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Epsilon toxin (ETX) is an exotoxin produced by type B and D Clostridium perfringens that causes enterotoxemia or necrotic enteritis in animals such as goats, sheep, and cattle. Vaccination is a key method in preventing such diseases. In this study, we developed a new type of dissolving microneedle patch (dMN) with a nanoparticle adjuvant for enhanced immune response to deliver the rETXY196E-C protein vaccine. We chose FDA-approved poly(lactic-co-glycolic acid) (PLGA) to prepare nanospheres as the vaccine adjuvant and introduced dimethyldioctadecylammonium bromide (DDAB) to make the surface of PLGA nanoparticles (PLGA NPs) positively charged for antigen adsorption. PLGA NPs with a diameter of 100~200 nm, a surface ZETA potential of approximately +40 mV, and good safety were successfully prepared and could effectively adsorb rETXY196E-C protein. Using non-toxic and antibacterial fish gelatin as the microneedle (MN) matrix, we prepared a PLGA-DDAB dMN vaccine with good mechanical properties that successfully penetrated the skin. After immunization of subcutaneous (SC) and dMN, antibody titers of the PLGA and Al adjuvant groups were similar in both two immune ways. However, in vivo neutralization experiments showed that the dMN vaccines had a better protective effect. When challenged with 100 × LD50 GST-ETX, the survival rate of the MN group was 100%, while that of the SC Al group was 80%. However, a 100% protective effect was achieved in both immunization methods using PLGA NPs. In vitro neutralization experiments showed that the serum antibodies from the dMN and SC PLGA NPs groups both protect naive mice from 10 × LD50 GST-ETX attack after being diluted 20 times and could also protect MDCK cells from 20 × CT50 GST-ETX attack. In conclusion, the PLGA-DDAB dMN vaccine we prepared has good mechanical properties, immunogenicity, and protection, and can effectively prevent ETX poisoning. This provides a better way of delivering protein vaccines. [ABSTRACT FROM AUTHOR]

Background: As a Class A bioterrorism agent, botulinum neurotoxin serotype A (BoNT/A) carries the risk of being used by terrorists to cause mass poisoning. The microneedle (MN) patch has a great potential for application as a novel vaccine delivery method. The aim of this study is to develop a thermally stable, dissolving microneedle patch for the delivery of a recombinant protein vaccine using a recombinant C-terminal heavy chain of BoNT/A (Hc of BoNT/A, AHc) to prevent botulism. Methods: Fish gelatin, a natural non-toxic and bacteriostatic material, was selected as the microneedle matrix for the preparation of the dissolving microneedle vaccine. Subsequently, the mechanical performance, bacteriostatic properties, vaccination effect, and stability of the microneedle patches were evaluated using instruments such as the displacement-force test station and optical coherence tomography (OCT) scanner. Results: Fish gelatin matrix at high concentrations has good bacteriostatic properties, and excellent mechanical performance and vaccination effect, meeting the necessities of a vaccine. In both in vivo and in vitro neutralization experiments, MN vaccines containing different antigen doses achieved the same protective efficacy as subcutaneous vaccinations, protecting mice against 106 LD50 of BoNT/A injected intraperitoneally. Thermal stability analysis of the MN vaccines revealed that the fish gelatin matrix protected the AHc vaccine from protein denaturation even after 7 days of storage at 37 °C and enabled the vaccine patches to maintain good immunogenicity and protective efficacy even after 6 months of storage at room temperature. Conclusion: In this study, we successfully prepared a bacteriostatic MN patch using a fish gelatin matrix that not only has a good vaccination effect, but also obviates the need for a cold chain for the AHc vaccine, providing the possibility of rapid, painless, and large-scale vaccination. [ABSTRACT FROM AUTHOR]

The efficient recovery and conversion of energy in oily sludge has great prospects. In this article, the main objective is to investigate the impact of the addition of CO2 during the pyrolysis of oily sludge on energy recovery and conversion by thermogravimetric analysis (TGA) and compare the effect with the traditional pyrolysis effect from the perspective of thermal conversion behavior, products composition, and kinetics analysis. The results of the experiment showed that in the CO2 atmosphere, the main weight loss temperature of oily sludge was mainly concentrated in the range of 300–500 °C, which is lower than the reaction temperature range of traditional pyrolysis. The yields of CO and H2 in the products have been greatly improved, and the highest proportion in the gas products can reach 19.29% and 22.38%, respectively. The Ea (activation energy) values of oily sludge were determined to be in the range of 40–120 kJ·mol−1 with the conversion between 0.2 and 0.8 via DAEM, KAS, Starink, and FWO methods, respectively, in which the FWO method has shown the strongest adaptability. The results of this study provide reference values for practical engineering applications. [ABSTRACT FROM AUTHOR]

Ricin and abrin are phytotoxins that can be easily used as biowarfare and bioterrorism agents. Therefore, developing a rapid detection method for both toxins is of great significance in the field of biosecurity. In this study, a novel nanoforest silicon microstructure was prepared by the micro-electro-mechanical systems (MEMS) technique; particularly, a novel microfluidic sensor chip with a capillary self-driven function and large surface area was designed. Through binding with the double antibodies sandwich immunoassay, the proposed sensor chip is confirmed to be a candidate for sensing the aforementioned toxins. Compared with conventional immunochromatographic test strips, the proposed sensor demonstrates significantly enhanced sensitivity (≤10 pg/mL for both toxins) and high specificity against the interference derived from juice or milk, while maintaining good linearity in the range of 10–6250 pg/mL. Owing to the silicon nanoforest microstructure and improved homogeneity of the color signal, short detection time (within 15 min) is evidenced for the sensor chip, which would be helpful for the rapid tracking of ricin and abrin for the field of biosecurity. [ABSTRACT FROM AUTHOR]

A new algorithm for solving S. Geetha and K.P.K. Nair's proposed variant of the n-by-n bottleneck assignment problem is introduced. The procedure is based on the application of M.L. Balinski's signature method or dual simplex method to the assignment problem at each stage. Unlike Geetha and Nair's O(n5) algorithm, it is simpler, having a complexity of 'O m(m + n log n)' where m is the number of arcs.

Diffusion plays a critical role in establishing functional bio/solid soft interfaces for bioassays, biosensors, and biofuel cells. An understanding of micro-diffusion near the interface is significant for developing high-performance bioassays, biosensors, and biofuel cells. Herein, we explored micro-diffusion behavior at different temperature gaps in microfluidic chip and enzyme-linked immunosorbent assay (ELISA) microplates. It exhibited that temperature gap could distinctly promote directional diffusion of molecules in microfluidic chip from high concentration zone to low concentration zone. In addition, experimental results of specially designed ELISA also partly confirmed that the temperature gap could effectively improve the performance of ELISA by 54.5%. In compared with conventional ELISA, the as-prepared temperature-gap functional module in microfluidic chip has obvious advantages of miniaturization, integration, customization, and low cost. All in all, the current work indicates that temperature-gap function has great potential in biomedical detection, food safety, and so on. It can be utilized to develop novel biological detection approaches and related instruments or microfluidic chips. [ABSTRACT FROM AUTHOR]