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Heat stress is known to negatively impact the reproductive process of livestock, which inevitably leads to a decline in animal fertility. Nuclear factor E2-related factor 2 (Nrf2) is an inducible transcription factor, which is essential for maintaining redox signal transmission against oxidative stress. However, there is no reliable research on the response mechanism of Sertoli Cells (SCs) against heat stress and the activation of Nrf2 when SCs are exposed to heat stress. Here, we used primary mouse SCs and SCs line TM4, along with Nrf2 specific inhibitor to determine the reaction mechanism of SCs to maintain intracellular redox homeostasis and self-survival by activating Nrf2. We found that acute heat stress only affected the vitality of SCs and the expression of functional molecules (tight junction-associated proteins and lactate dehydrogenase A [LDHA]) but did not cause cell apoptosis. When Nrf2 was inhibited, more cell death occurred in TM4 cells post heat stress treatment, along with a greater decrease in cell viability and a significant increase in intracellular ROS levels. Our study clarified for the first time the protective effect of Nrf2 activation on heat stress-induced SCs damage. It explained the possible reasons or mechanisms involved in the survival of SCs, the critical protective cells in the testis, which were not affected by heat stress. This study further improved the response mechanism of SCs in the reproductive injury caused by a high-temperature environment.

A direct, efficient, and versatile glycosylation methodology promises the systematic synthesis of oligosaccharides and glycoconjugates in a streamlined fashion like the synthesis of medium to long-chain nucleotides and peptides. The development of a generally applicable approach for the construction of 1,2-cis-glycosidic bond with controlled stereoselectivity remains a major challenge, especially for the synthesis of β-mannosides. Here, we report a direct mannosylation strategy mediated by ZnI2, a mild Lewis acid, for the highly stereoselective construction of 1,2-cis-β linkages employing easily accessible 4,6-O-tethered mannosyl trichloroacetimidate donors. The versatility and effectiveness of this strategy were demonstrated with successful β-mannosylation of a wide variety of alcohol acceptors, including complex natural products, amino acids, and glycosides. Through iteratively performing ZnI2-mediated mannosylation with the chitobiosyl azide acceptor followed by site-selective deprotection of the mannosylation product, the novel methodology enables the modular synthesis of the key intermediate trisaccharide with Man-β-(1 → 4)-GlcNAc-β-(1 → 4)-GlcNAc linkage for N-glycan synthesis. Theoretical investigations with density functional theory calculations delved into the mechanistic details of this β-selective mannosylation and elucidated two zinc cations' essential roles as the activating agent of the donor and the principal mediator of the cis-directing intermolecular interaction. We thank the Fundamental Research Funds for the Central Universities (no. 31610011 to F.D.), the Province Natural Science Fund of Guangdong (no. 42030015 to F.D.), and the Sun Yat-sen University Startup Fund (no.18841224 to F.D.) for financial support and the grant-in-aid for Specially Promoted Research (no. 16H06290 to Y.I. and A.I.) and for Scientific Research (no.18K05345 and 19H00929 to A.I.) from the Japan Society for the Promotion of Science.

As a type of thermal structure ceramic, vanadium nitride (VN) ceramics are widely used in harsh conditions, such as high temperatures, variable temperatures, or oxidizing atmospheres. However, high...

Facemasks are considered safe and wearable devices that cover the human mouth and nose for filtering exhaled aerosols and inhaled environmental exposures; various chemical and environmental residues thus can remain in facemasks. Therefore, direct analysis of residues in facemasks can be used to investigate the wearer's health and behavior. Here, we developed a simple paper-in-facemask sampling method for adsorbing a wearer's respiratory aerosol and environmental exposures by fixing paper strips at the outside and inside surfaces of facemasks, and the paper strips were then analyzed by paper spray mass spectrometry (PSMS) for directly detecting adsorbed analytes without any sample pretreatment. The applicability of this device was demonstrated by directly analyzing exhaled aerosolized saliva, breath metabolites, and inhalable environmental exposures. The technical aspects, including sampling time, sampling position, paper property, and spray solvent, were investigated. The sampling process was revealed to involve a continuous-flow adsorptive mechanism. These findings motivated us to extend this work and build a wearable sampling device that is capable of simultaneously monitoring both exhaled and inhaled biomarkers in situ to investigate human health and environmental exposure. This work highlights that facemasks are promising platforms for aerosol collection and direct MS analysis, which is expected to be a promising method for monitoring human health, diseases, and behaviors.

Objective To evaluate the quality of Arnebiae Radix (AR) and Dictamni Cortex (DC) and study the efficacy of herbal extracts of these two herbs on the treatment of allergic contact dermatitis (ACD). Methods Qualitative and quantitative analysis of effective components was performed using High Performance Thin Layer Chromatography (HPTLC), High Performance Liquid Chromatography (HPLC), and HPLC-Quadrupole Time of Flight-Mass Spectrometry (HPLC-QTOF-MS). In vitro allergic ACD 3D model was established by incubating 3D reconstructed human epidermis (RHE) with skin sensitizer, potassium dichromate. A total of 65 gene expression that were associated with ACD, which included 24 antioxidant responsive element (ARE) and 41 SENS-IS genes were quantified by qRT-PCR. More than or equal to 10 ARE genes and 18 SENN-IS genes were induced by 1.3-fold, demonstrating the successful establishment of in vitro ACD model. Oil extracts of AR and DC were applied on the in vitro ACD model to study the efficacy. Results Batch 3 of AR and batch 2 of DC showed presence of all active ingredients with the highest concentrations. Active ingredients of the herbs were extracted using a special oil and formulated into herbal oil extracts. The herbal oil extracts were able to down regulate the induced genes in the in-vitro ACD skin model, bringing the tissue back to homeostatic status. Conclusion The oil extracts showed the potent efficacy of using AR and DC in ACD treatment. The combination study will be done to optimize the formulation ratio which will be developed into a topical cream.

Amorphous solid dispersions (ASD) are one of most commonly used supersaturating drug delivery systems (SDDS) to formulate insoluble active pharmaceutical ingredients. However, the development of polymer-guided stabilization of ASD systems faces many obstacles. To overcome these shortcomings, co-amorphous supersaturable formulations have emerged as an alternative formulation strategy for poorly soluble compounds. Noteworthily, current researches around co-amorphous system (CAS) are mostly focused on preparation and characterization of these systems, but more detailed investigations of their supersaturation (“spring-parachute“ process), stability, in vivo bioavailability and molecular mechanisms are inadequate and need to be clarified. In present study, we chose pharmacological relevant BCS II drugs to fabricate and characterize “felodipine-indomethacin” CAS. To enrich the current inadequate but key knowledge on CAS studies, we carried out following highlighted investigations including dissolution/solubility, semi-continuous “spring-parachute” process, long-term stability profile of amorphous state, in vivo bioavailability and underlying molecular mechanisms (molecular interaction, molecular miscibility and crystallization inhibition). Generally, the research provides some key information in the field of current “drug-drug” CAS supersaturable formulations.

Here we report a glucosylation strategy mediated by ZnI2, a cheap and mild Lewis acid, for the highly stereoselective construction of 1,2-cis-O-glycosidic linkages using easily accessible and common 4,6-O-tethered glucosyl donors. The versatility and effectiveness of the α-glucosylation strategy were demonstrated successfully with various acceptors, including complex alcohols. This approach demonstrates the feasibility of the modular synthesis of various α-glucans with both linear and branched backbone structures.

Vascular calcification (VC) is characterized by pathological depositions of calcium and phosphate in the arteries and veins via an active cell-regulated process, in which vascular smooth muscle cells (VSMCs) transform into osteoblast/chondrocyte-like cells as in bone formation. VC is associated with significant morbidity and mortality in chronic kidney disease (CKD) and cardiovascular disease, but the underlying mechanisms remain unclear. In this study we investigated the role of large-conductance calcium-activated potassium (BK) channels in 3 experimental VC models. VC was induced in vascular smooth muscle cells (VSMCs) by β-glycerophosphate (β-GP), or in rats by subtotal nephrectomy, or in mice by high-dosage vitamin D3. We showed that the expression of BK channels in the artery of CKD rats with VC and in β-GP-treated VSMCs was significantly decreased, which was functionally confirmed by patch-clamp recording. In β-GP-treated VSMCs, BK channel opener NS1619 (20 μM) significantly alleviated VC by decreasing calcium content and alkaline phosphatase activity. Furthermore, NS1619 decreased mRNA expression of ostoegenic genes OCN and OPN, as well as Runx2 (a key transcription factor involved in preosteoblast to osteoblast differentiation), and increased the expression of α-SMA protein, whereas BK channel inhibitor paxilline (10 μM) caused the opposite effects. In primary cultured VSMCs from BK(−/−) mice, BK deficiency aggravated calcification as did BK channel inhibitor in normal VSMCs. Moreover, calcification was more severe in thoracic aorta rings of BK(−/−) mice than in those of wild-type littermates. Administration of BK channel activator BMS191011 (10 mg· kg(−1) ·d(−1)) in high-dosage vitamin D3-treated mice significantly ameliorated calcification. Finally, co-treatment with Akt inhibitor MK2206 (1 μM) or FoxO1 inhibitor AS1842856 (3 μM) in calcified VSMCs abrogated the effects of BK channel opener NS1619. Taken together, activation of BK channels ameliorates VC via Akt/FoxO1 signaling pathways. Strategies to activate BK channels and/or enhance BK channel expression may offer therapeutic avenues to control VC.

Banana fusarium wilt caused by Fusarium. oxysporum f.sp. cubense race 4, has already become one of the main diseases in banana production. Previous research suggests that Foc can secrete two phytotoxins, respectively, fusaric acid and beauvericin, to contribute to its pathogenicity when it infects bananas. A gene cluster FUB consisting of 12 genes has been identified for fusaric acid biosynthesis in several Fusarium spp, including Fusarium. oxysporum f.sp. cubense race 4, F. oxysporum f.sp. lycopersici etc. In the present study, using reverse genetic research methods, we report the evidence for the kynurenine pathway as alternative biosynthetic pathways of fusaric acid in F. oxysporum f.sp. cubense 4. In the meanwhile, our experiments suggests that the beauvericin might not directly contribute to the pathogenicity to banana of F. oxysporum f.sp. cubense 4.

BACKGROUND: Trimethylamine N-oxide (TMAO), a diet-derived, gut microbial-host cometabolite, has been linked to cardiometabolic diseases. However, the relations remain unclear between diet, TMAO, and cardiometabolic health in general populations from different regions and ethnicities. OBJECTIVES: To examine associations of circulating TMAO with dietary and cardiometabolic factors in a pooled analysis of 16 population-based studies from the United States, Europe, and Asia. METHODS: Included were 32,166 adults (16,269 white, 13,293 Asian, 1247 Hispanic/Latino, 1236 black, and 121 others) without cardiovascular disease, cancer, chronic kidney disease, or inflammatory bowel disease. Linear regression coefficients (β) were computed for standardized TMAO with harmonized variables. Study-specific results were combined by random-effects meta-analysis. A false discovery rate

Multiple clinical trials have shown that monoclonal antibodies (mAbs) against programmed death‐ligand 1 (PD‐1/PD‐L1) can benefit patients with lung cancer by increasing their progression‐free survival and overall survival. However, a significant proportion of patients do not respond to anti‐PD‐1/PD‐L1 mAbs. In the present study, we investigated whether galectin (Gal)‐3 inhibitors can enhance the antitumor effect of PD‐L1 blockade. Using the NSCLC‐derived cell line A549, we examined the expression of Gal‐3 in lung cancer cells under hypoxic conditions and investigated the regulatory effect of Gal‐3 on PD‐L1 expression, which is mediated by the STAT3 pathway. We also explored whether Gal‐3 inhibition can facilitate the cytotoxic effect of T cells induced by PD‐L1 blockade. The effects of combined use of a Gal‐3 inhibitor and PD‐L1 blockade on tumor growth and T‐cell function were also investigated, and we found that hypoxia increased the expression and secretion of Gal‐3 by lung cancer cells. Gal‐3 increased PD‐L1 expression via the upregulation of STAT3 phosphorylation, and administration of a Gal‐3 inhibitor enhanced the effect of PD‐L1 blockade on the cytotoxic activity of T cells against cancer cells in vitro. In a mouse xenograft model, the combination of a Gal‐3 inhibitor and PD‐L1 blockade synergistically suppressed tumor growth. Furthermore, the administration of a Gal‐3 inhibitor enhanced T‐cell infiltration and granzyme B release in tumors. Collectively, our results show that Gal‐3 increases PD‐L1 expression in lung cancer cells and that the administration of a Gal‐3 inhibitor as an adjuvant enhanced the antitumor activity of PD‐L1 blockade., Galectin (Gal)‐3 is involved in the immune response, and highly overexpressed and secreted by lung cancer cells. This study showed that the increased expression of Gal‐3 was induced by hypoxia, which then contributed to immune suppression through the STAT3 pathway in lung cancer. Furthermore, a Gal‐3 inhibitor enhanced the antitumor effect by PD‐L1 blockade through promoting T‐cell infiltration in lung cancer.

Evidence from animal models suggests that dietary fatty acids have both anticancer and tumor-promoting effects. Whether dietary fatty acids are associated with colorectal cancer (CRC) in humans remains inconclusive. We investigated associations between dietary fatty acids and risk of CRC among 59,986 men who participated in the Shanghai Men's Health Study (SMHS), an ongoing population-based prospective cohort study. We identified 876 incident CRC cases in the SMHS during a mean follow-up of 9.8 years. Associations between dietary fatty acid intake and CRC risk were evaluated by Cox proportional hazard regression analyses. Consumption of saturated (SFA), monounsaturated (MUFA) and polyunsaturated (PUFA) fatty acids was not significantly associated with CRC risk. Multivariate hazard ratios (HR) and respective 95% confidence intervals (CI) for quartile 4 vs. quartile 1 were 0.92 (0.74-1.14; Ptrend =0.47) for SFA, 0.95 (0.79-1.16; Ptrend =0.74) for MUFA and 1.18 (0.95-1.46; Ptrend =0.21) for PUFA. No significant association was found for total n-6 PUFA and total n-3 PUFA. Additionally, we performed a meta-analysis to summarize results from the present study and 28 reports from 26 additional cohorts, which supported the overall null association between dietary fatty acid intake and CRC risk among men. Docosahexanoic acid (DHA) and eicosapentaenoic acid (EPA) were associated with 11-12% reduced risk, and linoleic acid (LA) a 19% increased risk, of CRC in the meta-analysis of combined sexes. In conclusion, this population-based prospective study and meta-analysis of cohort studies found little evidence that dietary fatty acid intake was associated with risk of CRC in men. This article is protected by copyright. All rights reserved.

Aconitine (ACO), a main active ingredient of Aconitum, is well-known for its cardiotoxicity. However, the mechanisms of toxic action of ACO remain unclear. In the current study, we investigated the cardiac effects of ACO and mesaconitine (MACO), a structurally related analog of ACO identified in Aconitum with undocumented cardiotoxicity in guinea pigs. We showed that intravenous administration of ACO or MACO (25 μg/kg) to guinea pigs caused various types of arrhythmias in electrocardiogram (ECG) recording, including ventricular premature beats (VPB), atrioventricular blockade (AVB), ventricular tachycardia (VT), and ventricular fibrillation (VF). MACO displayed more potent arrhythmogenic effect than ACO. We conducted whole-cell patch-clamp recording in isolated guinea pig ventricular myocytes, and observed that treatment with ACO (0.3, 3 μM) or MACO (0.1, 0.3 μM) depolarized the resting membrane potential (RMP) and reduced the action potential amplitude (APA) and durations (APDs) in a concentration-dependent manner. The ACO- and MACO-induced AP remodeling was largely abolished by an INa blocker tetrodotoxin (2 μM) and partly abolished by a specific Na+/K+ pump (NKP) blocker ouabain (0.1 μM). Furthermore, we observed that treatment with ACO or MACO attenuated NKP current (INa/K) and increased peak INa by accelerating the sodium channel activation with the EC50 of 8.36 ± 1.89 and 1.33 ± 0.16 μM, respectively. Incubation of ventricular myocytes with ACO or MACO concentration-dependently increased intracellular Na+ and Ca2+ concentrations. In conclusion, the current study demonstrates strong arrhythmogenic effects of ACO and MACO resulted from increasing the peak INa via accelerating sodium channel activation and inhibiting the INa/K. These results may help to improve our understanding of cardiotoxic mechanisms of ACO and MACO, and identify potential novel therapeutic targets for Aconitum poisoning.

14-3-3γ is a small protein regulating its target proteins through binding to phosphorylated serine/threonine residues. Sequence analysis of large-conductance Ca2+-activated K+ (BK) channels revealed a putative 14-3-3 binding site in the COOH-terminal region. Our previous data showed that 14-3-3γ is widely expressed in the mouse kidney. Therefore, we hypothesized that 14-3-3γ has a novel role in the regulation of BK channel activity and protein expression. We used electrophysiology, Western blot analysis, and coimmunoprecipitation to examine the effects of 14-3-3γ on BK channels both in vitro and in vivo. We demonstrated the interaction of 14-3-3γ with BK α-subunits (BKα) by coimmunoprecipitation. In human embryonic kidney-293 cells stably expressing BKα, overexpression of 14-3-3γ significantly decreased BK channel activity and channel open probability. 14-3-3γ inhibited both total and cell surface BKα protein expression while enhancing ERK1/2 phosphorylation in Cos-7 cells cotransfected with flag-14-3-3γ and myc-BK. Knockdown of 14-3-3γ by siRNA transfection markedly increased BKα expression. Blockade of the ERK1/2 pathway by incubation with the MEK-specific inhibitor U0126 partially abolished 14-3-3γ-mediated inhibition of BK protein expression. Similarly, pretreatment of the lysosomal inhibitor bafilomycin A1 reversed the inhibitory effects of 14-3-3γ on BK protein expression. Furthermore, overexpression of 14-3-3γ significantly increased BK protein ubiquitination in embryonic kidney-293 cells stably expressing BKα. Additionally, 3 days of dietary K+ challenge reduced 14-3-3γ expression and ERK1/2 phosphorylation while enhancing renal BK protein expression and K+ excretion. These data suggest that 14-3-3γ modulates BK channel activity and protein expression through an ERK1/2-mediated ubiquitin-lysosomal pathway.

The micro-arc oxidization (MAO) ceramic layer on an LA103Z Mg-Li alloy substrate was treated with a hydrothermal treatment at 90 °C for various times. The effect of the hydrothermal treatment time on the microstructure and corrosion behavior of MAO/LDH composite coatings in 3.5 wt.% NaCl solution was investigated, and the mechanism of hydrothermal film formation and corrosion was discussed. The results show that MgO on the surface of the MAO ceramic coating was partially dissolved during the hydrothermal treatment, and the released Mg2+ ions combined with OH− ions in the hydrothermal solution to form Mg(OH)2 nanosheets, which were deposited on the surface of the ceramic coating and its pores. The hydrogen evolution rates of the MAO/LDH composite coatings rank as MAO/LDH-24 h

Modeling age‐related macular degeneration (AMD) is challenging, because it is a multifactorial disease. To focus on interactions between the retinal pigment epithelium (RPE) and Bruch's membrane, we generated RPE from AMD patients and used an altered extracellular matrix (ECM) that models aged Bruch's membrane. Induced pluripotent stem cells (iPSCs) were generated from fibroblasts isolated from AMD patients or age‐matched (normal) controls. RPE derived from iPSCs were analyzed by morphology, marker expression, transepithelial electrical resistance (TER), and phagocytosis of rod photoreceptor outer segments. Cell attachment and viability was tested on nitrite‐modified ECM, a typical modification of aged Bruch's membrane. DNA microarrays with hierarchical clustering and analysis of mitochondrial function were used to elucidate possible mechanisms for the observed phenotypes. Differentiated RPE displayed cell‐specific morphology and markers. The TER and phagocytic capacity were similar among iPSC‐derived RPE cultures. However, distinct clusters were found for the transcriptomes of AMD and control iPSC‐derived RPE. AMD‐derived iPSC‐RPE downregulated genes responsible for metabolic‐related pathways and cell attachment. AMD‐derived iPSC‐RPE exhibited reduced mitochondrial respiration and ability to attach and survive on nitrite‐modified ECM. Cells that did attach induced the expression of complement genes. Despite reprogramming, iPSC derived from AMD patients yielded RPE with a transcriptome that is distinct from that of age‐matched controls. When challenged with an AMD‐like modification of Bruch's membrane, AMD‐derived iPSC‐RPE activated the complement immune system., Bruch's membrane is the extracellular matrix (ECM) that underlies the retinal pigment epithelium (RPE). ECM cross‐linked by nitrosylation is a typical attribute of aging. Using a model of aged Bruch's membrane, we demonstrate that iPSC‐derived RPE from AMD patients are inherently less able to attach and survive on nitrosylated ECM compared to controls.

Pregnancy loss (PL) is one of the common complications that women can experience during pregnancy, with an occurrence rate of 1 to 5%. The potential causes of pregnancy loss are unclear, with no effective treatment modalities being available. It has been previously reported that the level of miR-125b was significantly increased in placentas of PL patients. However, the role of miR-125b in the development of PL still remains unknown. In the current study, an miR-125b placenta-specific over-expression model was constructed by lentiviral transfecting zona-free mouse embryos followed by embryo transfer. On gestation day 15, it was observed that the placenta was significantly smaller in the miR-125b placenta-specific overexpression group than the control group. Additionally, the abortion rate of the miR-125b placenta-specific overexpression group was markedly higher than in the control group. The blood vessel diameter was larger in the miR-125b-overexpressing specific placenta. In addition, miR-125b-overexpressing HTR8 and JEG3 cell lines were also generated to analyze the migration and invasion ability of trophoblasts. The results showed that miR-125b overexpression significantly suppressed the migration and invasion ability of HTR8 and JEG3 cells. Overall, our results demonstrated that miR-125b can affect embryo implantation through modulating placenta angiogenesis and trophoblast cell invasion capacity that can lead to PL.

Purpose: The combination therapy of rosuvastatin (RSV) and the platelet inhibitor clopidogrel (CP) is widely accepted in the management of cardiovascular diseases. The objective of the present study was to identify the mechanism of RSV–CP DDI and evaluate the risk of hepatotoxicity associated with the concomitant use of CP.Methods: We first studied the effect of CP and its major circulating metabolite, carboxylic acid metabolite (CPC), on RSV transport by overexpressing cells and membrane vesicles. Second, we investigated whether a rat model could replicate this DDI and then be used to conduct mechanistic studies and assess the risk of hepatotoxicity. Then, cytotoxicity assay in hepatocytes, biochemical examination, and histopathology were performed to measure the magnitude of liver injury in the presence and absence of DDI.Results: CP inhibited OATP1B1-mediated transport of RSV with an IC50 value of 27.39 μM. CP and CPC inhibited BCRP-mediated RSV transport with IC50 values of 0-infinity by 76.29% and decreased intravenous RSV CL by 42.62%. Similarly, multiple p.o. doses of CP significantly increased p.o. RSV plasma AUC0-infinity by 87.48% and decreased p.o. RSV CL by 43.27%. CP had no effect on cell viability, while RSV exhibited dose-dependent cytotoxicity after 96 h incubation. Co-incubation of 100 μM CP and RSV for 96 h significantly increased intracellular concentrations and cell-to-medium concentration ratios of RSV and reduced hepatocyte viability. Histological evaluation of liver specimens showed patterns of drug-induced liver injury. Cholestasis was found in rats in the presence of DDI.Conclusion: CP is not a clinically relevant inhibitor for OATP1B1 and OATP1B3. The primary mechanism of RSV–CP DDI can be attributed to the inhibition of intestinal BCRP by CP combined with the inhibition of hepatic BCRP by CPC. The latter is likely to be more clinically relevant and be a contributing factor for increased hepatotoxicity in the presence of DDI.

BackgroundOur previous study found that acupuncture with low frequency electrical stimulation (Acu/LFES) prevents muscle atrophy by attenuation of protein degradation in mice. The current study examines the impact of Acu/LFES on protein synthesis.MethodC57/BL6 mice received Acu/LFES treatment on hindlimb for 30 min once. Acu/LFES points were selected by WHO Standard Acupuncture Nomenclature and electric stimulation applied using an SDZ-II Electronic acupuncture instrument. Muscle protein synthesis was measured by the surface-sensing of translation (SUnSET) assay. Exosomes were isolated using serial centrifugation and concentration and size of the collected exosomes were measured using a NanoSight instrument. The mature microRNA library in serum exosomes was validated using a High Sensitivity DNA chip.ResultsProtein synthesis was enhanced in the both hindlimb and forelimb muscles. Blocking exosome secretion with GW4869 decreased the Acu/LFES-induced increases in protein synthesis. MicroRNA-deep sequencing demonstrated that four members of the Let-7 miRNA family were significantly decreased in serum exosomes. Real time qPCR further verified Acu/LFES-mediated decreases of let-7c-5p in serum exosomes and skeletal muscles. In cultured C2C12 myotubes, inhibition of let-7c not only increased protein synthesis, but also enhanced protein abundance of Igf1 and Igf1 receptors. Using a luciferase reporter assay, we demonstrated that let-7 directly inhibits Igf1.ConclusionAcu/LFES on hindlimb decreases let-7-5p leading to upregulation of the Igf1 signaling and increasing protein synthesis in both hindlimb and forelimb skeletal muscles. This provides a new understanding of how the electrical acupuncture treatment can positively influence muscle health.

Layered semiconductor gallium telluride (GaTe) undergoes a rapid structural transition to a degraded phase in ambient conditions, limiting its utility in devices such as optical switches. In this work, we demonstrate that the degradation process in GaTe flakes can be slowed down dramatically via encapsulation with graphene. Through examining Raman signatures of degradation, we show that the choice of substrate significantly impacts the degradation rate and that the process is accelerated by the transfer of GaTe to hydrophilic substrates such as SiO2/Si. We find that double encapsulation with both top and bottom graphene layers can extend the lifetime of the material for several weeks. The photoresponse of flakes encapsulated in this way is only reduced by 17.6 ± 0.4% after 2 weeks, whereas unencapsulated flakes display no response after this time. Our results demonstrate the potential for alternative, van der Waals material-based passivation strategies in unstable layered materials and highlight the need for careful selection of substrates for 2D electronic devices.

We elucidated the significance of partial substitution of C by N in three high nitrogen Fe-15Cr-1Mo-C-N martensitic stainless steels (MSSs), and particularly its influence on microstructural features and the strength-toughness balance. The results show that partial substitution of C by N avoided the formation of coarse intergranular carbides and caused significant changes in the type of carbonitrides from M23C6 to M23C6+M2N and finally to M2N. Meanwhile, partially replacing C by N first increased and then decreased the effective grain size, the fraction of retained austenite, and the plate martensite, while the variations of the amount of carbonitrides and dislocation density were just the opposite. The microstructural evolutions with varying (C+N) contents played a crucial role in determining the overall properties: the 0.35C-0.37N steel exhibited an improved impact toughness (~ 86.1 J), which was 2 or 3 times higher than those of 0.50C-0.16N and 0.20C-0.54N steels, even at a strength level of over 2 GPa. The contributions of partial substitution of C by N on the strengthening mechanisms of MSSs were also systematically revealed by combining the experimental and theoretical data.

To explore the effect of 24-epibrassinolide (EBR) on the postharvest yellowing of broccoli (Brassica oleracea var. italica) florets, these were treated with 0–20 μM EBR and kept at 4 °C for 24 days (shelf life). Changes in color, chlorophyll fluorescence, chloroplast ultrastructure, and in the activities of enzymes and expression of genes related to ethylene synthesis and chlorophyll degradation were determined. Low EBR concentrations (2–5 μM) effectively inhibited the yellowing of broccoli, while high EBR concentrations (15–20 μM) accelerated it. On the 24th day, the grana thylakoids of the chloroplasts of 2 μM EBR-treated florets were neatly arranged and clearly visible, while the chloroplast membrane and grana thylakoids of the chloroplast of 20 μM EBR-treated florets were dissolved. Chlorophyll fluorescence was significantly higher in broccoli florets treated with low EBR concentrations than in control broccoli; the opposite was found for broccoli treated with high EBR concentrations. In 20 μM EBR- treated broccoli, the expressions of the genes coding for 1-aminocyclopropane-1-carboxylic acid (ACC) synthase 4 (BoACS4), ACC oxidase 3 (BoACO3), pheophytinase (PPH), and polyamine oxidase (PaO) were increased, accelerating the degradation of chlorophyll, while in 2 μM EBR-treated broccoli the expressions of BoACS4 and BoACO3 were lower than in control broccoli, thereby reducing the production of endogenous ethylene and delaying yellowing. Thus, treatment with 2 μM EBR extends the shelf life of broccoli.

Aquaporin 1 (AQP1) plays an important role in endothelial functions and is regulated by MEF2C. However, how AQP1 level is stabilized to maintain endothelial water homeostasis is still not clear. Here, we show that AQP1 expression is significantly upregulated by MEF2C transcriptionally and inhibited by miR-133a-3p.1 post-transcriptionally. Meanwhile, MEF2C activates the expression of miR-133a1. Simultaneous overexpression of MEF2C and miR-133a-3p.1 suppresses the aptitude of changes in AQP1 expression caused by either MEF2C or miR-133a-3p.1. Accordingly, the changes in migration and tube formation of human umbilical vein endothelial cells (HUVECs) caused by MEF2C or miR133a-3p.1 are blunted by coexpression of both of them. These data demonstrate that the homeostasis and physiological function of AQP1 in endothelial health are maintained by the MEF2C and miR-133a-3p.1 regulatory circuit.

Brain edema is a major traumatic brain injury (TBI)-related neurological complication. In the initiation stage of TBI, brain edema is characterized by astrocyte swelling (cytotoxic edema). We studied the impact of a long noncoding RNA, Malat1, on the TBI-induced astrocyte swelling and brain edema. Our results showed that Malat1 was downregulated in both the TBI rat model and the astrocyte fluid percussion injury (FPI) model, which concurred with brain edema and astrocyte swelling. Overexpression of Malat1 significantly inhibited rat brain edema, meanwhile reducing interleukin-6 (IL-6), nuclear factor-κB (NF-κB), and aquaporin 4 (AQP4) expression after TBI. In addition, overexpression of Malat1 ameliorated FPI-induced astrocyte swelling and reduced IL-6 release. Quantitative real-time polymerase chain reaction and Western blot analysis also corroborated the inhibitory effects of Malat1 on NF-κB and AQP4 expression after FPI. Our results highlighted the protective effects of Malat1 on the TBI-induced brain edema, which were mediated through regulating IL-6, NF-κB, and AQP4 expression. Our study could provide a novel approach for TBI treatment.

For the better mechanical and thermal properties of polypropylene (PP), PP sea-island structured composites with elastomer ethylene-propylene-diene monomer (EPDM) and coal powder were prepared by melt-blending method. Coupling agent γ-methacryloxypropyltrimethoxysilane (KH-570) was used as modifier to improve the superficial capacity of coal powder. The structural changes and properties of modified coal powder and composites were characterized and analyzed by Fourier Transform Infrared Spectroscopy (FTIR), surface contact angle analysis, scanning electron microscope (SEM), thermogravimetric analysis (TGA) and mechanical testing. Results show that the modified coal powder has good hydrophobicity and sea-island structure is beneficial in improving the comprehensive performance of composites. The contact angle increases from 33.8° to 91.6° after modification. The initial decomposition temperature (T0) and largest weight loss temperature (Tmd) both have an increase with coal powder content and the maximum are 431.02 °C and 465.33 °C. The mechanical properties tend to go up first and then down with the addition of coal powder. PP/26EPDM/4coal composite has best mechanical properties as well as the impact strength and elongation at break are 60.9 MPa, 615.0%, respectively.

New cancer vaccine strategies are required to vanquish the self-tolerance and elicit robust immune responses against tumor-associated antigens and/or neoantigens. Contemporary approaches in nanomedicine center on the use of a single nanocarrier modified with multiple copies of multiple different functional domains, e.g., epitopes for vaccines. Therefore, we set out to develop a combinatorial approach toward the next-generation concept of epitope delivery: a prime-boost strategy in which the same epitope is delivered using different nanocarriers. We tested this concept in the setting of HER2(+) breast cancer. We synthesized HER2-based cancer vaccines using three icosahedral plant viruses as carriers and evaluated the immune response as a result of repetitive, homologous immunization using BALB/c mice. Two of the vaccines induced a Th2-predominant response and the other a Th1-predominant response. To enhance the immunogenicity of the vaccines, we developed a heterologous prime-boost strategy with each of the vaccines administered only once, yielding higher titers of HER2-specific immunoglobulins and increasing the toxicity of the antisera toward cancer cells. The prime-boost also induced a Th1-predominant response. An in vivo tumor challenge showed that the prime-boost regimen reduced tumor growth and improved survival in mice. This novel strategy to elicit robust immune responses against weakly immunogenic antigens in principle could be broadly applicable to cancers and other diseases.

Ethylene and ethylene absorber treatments were used to study the effects of ethylene content on the morphology and pigmentation of broccoli. Results showed that untreated broccoli began to turn yellow 8 days after being harvested at 10 °C. Ethylene treatment caused yellowing to occur 2 days earlier, and ethylene absorber treatment delayed and significantly reduced the degree of yellowing. Yellowing first occurred in the base of the bud; at that time, the shape of the bud had not changed significantly and there was no blooming of the florets. Scanning electron microscopy revealed that the increase in ethylene content caused the calyx cells of broccoli to shrink and that the cell arrangement had become disordered. Ethylene treatment caused increases in the content of chlorophyllide b (Chlide b), pheophorbide b (Pheide b), β-cryptoxanthin, lutein, and β-carotene. The increased degree of yellowing resulted in carotenoids rather than chlorophyll becoming the key pigment (β-cryptoxanthin, β-carotene, and lutein were very significantly correlated with h°). During this change, the expression levels of the BoChl2, BoPaO, BoRCCR, BoPSY, and BoLCYB genes were affected to a greater extent by ethylene. Ethylene absorber treatment did not reduce gene expression, but delayed it. These results indicate that the ethylene absorber only absorbed the ethylene produced and did not directly act on the degradation of chlorophyll. Ethylene also stimulated the expression of BoCCD1 and BoCCD7, resulting in the production of an unknown yellow substance that increased the yellow color of broccoli. In comparison, BoCCD7 expression was more sensitive than BoCCD1.

Owing to their low dimensionality, two-dimensional semiconductors, such as monolayer molybdenum disulfide, have a range of properties that make them valuable in the development of nanoelectronics. For example, the electronic bandgap of these semiconductors is not an intrinsic physical parameter and can be engineered by manipulating the dielectric environment around the monolayer. Here we show that this dielectric-dependent electronic bandgap can be used to engineer a lateral heterojunction within a homogeneous MoS2 monolayer. We visualize the heterostructure with Kelvin probe force microscopy and examine its influence on electrical transport experimentally and theoretically. We observe a lateral heterojunction with an approximately 90 meV band offset due to the differing degrees of bandgap renormalization of monolayer MoS2 when it is placed on a substrate in which one segment is made from an amorphous fluoropolymer (Cytop) and another segment is made of hexagonal boron nitride. This heterostructure leads to a diode-like electrical transport with a strong asymmetric behaviour. A lateral heterojunction with diode-like electrical transport can be created in a homogeneous MoS2 monolayer by using a substrate in which one segment is made from an amorphous fluoropolymer and another segment from hexagonal boron nitride.

In the present work, coal fly ash-derived mesoporous silica material (CFA-MS) has been successfully fabricated without employing any extra silica source. The obtained CFA-MS was characterized by Fourier transform infrared spectroscopy, nitrogen adsorption–desorption measurement, powder X-ray diffraction and transmission electron microscopy. Nitrogen adsorption–desorption measurement disclosed that CFA-MS possesses Brunauer–Emmett–Teller-specific surface area of 497 m2·g−1 and pore volume of 0.49 cm3·g−1, respectively. Furthermore, CFA-MS was evaluated for the adsorptive removal of methylene blue from aqueous solution. Several influence parameters on the removal of methylene blue including contact time, pH, initial concentration and temperature were studied in detail. Moreover, Langmuir, Freundlich, Temkin and Dubinin-Radushkevich isotherm models were employed for interpretation of the adsorption process, while the pseudo-first-order and pseudo-second-order kinetics equations were applied to investigate the adsorption kinetics. Results in the current work demonstrate that CFA-MS can be used as an efficient adsorbent for methylene blue removal.

Normal spermatogenic processes require the scrotal temperature to be lower than that of the body as excessive heat affects spermatogenesis in the testes, reduces sperm quality and quantity, and even causes infertility. Endoplasmic reticulum stress (ERS) is a crucial factor in many pathologies. Although several studies have linked ERS to heat stress, researchers have not yet determined which ERS signaling pathways contribute to heat-induced testicular damage. Melatonin activates antioxidant enzymes, scavenges free radicals, and protects the testes from inflammation; however, few studies have reported on the influence of melatonin on heat-induced testicular damage. Using a murine model of testicular hyperthermia, we observed that heat stress causes both ERS and apoptosis in the testes, especially in the spermatocytes. These observations were confirmed using the mouse spermatocyte cell line GC2, where the睾丸是精子发生的场所，哺乳动物的睾丸位于体腔外的阴囊中，成年哺乳动物包括人类，睾丸的正常温度均比身体核心温度要低。如果外界环境温度过高会影响睾丸中的精子生成，降低精子的质量和数量，甚至导致不育。当机体受到外界刺激时，内质网内会产生大量错误折叠或未折叠蛋白，使得内质网对蛋白的加工能力与载量失衡，从而导致内质网功能紊乱，即发生内质网应激（endoplasmic reticulum stress，ERS），ERS被认为是导致大量病理的关键因素。目前已有研究报道称热应激会引发ERS，但哪条ERS信号通路在热应激诱导的睾丸损伤中发挥主要作用尚不明确。褪黑激素（褪黑素）是一种具有抗氧化、抗凋亡能力的神经内分泌激素，它的抗氧化活性有助于防止应激、炎症引起的睾丸损伤，然而，褪黑素能否能缓解热应激所致的睾丸损伤尚未见报道。该研究通过建立小鼠睾丸热应激模型，并利用组织学、分子生物学检测发现，热应激会导致睾丸曲细精管内的生殖细胞数量减少、生精上皮紊乱及凋亡的产生，TUNEL染色结果显示凋亡主要发生在睾丸精母细胞中，通过实时荧光定量PCR（qRT-PCR）和免疫荧光技术发现热应激也会刺激小鼠睾丸发生ERS。在随后体外（GC2细胞系）试验中，我们通过qRT-PCR和Western blotting从RNA与蛋白水平进一步证实了上述结果，并发现ATF6和PERK信号通路在热诱导后的细胞中均被激活，而IRE1信号通路在该过程变化不明显。通过si-RNA干扰上述基因均可有效减少由热应激引起的精母细胞损伤。在此基础上继续探索褪黑素对热应激引起的精母细胞凋亡和ERS的影响，通过免疫荧光、流式细胞术、Western blotting等实验，发现褪黑素能通过褪黑素受体（MT1/MT2）抑制Atf6、Perk和凋亡相关基因Caspase12、Caspase3及Chop的表达。最后通过小鼠体内试验证实褪黑素可以缓解热应激引起的曲细精管生殖细胞数量减少和精母细胞凋亡。综上，该研究表明热应激能引起睾丸损伤并诱导精母细胞的ERS与凋亡的发生，褪黑素可以通过ATF6和PERK信号通路缓解热应激引起的精母细胞的凋亡，并对损伤的睾丸起到修复作用。该研究证明褪黑素可在一定程度上预防高温引起的精母细胞凋亡，并保护小鼠睾丸免受热应激的损害。.

We examined the associations of dietary cholesterol and egg intakes with cardiometabolic and all-cause mortality among Chinese and low-income Black and White Americans. Included were 47,789 Blacks, 20,360 Whites, and 134,280 Chinese aged 40–79 years at enrollment. Multivariable Cox models with restricted cubic splines were applied to estimate hazard ratios (HRs) and 95% confidence intervals (CIs) for mortality outcomes using intakes of 150 mg cholesterol/day and 1 egg/week as the references. Cholesterol intake showed a nonlinear association with increased all-cause mortality and a linear association with increased cardiometabolic mortality among Black Americans: HRs (95% CIs) associated with 300 and 600 mg/day vs. 150 mg/day were 1.07 (1.03–1.11) and 1.13 (1.05–1.21) for all-cause mortality (P-linearity = 0.04, P-nonlinearity = 0.002, and P-overall &lt, 0.001) and 1.10 (1.03–1.16) and 1.21 (1.08–1.36) for cardiometabolic mortality (P-linearity = 0.007, P-nonlinearity = 0.07, and P-overall = 0.005). Null associations with all-cause or cardiometabolic mortality were noted for White Americans (P-linearity ≥ 0.13, P-nonlinearity ≥ 0.06, and P-overall ≥ 0.05 for both). Nonlinear inverse associations were observed among Chinese: HR (95% CI) for 300 vs. 150 mg/day was 0.94 (0.92–0.97) for all-cause mortality and 0.91 (0.87–0.95) for cardiometabolic mortality, but the inverse associations disappeared with cholesterol intake &gt, 500 mg/day (P-linearity ≥ 0.12, P-nonlinearity ≤ 0.001, P-overall &lt, 0.001 for both). Similarly, we observed a positive association of egg intake with all-cause mortality in Black Americans, but a null association in White Americans and a nonlinear inverse association in Chinese. In conclusion, the associations of cholesterol and egg intakes with cardiometabolic and all-cause mortality may differ across ethnicities who have different dietary patterns and cardiometabolic risk profiles. However, residual confounding remains possible.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is continuing to evolve around the world, generating new variants that are of concern on the basis of their potential for altered transmissibility, pathogenicity, and coverage by vaccines and therapeutic agents1–5. Here we show that serum samples taken from twenty human volunteers, two or four weeks after their second dose of the BNT162b2 vaccine, neutralize engineered SARS-CoV-2 with a USA-WA1/2020 genetic background (a virus strain isolated in January 2020) and spike glycoproteins from the recently identified B.1.617.1, B.1.617.2, B.1.618 (all of which were first identified in India) or B.1.525 (first identified in Nigeria) lineages. Geometric mean plaque reduction neutralization titres against the variant viruses—particularly the B.1.617.1 variant—seemed to be lower than the titre against the USA-WA1/2020 virus, but all sera tested neutralized the variant viruses at titres of at least 1:40. The susceptibility of the variant strains to neutralization elicited by the BNT162b2 vaccine supports mass immunization as a central strategy to end the coronavirus disease 2019 (COVID-19) pandemic globally. Samples of serum from individuals immunized with the BNT162b2 vaccine show neutralization activity against engineered SARS-CoV-2s bearing the spike mutations from B.1.617 and other variants.

Colchicine (COL), an ancient and well-known drug, has been used in clinical practice for centuries. On the other hand, COL has also attracted extensive concerns for its potent toxic effects, especially gastrointestinal adverse reactions (nausea, vomiting, and diarrhea) before clinical symptoms relief. In this study, we used a rodent model to study the effects of COL on gastric mucosa and associated microbiota. The mice were exposed to various concentrations of COL (0.1, 0.5, and 2.5 mg kg−1 body weight per day) for 7 days, and the results showed that COL treatment caused severe gastric mucosal damage, accompanied by a significant decrease in gastric mucosal proinflammatory cytokines (IL-1β, IL-6, and TNF-α). The 16S rRNA gene sequencing revealed that COL significantly perturbed the gastric microbiota composition and reduced the gastric microbiota diversity in mice. Also, we identified bacterial biomarkers associated with diarrhea, including phylum Firmicutes, class Bacilli, order Lactobacillales, family Lactobacillaceae, genu Lactobacillus, and genu Blautia, suggesting that COL-triggered adverse reactions are closely related to gastric microbial perturbations. Our findings open new paths for understanding the mechanism of COL-related adverse gastrointestinal reactions, broadening the scientific view on the interaction between drugs and host gastrointestinal microbiota.

BACKGROUND: Choline is an essential nutrient; however, the associations of choline and its related metabolites with cardiometabolic risk remain unclear. OBJECTIVE: We examined the associations of circulating choline, betaine, carnitine, and dimethylglycine (DMG) with cardiometabolic biomarkers and their potential dietary and nondietary determinants. METHODS: The cross-sectional analyses included 32,853 participants from 17 studies, who were free of cancer, cardiovascular diseases, chronic kidney diseases, and inflammatory bowel disease. In each study, metabolites and biomarkers were log-transformed and standardized by means and SDs, and linear regression coefficients (β) and 95% CIs were estimated with adjustments for potential confounders. Study-specific results were combined by random-effects meta-analyses. A false discovery rate

Therapeutic mRNAs and vaccines are being developed for a broad range of human diseases, including COVID-19. However, their optimization is hindered by mRNA instability and inefficient protein expression. Here, we describe design principles that overcome these barriers. We develop an RNA sequencing-based platform called PERSIST-seq to systematically delineate in-cell mRNA stability, ribosome load, as well as in-solution stability of a library of diverse mRNAs. We find that, surprisingly, in-cell stability is a greater driver of protein output than high ribosome load. We further introduce a method called In-line-seq, applied to thousands of diverse RNAs, that reveals sequence and structure-based rules for mitigating hydrolytic degradation. Our findings show that highly structured “superfolder” mRNAs can be designed to improve both stability and expression with further enhancement through pseudouridine nucleoside modification. Together, our study demonstrates simultaneous improvement of mRNA stability and protein expression and provides a computational-experimental platform for the enhancement of mRNA medicines.

Calcineurin inhibitors (CNIs) are vital immunosuppressive therapies in the management of inflammatory conditions. A long-term consequence is nephrotoxicity. In the kidneys, the primary, catalytic calcineurin (CnA) isoforms are CnAα and CnAβ. Although the renal phenotype of CnAα(−/−) mice substantially mirrors CNI-induced nephrotoxicity, the mechanisms downstream of CnAα are poorly understood. Since NADPH oxidase-2 (Nox2)-derived oxidative damage has been implicated in CNI-induced nephrotoxicity, we hypothesized that CnAα inhibition drives Nox2 upregulation and promotes oxidative stress. To test the hypothesis, Nox2 regulation was investigated in kidneys from CnAα(−/−), CnAβ(−/−), and wild-type (WT) littermate mice. To identify the downstream mediator of CnAα, nuclear factor of activated T cells (NFAT) and NF-κB regulation was examined. To test if Nox2 is transcriptionally regulated via a NF-κB pathway, CnAα(−/−) and WT renal fibroblasts were treated with the NF-κB inhibitor caffeic acid phenethyl ester. Our findings showed that cyclosporine A treatment induced Nox2 upregulation and oxidative stress. Furthermore, Nox2 upregulation and elevated ROS generation occurred only in CnAα(−/−) mice. In these mice, NF-κB but not NFAT activity was increased. In CnAα(−/−) renal fibroblasts, NF-κB inhibition prevented Nox2 upregulation and reactive oxygen species (ROS) generation. In conclusion, these findings indicate that 1) CnAα loss stimulates Nox2 upregulation, 2) NF-κB is a novel CnAα-regulated transcription factor, and 3) NF-κB mediates CnAα-induced Nox2 and ROS regulation. Our results demonstrate that CnAα plays a key role in Nox2 and ROS generation. Furthermore, these novel findings provide evidence of divergent CnA isoform signaling pathways. Finally, this study advocates for CnAα-sparing CNIs, ultimately circumventing the CNI nephrotoxicity. NEW & NOTEWORTHY A long-term consequence of calcineurin inhibitors (CNIs) is oxidative damage and nephrotoxicity. This study indicates that NF-κB is a novel calcineurin-regulated transcription factor that is activated with calcineurin inhibition, thereby driving oxidative damage in CNI nephropathy. These findings provide additional evidence of divergent calcineurin signaling pathways and suggest that selective CNIs could improve the long-term outcomes of patients by mitigating renal side effects.

Background Diet is known to affect human gut microbiome composition; yet, how diet affects gut microbiome functionality remains unclear. Objective We compared the diversity and abundance/presence of fecal microbiome metabolic pathways among individuals according to their long-term diet quality. Methods In 2 longitudinal cohorts, we assessed participants’ usual diets via repeated surveys during 1996–2011 and collected a stool sample in 2015–2018. Participants who maintained a healthy or unhealthy diet (i.e., stayed in the highest or lowest quintile of a healthy diet score throughout follow-up) were selected. Participants were excluded if they reported a history of cancer, cardiovascular disease, diabetes, or hypertension; had diarrhea or constipation in the last 7 d; or used antibiotics in the last 6 mo before stool collection. Functional profiling of shotgun metagenomics was performed using HUMAnN2. Associations of dietary variables and 420 microbial metabolic pathways were evaluated via multivariable-adjusted linear or logistic regression models. Results We included 144 adults (mean age = 64 y; 55% female); 66 had an unhealthy diet and 78 maintained a healthy diet. The healthy diet group had higher Shannon α-diversity indexes of microbial gene families and metabolic pathways (both P 0.50). At P

Recently, a new SARS-CoV-2 lineage called B.1.1.7 has emerged in the United Kingdom that was reported to spread more efficiently than other strains. This variant has an unusually large number of mutations with 10 amino acid changes in the spike protein, raising concerns that its recognition by neutralizing antibodies may be affected. Here, we investigated SARS-CoV-2-S pseudoviruses bearing either the Wuhan reference strain or the B.1.1.7 lineage spike protein with sera of 16 participants in a previously reported trial with the mRNA-based COVID-19 vaccine BNT162b2. The immune sera had equivalent neutralizing titers to both variants. These data, together with the combined immunity involving humoral and cellular effectors induced by this vaccine, make it unlikely that the B.1.1.7 lineage will escape BNT162b2-mediated protection.

Human respiratory syncytial virus (RSV) is the leading viral cause of lower respiratory tract disease in infants and children worldwide. Currently, there are no FDA-approved vaccines to combat this virus. The large (L) polymerase protein of RSV replicates the viral genome and transcribes viral mRNAs. The L protein is organized as a core ring-like domain containing the RNA-dependent RNA polymerase and an appendage of globular domains containing an mRNA capping region and a cap methyltransferase region, which are linked by a flexible hinge region. Here, we found that the flexible hinge region of RSV L protein is tolerant to amino acid deletion or insertion. Recombinant RSVs carrying a single or double deletion or a single alanine insertion were genetically stable, highly attenuated in immortalized cells, had defects in replication and spread, and had a delay in innate immune cytokine responses in primary, well-differentiated, human bronchial epithelial (HBE) cultures. The replication of these recombinant viruses was highly attenuated in the upper and lower respiratory tracts of cotton rats. Importantly, these recombinant viruses elicited high levels of neutralizing antibody and provided complete protection against RSV replication. Taken together, amino acid deletions or insertions in the hinge region of the L protein can serve as a novel approach to rationally design genetically stable, highly attenuated, and immunogenic live virus vaccine candidates for RSV. IMPORTANCE Despite tremendous efforts, there are no FDA-approved vaccines for human respiratory syncytial virus (RSV). A live attenuated RSV vaccine is one of the most promising vaccine strategies for RSV. However, it has been a challenge to identify an RSV vaccine strain that has an optimal balance between attenuation and immunogenicity. In this study, we generated a panel of recombinant RSVs carrying a single and double deletion or a single alanine insertion in the large (L) polymerase protein that are genetically stable, sufficiently attenuated, and grow to high titer in cultured cells, while retaining high immunogenicity. Thus, these recombinant viruses may be promising vaccine candidates for RSV.

Removal of naturally occurring strontium (Sr) from groundwater is vital as excessive exposure may lead to bone growth problems in children. Nanofiltration/reverse osmosis (NF/RO) is commonly used in groundwater treatment due to the high effectiveness and simple maintenance of these pressure driven membrane processes. In this research, a pilot-scale NF/RO system was used to desalinate a natural groundwater sample containing high Sr concentration (10.3 mg/L) and &ldquo, old&rdquo, groundwater organic matter (70.9 mg/L) from Esilalei in northern Tanzania to understand the removal of strontium by NF/RO. The impact of applied pressure (10&ndash, 15 bar) and groundwater pH (3&ndash, 12) on the membrane performance including permeate flux, strontium and total organic carbon (TOC) flux and removal was investigated. Increasing applied pressure was found to enhance the flux by increasing the driving force and enhance Sr and TOC removal by dilution effect (water flux higher than Sr passage). The alkaline pH caused severe flux decline likely due to membrane fouling and scaling, while it slightly enhanced Sr removal of RO membranes, but weakened the TOC removal. In contrast, acidic and neutral pH of groundwater enhanced TOC removal. These findings suggest that appropriately high applied pressure and acidic pH condition of groundwater are recommended to apply to the NF/RO membrane system in groundwater desalination to achieve better membrane performance.

A method for quantitative analysis of methotrexate (MTX) from formulated gels/creams and from tissue/solution samples after percutaneous permeation studies is described. High performance liquid chromatography (I-IPLC) with a MicroBondapak C18, reverse-phase column was used. A calibration curve of doses versus peak areas for MTX showed linearity over the range of 0.010-14.056 μg with a very high correlation coefficient (r) of 0.99998. Replicate analysis of μg of MTX showed that the peak area was 9627424±22111 counts (mean ± SD; one count is 1/8 μV-sec, 1μY is 2×10^(-6)a.u.) with a relative standard deviation of 0.23%. The resolution factor between MTX and folic acid was 33.35 when the applied doses of both were 1 μg each. Results demonstrated that the photolytic products of MTX, samples from gel and samples extracted from hairless guinea pig skin after percutaneous permeation and epidermis/dermis separation by microwave technique, could be well detected by this method. Preliminary results are satisfactory and meet the requirements of U.S. Pharmacopeia XXII. This report decribes novel approaches for determination of methotrexate content in the formulations and skin specimens. These approaches include the separation of epidermis and dermis by microwaving, the pretreatment procedures employed before analysis, and the use of guard column during HPLC analysis.