Application of aptamer-conjugated graphene oxide for specific enrichment of microcystin-LR in Achatina fulica prior to matrix-assisted laser desorption ionization mass spectrometry.

Microcystin-LR (MC-LR), as a hepatotoxin, can cause liver swelling, hepatitis, and even liver cancer. In this study, MC-LR aptamer (Apt-3) modified graphene oxide (GO) was designed to enrich MC-LR in white jade snail (Achatina fulica) and pond water, followed by matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) analysis. Results indicated that the Apt-3/PEG/GO nanocomposites were highly specific to MC-LR, and the detection limit of MALDI-MS was 0.50 ng/mL. Moreover, the MC-LR can be released from nanocomposites at 75°C, thus, the reuse of Apt-3/PEG/GO is realized. Real sample analysis indicated that the Apt-3/PEG/GO nanocomposites coupled with MALDI-MS were efficient in detecting trace amounts of MC-LR in real samples. With the merits of being low cost, reusable, and easy to besynthesized, this Apt-3/PEG/GO MALDI-MS is expected to be comprehensively applied by anchoring suitable aptamers for different targets.

Neuroprotective effect of plasmalogens on AlCl3-induced Alzheimer's disease zebrafish via acting on the regulatory network of ferroptosis, apoptosis and synaptic neurotransmission release with oxidative stress as the center.

Plasmalogens (Pls) are considered to play a potential role in the treatment of neurodegenerative diseases. In the present study, an Alzheimer's disease (AD) model of zebrafish induced by AlCl3 was established to investigate whether the marine-derived Pls could alleviate cognitive impairments of AD zebrafish. Behavioral tests were carried out to assess the athletic ability. The transcriptional profiles of zebrafish in the control, AD model and AD_PLS group were compared and analyzed to determine the potential mechanisms of dietary Pls on AD. The study found that Pls could reverse athletic impairment in the AD zebrafish model, and the expression levels of genes related to ferroptosis, synaptic dysfunction and apoptosis were significantly altered between experimental groups. Further analysis showed that all of these genes were associated with oxidative stress (OS). These data suggest that healthy protective role of marine-derived Pls on AD zebrafish may result from inhibition of ferroptosis and neuronal apoptosis, restoring synaptic neurotransmission release, and reducing neuroinflammation. Among them, Oxidative stress is acted as the center to connect different regulation pathways. This study provides evidence to support the essential roles of OS in pathogenesis of AD, and the application of Pls in relieving AD.

Reversing stage III oral adenocarcinoma in a dog treated with anti-canine PD-1 therapeutic antibody: a case report.

Monoclonal antibody targeting programmed cell death-1 (PD-1) is one of the most promising treatment therapies for human cancers. Canine PD-1 antibodies used in clinical trials have also shown efficacy in treating canine cancers. An 11-year-old male intact border collie presented to us for evaluation of left cervical mass. Computed tomography (CT) examination revealed an irregular pharyngeal mass invading the surrounding soft tissue. Histological and immunohistochemical results were consistent with a diagnosis of adenocarcinoma, most likely originating from the minor salivary gland. An anti-canine PD-1 monoclonal antibody was administered. Two months after the initial treatment, the tumor reached partial remission and maintained as such for 6 months. Finally, the patient was euthanized due to reasons unrelated to cancer, with a survival time of 316 days. To our knowledge, this is the first report of response to PD-1 blockade treatment in canine adenocarcinoma.

Transcriptomics integrated with metabolomics reveals the ameliorating effect of mussel-derived plasmalogens on high-fat diet-induced hyperlipidemia in zebrafish.

Plasmalogens (Pls), a special group of phospholipids, are effective in ameliorating neurodegenerative disease. In the present study, the metabolic effects of seafood-derived Pls on high fat diet (HFD)-induced hyperlipidemia in zebrafish were evaluated, and the underlying mechanisms of dietary Pls against hyperlipidemia were explored through integrated analyses of hepatic transcriptomics and metabolomics. The results demonstrated that Pls supplementation could effectively alleviate HFD-induced obesity symptoms, such as body weight gain, and decrease total hepatic cholesterol and triglyceride levels. Integrated hepatic transcriptome and metabolome data suggested that Pls mainly altered lipid metabolism pathways (FA metabolism, primary bile acid biosynthesis, steroid hormone biosynthesis, and glycerolipid and glycerophospholipid metabolism) and the TCA cycle, induced the overexpression of anti-oxidation enzymes (Cat, Gpx4, Sod3a and Xdh), reduced disease biomarkers (such as glutarylcarnitine, gamma-glutamyltyrosine, and 11-prostaglandin f2) and gut microbiota-derived metabolites, and increased (±)12(13)-diHOME, EPA, lysoPC and PC levels. Moreover, 5 abnormally regulated metabolites were identified as potential biomarkers associated with hyperlipidemia according to the metabolomics results and suggested the involvement of gut microbiota in the anti-hyperlipidemic effects of Pls. Collectively, these findings suggest that the protective role of Pls is mainly associated with the promotion of unsaturated fatty acid biosynthesis and cholesterol efflux, lipid and phospholipid PUFA remodeling, and anti-oxidation and anti-inflammatory capabilities. This study provides valuable information for reasonably explaining the beneficial effects of seafood-derived Pls in alleviating hyperlipidemia and thus may contribute to the development and application of Pls as functional foods or dietary supplements to protect against obesity and hyperlipidemia.

Effect of Graphene Nanosheets on the Microstructure and Mechanical Properties of Sn-20Bi Solder.

The application of Sn-Bi series solder is limited due to the brittleness of Bi phase. Sn-20Bi solder with less Bi element content has great research prospects, but it needs modification to make it a substitute for traditional Sn-Pb solder. In this article, we mixed graphene nanosheets with nanometer Sn powder by means of ultrasonic oscillation, and Sn-20Bi-qGNS (q = 0.01, 0.02, 0.04, 0.06, and 0.1 wt.%) solder alloys were prepared by the melt-casting method. The effects of graphene nanosheets (GNSs) on the microstructure, physical properties, mechanical properties, and corrosion resistance of solder alloys were investigated. Scanning electron microscopy, energy dispersive spectroscopy, X-ray diffraction, and X-ray photoelectron spectroscopy were used to determine the microstructural morphology and composition. The results showed that the melting point, density, and wettability of the solder decreased slightly with the addition of GNSs. The addition of GNSs as a second phase refined the solder structure and improved the tensile strength of the molten Sn-20Bi composite solder to 99.6 MPa, while elongation decreased with the addition of GNSs. Furthermore, GNSs prevented the MC Sn-20Bi-qGNSs/Cu intermetallic compound layers' growth by interfering with atomic diffusion and grain boundary movement. In addition, the addition of 0.02 wt.% GNSs enhanced the shear strength of MC Sn-20Bi solder joints to 46.3 MPa. The electrochemical experimental results show that the surface corrosion products of MC Sn-20Bi-qGNSs under 3.5% NaCl solution were Sn3O(OH)2Cl2, with MC Sn-20Bi-0.01GNSs exhibiting the best corrosion resistance.

Aptamer-conjugated graphene oxide-based surface assisted laser desorption ionization mass spectrometry for selective extraction and detection of Aß1-42 in an Alzheimer's disease SH-SY5 cell model.

The generation and accumulation of amyloid-beta peptide (Aß1-42) in amyloid plaques are key characteristics of Alzheimer's disease (AD); thus, specific detection of Aß1-42 is essential for the diagnosis and treatment of AD. Herein, an aptamer-conjugated graphene oxide (Apt-GO) sensor was synthesized by π-π and hydrophobic interactions using thiol poly (ethylene glycol) amine (SH-PEG-NH2) as a spacer unit. Then, it was applied to selective capture of Aß1-42, and the resulting complex was directly analyzed by surface-assisted laser desorption ionization mass spectrometry (SALDI-MS). The results revealed that the Apt-GO could enhance the detection specificity and reduce non-specific adsorption. This method was validated to be sensitive in detecting Aß1-42 at a low level in human serum (ca. 0.1 µM) within a linear range from 0.1 to 10 µM. The immobilizing amount of aptamer on the GO was calculated to be 36.1 nmol/mg (RSD = 11.5%). In conclusion, this Apt-GO-based SALDI-MS method was sensitive and efficient in selective extraction and detection of Aß1-42, which proved to be a good option for early AD diagnosis.

Characterization of a vaccine-elicited human antibody with sequence homology to VRC01-class antibodies that binds the C1C2 gp120 domain.

Broadly HIV-1-neutralizing VRC01-class antibodies bind the CD4-binding site of Env and contain VH1-2*02-derived heavy chains paired with light chains expressing five-amino acid-long CDRL3s. Their unmutated germline forms do not recognize HIV-1 Env, and their lack of elicitation in human clinical trials could be due to the absence of activation of the corresponding naïve B cells by the vaccine immunogens. To address this point, we examined Env-specific B cell receptor sequences from participants in the HVTN 100 clinical trial. Of all the sequences analyzed, only one displayed homology to VRC01-class antibodies, but the corresponding antibody (FH1) recognized the C1C2 gp120 domain. For FH1 to switch epitope recognition to the CD4-binding site, alterations in the CDRH3 and CDRL3 were necessary. Only germ line-targeting Env immunogens efficiently activated VRC01 B cells, even in the presence of FH1 B cells. Our findings support the use of these immunogens to activate VRC01 B cells in humans.

The Stability and Electronic Structure of Cu(200)/AuCu(200) Interface: An Insight from First-Principle Calculation.

AuCu phase had a significant effect on the bonding strength of Au80Sn20 alloy and Cu substrate. The formation of the AuCu(200)/Cu(200) interface significantly improves the shear strength of solder joints. Therefore, it is particularly important to analyze the strengthening mechanism of the AuCu phase in the Cu matrix. The atomic structure, interfacial stability, and interfacial bonding properties of the Cu(200)/AuCu(200) interface were investigated using first-principle calculation. The layer spacing convergence results show that seven layers of Cu(200) surface and seven layers of AuCu(200) surface are enough thick to be chosen for the interface model. The calculation shows that the surface energies are 1.463 J/m2 and 1.081 J/m2 for AuCu(200) surface and Cu(200) surface, respectively. Four interface combinations of Top sit, Long bridge, Short bridge, and Hollow were investigated by considering four stacking methods of AuCu(200). It is shown that the interfacial configuration of the Long bridge is the most stable and favorable structure, which has the largest adhesion work, the smallest interfacial energy, and the smallest interfacial spacing. The density of states and electron difference density were calculated for the four interfacial configurations, and the results showed that the main bonding mode of the Long bridge interface is composed of both Cu-Cu covalent bonds and Au-Cu covalent bonds.

Selective separation and recovery of lithium, nickel, MnO2, and Co2O3 from LiNi0.5Mn0.3Co0.2O2 in spent battery.

The recovery of valuable metals from the LiNi0·5Mn0·3Co0·2O2 in spent batteries deserves more attention. We report a series of feasible procedures to selectively recover the four metals (Li, Ni, Mn, and Co) using a combination of hydrometallurgical and pyrometallurgyical processes. Firstly, oxalic acid is used to dissolve Li and precipitate the other three metals in oxalate forms. It is found that under the optimal condition, about 98% of the Li is dissolved, and on average 93% of the other three metals are transformed to precipitated oxalates. The oxalates are then transformed to NiO·Mn2O3·Co3O4 by being calcinated at 723 K under atmospheric environment. The selective recovery of NiO·Mn2O3·Co3O4 can be achieved by using H2SO4 under three different conditions. The first step is to use H2SO4 to selectively dissolve CoO from the Co3O4. Then the combination of H2SO4 and ultrasound is adopted to dissolve NiO, during which the ultrasound destroys the surficial oxide film on the NiO. Afterwards, the Mn2O3 is transformed to MnO2 and Mn2+ in heated H2SO4. The Co, Ni and Mn ions are dissolved in a sequence, which facilitates their separation and recovery. As the main components of the final residual solids, Co2O3 and MnO2 present in distinctly different sizes and shapes, which are beneficial for their separation and direct usage.

Protect Effects of Seafood-Derived Plasmalogens Against Amyloid-Beta (1-42) Induced Toxicity via Modulating the Transcripts Related to Endocytosis, Autophagy, Apoptosis, Neurotransmitter Release and Synaptic Transmission in SH-SY5Y Cells.

To investigate the underlying mechanisms of decreased plasmalogens (Pls) levels in neurodegenerative diseases, here the effects of seafood-derived Pls on undifferentiated and differentiated human SH-SY5Y neuroblastoma cells exposed to amyloid-ß1-42 was analyzed. Transcriptional profiles indicated that a total of 6,581 differentially expressed genes (DEGs) were significantly identified among different experimental groups, and KEGG analysis indicated that these DEGs were related to AD, endocytosis, synaptic vesicle cycle, autophagy and cellular apoptosis. After Pls treatment, the striking expression changes of ADORA2A, ATP6V1C2, CELF6, and SLC18A2 mRNA strongly suggest that Pls exerts a beneficial role in alleviating AD pathology partly by modulating the neurotransmitter release and synaptic transmission at the transcriptional level. Besides these, GPCRs are also broadly involved in Pls-signaling in neuronal cells. These results provide evidence for supporting the potential use of Pls as an effective therapeutic approach for AD.

In Situ Quality Assessment of Dried Sea Cucumber (Stichopus japonicus) Oxidation Characteristics during Storage by iKnife Rapid Evaporative Ionization Mass Spectrometry.

Sea cucumber (Stichopus japonicus) is one of the most luxurious and nutritious seafoods in Asia. It is always processed into dried products to prevent autolysis, but its quality is easily destructed during storage. Herein, an extremely simplified workflow was established for real-time and in situ quality assessment of dried sea cucumbers (DSCs) during storage based on the lipid oxidation characteristics using an intelligent surgical knife (iKnife) coupled with rapid evaporative ionization mass spectrometry (REIMS). The lipidomic phenotypes of DSCs at different storage times were acquired successfully, which were then processed by multivariate statistical analysis. The results showed that the discrepancy in the characteristic ions in different DSCs was significant (p < 0.05) with high R2(Y) and Q2 values (0.975 and 0.986, respectively). The receiver operating characteristic curve revealed that the ions of m/z 739.5, m/z 831.5, m/z 847.6, and m/z 859.6 were the most specific and characteristic candidate biomarkers for quality assessment of DSCs during accelerated storage. Finally, this method was validated to be qualified in precision (RSDintraday ≤ 9.65% and RSDinterday ≤ 9.36%). In conclusion, the results showed that the well-established iKnife-REIMS method was high-throughput, rapid, and reliable in the real-time quality assessment of DSCs.

Plasmalogens improve swimming performance by modulating the expression of genes involved in amino acid and lipid metabolism, oxidative stress, and ferroptosis in an Alzheimer's disease zebrafish model.

Plasmalogens (PLs) are critical to human health. Studies have reported a link between the downregulation of PLs levels and cognitive impairments in patients with Alzheimer's disease (AD). However, the underlying mechanisms remain to be clarified. In the present study, an AlCl3-induced AD zebrafish model was established, and the model was used to elucidate the neuroprotective effects of PLs on AD by analysing the transcriptional profiles of zebrafish in the control, AD model, AD_PL, and PL groups. Chronic AlCl3 exposure caused swimming performance impairments in the zebrafish, yet PLs supplementation could improve the dyskinesia recovery rate in the AD zebrafish model. Through transcriptional profiling, a total of 5413 statistically significant differentially expressed genes (DEGs) were identified among the groups. In addition to the DEGs involved in amino acid metabolism, we found that the genes related to iron homeostasis, lipid peroxidation, and oxidative stress, all of which contribute to ferroptosis, were dramatically altered among different groups. These results suggest that seafood-derived PLs, in addition to their role in eliminating oxidative stress, can improve the swimming performance in AlCl3-exposed zebrafish partly by suppressing neuronal ferroptosis and accelerating synaptic transmission at the transcriptional level. This study provides evidence for PLs to be developed as a functional food supplement to relieve AD symptoms.

Characterization of Metabolites in a Zebrafish Model of Alzheimer's Disease Supplemented with Mussel-Derived Plasmalogens by Ultraperformance Liquid Chromatography Q-Exactive Orbitrap Mass Spectrometry-Based Unbiased Metabolomics.

Plasmalogens (Pls) are bioactive substances enriched in the brain with a regulatory effect on Alzheimer's disease (AD), while their metabolomic influence accompanying AD and the underlying mechanisms remain unclear. Here, we extracted and purified Pls (purity of ≥90%) from mussels and applied unbiased metabolomics using ultraperformance liquid chromatography Q-Exactive Orbitrap mass spectrometry to analyze the variation of metabolites in the major metabolic pathways of AD and revealed the cognitive improvement effect of Pls using an experimental AD zebrafish model. The results showed that 37 differential endogenous metabolites were identified, among which glycerophosphocholine, choline, S-adenosylmethionine (SAM), l-glutamine, linoleic acid, 9(S)-HPODE, methionine, and creatine were the major abnormally regulated metabolites, and the first four metabolites were viewed as potential endogenous markers. This study suggested that systemic metabolic profiling could reveal the potential metabolic networks of AD and illuminated the protective effect of Pls on AD through biochemistry mechanisms and metabolic pathways.

Isolation and characterization of cross-neutralizing coronavirus antibodies from COVID-19+ subjects.

SARS-CoV-2 is one of three coronaviruses that have crossed the animal-to-human barrier and caused widespread disease in the past two decades. The development of a universal human coronavirus vaccine could prevent future pandemics. We characterize 198 antibodies isolated from four COVID-19+ subjects and identify 14 SARS-CoV-2 neutralizing antibodies. One targets the N-terminal domain (NTD), one recognizes an epitope in S2, and 11 bind the receptor-binding domain (RBD). Three anti-RBD neutralizing antibodies cross-neutralize SARS-CoV-1 by effectively blocking binding of both the SARS-CoV-1 and SARS-CoV-2 RBDs to the ACE2 receptor. Using the K18-hACE transgenic mouse model, we demonstrate that the neutralization potency and antibody epitope specificity regulates the in vivo protective potential of anti-SARS-CoV-2 antibodies. All four cross-neutralizing antibodies neutralize the B.1.351 mutant strain. Thus, our study reveals that epitopes in S2 can serve as blueprints for the design of immunogens capable of eliciting cross-neutralizing coronavirus antibodies.

Impact of air-frying on the plasmalogens deterioration and oxidation in oyster revealed by mild acid hydrolysis and HILIC-MS-based lipidomics analysis.

Oyster is rich in plasmalogens that are ether phospholipids with biological functions to human body. Air-frying is a popular technique for preparing delicious oyster but makes the plasmalogens vulnerable to oxidation. In this study, the effect of air-frying processing on plasmalogens oxidation was studied by lipidomic approach. Plasmalogens were always mixed with normal phospholipids, thus the lipid extract was treated with mild acid hydrolysis to rapidly degrade plasmalogens owing to the acid lability of vinyl ether linkage at sn-1 position. After hydrophilic interaction chromatography MS/MS analysis, there were three plasmalogen classes, plasmanylcholine, plasmanylethanolamine, and plasmanylinositol, completely separated, and each plasmalogen molecular species was identified and quantified. It indicated that the content of plasmalogens underwent an obvious decrease during the air-frying process. To weaken such effect, the influence of air-frying temperature was further inspected by multivariate statistical analyses. The main variables, including the ions of m/z 756.4927, 784.5486, 828.5812, etc., were revealed by unsupervised principle component analysis, supervised orthogonal partial least-square analysis, and variable importance in projection plot. As a conclusion, air-frying has health benefits in reducing fat content but destructive to plasmalogens, thus interventions are recommended to prevent the degradation of plasmalogens.

Development of a VRC01-class germline targeting immunogen derived from anti-idiotypic antibodies.

An effective HIV-1 vaccine will likely need to elicit broadly neutralizing antibodies (bNAbs). Broad and potent VRC01-class bNAbs have been isolated from multiple infected individuals, suggesting that they could be reproducibly elicited by vaccination. Several HIV-1 envelope-derived germline-targeting immunogens have been designed to engage naive VRC01-class precursor B cells. However, they also present off-target epitopes that could hinder development of VRC01-class bNAbs. We characterize a panel of anti-idiotypic monoclonal antibodies (ai-mAbs) raised against inferred-germline (iGL) VRC01-class antibodies. By leveraging binding, structural, and B cell sorting data, we engineered a bispecific molecule derived from two ai-mAbs; one specific for VRC01-class heavy chains and one specific for VRC01-class light chains. The bispecific molecule preferentially activates iGL-VRC01 B cells in vitro and induces specific antibody responses in a murine adoptive transfer model with a diverse polyclonal B cell repertoire. This molecule represents an alternative non-envelope-derived germline-targeting immunogen that can selectively activate VRC01-class precursors in vivo.

Isolation and Characterization of Cross-Neutralizing Coronavirus Antibodies from COVID-19+ Subjects.

SARS-CoV-2 is one of three coronaviruses that have crossed the animal-to-human barrier in the past two decades. The development of a universal human coronavirus vaccine could prevent future pandemics. We characterized 198 antibodies isolated from four COVID19+ subjects and identified 14 SARS-CoV-2 neutralizing antibodies. One targeted the NTD, one recognized an epitope in S2 and twelve bound the RBD. Three anti-RBD neutralizing antibodies cross-neutralized SARS-CoV-1 by effectively blocking binding of both the SARS-CoV-1 and SARS-CoV-2 RBDs to the ACE2 receptor. Using the K18-hACE transgenic mouse model, we demonstrate that the neutralization potency rather than the antibody epitope specificity regulates the in vivo protective potential of anti-SARS-CoV-2 antibodies. The anti-S2 antibody also neutralized SARS-CoV-1 and all four cross-neutralizing antibodies neutralized the B.1.351 mutant strain. Thus, our study reveals that epitopes in S2 can serve as blueprints for the design of immunogens capable of eliciting cross-neutralizing coronavirus antibodies.

mRNA vaccination boosts cross-variant neutralizing antibodies elicited by SARS-CoV-2 infection.

Emerging SARS-CoV-2 variants have raised concerns about resistance to neutralizing antibodies elicited by previous infection or vaccination. We examined whether sera from recovered and naïve donors collected prior to, and following immunizations with existing mRNA vaccines, could neutralize the Wuhan-Hu-1 and B.1.351 variants. Pre-vaccination sera from recovered donors neutralized Wuhan-Hu-1 and sporadically neutralized B.1.351, but a single immunization boosted neutralizing titers against all variants and SARS-CoV-1 by up to 1000-fold. Neutralization was due to antibodies targeting the receptor binding domain and was not boosted by a second immunization. Immunization of naïve donors also elicited cross-neutralizing responses, but at lower titers. Our study highlights the importance of vaccinating both uninfected and previously infected persons to elicit cross-variant neutralizing antibodies.

Lipidomics phenotyping of clam (Corbicula fluminea) through graphene/fibrous silica nanohybrids based solid-phase extraction and HILIC-MS analysis.

Polyunsaturated phospholipids are abundant in clam (Corbicula fluminea) but difficult to be fully extracted. Herein, graphene/fibrous silica (G/KCC-1) nanohybrids were synthesized, characterized, and applied for solid-phase extraction (SPE) of phospholipids in clam. The effectiveness of G/KCC-1 SPE was verified by hydrophilic interaction chromatography mass spectrometry (HILIC-MS) based lipidomics and statistical analysis. The ions of PE 16:0/18:1 (m/z 716.4), PC 16:0/20:5 (m/z 824.6) and etc. were regarded as the main difference among the crude lipids, acetone washed extract, and eluate of G/KCC-1 SPE. Finally, this method was validated in terms of linearity (R2 0.9965 to 0.9981), sensitivity (LOD 0.19-0.51 µg·mL-1 and LOQ 0.48 - 1.47 µg·mL-1), and precision (RSDintra-day ≤ 7.16% and RSDinter-day ≤ 7.30%). In conclusion, the G/KCC-1 SPE and HILIC-MS method was shown to be accurate and efficient in selective extracting and phenotyping phospholipids in C. fluminea.