Metallothioneins (MTs) are metal-binding proteins that are overexpressed in various human cancers and are thought to be associated with resistance to cytotoxic drugs. The knowledge on MT expression, regulation, and function in human gliomas is limited. We found that MT3 mRNA was highly expressed in cell lines derived from grade IV gliomas (i.e., A172 and U87 cells), as compared to grade II astrocytoma cells (i.e., 1321N1). Different from 1321N1, U87 cells were partly resistant to the alkylating drug, temozolomide (TMZ) (100 μM for 96 h), which induced a massive accumulation of U87 into the S and G2 fractions of the cell cycle but not apoptotic death. Silencing of MT3 did not significantly affect U87 cell proliferation and survival, but it delayed G1/S transition and favored the occurrence of apoptosis in TMZ-treated cells. Accordingly, the combination of MT3 silencing and TMZ treatment increased the protein levels of checkpoint kinase-1, which was ultimately responsible for the lasting G1 arrest and death of double treated U87 cells.

Increasing resistance to presently available antimalarial drugs urges the need to look for new promising compounds. The β-carboline moiety, present in several biologically active natural products and drugs, is an important scaffold for antimalarial drug discovery. The present study explores the antimalarial activity of a β-carboline derivative (1R,3S)-methyl 1-(benzo[d][1,3]dioxol-5-yl)-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole-3-carboxylate (9a) alone in vitro against Plasmodium falciparum and in vivo in combination therapy with the standard drug artesunate against Plasmodium berghei. Compound 9a inhibited both 3D7 and RKL-9 strains of P. falciparum with half-maximal inhibitory concentration (IC50) < 1 μg/mL, respectively. The compound was nontoxic (50% cytotoxic concentration (CC50) > 640 μg/mL) to normal dermal fibroblasts. Selectivity index was >10 against both the strains. The compound exhibited considerable in vivo antimalarial activity (median effective dose (ED50) = 27.74 mg/kg) in monotherapy. The combination of 9a (100 mg/kg) and artesunate (50 mg/kg) resulted in 99.69% chemosuppression on day 5 along with a mean survival time of 25.8 ± 4.91 days with complete parasite clearance. Biochemical studies indicated the safety of the HIT compound to hepatic and renal functions of mice. Molecular docking also highlighted the suitability of 9a as a potential antimalarial candidate.

Flavonoids are a large class of polyphenols widely distributed in plants in the free form or as glycosides, and they have antioxidation, antibacterial, antitumor growth, and other pharmacological effects. As an important active component of traditional Chinese medicine, they have high medicinal value and development prospects. In this work, the biomolecular properties of 10 common flavonoids, including baicalein, baicalin, apigenin, quercetin, naringenin, hesperetin, daidzein, genistein, puerarin, and gastrodin, are studied by terahertz time-domain spectroscopy (THz-TDS) in the range of 0.2-2.5 THz. The results reveal that these flavonoids have different characteristic absorption peaks in the terahertz band. Moreover, the terahertz absorption characteristics of samples in the temperature range of 78-320 K are studied. The results show that the characteristic absorption peaks gradually increase with the decrease in temperature, and the frequency position of the absorption peak has a slight blue shift. Furthermore, qualitative identification and quantitative analysis of flavonoids are carried out by terahertz spectra combined with chemometrics. Specifically, a series of mixtures of three flavonoids with similar molecular structures under various concentrations are analyzed. The partial least-squares regression (PLSR) model and the artificial neural network (ANN) model are applied to quantitatively analyze the ternary mixture. The results confirm that the ANN model obtains the best predicted value, with the root-mean-square errors in the prediction set (RMSEP) of 1.27% for daidzein. In summary, the biomolecular properties of flavonoids are studied by the THz-TDS technique, and a rapid, effective, and nondestructive method for qualitative identification and quantitative analysis of flavonoids is provided. The results demonstrate that this method has potential application value in the detection of Chinese herbal medicine and has better referential significance for the study of other biomolecules, especially for isomers or similar molecular structures.

In this study, we prepared a monoclonal antibody (mAb) against metalaxyl (Met) with a half-maximum inhibitory concentration (IC50) of 0.54 ng/mL based on a new hapten, and a gold nanoparticle-based immunochromatographic assay (GICA) was developed for the rapid detection of Met residues in tobacco. Under optimal conditions, even with the naked eye, one can see the semiquantitative analysis results. The naked eye detection limit of Met in tobacco is 25 μg/kg, and the detection threshold is 100 μg/kg. In addition, the cross-reactivity test shows that the mAb has good specificity for Met, and the GICA results have a good correlation with the indirect competitive enzyme-linked immunosorbent assay and liquid chromatography with tandem mass spectrometry test results, which show that the method is feasible and reliable and are more convenient and quicker than the methods using instrumentation for detection. Therefore, GICA may provide a useful tool for the rapid screening and detection of Met residues in tobacco.

Oral administration of vaccines has been limited due to low immune response compared to parenteral administration. Antigen degradation in the acidic gastrointestinal environment (GI), mucus barriers, and inefficient cellular uptake by immune cells are the major challenges for oral vaccine delivery. To solve these issues, the current study investigates calcium phosphate nanoparticles (CaP NPs) coated with polysaccharides as nanocarriers for oral protein antigen delivery. In this design, the CaP NP core had an optimized antigen encapsulation capacity of 90 mg (BSA-FITC)/g (CaP NPs). The polysaccharides chitosan and alginate were coated onto the CaP NPs to protect the antigens against acidic degradation in the GI environment and enhance the immune response in the small intestine. The antigen release profiles showed that alginate-chitosan-coated CaP NPs prevented antigen release in a simulated gastric fluid (pH 1.2), followed by sustained release in simulated intestinal (pH 6.8) and colonic (pH 7.4) fluids. Cellular uptake and macrophage stimulation data revealed that the chitosan coating enhanced antigen uptake by intestine epithelia cells (Caco-2) and macrophages and improved surface expression of costimulatory molecules on macrophages. In vivo test further demonstrated that oral administration of alginate-chitosan-coated CaP@OVA NPs significantly enhanced the mucosal IgA and serum IgG antibody responses as compared to naked OVA, indicating that the CaP-Chi-Alg nanoparticle can potentially be used as a promising oral vaccine delivery system.

NASICON-type Li1+x Al x Ge2-x (PO4)3 (LAGP) is a promising electrolyte with high ionic conductivity (>10-4 S cm-1), excellent oxidation stability, and moderate sintering temperature. However, preparing dense LAGP pellets with high ionic conductivity is still challenging because of the hazards of dopant loss and partial decomposition on conventional sintering. Here, spark plasma sintering (SPS) of LAGP membranes is explored as a promising ultrarapid manufacturing technique, yielding dense electrolyte membranes. Optimizing the SPS temperature is important to achieve desirable density and hence ionic conductance. Our results show that LAGP samples spark plasma-sintered at 750 °C exhibit the highest total ionic conductivity of 3.9 × 10-4 S cm-1 with a compactness of 97% and nearly single-crystalline particles. Our solid-state NMR results, X-ray diffraction studies, and scanning electron microscopy micrographs confirm that the achievable ionic conductivity is controlled by the retention of the Al dopant within the LAGP phase, necking between particles, and the minimization of grain boundaries between crystallites within a particle. To benchmark the performance of our spark plasma-sintered solid electrolyte membranes over conventionally prepared LAGP, we demonstrate their favorable performance in hybrid Li-air batteries. The highest energy efficiency is achieved for the fastest ion-conducting membrane sintered at 750 °C.

Threatened by the energy crisis and environmental pollution, most countries in the world are vigorously developing new energy vehicles to promote low-carbon environmental protection and boost a green transportation system. Based on the intelligent manufacturing standard system, this study constructed a new energy vehicle intelligent manufacturing development-influencing factor model. Taking the intelligent manufacturing development ability as the dependent variable, taking external environment factors, commonalities among the new energy vehicle enterprises, and industry progress as independent variables, five hypotheses are proposed. This study used a structural equation model to test the hypotheses and reveal the mechanism of how factors influence the new energy vehicle intelligent manufacturing. The results show that external environment factors and industry progress directly and positively affect the development capability of intelligent manufacturing of new energy vehicles, while the commonalities among the NEV enterprises have an indirect effect through industry progress on intelligent manufacturing of new energy vehicles. Based on the analysis, this study puts forward some suggestions for better development of new energy vehicle intelligent manufacturing.

A new denaturation agent is the mixture of 4,5-dihydroxy-1,3-bis(methoxymethyl)imidazolidin-2-one (m-DMDHEU)/choline chloride (CC) introduced to modify three kinds of lignocellulosic materials containing different lignin contents in the following order: cotton used in medicine < sawdust from acacia auriculiformis wood < powder from the coconut shell. The modification process is carried out through two main steps: 0.2 N NaOH solution with 70% v/v ethanol and 30% v/v water was applied to remove lignin and activate the initial raw materials, and then delignified materials were modified with m-DMDHEU/CC by using a parched heat supply method after chemical impregnation. Structural characterictics and physicochemical properties of modified materials were tested and dissected by scanning electron microscopy, Fourier transform infrared spectroscopy, solid-state 13C nuclear magnetic resonance spectroscopy (solid-state 13C CP-MAS NMR), specific surface area, and pH at the point of zero charge (pHPZC). The ability to adsorb and exchange anions of modified materials was referred and examined by using aqueous solutions containing CrO4 2-, NO3 -, and H2AsO4 - anions in different conditions. The results revealed that anionite lignocellulosic materials could separate these anions with very good efficiency and better than strong anion exchange resin (GA-13) in the same conditions; outlet water could meet the permissible drinking and living water standards; and the m-DMDHEU cross-link bridge also was a good bridge to connect CC to cellulose chain beside other common urea cross-link bridges.

The current COVID-19 outbreak warrants the design and development of novel anti-COVID therapeutics. Using a combination of bioinformatics and computational tools, we modelled the 3D structure of the RdRp (RNA-dependent RNA polymerase) of SARS-CoV2 (severe acute respiratory syndrome coronavirus-2) and predicted its probable GTP binding pocket in the active site. GTP is crucial for the formation of the initiation complex during RNA replication. This site was computationally targeted using a number of small molecule inhibitors of the hepatitis C RNA polymerase reported previously. Further optimizations suggested a lead molecule that may prove fruitful in the development of potent inhibitors against the RdRp of SARS-CoV2.

Retinol stability has been reported to be improved by encapsulation in liposomes, both with and without cholesterol. However, this improvement is limited because of lipid peroxidation. In this study, we compare the stability of retinol in phosphatidylcholine liposomes under ultraviolet (UV) light or standard room air, with and without the addition of antioxidants. Both butylated hydroxytoluene (BHT) and a proprietary mix (StoppOx) improved the shelf stability from <10 to over 30 d. The addition of cholesterol had no effect. Fluorescence imaging showed a heterogeneous distribution of retinol within the vesicles, including within the aqueous layer. Fluorescence lifetimes were equally heterogeneous. Under UV irradiation, StoppOx protected retinol for significantly longer than BHT and via different mechanisms. This suggests that natural antioxidants work well to improve the retinol stability, but that further work to determine the optimal vesicle structure remains to be performed.

Head and neck cancers are mostly composed of head and neck squamous cell carcinoma (HNSCC). The incidence and mortality of HNSCC are higher in countries with emerging health care systems, particularly Africa. Given that they are more genetically diverse, characterization of polymorphism in African HNSCC may result in the identification of distinct molecular targets as compared with the known HNSCC candidate genes. This study objective is to review the current evidence of genetic data on HNSCC among African populations as well as to demonstrate any distinctions as compared with known candidate genes and to appraise any research gaps.

Advance care planning (ACP) allows patients to declare their preferences for life-sustaining and hospice palliative care. However, the perception of ACP remains low in Korea. The present study assessed the attitudes and status of medical professionals in relation to end-of-life care decisions in older and noncancerous patients.

The Lawton Instrumental Activities of Daily Living (IADL) scale is the most widely used scale for the assessment of IADL in the elderly population. The aim of this study was to adapt the Lawton IADL Scale in Turkish and to investigate the validity and the reliability of the scale in older adults.

Rheumatoid arthritis (RA) is a chronic, potentially debilitating, inflammatory disease that primarily affects synovial joints. While the etiology of RA remains incompletely understood, it is clear that the disease is autoimmune in nature. A hallmark of RA is that the specific epitopes on self-antigens that are targeted by the immune system are often modified by arginine deimination, also referred to as citrullination. In fact, anti-citrullinated protein autoantibodies (ACPA) at high enough titers are diagnostic of RA and appear to have many different targets. Here, we report that RA patients have IgG autoantibodies that react with human serum albumin (HSA) when it had been citrullinated by protein arginine deiminase (PAD) 4, but not by PAD2. Unmodified albumin was not recognized by autoantibodies. In a cohort of 79 RA patients, 38% had anti-citrullinated HSA (anti-cit-HSA) reactivity above the cut-off of the average plus two standard deviations in a healthy subject cohort (n = 16). The titers of these autoantibodies correlated with ACPA status and seropositivity. There was also a trend toward correlation with the presence of radiographic joint erosions, but this did not reach statistical significance. Finally, patients with anti-cit-HSA were more frequently treated with biologics and combination regimens than patients without these autoantibodies. We conclude that ACPA directed against citrullinated albumin exist in a subset of RA patients. Because of the abundance of albumin, its modification by citrullination, as well as autoantibodies binding to it, may have deleterious consequences for the health of affected RA patients.

Active rheumatoid arthritis (RA) is accompanied by increased appendicular and axial bone loss, closely associated to the degree of inflammation. The programmed death-1 (PD-1) pathway is important for maintaining peripheral tolerance, and its ligand PD-L2 has recently been associated with bone morphogenetic protein activity. Here, we report that PD-L2 plays a central role in RA osteoimmunology.

Investigation of the fungal communities in animal models of Inflammatory Bowel Diseases (IBD) showed a controversial role of Saccharomyces cerevisiae and Candida spp. In health and disease. These conflicting observations could be ascribed to immunogenic differences among co-specific strains. To assess the relevance of intra-strains differences on yeast immunogenicity and impact on the microbiota, we screened S. cerevisiae and Candida spp. Strains isolated from fecal samples of IBD patients. We compared the cytokine profiles, obtained upon stimulation of Peripheral Blood Mononuclear Cells (PBMCs) and Dendritic Cells with different yeast strains, and evaluated the relationship between strain's cell wall sugar amount and immune response. Moreover, the gut microbiota composition was explored in relation to fungal isolation from fecal samples by metabarcoding analysis. The comparison of cytokine profiles showed strain dependent rather than species-dependent differences in immune responses. Differences in immunogenicity correlated with the cell wall composition of S. cerevisiae intestinal strains. Stimulation of human healthy PBMCs with different strains showed a pro-inflammatory IL-6 response counterbalanced by IL-10 production. Interestingly, Crohn's (CD) patients responded differently to "self" and "non-self" strains, eliciting pure Th1 or Th17 cytokine patterns. The differences observed in vitro were recapitulated in vivo, where different strains contributed in dramatically different ways to local epithelial activity and to the inflammation of wild type and Interleukin-deficient mice. Furthermore, we observed that the gut microbiota profiles significantly differentiated according to the presence of Saccharomyces or Candida spp. or the absence of fungal isolates in fecal samples. Our results show the importance to deepen metagenomics and immunophenotyping analyses to the strain level, to elucidate the role of fungal and bacterial communities in health and disease.

Improved knowledge of different biomarkers is crucial for early diagnosis of rheumatic diseases and to provide important insights for clinical management. In this study, we evaluated the seroreactivity of patients with different connective tissue diseases (CTDs) (rheumatoid arthritis, RA; systemic lupus erythematosus, SLE; systemic sclerosis, SSc; and Sjogren's syndrome, SSj) to interferon regulatory factor 5 (IRF5) peptide and homologs derived from Epstein-Barr virus (EBV) and Mycobacterium avium subsp. paratuberculosis (MAP). Antigen-induced arthritis (AIA) experiments have been performed in control and IRF5 conditional knockout mice to reinforce the hypothesis that antibodies generated against the three homologous peptides are cross-reactive.

Narcolepsy type 1, likely an immune-mediated disease, is characterized by excessive daytime sleepiness and cataplexy. The disease is strongly associated with human leukocyte antigen (HLA) DQB1∗06:02. A significant increase in the incidence of childhood and adolescent narcolepsy was observed after a vaccination campaign with AS03-adjuvanted Pandemrix influenza vaccine in Nordic and several other countries in 2010 and 2011. Previously, it has been suggested that a surface-exposed region of influenza A nucleoprotein, a structural component of the Pandemrix vaccine, shares amino acid residues with the first extracellular domain of the human OX2 orexin/hypocretin receptor eliciting the development of autoantibodies. Here, we analyzed, whether H1N1pdm09 infection or Pandemrix vaccination contributed to the development of autoantibodies to the orexin precursor protein or the OX1 or OX2 receptors. The analysis was based on the presence or absence of autoantibody responses against analyzed proteins. Entire OX1 and OX2 receptors or just their extracellular N-termini were transiently expressed in HuH7 cells to determine specific antibody responses in human sera. Based on our immunofluorescence analysis, none of the 56 Pandemrix-vaccinated narcoleptic patients, 28 patients who suffered from a laboratory-confirmed H1N1pdm09 infection or 19 Pandemrix-vaccinated controls showed specific autoantibody responses to prepro-orexin, orexin receptors or the isolated extracellular N-termini of orexin receptors. We also did not find any evidence for cell-mediated immunity against the N-terminal epitopes of OX2. Our findings do not support the hypothesis that the surface-exposed region of the influenza nucleoprotein A would elicit the development of an immune response against orexin receptors.

Several autoimmune conditions have adverse effects on reproductive outcomes, but the relationship between family history of autoimmune disease in women without these conditions and pregnancy is uncertain. The objective of this study was to determine if there is an association between a family history of an autoimmune condition and time-to-pregnancy (TTP), pregnancy loss, and live birth. This was a prospective cohort study from a RCT of 1228 adult women ages 18-40, who were healthy, had no history of infertility, were actively attempting to conceive, and had one or two prior pregnancy losses. Of these, 1172 women had data available regarding family history of autoimmune conditions. Women with an affected first-degree relative had similar TTP when compared to those without a FHx (fecundability odds ratio 0.90, 95% confidence interval [CI] 0.70, 1.15). Women with an affected first-degree relative had a lower likelihood of live birth (relative risk [RR] 0.83, 95% CI 0.69, 0.99). Among women who achieved pregnancy, FHx of autoimmune disease was associated with a higher likelihood of pregnancy loss (RR 1.49, 95% CI 1.10, 2.03). Women who had a first-degree relative with an autoimmune disease had a similar TTP as unaffected women but a lower likelihood of live birth and higher risk of pregnancy loss. This information may encourage clinicians to evaluate women with a family history of autoimmune conditions prior to pregnancy and highlights the need for further studies to ascertain the effects of autoimmunity and pregnancy.

Cryo-electron microscopy (cryo-EM) has revolutionized structural biology by providing 3D density maps of biomolecules at near-atomic resolution. However, map validation is still an open issue. Despite several efforts from the community, it is possible to overfit 3D maps to noisy data. Here, we develop a novel methodology that uses a small independent particle set (not used during the 3D refinement) to validate the maps. The main idea is to monitor how the map probability evolves over the control set during the 3D refinement. The method is complementary to the gold-standard procedure, which generates two reconstructions at each iteration. We low-pass filter the two reconstructions for different frequency cutoffs, and we calculate the probability of each filtered map given the control set. For high-quality maps, the probability should increase as a function of the frequency cutoff and the refinement iteration. We also compute the similarity between the densities of probability distributions of the two reconstructions. As higher frequencies are included, the distributions become more dissimilar. We optimized the BioEM package to perform these calculations, and tested it over systems ranging from quality data to pure noise. Our results show that with our methodology, it possible to discriminate datasets that are constructed from noise particles. We conclude that validation against a control particle set provides a powerful tool to assess the quality of cryo-EM maps.