Polymers as advanced antibacterial and antibiofilm agents for direct and combination therapies

The growing prevalence of antimicrobial drug resistance in pathogenic bacteria is a critical threat to global health. Conventional antibiotics still play a crucial role in treating bacterial infections, but the...

Evaluation of Heterocyclic Carboxamides as Potential Efflux Pump Inhibitors in Pseudomonas aeruginosa

The ability to rescue the activity of antimicrobials that are no longer effective against bacterial pathogens such as Pseudomonas aeruginosa is an attractive strategy to combat antimicrobial drug resistance. Herein, novel efflux pump inhibitors (EPIs) demonstrating strong potentiation in combination with levofloxacin against wild-type P. aeruginosa ATCC 27853 are presented. A structure activity relationship of aryl substituted heterocyclic carboxamides containing a pentane diamine side chain is described. Out of several classes of fused heterocyclic carboxamides, aryl indole carboxamide compound 6j (TXA01182) at 6.25 µg/mL showed 8-fold potentiation of levofloxacin. TXA01182 was found to have equally synergistic activities with other antimicrobial classes (monobactam, fluoroquinolones, sulfonamide and tetracyclines) against P. aeruginosa. Several biophysical and genetic studies rule out membrane disruption and support efflux inhibition as the mechanism of action (MOA) of TXA01182. TXA01182 was determined to lower the frequency of resistance (FoR) of the partner antimicrobials and enhance the killing kinetics of levofloxacin. Furthermore, TXA01182 demonstrated a synergistic effect with levofloxacin against several multidrug resistant P. aeruginosa clinical isolates.

Efficient production of immunologically active Shigella invasion plasmid antigens IpaB and IpaH using a cell-free expression system

Shigella spp. invade the colonic epithelium and cause bacillary dysentery in humans. Individuals living in areas that lack access to clean water and sanitation are the most affected. Even though infection can be treated with antibiotics, Shigella antimicrobial drug resistance complicates clinical management. Despite decades of effort, there are no licensed vaccines to prevent shigellosis. The highly conserved invasion plasmid antigens (Ipa), which are components of the Shigella type III secretion system, participate in bacterial epithelial cell invasion and have been pursued as vaccine targets. However, expression and purification of these proteins in conventional cell-based systems have been challenging due to solubility issues and extremely low recovery yields. These difficulties have impeded manufacturing and clinical advancement. In this study, we describe a new method to express Ipa proteins using the Xpress+TM cell-free protein synthesis (CFPS) platform. Both IpaB and the C-terminal domain of IpaH1.4 (IpaH-CTD) were efficiently produced with this technology at yields > 200 mg/L. Furthermore, the expression was linearly scaled in a bioreactor under controlled conditions, and proteins were successfully purified using multimode column chromatography to > 95% purity as determined by SDS-PAGE. Biophysical characterization of the cell-free synthetized IpaB and IpaH-CTD using SEC-MALS analysis showed well-defined oligomeric states of the proteins in solution. Functional analysis revealed similar immunoreactivity as compared to antigens purified from E. coli. These results demonstrate the efficiency of CFPS for Shigella protein production; the practicality and scalability of this method will facilitate production of antigens for Shigella vaccine development and immunological analysis. Key points • First report of Shigella IpaB and IpaH produced at high purity and yield using CFPS • CFPS-IpaB and IpaH perform similarly to E. coli–produced proteins in immunoassays • CFPS-IpaB and IpaH react with Shigella-specific human antibodies and are immunogenic in mice. Graphical abstract

The Role of Staphylococcus aureus YycFG in Gene Regulation, Biofilm Organization and Drug Resistance

Antibiotic resistance is a serious global health concern that may have significant social and financial consequences. Methicillin-resistant Staphylococcus aureus (MRSA) infection is responsible for substantial morbidity and leads to the death of 21.8% of infected patients annually. A lack of novel antibiotics has prompted the exploration of therapies targeting bacterial virulence mechanisms. The two-component signal transduction system (TCS) enables microbial cells to regulate gene expression and the subsequent metabolic processes that occur due to environmental changes. The YycFG TCS in S. aureus is essential for bacterial viability, the regulation of cell membrane metabolism, cell wall synthesis and biofilm formation. However, the role of YycFG-associated biofilm organization in S. aureus antimicrobial drug resistance and gene regulation has not been discussed in detail. We reviewed the main molecules involved in YycFG-associated cell wall biosynthesis, biofilm development and polysaccharide intercellular adhesin (PIA) accumulation. Two YycFG-associated regulatory mechanisms, accessory gene regulator (agr) and staphylococcal accessory regulator (SarA), were also discussed. We highlighted the importance of biofilm formation in the development of antimicrobial drug resistance in S. aureus infections. Data revealed that inhibition of the YycFG pathway reduced PIA production, biofilm formation and bacterial pathogenicity, which provides a potential target for the management of MRSA-induced infections.

Nanotechnology as a Novel Approach in Combating Microbes Providing an Alternative to Antibiotics

The emergence of infectious diseases promises to be one of the leading mortality factors in the healthcare sector. Although several drugs are available on the market, newly found microorganisms carrying multidrug resistance (MDR) against which existing drugs cannot function effectively, giving rise to escalated antibiotic dosage therapies and the need to develop novel drugs, which require time, money, and manpower. Thus, the exploitation of antimicrobials has led to the production of MDR bacteria, and their prevalence and growth are a major concern. Novel approaches to prevent antimicrobial drug resistance are in practice. Nanotechnology-based innovation provides physicians and patients the opportunity to overcome the crisis of drug resistance. Nanoparticles have promising potential in the healthcare sector. Recently, nanoparticles have been designed to address pathogenic microorganisms. A multitude of processes that can vary with various traits, including size, morphology, electrical charge, and surface coatings, allow researchers to develop novel composite antimicrobial substances for use in different applications performing antimicrobial activities. The antimicrobial activity of inorganic and carbon-based nanoparticles can be applied to various research, medical, and industrial uses in the future and offer a solution to the crisis of antimicrobial resistance to traditional approaches. Metal-based nanoparticles have also been extensively studied for many biomedical applications. In addition to reduced size and selectivity for bacteria, metal-based nanoparticles have proven effective against pathogens listed as a priority, according to the World Health Organization (WHO). Moreover, antimicrobial studies of nanoparticles were carried out not only in vitro but in vivo as well in order to investigate their efficacy. In addition, nanomaterials provide numerous opportunities for infection prevention, diagnosis, treatment, and biofilm control. This study emphasizes the antimicrobial effects of nanoparticles and contrasts nanoparticles’ with antibiotics’ role in the fight against pathogenic microorganisms. Future prospects revolve around developing new strategies and products to prevent, control, and treat microbial infections in humans and other animals, including viral infections seen in the current pandemic scenarios.

Etiological significance of Corynebacterium spp. in the development of diseases of the respiratory tract

Corynebacterium spp. It is associated with inflammatory diseases of the respiratory tract (tracheitis, pharyngitis, rhinosinusitis, bronchitis, pneumonia, etc.). C. pseudodiphtheriticum can be the causative agent of bacterial coinfection in patients with a new coronavirus infection (COVID-19). The aim is to determine the pathogenic properties and resistance to antimicrobial drugs of Corynebacterium spp. strains to establish their etiological significance in the development of inflammatory diseases of the respiratory tract. Strains of Corynebacterium spp. isolated from patients with inflammatory diseases of the respiratory tract (43 pcs.) and practically healthy individuals (29 pcs.). Isolates were identified by mass spectrometric method (MALDI-TOF MS), their cytopathic effect in CHO-K1 cell culture, hemolytic, urease activity, antimicrobial drug resistance were determined. Strains of Corynebacterium spp. isolated from patients in the amount of 105 CFU/ml or more, practically healthy - 104 CFU/ml or less. Isolates of Corynebacterium spp. patients had a more pronounced cytopathic effect (83.7±11.1%) and were more often resistant to antimicrobial drugs than those isolated from practically healthy. To establish the etiological significance of Corynebacterium spp. isolates. in the development of inflammatory diseases of the respiratory tract, it is advisable to determine their amount in biological material (105 CFU/ml or more), the cytopathic effect on CHO-K1 cell culture, as well as the presence of multiple resistance to antimicrobial drugs. Differences in the characteristics of Corynebacterium spp. isolates. from patients with respiratory tract pathology and practically healthy individuals are associated with the strain, not the species, of corynebacteria.

Virtual Screening to Identify the Protein Network Interaction of Triclosan in Red Complex Pathogens

Background: Antimicrobial drug resistance is the major problem encountered world-wide. Novel therapeutic leads have been identified and are regularly tested for their activity against microbial pathogens. Aim: To identify the protein network interactions of triclosan in red complex pathogens. Materials and Methods: The present study follows an observational study design which aims to screen for the interaction of triclosan in red complex pathogens. The interaction was analyzed using the STITCH v.5 pipeline. The functional class of proteins identified were assessed using VICMPred and VirulentPred softwares. The microbial pathogens Treponema denticola ATCC 35405, Tannerella forsythia ATCC 43037, Porphyromonas gingivalis ATCC 33277 are the strains of red complex pathogens that are included in the present study. Results and Discussion: Several proteins were found to interact with triclosan. Among the protein interactions, interactions of triclosan with virulent proteins seems to have a greater impact. The NAD-dependent nucleotide-diphosphate-sugar epimerase [PGN_1370], Putative NAD dependent epimerase [PGN_1886], GDP-fucose synthetase [PGN_1079], Probable oxidoreductase [PGN_1360] of Porphyromonas gingivalis, Conserved hypothetical protein [TDE_2401], Epimerase/dehydratase family protein [TDE_1439] of Treponema denticola, NAD dependent epimerase/dehydratase family protein [BFO_2919], Hypothetical protein [BFO_1782], Nitroreductase family protein [BFO_1604] and Nitroreductase family protein [BFO_1516] Tannerella forsythia were found to be exhibit virulence nature. Conclusion: This study identifies the molecular targets of triclosan on red complex pathogens. As triclosan interacts with the red complex pathogens, in future it can be used as a primary medicine for periodontitis and some oral conditions.

ANTIMICROBIAL SUSCEPTIBILITY OF FOSFOMYCIN IN BACTERIAL ISOLATES FROM URINARY TRACT INFECTION : A STUDY FROM TERTIARY HEALTHCARE CENTER IN HIMACHAL PRADESH

BACKGROUND: Urinary tract infections (UTIs) are common with an annual global incidence of at least 250 million cases; and Escherichia coli is the most common pathogen. Many antibiotics once used to treat UTI are now ineffective due to the development of antimicrobial resistance. Fosfomycin, discovered in late 1960s, has rekindled clinical interests because of reported susceptibilities of current pathogens to the agent. This study was done with the objective to determine in vitro fosfomycin susceptibility of common uropathogens and determining the antimicrobial susceptibility pattern of these organisms. MATERIAL AND METHOD: Retrospective study was conducted in the Department of Microbiology for a duration of 2 years from October 2019 to September 2021. Urine samples were received and culture was done on MacConkey agar and AST was performed on signicant bacteriuria ≥105 CFU /ml with fosfomycin as per CLSI guidelines. Total of 9442 urine samples were collected duri RESULT: ng the duration of 2 years. Out of these 1657(17.5%) showed signicant growth. Male to female ratio was 1:1.9 in our study. Majority of the bacteria isolated were uropathogenic E.coli 960(57.9%) followed by Klebsiella spp 185(11.1%). Susceptibility to fosfomycin was seen majorly in all enterobacterales with 96.7% sensitivity to E.coli and 100% sensitivity towards Staphylococcus aureus and Enterococcus spp. In CONCLUSION: conclusion, our study indicates that fosfomycin is active in vitro against a considerable percentage of urinary isolates, which simultaneously exhibit high rates of antimicrobial drug resistance to the conventionally used antimicrobial agents.