were prospectively studied. Liver fibrosis were assessed according to the non-alcoholic fatty liver disease fibrosis score (NFS), which includes age, body mass index, impaired fasting glycemia or diabetes mellitus, aspartate aminotransferase /alanine aminotransferase ratio, platelets, and albumin. Echocardiographic response to CRT was defined by a ≥15% reduction in left ventricular end-systolic volume at six months at follow-up. RESULTS: Two hundred thirty-eight patients (71%) had CRT response after 6 months of follow-up. Receiver-operator characteristic curve analysis showed NFS cutoff value of < -1.12 for predicting CRT response with a sensitivity of 70.4% and a specificity of 52.9%. The patients were also divided into four groups according to the quartiles of NFS. The proportion of response to CRT was increased with lower level of NFS value. Multivariate logistic regression analysis demonstrated the NFS score < -1.12 and LVIDs were independent predictors of the CRT response. In the second model of analysis which included NFS, quartiles demonstrated that fourth NFS quartile and LVIDs were independent predictors of CRT response. CONCLUSION: Liver fibrosis assessed by NFS can provide valuable information to predict reverse remodeling in patients undergoing CRT. The present study supports monitoring of NFS to improve preoperative risk stratification of these patients.

Gaps in Confirmatory Fibrosis Risk Assessment in Primary Care Patients with Nonalcoholic Fatty Liver Disease.

BACKGROUND: As recommendations for non-invasive fibrosis risk assessment in nonalcoholic fatty liver disease (NAFLD) emerge, it is not known how often they are performed in primary care. AIMS: We investigated the completion of confirmatory fibrosis risk assessment in primary care patients with NAFLD and indeterminate-risk or greater Fibrosis-4 Index (FIB-4) and NAFLD Fibrosis Scores (NFS). METHODS: This retrospective cohort study of electronic health record data from a primary care clinic identified patients with diagnoses of NAFLD from 2012 through 2021. Patients with a diagnosis of a severe liver disease outcome during the study period were excluded. The most recent FIB-4 and NFS scores were calculated and categorized by advanced fibrosis risk. Charts were reviewed to identify the outcome of a confirmatory fibrosis risk assessment by liver elastography or liver biopsy for all patients with indeterminate-risk or higher FIB-4 (≥ 1.3) and NFS (≥ - 1.455) scores. RESULTS: The cohort included 604 patients diagnosed with NAFLD. Two-thirds of included patients (399) had a FIB-4 or NFS score greater than low-risk, 19% (113) had a high-risk FIB-4 (≥ 2.67) or NFS (≥ 0.676) score, and 7% (44) had high-risk FIB-4 and NFS values. Of these 399 patients with an indication for a confirmatory fibrosis test, 10% (41) underwent liver elastography (24) or liver biopsy (18) or both (1). CONCLUSIONS: Advanced fibrosis is a key indicator of future poor health outcomes in patients with NAFLD and a critical signal for referral to hepatology. Significant opportunities exist to improve confirmatory fibrosis risk assessment in patients with NAFLD.

Electrochemical nano-biosensor based on electrospun indium zinc oxide nanofibers for the determination of complement component 3 protein.

Age-related macular degeneration (AMD) is a progressive chronic neurodegenerative retinal disease leading to vision loss, irreversible blindness, and visual impairment in older adults worldwide. Complement component 3 (C3) protein has been identified as the most predominant biomarker towards early diagnosis of AMD; therefore, there is an utmost requirement for non-invasive detection of C3 protein in the tear fluids of AMD patients. Considering this, we report an insightful electrochemical sensor capable of detecting clinically relevant concentrations ranging from 10 fg/mL to 1 µg/mL using electrospun indium-doped zinc oxide (InZnO) nanofibers as the transducing layer. The InZnO nanofibers have facilitated high anti-C3 antibody loading of 3.42 × 10-9 mol/cm2 and enhanced the overall charge transport mechanism at the sensor interface. The biofunctionalization process of the biosensor was investigated thoroughly using X-ray photoelectron spectroscopy (XPS) as well as different electrochemical techniques. The target C3 proteins were captured on the fabricated biosensor surface and determined through changes in charge transfer resistance (RCT) while executing electrochemical impedance spectroscopy (EIS) and peak current (Ip) in the case of cyclic voltammetry (CV) and differential pulse voltammetry (DPV), respectively. The InZnO nanofiber-based nano-biosensor demonstrated a very low limit of detections (LODs) of 5.214 fg/mL and 0.241 fg/mL with an excellent sensitivity of 4.6709 (ΔR/R) (g/mL)-1 cm-2 and 54.4939 (ΔIp/Ip)% (g/mL)-1 cm-2 for EIS and DPV techniques, respectively. By virtue of high antibody loading, ultrasensitive and ultra-selective capability, the indium-doped ZnO nanofibers show huge potential to be used as a high-performance diagnostic platform for AMD diagnosis.

Clinical Implications of Cardiac Symptoms and Electrocardiographic Abnormalities for Advanced Liver Fibrosis in Patients with Nonalcoholic Fatty Liver Disease.

Background and Objectives: Advanced liver fibrosis in patients with nonalcoholic fatty liver disease (NAFLD) can be a major predictor of cardiovascular disease (CVD) events and cardiac complications. However, the clinical significance of cardiac symptoms and abnormal electrocardiography (ECG) findings in patients with NAFLD associated with advanced liver fibrosis is unclear. Therefore, our study was aimed to evaluate the clinical implications based on the association between cardiac symptoms with ECG abnormalities for advanced liver fibrosis in patients with NAFLD. Materials and Methods: Of 31,795 participants who underwent health checkups, 6293 were diagnosed with NAFLD using ultrasound and inclusion criteria in a retrospective cross-sectional study. Advanced liver fibrosis was assessed based on a low NAFLD fibrosis score (NFS) and fibrosis-4 index (Fib-4) cut-off values (COVs). Cardiac data were assessed using a cardiac symptom questionnaire and 12-lead electrocardiography (ECG). Results: Among 6293 NAFLD patients with NAFLD, 304 (4.8%) experienced cardiac symptoms. NFS and Fib-4 indicated higher rates of advanced fibrosis in the cardiac-symptomatic group than in the non-symptomatic group (NFS: 7.3 vs. 4.1%; Fib-4: 7.8 vs. 3.7%; both p < 0.001). Cardiac symptoms were independently associated with advanced liver fibrosis using a step-wise-adjusted model and NFS and Fib-4 (final adjusted odds ratio (aOR), 1.40; 95% CI, 1.06-1.85; p = 0.018 for NFS; aOR, 1.67; 95%, 1.30-2.15; p < 0.001 for Fib-4). Cardiac symptoms with abnormal ECG findings independently predicted advanced liver fibrosis (aOR, 2.43; 95% CI, 1.72-3.39; p < 0.001 for NFS; aOR, 3.02; 95% CI, 2.19-4.15; p < 0.001 for Fib-4). Conclusions: Patients who have had cardiac symptoms and some ECG abnormalities may have a higher association with advanced liver fibrosis.

Expanding the phenotypic and molecular spectrum of NFS1-related disorders that cause functional deficiencies in mitochondrial and cytosolic iron-sulfur cluster containing enzymes.

Iron-sulfur cluster proteins are involved in critical functions for gene expression regulation and mitochondrial bioenergetics including the oxidative phosphorylation system. The c.215G>A p.(Arg72Gln) variant in NFS1 has been previously reported to cause infantile mitochondrial complex II and III deficiency. We describe three additional unrelated patients with the same missense variant. Two infants with the same homozygous variant presented with hypotonia, weakness and lactic acidosis, and one patient with compound heterozygous p.(Arg72Gln) and p.(Arg412His) variants presented as a young adult with gastrointestinal symptoms and fatigue. Skeletal muscle biopsy from patients 1 and 3 showed abnormal mitochondrial morphology, and functional analyses demonstrated decreased activity in respiratory chain complex II and variably in complexes I and III. We found decreased mitochondrial and cytosolic aconitase activities but only mildly affected lipoylation of pyruvate dehydrogenase and 2-oxoglutarate dehydrogenase enzymes. Our studies expand the phenotypic spectrum and provide further evidence for the pathogenicity and functional sequelae of NFS1-related disorders with disturbances in both mitochondrial and cytosolic iron-sulfur cluster containing enzymes.

An Optical Sensing Platform for Beta-Glucosidase Activity Using Protein-Inorganic Hybrid Nanoflowers.

In this work, a convenient and dual-signal readout optical sensing platform for the sensitively and selectively determination of beta-glucosidase (ß-Glu) activity was reported using protein-inorganic hybrid nanoflowers [BSA-Cu3(PO4)2·3H2O] possessing peroxidase-mimicking activity. The nanoflowers (NFs) were facilely synthesized through a self-assembled synthesis strategy at room temperature. The as-prepared NFs could catalytically convert the colorless and non-fluorescent Amplex Red into colored and highly fluorescent resorufin in the presence of hydrogen peroxide via electron transfer process. ß-Glu could hydrolyze cyanogenic glycoside, using amygdalin (Amy) as a model, into cyanide ions (CN-), which can subsequently efficiently suppress the catalytic activity of NFs, accompanied with the fluorescence decrease and the color fading. The concentration of CN- was controlled by ß-Glu-triggered enzymatic reaction of Amy. Thus, a sensing system was established for fluorescent and visual determination of ß-Glu activity. Under the optimum conditions, the present fluorescent and visual bimodal sensing platform exhibited good sensitivity for ß-Glu activity assay with a detection limit of 0.33 U·L-1. The sensing platform was further applied to determinate ß-Glu in real samples and satisfactory results were attained. Additionally, the optical sensing system can potentially be a promising candidate for ß-Glu inhibitors screening.

Overexpression of TGF-ß1 and SDF-1 in cervical cancer-associated fibroblasts promotes cell growth, invasion and migration.

OBJECTIVE: The purpose of this study was to investigate the effect of overexpression of transforming growth factor ß1 (TGF-ß1) and stromal cell-derived factor 1 (SDF-1) in cervical cancer-associated fibroblasts (CAFs) on regulating cell growth, invasion and migration. METHODS: CAF cells and normal fibroblast cells (NFs) were obtained from patients with cervical squamous cell carcinoma and multiple uterine leiomyomas, respectively. Immunofluorescence assay and western blot were used to determine the expression of Vimentin and α-smooth muscle actin (α-SMA). CCK-8 assay was used to detect cell viability. Giemsa dyer was used to detect the colony formation. Flow cytometry was used to detect the growth state of cells. Transwell assays were used to detect the migration and invasion. RESULTS: Vimentin and α-SMA expression in CAFs were significantly increased than those in NFs. In addition, TGF-ß1 and SDF-1 expression were notably increased, and transforming growth factor beta receptor 2 (TßRII) expression was markedly decreased in CAF cells than those in NFs. Similarly, TGF-ß1 and SDF-1 expression in the co-culture of CAFs and Hela cells were significantly increased, and cell proliferation, migration, invasion, colony formation and cell cycle progression were also promoted, while cell apoptosis was decreased. Those phenomena were reversed in the co-culture system with neutralizing antibodies to TGF-ß1 and SDF-1. Furthermore, exogenous TGF-ß1 and SDF-1 enhanced proliferation, colony formation, cell cycle progression, migration and invasion while decreased apoptosis of cells. These phenomena were also reversed by the addition of neutralizing antibodies to TGF-ß1 and SDF-1. CONCLUSION: Overexpression of TGF-ß1 and SDF-1 in CAFs can promote the growth, invasion and migration of cervical cancer cells.

Efficient NFS Model for Risk Estimation in a Risk-Based Access Control Model.

Providing a dynamic access control model that uses real-time features to make access decisions for IoT applications is one of the research gaps that many researchers are trying to tackle. This is because existing access control models are built using static and predefined policies that always give the same result in different situations and cannot adapt to changing and unpredicted situations. One of the dynamic models that utilize real-time and contextual features to make access decisions is the risk-based access control model. This model performs a risk analysis on each access request to permit or deny access dynamically based on the estimated risk value. However, the major issue associated with building this model is providing a dynamic, reliable, and accurate risk estimation technique, especially when there is no available dataset to describe risk likelihood and impact. Therefore, this paper proposes a Neuro-Fuzzy System (NFS) model to estimate the security risk value associated with each access request. The proposed NFS model was trained using three learning algorithms: Levenberg-Marquardt (LM), Conjugate Gradient with Fletcher-Reeves (CGF), and Scaled Conjugate Gradient (SCG). The results demonstrated that the LM algorithm is the optimal learning algorithm to implement the NFS model for risk estimation. The results also demonstrated that the proposed NFS model provides a short and efficient processing time, which can provide timeliness risk estimation technique for various IoT applications. The proposed NFS model was evaluated against access control scenarios of a children's hospital, and the results demonstrated that the proposed model can be applied to provide dynamic and contextual-aware access decisions based on real-time features.

In Vitro Effect of Δ9-Tetrahydrocannabinol and Cannabidiol on Cancer-Associated Fibroblasts Isolated from Lung Cancer.

There is evidence that demonstrates the effect of cannabinoid agonists inhibiting relevant aspects in lung cancer, such as proliferation or epithelial-to-mesenchymal transition (EMT). Most of these studies are based on evidence observed in in vitro models developed on cancer cell lines. These studies do not consider the complexity of the tumor microenvironment (TME). One of the main components of the TME is cancer-associated fibroblasts (CAFs), cells that are relevant in the control of proliferation and metastasis in lung cancer. In this work, we evaluated the direct effects of two cannabinoid agonists, tetrahydrocannabinol (THC) and cannabidiol (CBD), used alone or in combination, on CAFs and non-tumor normal fibroblasts (NFs) isolated from adenocarcinoma or from healthy lung tissue from the same patients. We observed that these compounds decrease cell density in vitro and inhibit the increase in the relative expression of type 1 collagen (COL1A1) and fibroblast-specific protein 1 (FSP1) induced by transforming growth factor beta (TGFß). On the other hand, we studied whether THC and CBD could modulate the interactions between CAFs or NFs and cancer cells. We conditioned the culture medium with stromal cells treated or not with THC and/or CBD and cultured A549 cells with them. We found that culture media conditioned with CAFs or NFs increased cell density, induced morphological changes consistent with EMT, inhibited cadherin-1 (CDH1) gene expression, and induced an increase in the relative expression of cadherin-2 (CDH2) and vimentin (VIM) genes in A549 cells. These changes were inhibited or decreased by THC and CBD administered alone or in combination. In another series of experiments, we conditioned culture media with A549 cells treated or not with THC and/or CBD, in the presence or absence of TGFß. We observed that culture media conditioned with A549 in the presence of TGFß induced an increase in the expression of COL1A1 and VIM, both in CAFs and in non-tumor NFs. Both THC and CBD ameliorated these effects. In summary, the results presented here reinforce the usefulness of cannabinoid agonists for the treatment of some relevant aspects of lung cancer pathology, and demonstrate in a novel way their possible effects on CAFs as a result of their relationship with cancer cells. Likewise, the results reinforce the usefulness of the combined use of THC and CBD, which has important advantages in relation to the possibility of using lower doses, thus minimizing the psychoactive effects of THC.

Identification of the Wzx flippase, Wzy polymerase and sugar-modifying enzymes for spore coat polysaccharide biosynthesis in Myxococcus xanthus.

The rod-shaped cells of Myxococcus xanthus, a Gram-negative deltaproteobacterium, differentiate to environmentally resistant spores upon starvation or chemical stress. The environmental resistance depends on a spore coat polysaccharide that is synthesised by the ExoA-I proteins, some of which are part of a Wzx/Wzy-dependent pathway for polysaccharide synthesis and export; however, key components of this pathway have remained unidentified. Here, we identify and characterise two additional loci encoding proteins with homology to enzymes involved in polysaccharide synthesis and export, as well as sugar modification and show that six of the proteins encoded by these loci are essential for the formation of environmentally resistant spores. Our data support that MXAN_3260, renamed ExoM and MXAN_3026, renamed ExoJ, are the Wzx flippase and Wzy polymerase, respectively, responsible for translocation and polymerisation of the repeat unit of the spore coat polysaccharide. Moreover, we provide evidence that three glycosyltransferases (MXAN_3027/ExoK, MXAN_3262/ExoO and MXAN_3263/ExoP) and a polysaccharide deacetylase (MXAN_3259/ExoL) are important for formation of the intact spore coat, while ExoE is the polyisoprenyl-phosphate hexose-1-phosphate transferase responsible for initiating repeat unit synthesis, likely by transferring N-acetylgalactosamine-1-P to undecaprenyl-phosphate. Together, our data generate a more complete model of the Exo pathway for spore coat polysaccharide biosynthesis and export.

Long-term outcomes and predictive ability of non-invasive scoring systems in patients with non-alcoholic fatty liver disease.

BACKGROUND & AIMS: Non-invasive scoring systems (NSS) are used to identify patients with non-alcoholic fatty liver disease (NAFLD) who are at risk of advanced fibrosis, but their reliability in predicting long-term outcomes for hepatic/extrahepatic complications or death and their concordance in cross-sectional and longitudinal risk stratification remain uncertain. METHODS: The most common NSS (NFS, FIB-4, BARD, APRI) and the Hepamet fibrosis score (HFS) were assessed in 1,173 European patients with NAFLD from tertiary centres. Performance for fibrosis risk stratification and for the prediction of long-term hepatic/extrahepatic events, hepatocarcinoma (HCC) and overall mortality were evaluated in terms of AUC and Harrell's c-index. For longitudinal data, NSS-based Cox proportional hazard models were trained on the whole cohort with repeated 5-fold cross-validation, sampling for testing from the 607 patients with all NSS available. RESULTS: Cross-sectional analysis revealed HFS as the best performer for the identification of significant (F0-1 vs. F2-4, AUC = 0.758) and advanced (F0-2 vs. F3-4, AUC = 0.805) fibrosis, while NFS and FIB-4 showed the best performance for detecting histological cirrhosis (range AUCs 0.85-0.88). Considering longitudinal data (follow-up between 62 and 110 months), NFS and FIB-4 were the best at predicting liver-related events (c-indices>0.7), NFS for HCC (c-index = 0.9 on average), and FIB-4 and HFS for overall mortality (c-indices >0.8). All NSS showed limited performance (c-indices <0.7) for extrahepatic events. CONCLUSIONS: Overall, NFS, HFS and FIB-4 outperformed APRI and BARD for both cross-sectional identification of fibrosis and prediction of long-term outcomes, confirming that they are useful tools for the clinical management of patients with NAFLD at increased risk of fibrosis and liver-related complications or death. LAY SUMMARY: Non-invasive scoring systems are increasingly being used in patients with non-alcoholic fatty liver disease to identify those at risk of advanced fibrosis and hence clinical complications. Herein, we compared various non-invasive scoring systems and identified those that were best at identifying risk, as well as those that were best for the prediction of long-term outcomes, such as liver-related events, liver cancer and death.

Advanced liver fibrosis and the metabolic syndrome in a primary care setting.

AIMS: The fibrosis-4 index (FIB-4) and NAFLD fibrosis score (NFS) are noninvasive and accessible methods for assessing advanced liver fibrosis risk in primary care. We evaluated the distribution of FIB-4 and NFS scores in primary care patients with clinical signals for nonalcoholic fatty liver disease (NAFLD). MATERIALS AND METHODS: This retrospective cohort study of electronic record data between 2007 and 2018 included adults with at least one abnormal aminotransferase and no known (non-NAFLD) liver disease. We calculated patient-level FIB-4 and NFS scores, the proportion of patients with mean values exceeding advanced fibrosis thresholds (indeterminate risk: FIB-4 > 1.3, NFS > -1.455; high-risk: FIB-4 > 2.67, NFS > 0.676), and the proportion of patients with a NAFLD International Classification of Diseases-9/10 code. Logistic regression models evaluated the associations of metabolic syndrome (MetS) components with elevated FIB-4 and NFS scores. RESULTS: The cohort included 6506 patients with a median of 6 (interquartile range: 3-13) FIB-4 and NFS scores per patient. Of these patients, 81% had at least two components of MetS, 29% had mean FIB-4 and NFS scores for indeterminate fibrosis risk, and 11% had either mean FIB-4 or NFS scores exceeding the high advanced fibrosis risk thresholds. Regression models identified associations of low high-density lipoprotein, hyperglycemia, Black race and male gender with high-risk FIB-4 and NFS values. Only 5% of patients had existing diagnoses for NAFLD identified. CONCLUSIONS: Many primary care patients have FIB-4 and NFS scores concerning for advanced fibrosis, but rarely a diagnosis of NAFLD. Elevated FIB-4 and NFS scores may provide signals for further clinical evaluation of liver disease in primary care settings.