De novo protein sequencing of antibodies for identification of neutralizing antibodies in human plasma post SARS-CoV-2 vaccination.

The antibody response to vaccination and infection is a key component of the immune response to pathogens. Sequencing of peripheral B cells may not represent the complete B cell receptor repertoire. Here we present a method for sequencing human plasma-derived polyclonal IgG using a combination of mass spectrometry and B-cell sequencing. We investigate the IgG response to the Moderna Spikevax COVID-19 vaccine. From the sequencing data of the natural polyclonal response to vaccination, we generate 12 recombinant antibodies. Six derived recombinant antibodies, including four generated with de novo protein sequencing, exhibit similar or higher binding affinities than the original natural polyclonal antibody. Neutralization tests reveal that the six antibodies possess neutralizing capabilities against the target antigen. This research provides insights into sequencing polyclonal IgG antibodies and the potential of our approach in generating recombinant antibodies with robust binding affinity and neutralization capabilities. Directly examining the circulating IgG pool is crucial due to potential misrepresentations by B-cell analysis alone.

Synergistic Effect of Aluminum Nitride and Carbon Nanotube-Reinforced Silicon Rubber Nanocomposites.

Constructing a synergistic effect with different structural fillers is an important strategy for improving the comprehensive properties of polymeric composites. To improve the comprehensive properties of two-component additive liquid silicon rubber (SR) materials used in electronics packaging, the synergistic effect of granular aluminum nitride (AlN) and tubular carbon nanotube (CNT)-reinforced SR nanocomposites was investigated. AlN/CNT/SR composites with different AlN/CNT ratios were fabricated with two-component additive liquid SR via the thermal curing technique, and the influence of AlN/CNT hybrid fillers on the hardness, strength, elongation at break, surface resistivity, thermal conductivity, and thermal decomposition was investigated in detail. With the incorporation of AlN/CNT hybrid fillers, the comprehensive properties of the obtained AlN/CNT/SR composites are better than those of the AlN/SR and CNT/SR composites. The synergistic thermal conductive mechanism of AlN/CNT hybrid fillers was proposed and demonstrated with the fractural surface morphology of the obtained composites. The obtained AlN/CNT/SR composites show promising applications in electronic packaging, where necessary mechanical strength, electrical insulating, thermal conductivity, and thermal stable materials are needed.

Preparation and sustained release of diatomite incorporated and Eudragit L100 coated hydroxypropyl cellulose/chitosan aerogel microspheres.

The drug encapsulation efficiency, release rate and time, sustained release, and stimulus-response of carriers are very important for drug delivery. However, these always cannot obtained for the carrier with a single component. To improve the comprehensive performance of chitosan-based carriers for 5-Fu delivery, diatomite-incorporated hydroxypropyl cellulose/chitosan (DE/HPC/CS) composite aerogel microspheres were fabricated for the release of 5-fluorouracil (5-Fu), and the release performance was regulated with the content of diatomite, pH value, and external coating material. Firstly, the 5-Fu loaded DE/HPC/CS composite aerogel microspheres and Eudragit L100 coated microspheres were prepared with cross-linking followed by freeze-drying, and characterized by SEM, EDS, FTIR, XRD, DSC, TG, and swelling. The obtained aerogel microspheres have a diameter of about 0.5 mm, the weight percentage of F and Si elements on the surface are 0.55 % and 0.78 % respectively. The glass transition temperature increased from 179 °C to 181 °C and 185 °C with the incorporation of DE and coating of Eudragit, and the equilibrium swelling percentage of DE/HPC/CS (1.5:3:2) carriers are 101.52 %, 45.27 %, 67.32 % at pH 1.2, 5.0, 7.4, respectively. Then, the effect of DE content on the drug loading efficiency of DE/HPC/CS@5-Fu was investigated, with the increase of DE content, the highest encapsulation efficiency was 82.6 %. Finally, the release behavior of DE incorporated and Eudragit L100 Coated microspheres were investigated under different pH values, and evaluated with four kinetic models. The results revealed that the release rate of 5-Fu decreased with the increase of DE content, sustained release with extending time and pH-responsive were observed for the Eudragit-coated aerogel microspheres.

A novel lncRNA-hidden polypeptide regulates malignant phenotypes and pemetrexed sensitivity in A549 pulmonary adenocarcinoma cells.

The advance of high-throughput sequencing enhances the discovery of short ORFs embedded in long non-coding RNAs (lncRNAs). Here, we uncovered the production and biological activity of lncRNA-hidden polypeptides in lung adenocarcinoma (LUAD). In the present study, bioinformatics was used to screen the lncRNA-hidden polypeptides in LUAD. Analysis of protein expression was done by western blot or immunofluorescence assay. The functions of the polypeptide were determined by detecting its effects on cell viability, proliferation, migration, invasion, and pemetrexed (PEM) sensitivity. The protein interactors of the polypeptide were analyzed by mass spectrometry after Co-immunoprecipitation (Co-IP) assay. The results showed that the lncRNA LINC00954 was confirmed to encode a novel polypeptide LINC00954-ORF. The polypeptide had tumor-suppressor features in A549 cells by repressing cell growth, motility and invasion. Moreover, the polypeptide enhanced PEM sensitivity and suppressed growth in A549/PEM cells. The protein interactors of this polypeptide had close correlations with RNA processing, amide metabolic process, translation, RNA binding, RNA transport, and DNA replication. As a conclusion, the LINC00954-ORF polypeptide embedded in lncRNA LINC00954 possesses tumor-suppressor features in A549 and PEM-resistant A549 cells and sensitizes PEM-resistant A549 cells to PEM, providing evidence that the LINC00954-ORF polypeptide is a potential anti-cancer agent in LUAD.

Adsorption of Levofloxacin onto Graphene Oxide/Chitosan Composite Aerogel Microspheres.

The removal of pharmaceutical residues from water resources using bio-based materials is very important for human safety and health. Bio-based graphene oxide/chitosan (GO/CS) aerogel microspheres were fabricated with emulsification and cross-linking, followed by freeze drying, and were used for the adsorption of levofloxacin (LOF). The obtained GO/CS aerogel microspheres were characterized with scanning electron microscopy (SEM), Fourier-transform infrared (FTIR), and thermogravimetry (TG). The effects of GO content, pH value, and temperature on their adsorption capacity were investigated. With the incorporation of 40 wt% GO, the adsorption capacity increased from 9.9 to 45.6 mg/g, and the highest adsorption capacity, 51.5 mg/g, was obtained at pH = 8 and T = 25 °C. In addition, to obtain deeper insight into the adsorption process, the thermodynamics and kinetics of the process were also investigated with four different models of LOF adsorption. The thermodynamic modeling results revealed that LOF adsorption is exothermic, and the kinetic investigation demonstrated that LOF adsorption is generally consistent with a pseudo-first-order rate law.

Effect of Carbon Nanoparticles Morphology on the Properties of Poly(styrene-b-isoprene-b-styrene) Elastomer Composites.

Though nanomaterials based on carbon have been widely used for the preparation of high-performance polymeric nanocomposites, there are few works focused on the effect of carbon nanoparticle morphology on the performance of corresponding polymer nanocomposites. Therefore, four representative carbon nanoparticles, including fullerene, carbon nanotubes, graphene, and carbon black incorporated poly(styrene-b-isoprene-b-styrene) (SIS) elastomer nanocomposites were fabricated using the solvent casting method. In addition, the effect of carbon nanoparticle morphology on the rheological, mechanical, electrical, and thermal properties of the obtained polymeric nanocomposites was systematically investigated. The results showed that the shape of carbon nanoparticles has a different effect on the properties of the obtained elastomer nanocomposites, which lays the foundation of carbon nanoparticle screening for high-performance polymer nanocomposite construction.

Adsorption and Recognition Property of Tyrosine Molecularly Imprinted Polymer Prepared via Electron Beam Irradiation.

To realize the selective separation of L-tyrosine (L-Tyr) and avoid the drawbacks of traditional thermal polymerization, electron beam irradiation polymerization was developed for the fabrication of L-Tyr molecularly imprinted polymers (MIPs). Firstly, L-Tyr MIPs were prepared with methacrylic acid and ethylene glycol dimethacrylate and without an initiator. Then, the influence of absorbed dosage and temperature on the adsorption capacity of L-Tyr, as well as the thermodynamic behavior, were investigated. The maximum adsorption capacity of 10.96 mg/g for MIPs was obtained with an irradiation dosage of 340 kGy under 15 °C, and the ΔH0 and ΔS0 of the adsorption process are -99.79 kJ/mol and -0.31 kJ/mol·K, respectively. In addition, the effect of adsorption time on adsorption performance was evaluated under different initial concentrations, and the kinetic behavior was fitted with four different models. Finally, the recognition property of the obtained MIPs was investigated with L-Tyr and two analogues. The obtained MIPs have an imprinting factor of 5.1 and relatively high selective coefficients of 3.9 and 3.5 against L-tryptophan and L-phenylalanine, respectively. This work not only provided an L-Tyr MIP with high adsorption capacity and selectivity but also provided an effective and clean method for the synthesis of MIPs.

Equine Polyclonal Antibodies Prevent Acute Chikungunya Virus Infection in Mice.

Chikungunya virus (CHIKV) is a mosquito-transmitted pathogen that causes chikungunya disease (CHIK); the disease is characterized by fever, muscle ache, rash, and arthralgia. This arthralgia can be debilitating and long-lasting, seriously impacting quality of life for years. Currently, there is no specific therapy available for CHIKV infection. We have developed a despeciated equine polyclonal antibody (CHIKV-EIG) treatment against CHIKV and evaluated its protective efficacy in mouse models of CHIKV infection. In immunocompromised (IFNAR-/-) mice infected with CHIKV, daily treatment for five consecutive days with CHIKV-EIG administered at 100 mg/kg starting on the day of infection prevented mortality, reduced viremia, and improved clinical condition as measured by body weight loss. These beneficial effects were seen even when treatment was delayed to 1 day after infection. In immunocompetent mice, CHIKV-EIG treatment reduced virus induced arthritis (including footpad swelling), arthralgia-associated cytokines, viremia, and tissue virus loads in a dose-dependent fashion. Collectively, these results suggest that CHIKV-EIG is effective at preventing CHIK and could be a viable candidate for further development as a treatment for human disease.

Preparation and Application Progress of Imprinted Polymers.

Due to the specific recognition performance, imprinted polymers have been widely investigated and applied in the field of separation and detection. Based on the introduction of the imprinting principles, the classification of imprinted polymers (bulk imprinting, surface imprinting, and epitope imprinting) are summarized according to their structure first. Secondly, the preparation methods of imprinted polymers are summarized in detail, including traditional thermal polymerization, novel radiation polymerization, and green polymerization. Then, the practical applications of imprinted polymers for the selective recognition of different substrates, such as metal ions, organic molecules, and biological macromolecules, are systematically summarized. Finally, the existing problems in its preparation and application are summarized, and its prospects have been prospected.

Transgenerational inheritance of wing development defects in Drosophila melanogaster induced by cadmium.

The transgenerational inheritance of phenotype induced by environmental factors is a new focus in epigenetic research. In this study, Drosophila melanogaster (F0) was cultured in the medium containing cadmium (Cd, 4.5 mg/kg) from eggs to adults, and offspring (F1-F4) were continuously kept in standard medium (without cadmium). The phenotype analysis showed that cadmium induced developmental defects on wings and apoptosis in the wing disc cells of Drosophila (F0). The wing defects were transmitted for at least four generations even without Cd afterwards. And the effect on the mRNA expression of wing development related genes (shg, omb, F-actin, Mekk1) can be maintained for at least two or three generations. More importantly, under cadmium stress, the post-translational modification (PTM) on the histones H3K4me3 in the third instar larvae and ovaries or testes of adult flies increased significantly, while the levels of H3K9me3 and H3K27me3 decreased significantly. The expression of histone methylation related genes (dSet-1, ash1, Lsd1) increased significantly and these changes can be transmitted to offspring from one or two generations in ovaries or testes. These results suggest that the phenotypic defects of wings caused by cadmium can be inherited to the offspring, and this transgenerational inheritance effect may be related to the epigenetic regulation of histone methylation. Therefore, the adaptability of offspring should be considered when evaluating the toxicity and environmental risk of cadmium.

Adjuvant chemotherapy versus adjuvant concurrent chemoradiotherapy after radical surgery for early-stage cervical cancer: a randomized, non-inferiority, multicenter trial.

We conducted a prospective study to assess the non-inferiority of adjuvant chemotherapy alone versus adjuvant concurrent chemoradiotherapy (CCRT) as an alternative strategy for patients with early-stage (FIGO 2009 stage IB-IIA) cervical cancer having risk factors after surgery. The condition was assessed in terms of prognosis, adverse effects, and quality of life. This randomized trial involved nine centers across China. Eligible patients were randomized to receive adjuvant chemotherapy or CCRT after surgery. The primary end-point was progression-free survival (PFS). From December 2012 to December 2014, 337 patients were subjected to randomization. Final analysis included 329 patients, including 165 in the adjuvant chemotherapy group and 164 in the adjuvant CCRT group. The median follow-up was 72.1 months. The three-year PFS rates were both 91.9%, and the five-year OS was 90.6% versus 90.0% in adjuvant chemotherapy and CCRT groups, respectively. No significant differences were observed in the PFS or OS between groups. The adjusted HR for PFS was 0.854 (95% confidence interval 0.415-1.757; P = 0.667) favoring adjuvant chemotherapy, excluding the predefined non-inferiority boundary of 1.9. The chemotherapy group showed a tendency toward good quality of life. In comparison with post-operative adjuvant CCRT, adjuvant chemotherapy treatment showed non-inferior efficacy in patients with early-stage cervical cancer having pathological risk factors. Adjuvant chemotherapy alone is a favorable alternative post-operative treatment.

Progress of Polymer Application in Coated Proppant and Ultra-Low Density Proppant.

Design, synthesis and application of low-density proppant (LDP) are of great significance for efficient and clean exploitation of low permeability oil and gas. On the basis of a brief introduction of hydraulic fracturing and the application of traditional proppants, this review systematically summarized the polymer application progress in LDP, including coated sand, coated ceramics, coated nutshells, especially for polymer composites based ultra-low density proppant (ULDP). Finally, the existing problems and future development direction are also prospected.

Application Progress of PALS in the Correlation of Structure and Properties for Graphene/Polymer Nanocomposites.

Giving a deep insight into the microstructure, and realizing the correlation between microstructure and properties is very important to the precise construction of high-performance graphene/polymer nanocomposites (GPN). For the promising application in microstructure characterization, much attention has been focused on the effective technique of positron annihilation lifetime spectroscopy (PALS). Based on the introduction of the basic principle, this review summarized the application progress of PALS in the correlation of microstructure and properties for GPN, especially for the characterization of free volume and interfacial interaction, and the correlation of these microstructures and properties.

Green Preparation of Aminated Magnetic PMMA Microspheres via EB Irradiation and Its Highly Efficient Uptake of Ce(III).

The modification of polymers can significantly improve the ability to remove rare earth ions from wastewater, but so far few studies have focused on the irradiation-induced grafting method. In this study, a novel magnetic chelating resin for Ce(III) uptake was first synthesized by suspension polymerization of PMMA@Fe3O4 microspheres followed by irradiation-induced grafting of glycidyl methacrylate (GMA) and subsequent amination with polyethyleneimine (PEI). The FT-IR, SEM, TG and XRD characterization confirmed that we had successfully fabricated magnetic PMMA-PGMA-PEI microspheres with a well-defined structure and good thermal stability. The obtained adsorbent exhibited a satisfactory uptake capacity of 189.81 mg/g for Ce(III) at 318.15 K and an initial pH = 6.0. Additionally, the impact of the absorbed dose and GMA monomer concentration, pH, adsorbent dosage, contact time and initial concentration were thoroughly examined. The pseudo-second order and Langmuir models were able to describe the kinetics and isotherms of the adsorption process well. In addition, the thermodynamic data indicated that the uptake process was spontaneous and endothermic. Altogether, this research enriched the Ce(III) trapping agent and provided a new method for the removal rare earth pollutants.

Terahertz Vibrational Fingerprints Detection of Molecules with Particularly Designed Graphene Biosensors.

In this research, an arc I-shaped graphene sensing structure with multi-resonance characteristics is proposed for the simultaneous detection of vibrational fingerprints with spectral separation in the terahertz range. The resonant frequencies of the sensor can be dynamically tuned by changing the gate voltage applied to the graphene arrays. The two vibrational fingerprints of lactose molecules (0.53 THz and 1.37 THz) in the transmission spectrum can be enhanced simultaneously by strictly optimizing the geometrical parameters of the sensor. More importantly, these two resonant frequencies can be tuned precisely to coincide with the two standard resonances of the lactose molecule. The physical mechanism of the sensor is revealed by inspection of the electric field intensity distribution, and the advantage of the sensor, which is its ability to operate at a wide range of incident angles, has been demonstrated. The sensing performance of the structure as a refractive index sensor has also been studied. Finally, a double arc I-shaped graphene sensor is further designed to overcome the polarization sensitivity, which demonstrates excellent molecular detection performance under different polarization conditions. This study may serve as a reference for designing graphene biosensors for molecular detection.

Human papillomavirus integration perspective in small cell cervical carcinoma.

Small cell cervical carcinoma (SCCC) is a rare but aggressive malignancy. Here, we report human papillomavirus features and genomic landscape in SCCC via high-throughput HPV captured sequencing, whole-genome sequencing, whole-transcriptome sequencing, and OncoScan microarrays. HPV18 infections and integrations are commonly detected. Besides MYC family genes (37.9%), we identify SOX (8.4%), NR4A (6.3%), ANKRD (7.4%), and CEA (3.2%) family genes as HPV-integrated hotspots. We construct the genomic local haplotype around HPV-integrated sites, and find tandem duplications and amplified HPV long control regions (LCR). We propose three prominent HPV integration patterns: duplicating oncogenes (MYCN, MYC, and NR4A2), forming fusions (FGFR3-TACC3 and ANKRD12-NDUFV2), and activating genes (MYC) via the cis-regulations of viral LCRs. Moreover, focal CNA amplification peaks harbor canonical cancer genes including the HPV-integrated hotspots within MYC family, SOX2, and others. Our findings may provide potential molecular criteria for the accurate diagnosis and efficacious therapies for this lethal disease.

Fabrication and Evaluation of Graphene Oxide/Hydroxypropyl Cellulose/Chitosan Hybrid Aerogel for 5-Fluorouracil Release.

The incorporation of graphene oxide (GO) into a polymeric drug carrier can not only enhance the loading efficiency but also reduce the initial burst and consequently improve the controllability of drug release. Firstly, 5-fluorouracil (5-Fu)-loaded hydroxypropyl cellulose/chitosan (HPC/CS@5-Fu) and GO/HPC/CS@5-Fu aerogels were successfully fabricated through chemical cross-linking with glutaraldehyde. Then, the obtained aerogels were characterized using scanning electron microscopy (SEM), Fourier transform infrared (FITR), X-ray diffraction (XRD), differential scanning calorimetry (DSC), thermogravimetry (TG), and the effect of HPC and GO content on the drug loading (DL) and encapsulation efficiency (EE) for the two aerogels were investigated, respectively. Finally, the drug release behavior of the GO/HPC/CS@5-Fu aerogels with different GO content was evaluated at two different pH values, and four kinds of kinetic models were used to evaluate the release behavior.

Facile Synthesis of Sustainable Tannin/Sodium Alginate Composite Hydrogel Beads for Efficient Removal of Methylene Blue.

To meet the requirement of sustainable development, bio-based adsorbents were developed for the removal of dye contaminant. To improve the adsorption capacity of pure sodium alginate (SA) adsorbent for the removal of methylene blue (MB), aromatic bio-based tannin (Tan) was incorporated through the cross-linking with calcium ion. The obtained Tan/SA composite hydrogel beads were characterized with SEM, FTIR and TG, demonstrating that millimeter-sized beads were obtained through calcium cross-linking with enhanced thermal stability. The maximum capacity (247.2 mg/g) at optimal condition (pH = 12, T = 45 °C) was obtained for the 40%Tan/SA adsorbents, with a removal efficiency of 82.4%. This can be ascribed to the electrostatic attraction between SA and MB, as well as the formation of π-π stacking between Tan and MB. The adsorption process for MB is endothermic, and chemical adsorption, the removal efficiency was exceeded 90% after five cycles.

Efficient Removal of Methylene Blue by Bio-Based Sodium Alginate/Lignin Composite Hydrogel Beads.

Dye pollution is a serious issue in current environment protection, and bio-based adsorbents have been receiving much attention in wastewater treatment, due to their low cost, renewable, and environmentally friendly characteristics. Bio-based sodium alginate/lignin composite (SA/Lig) hydrogel beads were fabricated by a facile cross-linking with calcium ion and used for the removal of methylene blue (MB). The obtained SA/Lig microbeads were characterized with SEM, FTIR, and TG, and the effect of lignin content, pH, and temperature on the MB adsorption was investigated. The results indicated that the introduction of aromatic lignin can not only enhance thermal stability but also can improve the adsorption performance. Under optimal conditions, the maximum adsorption capacity (254.3 mg/g) was obtained for the SA/Lig-20% beads, with a removal efficiency of 84.8%. The adsorption process for MB is endothermic, and the rate-limiting step is chemical adsorption. The removal efficiency is higher than 90% after five cycles, revealing that the prepared beads show good regeneration ability.

Hyperimmune Targeting Staphylococcal Toxins Effectively Protect Against USA 300 MRSA Infection in Mouse Bacteremia and Pneumonia Models.

Staphylococcus aureus has been acquiring multiple drug resistance and has evolved into superbugs such as Methicillin/Vancomycin-resistant S. aureus (MRSA/VRSA) and, consequently, is a major cause of nosocomial and community infections associated with high morbidity and mortality for which no FDA-approved vaccines or biotherapeutics are available. Previous efforts targeting the surface-associated antigens have failed in clinical testing. Here, we generated hyperimmune products from sera in rabbits against six major S. aureus toxins targeted by an experimental vaccine (IBT-V02) and demonstrated significant efficacy for an anti-virulence passive immunization strategy. Extensive in vitro binding and neutralizing titers were analyzed against six extracellular toxins from individual animal sera. All IBT-V02 immunized animals elicited the maximum immune response upon the first boost dose against all pore-forming vaccine components, while for superantigen (SAgs) components of the vaccine, second and third doses of a boost were needed to reach a plateau in binding and toxin neutralizing titers. Importantly, both anti-staphylococcus hyperimmune products consisting of full-length IgG (IBT-V02-IgG) purified from the pooled sera and de-speciated F(ab')2 (IBT-V02-F(ab')2) retained the binding and neutralizing titers against IBT-V02 target toxins. F(ab')2 also exhibited cross-neutralization titers against three leukotoxins (HlgAB, HlgCB, and LukED) and four SAgs (SEC1, SED, SEK, and SEQ) which were not part of IBT-V02. F(ab')2 also neutralized toxins in bacterial culture supernatant from major clinical strains of S. aureus. In vivo efficacy data generated in bacteremia and pneumonia models using USA300 S. aureus strain demonstrated dose-dependent protection by F(ab')2. These efficacy data confirmed the staphylococcal toxins as viable targets and support the further development effort of hyperimmune products as a potential adjunctive therapy for emergency uses against life-threatening S. aureus infections.