Search Results (8,325)

In this study, the removal performances of a multi-pollutant elimination cartridge system (MPECS) composed of palm shell waste-based activated carbon (PSAC), magnesium (Mg)/silicon (Si)-doped PSAC (Mg/Si-PSAC), and silver (Ag)-doped PSAC (Ag-PSAC) for heavy metals, organic pollutants, and Escherichia coli were investigated. Mg/Si impregnation significantly improved heavy metal removal using PSAC by increasing the surface area and adding more sorption sites to the magnesium silicate nanolayer. Fixed-bed column experiments showed that the MPECS column outperformed PSAC and commercial activated carbon (DJAC), with a 1.5 to 2.0 times higher E. coli removal and a higher removal of organic pollutants and heavy metals. The MPECS column, with its disinfection ability and adsorption of heavy metals and organic matter, is a promising system for removing multiple pollutants from rainwater. Full article

The development of effective vaccines necessitates a delicate balance between maximizing immunogenicity and minimizing safety concerns. Subunit vaccines, while generally considered safe, often fail to elicit robust and durable immune responses. Nanotechnology presents a promising approach to address this dilemma, enabling subunit antigens to mimic critical aspects of native pathogens, such as nanoscale dimensions, geometry, and highly repetitive antigen display. Various expression systems, including Escherichia coli (E. coli), yeast, baculovirus/insect cells, and Chinese hamster ovary (CHO) cells, have been explored for the production of nanoparticle vaccines. Among these, E. coli stands out due to its cost-effectiveness, scalability, rapid production cycle, and high yields. However, the E. coli manufacturing platform faces challenges related to its unfavorable redox environment for disulfide bond formation, lack of post-translational modifications, and difficulties in achieving proper protein folding. This review focuses on molecular and protein engineering strategies to enhance protein solubility in E. coli and facilitate the in vitro reassembly of virus-like particles (VLPs). We also discuss approaches for antigen display on nanocarrier surfaces and methods to stabilize these carriers. These bioengineering approaches, in combination with advanced nanocarrier design, hold significant potential for developing highly effective and affordable E. coli-derived nanovaccines, paving the way for improved protection against a wide range of infectious diseases. Full article

Background: The worldwide misuse of antibiotics is one of the main factors in microbial resistance that is a serious threat worldwide. Alternative strategies are needed to overcome this issue. Objectives: In this study, a novel strategy was adopted to suppress the growth of resistant pathogens through immobilization of silver nanoparticles (AgNPs) in gum of Moringa oleifera. Methods: The AgNPs were prepared from the leaves of Moringa oleifera and subsequently characterized through UV-spectrophotometry, FTIR, SEM, and XRD. The differential ratios of characterized AgNPs were immobilized with gum of M. oleifera and investigated for antimicrobial potential against highly resistant pathogens. Results: The immobilized AgNPs displayed promising activities against highly resistant B. subtilis (23.6 mm; 50 µL:200 µL), E. coli (19.3 mm; 75 µL:200 µL), K. pneumoniae (22 mm; 200 µL:200 µL), P. mirabilis (16.3 mm; 100 µL:200 µL), P. aeruginosa (22 mm; 175 µL:200 µL), and S. typhi (19.3; 25 µL:200 µL) than either AgNPs alone or gum. The immobilized AgNPs released positive sliver ions that easily attached to negatively charged bacterial cells. After attachment and permeation to bacterial cells, the immobilized NPs alter the cell membrane permeability, protein/enzymes denaturation, oxidative stress (ROS), damage DNA, and change the gene expression level. It has been mechanistically considered that the immobilized AgNPs can kill bacteria by damaging their cell membranes, dephosphorylating tyrosine residues during their signal transduction pathways, inducing cell apoptosis, rupturing organelles, and inhibiting cell division, which finally leads to cell death. Conclusions: This study proposes a potential alternative drug for curing various infections. Full article

Aggregatibacter actinomycetemcomitans ApiA is a trimeric autotransporter outer membrane protein (Omp) that participates in multiple functions, enabling A. actinomycetemcomitans to adapt to a variety of environments. The goal of this study is to identify regions in the apiA gene responsible for three of these functions: auto-aggregation, buccal epithelial cell binding, and complement resistance. Initially, apiA was expressed in Escherichia coli. Finally, wild-type A. actinomycetemcomitans and an apiA-deleted version were tested for their expression in the presence and absence of serum and genes related to stress adaptation, such as oxygen regulation, catalase activity, and Omp proteins. Sequential deletions in specific regions in the apiA gene as expressed in E. coli were examined for membrane proteins, which were confirmed by microscopy. The functional activity of epithelial cell binding, auto-aggregation, and complement resistance were then assessed, and regions in the apiA gene responsible for these functions were identified. A region spanning amino acids 186–217, when deleted, abrogated complement resistance and Factor H (FH) binding, while a region spanning amino acids 28–33 was related to epithelial cell binding. A 13-amino-acid peptide responsible for FH binding was shown to promote serum resistance. An apiA deletion in a clinical isolate (IDH781) was created and tested in the presence and/or absence of active and inactive serum and genes deemed responsible for prominent functional activity related to A. actinomycetemcomitans survival using qRT-PCR. These experiments suggested that apiA expression in IDH781 is involved in global regulatory mechanisms that are serum-dependent and show complement resistance. This is the first study to identify specific apiA regions in A. actinomycetemcomitans responsible for FH binding, complement resistance, and other stress-related functions. Moreover, the role of apiA in overall gene regulation was observed. Full article

Desmoplasia is a hallmark feature of pancreatic ductal adenocarcinoma (PDAC) that contributes significantly to treatment resistance. Approaches to enhance drug delivery into fibrotic PDAC tumors continue to be an important unmet need. In this study, we have engineered a tumor-colonizing E. coli-based agent that expresses both collagenase and hyaluronidase as a strategy to reduce desmoplasia and enhance the intratumoral perfusion of anticancer agents. Overall, we observed that the tandem expression of both these enzymes by tumor-colonizing E. coli resulted in the reduced presence of intratumoral collagen and hyaluronan, which likely contributed to the enhanced chemotherapeutic efficacy observed when used in combination. These results highlight the importance of combination treatments involving the depletion of desmoplastic components in PDAC before or during treatment. Full article

In the context of revitalizing the use of traditional plant species as remarkable sources of bioactive compounds, the determination of their biological effects is of utmost importance. Among Lamiaceae species, Teucrium montanum (Mountain Germander) represents understudied Mediterranean plant species; it is rich in polyphenols, which are well-studied biologically active compounds for human disease prevention and the reduction of oxidative stress, i.e., phenolic acids, phenylethanoid glycosides, and flavonoids. For that purpose, the aim of this study was to investigate the antioxidant, cytotoxic, and genotoxic effects of Mountain Germander (MG) polyphenolic extract (0.025, 0.050, 0.150, and 0.500 mg extract mL⁻¹) on the hepatocellular (HepG2), tongue (CAL 27), gastric (AGS), and colorectal (Caco-2) continuous human cancer cell lines, as well as its bacteriostatic potential on representative members of human microbiota. In addition, the antioxidant potential of the MG polyphenolic extract was determined using bovine serum album and DNA plasmid as cellular model macromolecules. In vitro analysis revealed a significant cytotoxic effect of all MG extract concentrations on AGS and Caco-2 cell lines after prolonged treatment (24 h). In addition, treatment with 0.500 mg extract mL⁻¹ showed the most pronounced antioxidant effect under prolonged treatment (24 h) on CAL 27 and HepG2 cell lines. All of the applied MG extract concentrations seem to have a genoprotective effect on DNA plasmid. Furthermore, a significant inhibitory effect on E. coli was detected upon the treatment with 0.150 mg extract mL⁻¹, reducing the cell viability by 56%. Full article

The combination of carbon dots/polyvinyl alcohol (CDs/PVA) active film and radio frequency (RF) was performed to improve the storage quality of fried meatball samples. The microbicidal effect of RF, and the antioxidant and antibacterial activities of CDs/PVA film were investigated. The CDs/PVA film can effectively restrain the growth of B. subtilis, S. aureus, and E. coli, and eliminate DPPH and ABTS free radicals. RF exerts inhibitory effects on C. albicans, B. subtilis, and E. coli. For meatball samples, CDs/PVA+RF20 can extend the refrigerated shelf life from 2 w to 6 w. At the sixth week, the total bacterial count (TBC) in CDs/PVA+RF20 group (3.72 log CFU/g) was remarkably lower than those in polyethylene terephthalate (PET) group (7.78 log CFU/g) and CDs/PVA (6.41 log CFU/g) group. CDs/PVA+RF20 can also inhibit the increase in TBARS and POV values. The results manifest the feasibility of CDs/PVA+RF as a novel mild pasteurization or preservation technology. Full article

The use of antimicrobial peptides (AMPs) covalently grafted on surfaces has been recognized in recent years as a promising strategy to fight against biofilm formation. However, after grafting, the understanding of AMP–bacteria interactions is still debated in the literature. In this study, Nisin, a cyclic AMP, was grafted onto gold surfaces via an indirect grafting on acidic thiol self-assembled monolayers using succinimide linkers. The physical and chemical properties of these SAMs were then finely characterized by XPS and FT-IR to confirm the covalent grafting of Nisin. The antiadhesion and bactericidal effects were then studied for Escherichia coli ATCC25922, Staphylococcus aureus ATCC 25923, and Listeria ivanovii Li4(pVS2) by a posteriori analysis of the culture supernatants (i.e., indirect technique) and ex situ by optical microscopy following crystal violet staining (i.e., direct technique). Statistical analysis reveals that the Nisin coating has bactericidal and antiadhesive properties towards Gram-positive bacteria, while no significant results were obtained for Gram-negative bacteria. Full article

This study analysed the water quality of 156 samples collected from 13 different points within the Chief Albert Luthuli Local Municipality between January and December 2021, focusing on compliance with the South African National Standard 241 (SANS 241). Microbiological assessments revealed widespread contamination, with heterotrophic plate counts (HPCs) exceeding the recommended range in nearly all samples. The median HPC levels were particularly concerning, with the untreated source (RE000) exhibiting a median of 2555 CFU/mL. Total coliforms and Escherichia coli were present at alarming levels, indicating significant faecal contamination; the highest total coliform count, 2420 CFU/100 mL, was recorded at RE000. Physically, while all samples complied with electrical conductivity standards (0–70 µS/cm), turbidity levels showed variability, with nine samples exceeding the acceptable threshold of 1 NTU. The highest turbidity, 7.55 NTU, was recorded at the untreated source (RE000). Chemically, pH levels fell within safe ranges (6.0–9.0), with all samples being compliant with standards. However, only one sample (E001) met the free chlorine standard of 0.3–0.6 mg/L, suggesting inadequate disinfection practices. The untreated source was classified as ‘Unsuitable for drinking’ (WQI = 207.27). These findings highlight the critical need for enhanced monitoring and remediation strategies to improve drinking water safety in the region. Full article

Finding new molecules to prevent the growth of antimicrobial resistance is a hot topic for scientists worldwide. It has been reported that some raw bioproducts containing Monascus polyketides have antimicrobial activities, but extensive studies on this effect have not been conducted. In this context, our studies aimed to evaluate the antimicrobial properties of six raw bioproducts containing three classes of microbial polyketides biosynthesized by three Monascus strains through solid-state biosynthesis. As a methodology, we performed in silico predictions using programs such as PyMOL v3.0.4 and employed ESI-MS techniques to provide evidence of the presence of the six studied compounds in our bioproducts. The results obtained in silico were validated through in vitro studies using the Kirby-Bauer diffusion method on bacteria and fungi. The test performed in silico showed that Monascorubramine has the highest affinity for both Gram-positive and Gram-negative bacteria, followed by yellow polyketides such as Ankaflavin and Monascin. The estimated pharmacokinetic parameters indicated high gastrointestinal absorption and the potential to cross the blood-brain barrier for all studied compounds. However, the compounds also inhibit most enzymes involved in drug metabolism, presenting some level of toxicity. The best in vitro results were obtained for S. aureus, with an extract containing yellow Monascus polyketides. Predictions made for E. coli were validated in vitro for P. aeruginosa, S. enterica, and S. marcescens, as well as for fungi. Significant antibacterial properties were observed during this study for C. albicans, S. aureus, and fungal dermatophytes for crude bioproducts containing Monascus polyketides. In conclusion, the antimicrobial properties of Monascus polyketides were validated both in silico and in vitro. However, due to their potential toxicity, these bioproducts would be safer to use as topical formulations. Full article

Dye wastewater pollution, particularly from persistent and toxic polycyclic organic pollutants, such as aniline blue, poses a significant environmental challenge. Aniline blue, a triphenylmethane dye widely used in the textile, leather, paper, and pharmaceutical industries, is notoriously difficult to treat owing to its complex structure and potential for bioaccumulation. In this study, we explored the capacity of Comamonas testosteroni (CT1) to efficiently degrade aniline blue, focusing on the underlying enzymatic mechanisms and degradation pathways. Through prokaryotic transcriptome analysis, we identified a significantly upregulated short-chain dehydrogenase (SDRz) gene (log₂FC = 2.11, p < 0.05) that plays a crucial role in the degradation process. The SDRz enzyme possessed highly conserved motifs and a typical short-chain dehydrogenase structure. Functional validation using an SDRz-knockout strain (CT-ΔSDRz) and an SDRz-expressioning strains (E-SDRz) confirmed that SDRz is essential for aniline blue degradation. The knockout strain CT-ΔSDRz exhibited a 1.27-fold reduction in the degradation efficiency, compared to CT1 strain after 12 h; while the expression strain E-SDRz showed a 1.24-fold increase compared to Escherichia coli DH5α after 12 h. Recombinant SDRz (rSDRz) was successfully produced, showing significant enzymatic activity (1.267 ± 0.04 mmol·L⁻¹·min⁻¹ protein), with kinetic parameters Vmax = 2.870 ± 0.0156 mmol·L⁻¹·min⁻¹ protein and Km = 1.805 ± 0.0128 mM·mL⁻¹. Under optimal conditions, the rSDRz achieved a degradation efficiency of 62.17% for aniline blue. Gas chromatography–mass spectrometry (GC-MS) analysis identified several intermediate metabolites in the degradation pathway, including benzeneacetaldehyde, a, a-diphenyl, 2-amino-4-methylbenzophenone, benzene, 1-dimethylamino-4-phenylmethyl, benzenesulfonic acid, methyl ester, further elucidating the biodegradation mechanism. These findings highlight SDRz as a critical enzyme in the biodegradation of aniline blue, offering valuable insights and a robust theoretical foundation for developing advanced bioremediation strategies to address dye wastewater pollution. Full article

Peptidoglycan recognition proteins (PGRPs) are a class of pattern recognition receptors (PRRs) that activate the innate immune system in response to microbial infection by detection of peptidoglycan, a distinct component of bacterial cell walls. Bioinformatic studies have revealed four PGRPs in the red imported fire ant Solenopsis invicta; nonetheless, the mechanism of the immune response of S. invicta induced by pathogens is still poorly understood. The peptidoglycan recognition protein full-length cDNA (designated as SiPGRP-S1/S2/S3/L) from S. invicta was used in this investigation. According to the sequencing analysis, there was a significant degree of homology between the anticipated amino acid sequence of SiPGRPs and other members of the PGRPs superfamily. Molecular docking studies demonstrated that SiPGRPs show strong binding affinity for a variety of PGN substrates. Additionally, tissue distribution analysis indicated that SiPGRPs are primarily expressed in several tissues of naïve larvae, including fat body, hemocytes, head, and thorax, as detected by quantitative real-time PCR (RT-qPCR). Microbial challenges resulted in variable changes in mRNA levels across different tissues. Furthermore, the antibacterial effects of antimicrobial peptides (AMPs) produced by major ants infected with Metarhizium anisopliae were assessed. These AMPs demonstrated inhibitory effects against M. anisopliae, Staphylococcus aureus, and Escherichia coli, with the most pronounced effect observed against E. coli. In conclusion, SiPGRPs act as pattern recognition receptors (PRRs) that identify pathogens and initiate the expression of AMPs in S. invicta, this mechanism contributes to the development of biopesticides designed for the targeted control of invasive agricultural pests. Full article

The urgent need for an effective COVID-19 therapy has propelled the exploration of innovative strategies to combat the fast-mutating SARS-CoV-2 virus. This study attempted to develop nanobodies (Nbs) against the SARS-CoV-2 Omicron variants by redirecting the 1.29 neutralizing Nb, a receptor-binding domain (RBD)-specific Nb that can protect against various SARS-CoV-2 variants other than Omicron, to target SARS-CoV-2 Omicron subvariant BA.5, the variant used for the development of the bivalent vaccine. Error-prone libraries of the 1.29 Nb were constructed. Following two rounds of selection using the functional ligand-binding identification by Tat-based recognition of associating proteins (FLI-TRAP) technique, we rapidly identified two Nbs, namely, C11 and K9, that could target the RBD of the Omicron subvariant BA.5, XBB.1.5, and XBB.1.16 subvariants. Molecular docking provided insights into how these Nbs interact with the RBD of the BA.5 and JN.1 variants. The application of directed evolution via utilization of error-prone PCR and the synthetic E. coli applied in the FLI-TRAP selection method may be a powerful tool for facilitating simple, fast and economical selection to redirect existing antibodies and to generate antibody fragments to target proteins susceptible to autonomous mutation, not only for viral infection but also other diseases, such as cancer. Full article

African horse sickness (AHS) is an acute, fatal, contagious disease of animals of the family Equidae and is caused by infection with the African horse sickness virus (AHSV). Based on the outer capsid protein VP2, AHSV is classified into nine serotypes (AHSV−1 to −9) with little or no serological cross-reactivity between them. In 2020, AHS outbreaks caused by AHSV−1 were reported in Thailand and Malaysia, marking the first occurrences of AHS in Southeast Asia. However, little is known about the antigenic profile of AHSV−1 VP2. In this study, a recombinant VP2 protein was expressed in Escherichia coli and used as an immunogen, and three monoclonal antibodies (mAbs), designated 7D11, 10A9, and 9E7, against AHSV−1 VP2, were generated. These three mAbs were then successfully used in IFA, WB, and ELISA for the detection of AHSV−1 VP2. Two overlapping linear epitopes, ⁶⁷⁰NEFDFE⁶⁷⁵ (E670–675) recognized by 9E7 and ⁶⁷⁰NEFDF⁶⁷⁴ (E670–674) recognized by 7D11 and 10A9, were identified through truncation of GST-fused VP2. Amino acid sequence alignment shows that the ⁶⁷⁰NEFDFE⁶⁷⁵ motif is completely conserved within AHSV−1 but is highly divergent in other AHSV serotypes. Our studies provide an important tool for basic research into AHSV−1 and for the diagnosis of AHSV−1. Full article

Escherichia coli O157:H7, a Shiga-toxin-producing E. coli (STEC), is an important pathogen related to foodborne disease that is responsible for a growing number of outbreaks worldwide and has been detected in processed meats, dairy, and fresh vegetables. Although culturing is the gold standard method for detection of this bacterium, molecular methods based on nucleic acid amplification techniques such as PCR are becoming more common because of their rapidity, sensitivity, and specificity. However, to ensure reliable results among the several alternative PCR protocols (e.g., commercial kits and reference methods), different measurement assurance tools, including validated methods, reference materials, and proficiency tests, among others, are required. Herein, we present a digital PCR method validation for E. coli O157:H7 detection and quantification using seven specific gene sequences; this method quantified nucleic acids from different E. coli serotypes, with a detection range of 6.6 to 7900 copies/µL and a repeatability standard deviation over the concentration range of 1% to 13.6%. The relative standard uncertainty was 3.5–14.6%, and the detection limit was 0.27 copies/µL. Subsequently, two batches of a candidate reference material based on E. coli O157:H7 genomic DNA were then produced and characterized for evaluation of copy number concentration with the validated ddPCR method, with assigned values of 164,770 ± 9251 and 172 ± 9 copies/μL. Thus, this study demonstrated the development of a validated method and reference material for dPCR and qPCR detection of E. coli O157:H7, a key STEC responsible for food poisoning. Full article