recombinant human interferon alfa-2a

A bireporter recombinant SARS-CoV-2 Omicron BA.5 for in vitro and in vivo studies.

Castro Esteban M EM, Barre Ramya S RS, Ye Chengjin C, Imbiakha Brian B et al.

The continuous emergence of variants of concern (VoCs) represents a significant challenge to effectively control severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Although FDA-approved vaccines and antivirals have been successfully developed and implemented for the prophylactic and therapeutic intervention of SARS-CoV-2 infection, recent VoCs could escape protection garnered by previous vaccine and antiviral approaches. Determining the efficacy of prophylactics and/or therapeutics against recent VoCs will assist in efficiently controlling currently circulating SARS-CoV-2 strains. We used our previously described bacterial artificial chromosome-based reverse genetics approach for Omicron BA.5 to generate a recombinant SARS-CoV-2 BA.5 encoding a fusion of ZsGreen to Nanoluciferase (rBA.5 ZsG-Nluc) from the locus of the viral nucleocapsid (N) protein separated by the porcine teschovirus-1 2A proteolytic cleavage site. The rBA.5 ZsG-Nluc replicates to levels comparable to recombinant BA.5 wild type (rBA.5 WT) and expresses high levels of ZsG and Nluc in cultured cells. This facilitates tracking viral infection and the identification of antivirals and neutralizing antibodies with EC50 and NT50 values, respectively, similar to those obtained with rBA.5 WT. Importantly, in Keratin-18 human angiotensin-converting enzyme-2 mice, rBA.5 ZsG-Nluc retains the same pathogenicity and ability to replicate in the lungs of infected mice as rBA.5 WT. Using rBA.5 ZsG-Nluc, we detected Nluc activity systemically and Nluc and ZsG expression in the lungs of infected mice using an in vivo imaging system. Our results demonstrate the feasibility of using rBA.5 ZsG-Nluc to track viral infections and identify prophylactics and therapeutics against recent SARS-CoV-2 VoCs in vitro, ex vivo, and in vivo.IMPORTANCESevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative virus of the coronavirus disease 2019 pandemic, is continually evolving to escape immunity acquired by previous natural infections or vaccinations. Moreover, recent SARS-CoV-2 variants of concern (VoCs) have acquired antiviral-resistant mutations to FDA-approved drugs. The emergence of these VoCs highlights the importance of identifying new prophylactics and therapeutics against currently circulating SARS-CoV-2 strains. We generated a recombinant bireporter Omicron BA.5 SARS-CoV-2 (rBA.5 ZsG-Nluc) that expresses reporter proteins, which are useful for cellular and whole animal studies, and has similar viral replication and pathogenicity to a wild-type recombinant Omicron BA.5 SARS-CoV-2. In Keratin-18 human angiotensin-converting enzyme-2 mice, rBA.5 ZsG-Nluc infection can be tracked systemically or in the lungs of infected mice using an in vivo imaging system. We establish a proof-of-concept platform of rBA.5 ZsG-Nluc in combination with an ancestral SARS-CoV-2 strain expressing mCherry to simultaneously identify antivirals and neutralizing antibodies against original and recent SARS-CoV-2 strains.

Clean Synthesis of N-4-Alkylamino-7-chloroquinoline Cyclic Imides: Influence of Lipophilicity on Antibacterial and Ecotoxicological Properties.

Luz-Filho Agenor P AP, Silva Thais C TC, Silva Matheus B MB, Sousa Abraão P AP et al.

Motivated by the promising antimicrobial and ecotoxicological effects of 4-alkylamino-7-chloroquinoline and N-alkylphthalimide derivatives, these privileged nuclei were linked to investigate how lipophilicity influences antibacterial activity and ecotoxicity. For this purpose, seven 4-alkylamino-7-chloroquinolines (2a-g) were prepared, of which four were used as precursors (2a-d) and reacted with different anhydrides. Using a clean synthetic approach, 4-alkylamino-7-chloroquinoline-succinimides (3a-d) and -phthalimides (5a-d) were obtained in yields of 60%-99% after only filtration with distilled water. In vitro assays were performed to determine the minimum inhibitory/bactericidal concentration (MIC/MBC) against Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus mutans. Lipophilicity (log P) and toxicity were predicted using the pkCSM program. In vitro toxicity tests on Artemia salina larvae were used as an indicator of ecotoxicity. 4-Alkylamino-7-chloroquinolines (2b, 2f, 2g) revealed a moderate MIC of 156.25 µg·mL-1 against Gram-(-) and Gram-(+) bacteria, whose predicted toxicity does not pose a potential risk to human health. Results indicate a correlation between increased lipophilicity and higher ecotoxicity, where less lipophilic compounds were less toxic to A. salina (LC50 133.01-777.50 µg·mL-1) than 4-alkylamino-7-chloroquinoline-phthalimides, suggesting further investigation of their cytotoxicity.

Correction: Postoperative adding-on phenomenon in Lenke 1A/B and 2A/B adolescent idiopathic scoliosis: risk factors and predictive index.

Zhang Hongqi H, Li Tao T, Zhang Gengming G, Deng Ang A et al.

Development of an inactivated, oral immunogenic product against post-weaning colibacillosis caused by enterotoxigenic Escherichia coli in piglets.

Inglesi Alessia A, Filipe Joel J, Valli Giulia G, Riva Federica F et al.

Post-weaning diarrhea (PWD) caused by enterotoxigenic Escherichia coli (ETEC) expressing F4 and F18 fimbriae remains a major challenge in pig production, contributing to economic losses and increased antimicrobial use. This study aimed to preliminary evaluate the immunogenic potential of an orally administered formulation comprising heat-inactivated ETEC strains expressing F4 and F18 fimbriae, combined with low-dose recombinant human interferon-alpha (IFN-α) as a mucosal adjuvant, in piglets. Piglets from two different litters were allocated into two experimental groups: a treated group (T) receiving the inactivated ETEC formulation with IFN-α for 26 days, and a control group (C) receiving only IFN-α. According to the farmer, the sows had not been vaccinated. Immune responses were evaluated in the sows colostrum and piglet serum, saliva, and feces by ELISA. In mesenteric lymph nodes anti-F4/F18 IgA and IgG antibodies were quantified by ELISPOT. The inactivated product preserved fimbrial antigenicity and remained sterile. Colostrum from both sows displayed elevated levels of fimbriae-specific IgA and IgG despite neither sow being vaccinated against E. coli. Treated piglets showed a transient serum IgA increase against F4 whereas serum IgG levels were often comparable to the controls. Notably, elevated mucosal IgA responses were observed in saliva and feces against both F4 and F18 (P < 0.01) in T group, accompanied by enhanced IgA-secreting B-cell activity in mesenteric lymph nodes. ELISA validation confirmed high assay reproducibility (R² > 0.98; CV < 10%). ELISPOT analysis underscored the adjuvant role of F4 and IFN-α in stimulating mucosal immunity. Our preliminary findings may represent a promising strategy to control PWD and reduce antimicrobial use in pig production.