F-box protein 43 promoter methylation as a novel biomarker for hepatitis B virus-associated hepatocellular carcinoma.

Background: Hepatocellular carcinoma (HCC) has a high prevalence and poor prognosis worldwide. Therefore, it is urgent to find effective and timely diagnostic markers. The objective of this study was to evaluate the diagnostic value of F-box protein 43 promoter methylation in peripheral blood mononuclear cells (PBMCs) for HCC. Method: A total of 247 participants were included in this study, comprising individuals with 123 hepatitis B virus-associated HCC, 79 chronic hepatitis B, and 45 healthy controls. F-box protein 43 methylation and mRNA levels in PBMCs were detected by MethyLight and quantitative real-time PCR. Result: F-box protein 43 promoter methylation levels were significantly lower in HCC PBMCs than the chronic hepatitis B (P < 0.001) and healthy control PBMCs (P < 0.001). Relative mRNA expression levels of F-box protein 43 in HCC PBMCs were significantly higher than those in chronic hepatitis B (P < 0.001) and healthy control PBMCs (P < 0.001). Receiver operating characteristic analysis of F-box protein 43 promoter methylation levels yielded an area under curve (AUC) of 0.793 with 76.42% sensitivity and 68.35% specificity when differentiating HCC from chronic hepatitis. These values for the F-box protein 43 promoter methylation level were superior to those of the alpha-fetoprotein serum (AFP) level (AUC: 0.780, sensitivity: 47.97%, and specificity: 96.20%), with increments in values for the combination of F-box protein 43 promoter methylation AFP levels (AUC: 0.888, sensitivity: 76.42%, and specificity: 86.08%). Conclusion: Hypomethylation of the F-box protein 43 promoter in PBMCs is a promising biochemical marker for HBV-associated HCC.

Hypomethylation of Tumor necrosis factor-like cytokine 1A(TL1A) and its decoy receptor 3 expressive level increase has diagnostic value in HBV-associated cirrhosis.

For patients with cirrhosis, early diagnosis is the key to delaying the development of liver fibrosis and improving prognosis. This study aimed to investigate the clinical significance of TL1A, which is a susceptibility gene for hepatic fibrosis, and DR3 in the development of cirrhosis and fibrosis. We analyzed the expression of TL1A, DR3, and other inflammatory cytokines associated with liver fibrosis in serum and PBMCs in 200 patients.TL1A methylation level was lower in patients with HBV-associated LC than in the other groups. In addition, the mRNA level and serum of TL1A and DR3 expression levels were found to increase in the LC. Hypomethylation of the TL1A promoter is present in HBV-associated LC, and TL1A and DR3 are highly expressed in HBV-associated cirrhosis. These results indicate that TL1A and DR3 may play an important role in the pathogenesis of LC and TL1A methylation levels may serve as a noninvasive biomarker for early diagnosis and progression of LC.

Correction: Heller et al. Characterization Methods along the Process Chain of Electrical Steel Sheet-From Best Practices to Advanced Characterization. Materials 2022, 15, 32.

In the original publication [...].

Superoxide dismutase 2 as a predictor in patients with hepatitis B virus-associated acute-on-chronic liver failure.

The prognosis of hepatitis B virus-associated acute-on-chronic liver failure (HBV-ACLF) is critical in clinical management. We aimed to assess the prognostic efficacy of superoxide dismutase 2 (SOD2) for 90-day mortality in HBV-ACLF patients. The expression patterns of SOD2 in peripheral blood mononuclear cells (PBMCs) were examined in a derivation set (n = 82) by quantitative real-time polymerase chain reaction (RT-qPCR). The results were further validated in a validation set (n = 35). The expression levels of SOD2 were significantly decreased in the derivation set compared to those with chronic hepatitis B (CHB) or the healthy controls (HCs) (P < 0.001). In HBV-ACLF patients, SOD2 levels were negatively correlated with serum total bilirubin (TBIL) (rs = - 0.43, P < 0.001) and model for end-stage liver disease (MELD) scores (rs = - 0.22, P = 0.047), but positively correlated with alkaline phosphatase (AKP) (rs = 0.23, P = 0.034). SOD2 was identified as an independent risk factor for 90-day mortality in HBV-ACLF patients (hazard ratio: 0.124, 95% confidence interval: 0.059-0.261, P < 0.001). SOD2 yielded a larger area under the receiver operating characteristic curve (AUROC) than the MELD score in predicting 90-day mortality (0.914 vs. 0.712, P < 0.001). Kaplan-Meier analysis revealed a favorable overall survival (OS) for the SOD2 high expression group compared with the SOD2 low expression group in both the derivation and validation sets (P < 0.001). SOD2 has promising potential as a predictor of 90-day mortality in patients with HBV-ACLF.

Efficient preparation of red mud-based geopolymer microspheres (RM@GMs) and adsorption of fluoride ions in wastewater.

In this paper, red mud-based geopolymer microspheres (RM@GMs: 75-150 µm) was prepared by dispersion-suspension-solidification method to remove fluoride ions (F-). It was found that RM@GMs still had good mechanical properties and better F- removal effect at RM content reached 80 % of the total solid mass. The batch adsorption experiment results showed that the F- concentration (< 1.5 mg/L) reached the drinking water standard in 45 min at pH = 2 and RM@GMs dosage was 1 g/L. RM@GMs showed maximum adsorption capacity of 76.57 mg/g for F-, and the adsorption kinetics and isotherm fitted the pseudo-second-order kinetic and Langmuir isotherm model, respectively. RM@GMs exhibited excellent dynamic separation effect at the flow rate of 4 mL/min and column height of 1 cm. In addition, RM@GMs had good selectivity for F- in the competitive adsorption experiments and followed an order of: PO43- > > SO42- ≈ NO3- ≈ Cl-. In real seawater, natural surface water and tap water, RM@GMs still had excellent F- removal effect. The adsorption mechanism revealed that RM@GMs removed F- mainly through the synergistic effect of adsorption and ion exchange. Therefore, this paper provides the potential value for the large-scale utilization of RM in the application of F--containing wastewater.

Prophenoloxidase-positive tubes derived from the hindguts may be the doorkeeper to detoxify the waste metabolites collected by Malpighian tubules in Lepidoptera insects.

Prophenoloxidase (PPO), an important immunity protein in insects, is mainly produced by hemocytes and released into the hemolymph upon cell lysis. In addition, PPO can also be produced by epidermal cells in the foregut to detoxify the toxic plant secondary metabolites and in the hindgut to kill pathogens through PPO-induced melanization. Previously, we noticed a pair of tubes extended from the larval hindgut became melanized upon staining in dopamine dissolved in 30% ethanol. However, the structure and function of these tubes are largely unknown. In this study, we performed staining of the tubes and the neighboring Malpighian tubule for further confirmation. Eventually, we detected PPO inside epidermal cells of the tubes, and called them as PPO-positive tubes. We observed that the PPO-positive tubes are physically derived from the hindgut but strongly adhere to the Malpighian tubule. Inside the PPO-positive tubes, there is an acellular peritrophic membrane to protect the epidermal cells. Furthermore, the PPO-positive tubes act like a doorkeeper to firstly detoxify the metabolite wastes collected by the Malpighian tubule from the hemolymph.

Activation of microlipophagy during early infection of insect hosts by Metarhizium robertsii.

The requirement of macroautophagic/autophagic machinery for filamentous fungal development and pathogenicity has been recognized, but the underlying effects and mechanisms remain elusive. The insect pathogenic fungus Metarhizium robertsii infects hosts by cuticular penetration through the formation of the infection structure appressoria. Here, we show that autophagic fluxes were highly activated during the appressorial formation of M. robertsii. Genome-wide deletion of the autophagy-related genes and insect bioassays identified 10 of 23 encoded MrATG genes with requirements for topical fungal infection of insect hosts. Besides the defect in forming appressoria on insects (two null mutants), these virulence-reduced mutants were largely impaired in penetrating cellophane membrane and insect cuticles, suggesting their failures in generating proper appressorium turgor. We found that the conidial storage of lipid droplets (LDs) had no obvious difference between strains, but autophagic LD degradation was impaired in different mutants. After induction of cell autophagy by nitrogen starvation, we found that LD entry into vacuoles was unaffected in the selected mutant cells with potential failures in forming autophagosomes. The finding therefore reveals a microlipophagy machinery employed in this fungus and that the direct engulfment of LDs occurs without inhibition by the downstream defective lipolysis. Our data first unveil the activation and contribution of microlipophagy to fungal infection biology. The obtained technique may benefit future detection of microlipophagy in different organisms by examining vacuolar or lysosomal engulfment of LDs in core autophagic gene deletion mutants.Abbreviations: AIM: Atg8-family interacting motif; ATG: autophagy-related; CM: complete medium; CMAC: 7-amino-4-chloromethylcoumarin; DTT: dithiothreitol; ER: endoplasmic reticulum; GFP: green fluorescent protein; LD, lipid droplet; MM: minimum medium; MM-N: minimum medium without nitrogen source; PDA: potato dextrose agar; PMSF: phenylmethylsulfonyl fluoride; RFP: red fluorescent protein; SDB: Sabouraud dextrose broth; SDS-PAGE: sodium dodecyl sulfate-polyacrylamide gel electrophoresis; TAG: triacylglycerol; TEM: transmission electron microscopy; WT, wild type.

Material Design for Low-Loss Non-Oriented Electrical Steel for Energy Efficient Drives.

Due to the nonlinear material behavior and contradicting application requirements, the selection of a specific electrical steel grade for a highly efficient electrical machine during its design stage is challenging. With sufficient knowledge of the correlations between material and magnetic properties and capable material models, a material design for specific requirements can be enabled. In this work, the correlations between magnetization behavior, iron loss and the most relevant material parameters for non-oriented electrical steels, i.e., alloying, sheet thickness and grain size, are studied on laboratory-produced iron-based electrical steels of 2.4 and 3.2 wt % silicon. Different final thicknesses and grain sizes for both alloys are obtained by different production parameters to produce a total of 21 final material states, which are characterized by state-of-the-art material characterization methods. The magnetic properties are measured on a single sheet tester, quantified up to 5 kHz and used to parametrize the semi-physical IEM loss model. From the loss parameters, a tailor-made material, marked by its thickness and grain size is deduced. The influence of different steel grades and the chance of tailor-made material design is discussed in the context of an exemplary e-mobility application by performing finite-element electrical machine simulations and post-processing on four of the twenty-one materials and the tailor-made material. It is shown that thicker materials can lead to fewer iron losses if the alloying and grain size are adapted and that the three studied parameters are in fact levers for material design where resources can be saved by a targeted optimization.

Integrated Process Simulation of Non-Oriented Electrical Steel.

A tailor-made microstructure, especially regarding grain size and texture, improves the magnetic properties of non-oriented electrical steels. One way to adjust the microstructure is to control the production and processing in great detail. Simulation and modeling approaches can help to evaluate the impact of different process parameters and finally select them appropriately. We present individual model approaches for hot rolling, cold rolling, annealing and shear cutting and aim to connect the models to account for the complex interrelationships between the process steps. A layer model combined with a microstructure model describes the grain size evolution during hot rolling. The crystal plasticity finite-element method (CPFEM) predicts the cold-rolling texture. Grain size and texture evolution during annealing is captured by the level-set method and the heat treatment model GraGLeS2D+. The impact of different grain sizes across the sheet thickness on residual stress state is evaluated by the surface model. All models take heterogeneous microstructures across the sheet thickness into account. Furthermore, a relationship is established between process and material parameters and magnetic properties. The basic mathematical principles of the models are explained and demonstrated using laboratory experiments on a non-oriented electrical steel with 3.16 wt.% Si as an example.

Influence of Process Parameters on Grain Size and Texture Evolution of Fe-3.2 wt.-% Si Non-Oriented Electrical Steels.

The magnetic properties of non-oriented electrical steel, widely used in electric machines, are closely related to the grain size and texture of the material. How to control the evolution of grain size and texture through processing in order to improve the magnetic properties is the research focus of this article. Therefore, the complete process chain of a non-oriented electrical steel with 3.2 wt.-% Si was studied with regard to hot rolling, cold rolling, and final annealing on laboratory scale. Through a comprehensive analysis of the process chain, the influence of important process parameters on the grain size and texture evolution as well as the magnetic properties was determined. It was found that furnace cooling after the last hot rolling pass led to a fully recrystallized grain structure with the favorable ND-rotated-cube component, and a large portion of this component was retained in the thin strip after cold rolling, resulting in a texture with a low γ-fiber and a high ND-cube component after final annealing at moderate to high temperatures. These promising results on a laboratory scale can be regarded as an effective way to control the processing on an industrial scale, to finally tailor the magnetic properties of non-oriented electrical steel according to their final application.

Grain Size Influence on the Magnetic Property Deterioration of Blanked Non-Oriented Electrical Steels.

Non-oriented electrical steel sheets are applied as a core material in rotors and stators of electric machines in order to guide and magnify their magnetic flux density. Their contouring is often realized in a blanking process step, which results in plastic deformation of the cut edges and thus deteriorates the magnetic properties of the base material. This work evaluates the influence of the material's grain size on its iron losses after the blanking process. Samples for the single sheet test were blanked at different cutting clearances (15 µm-70 µm) from sheets with identical chemical composition (3.2 wt.% Si) but varying average grain size (28 µm-210 µm) and thickness (0.25 mm and 0.5 mm). Additionally, in situ measurements of blanking force and punch travel were carried out. Results show that blanking-related iron losses either increase for 0.25 mm thick sheets or decrease for 0.5 mm thick sheets with increasing grain size. Although this is partly in contradiction to previous research, it can be explained by the interplay of dislocation annihilation and transgranular fracturing. The paper thus contributes to a deeper understanding of the blanking process of coarse-grained, thin electrical steel sheets.

Involvement of Epidermis Cell Proliferation in Defense Against Beauveria bassiana Infection.

Entomopathogenic fungi Beauveria bassiana can infect many species of insects and is used as a biological pesticide world-wide. Before reaching the hemocoel, B. bassiana has to penetrate the integument which is composed of a thick chitin layer and epidermal cells. Some chitinase, protease and lipase secreted by B. bassiana are probably involved in the fungal penetration of the integument. While microscopic proof is needed, it is difficult to locate the precise infection sites following the traditional method of immersion infection. Consequently, we developed a new method to inoculate conidia solution into a single fixed-site on the back of one segment. This fixed-site infection method is pathogenic but it is also dose dependent. Using the fixed-site infection protocol, it is also very convenient to track hyphae inside the cuticle layer by light and transmission electron microscopy. The fact that few hyphae were detected inside the chitin layer after fixed-site infection with mutant ΔBPS8, a protease secreted during fungi germination, indicates that this method is suitable for screening genes involved in penetrating the integument in large scale. We also found that melanization occurs before new hyphae penetrate the chitin layer. Most importantly, we discovered that fungal infection can induce epidermal cell proliferation through DNA duplication and cell division, which is essential for the host to defend against fungal infection. Taken together the fixed-site infection method may be helpful to determine the mechanism of fungal and host interaction in the integument so as to effectively exert fungal biological virulence.

A Reference chart for clinical biochemical tests of hemolyzed serum samples.

BACKGROUND: Although the effect of hemolysis has been extensively evaluated on clinical biochemical tests, a practical guidance for laboratory staff to rapidly determine whether a hemolyzed blood sample is acceptable and how to interpret the results is lacking. Here, we introduce a chart as a convenient reference for dealing with such samples. METHODS: Serum samples with 0.1%, 0.3%, 1%, 3%, and 10% hemolysis were prepared from sonicated endogenous red blood cells and received 35 wet and 22 dry clinical biochemical tests, respectively. The contributing part in the biochemical test result at each hemolysis condition was derived by subtracting the original test result of this sample with no hemolysis. The net results were used for analyses and preparation of the reference chart. RESULTS: The reference chart displayed the analytically calculated hemolysis interference and related statistical analyses. The chart also provided the color appearance of serum samples at each hemolysis condition for clinical staffs to determine whether a hemolyzed sample could be accepted. CONCLUSION: In clinical laboratories, preparation of such a reference chart is extremely useful in dealing with hemolyzed blood samples for clinical biochemical tests.

Characterization Methods along the Process Chain of Electrical Steel Sheet-From Best Practices to Advanced Characterization.

Non-oriented (NO) electrical steel sheets find their application in rotating electrical machines, ranging from generators for wind turbines to motors for the transportation sector and small motors for kitchen appliances. With the current trend of moving away from fossil fuel-based energy conversion towards an electricity-based one, these machines become more and more important and, as a consequence, the leverage effect in saving energy by improving efficiency is huge. It is already well established that different applications of an electrical machine have individual requirements for the properties of the NO electrical steel sheets, which in turn result from the microstructures and textures thereof. However, designing and producing tailor-made NO electrical steel sheet is still challenging, because the complex interdependence between processing steps, the different phenomena taking place and the resulting material properties are still not sufficiently understood. This work shows how established, as well as advanced and newly developed characterization methods, can be used to unfold these intricate connections. In this context, the respective characterization methods are explained and applied to NO electrical steel as well as to the typical processing steps. In addition, several experimental results are reviewed to show the strengths of the different methods, as well as their (dis)advantages, typical applications and obtainable data.

Stable Plasma Sample Storage in Acetonitrile for Angiotensin and Aldosterone Analysis.

BACKGROUND: Angiotensin I, II (AI, AII) and aldosterone are unstable in plasma specimens at room temperature, making it difficult for collect samples for remote regions in centralized and collaborative studies. Here we introduce a stable storage method which do not require cold conditions.. METHODS: Acetonitrile was added to the plasma to 60%, and then the supernatants were kept at 4°C and room temperature for 0, 1, 2, 3, 10 and 30 days. AI, AII and aldosterone were extracted and analyzed by chemiluminescence immunoassays. RESULTS: AI, AII and aldosterone were well retained in the supernatant under this method. The intra- and inter-day CVs of this method were all below 10%. The levels of AI, AII and aldosterone by this method remained stable for 30 days at room temperature. CONCLUSION: Addition of 60% acetonitrile in the plasma provides a stable storage method for clinical AI, AII and aldosterone.

Effects of U1 Small Nuclear Ribonucleoprotein Inhibition on the Expression of Genes Involved in Alzheimer's Disease.

Deposition and dysfunction of U1 small nuclear ribonucleoprotein (snRNP) have been revealed in Alzheimer's disease (AD), but whether U1 is involved in the amyloid precursor protein (APP) and Tau pathways remains unclear. Here, we investigate this by inhibiting the U1 components in cultured cells and examining the expression changes of AD-related genes to these two canonic pathways. We find that knockdown of U1-70K and U1C increases the protein expressions of APP and GSK-3ß while reduces that of Nicastrin in a dose-dependent manner. Knockdown of U1A shows no effects on the expression of these proteins. The real-time PCR results show that the mRNA expression levels of APP, Nicastrin and GSK-3ß are unchanged, decreased, and increased, respectively. In addition, U1-70K knockdown suppresses Tau phosphorylation and causes altered splicing of Tau exon 10. This study suggests that the effect of U1 snRNP knockdown is component-specific and more likely involved in APP deregulation in AD.

Magnetic Nanoparticle-Based Automatic Chemiluminescent Enzyme Immunoassay for Golgi Protein 73 and the Clinical Assessment.

Golgi protein 73 (GP73) is an independent diagnostic indicator of cirrhosis. However, it lacks automatic detection techniques to meet the large-scale clinical requirement in physical examination. In this paper, an automatic approach was established based on the ACL2800 automatic chemiluminescent analyzer, using magnetic nanoparticles (MNPs) and chemiluminescence. This method depended on sandwich strategy among biotin-labeled capture monoclonal antibody, target GP73 and acridinium ester (AE)-labeled reporter monoclonal antibody, conjugation of streptavidin-labeled MNPs to biotin-labeled capture monoclonal antibody, and chemiluminescent detection of AE-linked targets. Optimal conditions were investigated and clinical assessment was processed. Detection of GP73 demonstrated a high sensitivity of 1.19 ng/mL with a wide range from 1.34 ng/mL to 684.38 ng/mL. The quantitative detection was achieved with the repeatability of 2.69%, the coefficient of variation of 3.55% and the percent recovery of 93.46%-107.82%. Therefore, an automatic quantitative detection method was successfully developed, which was a potential screening method in physical examination.

HRD1 sensitizes breast cancer cells to Tamoxifen by promoting S100A8 degradation.

Estrogen receptor alpha positive (ER+) of breast cancer could develop resistance to antiestrogens including Tamoxifen. Our previous study showed that the E3 ubiquitin ligase HRD1 played an important role in anti-breast cancer. However, its role in chemotherapy resistance hasn't been reported. In this study, we found that HRD1 expression was downregulated in Tamoxifen-resistant breast cancer cell line MCF7/Tam compared to the Tamoxifen sensitive cell line MCF7. Moreover, S100A8 is the direct target of HRD1 by proteome analysis. Our data showed that HRD1 decreased the protein level of S100A8 through ubiquitination while HRD1 was regulated by acetylation of histone. More importantly, HRD1 knockdown significantly increased the cell survival of MCF7 cells to the Tamoxifen treatment. HRD1 overexpression sensitized MCF7/Tam cells to the Tamoxifen treatment in vitro and in vivo. In conclusion, the decrease of HRD1 expression contributed to Tamoxifen resistance in breast cancer.

[The characteristics of enhanced gadolinium DTPA brain magnetic resonance images in 298 multiple sclerosis patients].

OBJECTIVE: To investigate the characteristics of enhanced brain magnetic resonance imaging (MRI) with gadolinium-DTPA (GD) in patients with multiple sclerosis (MS). METHODS: Brain enhanced MRI were studied in 186 patients with clinically definite multiple sclerosis. 298 MRI enhanced lesions were followed every 4 weeks and the study lasted 3 months to 2 years. RESULTS: Of the 298 enhanced MRI lesions 207 (69.5%) lasted less than 4 weeks, 251 (84.2%) 8 weeks, and 272 (91.3%) 12 weeks. There were 3 lesions lasting more than 2 years. Round, oval, ring form, arcuate, spotty, or irregular MRI GD enhanced lesions were found in our MS data. CONCLUSIONS: Most of the MRI GD enhanced lesions disappear in 4 weeks. Very few can last more than 2 years. So it is necessary to perform the GD enhanced MRI in 4 weeks for studying MS intracranial GD enhanced lesions. The DA enhanced MRI lesions in MS may be round, oval, ring form, arcuate, spotty or irregular.