Search Results (24)

Acanthamoeba, a free-living amoeba ubiquitous in environmental water, has been considered as the environmental reservoir of certain bacterial pathogens, including Campylobacter jejuni, an intracellular human pathogen causing self-limiting gastroenteritis. Acanthamoeba-C. jejuni interaction mechanisms may help clarify how the otherwise fastidious bacterium C. jejuni survives in environmental waters. In this study, we constructed single deletion mutants of C. jejuni strain 81–176 for the 10 selected genes (motAB, ciaB, kpsE, virB11, cheY, flaAB, cstII, docB, sodB, and cadF) previously shown to be important for the interaction (invasion and intracellular survival) of C. jejuni with mammalian hosts. We used a modified gentamicin protection assay to quantify the internalization and intracellular survival of these mutants and the wild type with the two species of Acanthamoeba (A. castellanii and A. polyphaga). Both internalization and intracellular survival were significantly lower for all mutants compared to the wild type with both amoeba strains, except for ΔcstII in the internalization assay with A. castellanii (p < 0.05). The results of this study highlight that the mechanisms used by C. jejuni to interact with mammalian hosts are conserved in its interactions with amoeba hosts. This understanding may be useful in developing effective strategies to reduce the transmission of C. jejuni to chickens through drinking water. Full article

Background/Objectives: Identification and characterization of broadly neutralizing monoclonal antibodies from individuals exposed to SARS-CoV-2, either by infection or vaccination, can inform the development of next-generation vaccines and antibody therapeutics with pan-SARS-CoV-2 protection. Methods: Through single B cell sorting and RT-PCR, monoclonal antibodies (mAbs) were isolated from a donor who experienced a BA.5 or BF.7 breakthrough infection after three doses of inactivated vaccines. Their binding and neutralizing capacities were measured with ELISA and a pseudovirus-based neutralization assay, respectively. Their epitopes were mapped by competition ELISA and site-directed mutation. Results: Among a total of 67 spike-specific mAbs cloned from the donor, four mAbs (KXD643, KXD652, KXD681, and KXD686) can neutralize all tested SARS-CoV-2 variants from wild-type to KP.3. Moreover, KXD643, KXD652, and KXD681 belong to a clonotype encoded by IGHV5-51 and IGKV1-13 and recognize the cryptic and conserved RBD-8 epitope on the receptor-binding domain (RBD). In contrast, KXD686 is encoded by IGHV1-69 and IGKV3-20 and targets a conserved epitope (NTD Site iv) outside the antigenic supersite (NTD Site i) of the N-terminal domain (NTD). Notably, antibody cocktails containing these two groups of mAbs can neutralize SARS-CoV-2 more potently due to synergistic effects. In addition, bispecific antibodies derived from KXD643 and KXD686 demonstrate further improved neutralizing potency compared to antibody cocktails. Conclusions: These four mAbs can be developed as candidates of pan-SARS-CoV-2 antibody therapeutics through further antibody engineering. On the other hand, vaccines designed to simultaneously elicit neutralizing antibodies towards RBD-8 and NTD Site iv have the potential to provide pan-SARS-CoV-2 protection. Full article

Background: Bacterial superinfections are common complications during viral infections, but the impact of multidrug-resistant (MDR) pathogens in critically ill patients affected by coronavirus disease 2019 (COVID-19) is still debated. Methods: This is an observational, monocentric, and prospective study designed to investigate the incidence, risk factors, and outcomes of MDR bacterial superinfections in COVID-19 patients admitted to the intensive care unit (ICU). Results: A high incidence of superinfections (66%, 159/241) was observed: ventilator-associated pneumonia (VAP) (65%, 104/159) and bloodstream infection (BSI, 32%, 51/159) were the most common. Superinfections, Extra-Corporeal Membrane Oxygenation (ECMO) support, and prone positioning increased the risk of death five, four, and more-than-two times, respectively (OR = 5.431, IC 95%: 1.637–18.014; 4.462, IC 95%: 1.616–12.324 and 2.346, IC 95%: 1.127–4.883). MDR bacteria were identified in 61% of patients with superinfection, with a cumulative incidence of 37.2% at day 14. Carbapenem-resistant Acinetobacter baumannii (CR-AB) and CR-Klebsiella pneumoniae (CR-KP) were the most common causative agents (24.3% and 13.7%). CR-AB was found to significantly increase both ICU and in-hospital mortality (76.4% and 78.2%), whereas CR-KP had no direct impact on mortality. Prior rectal colonization (p < 0.0001), mechanical ventilation (p = 0.0017), a prolonged ICU stay (p < 0.0001), the use of iNO (p = 0.0082), vasopressors (p = 0.0025), curarization (p = 0.0004), and prone positioning (p = 0.0084) were found to be risk factors for CR-AB. Conclusions: Critically ill COVID-19 patients are at high risk of developing MDR superinfection. While CR-KP had no direct impact on mortality, CR-AB appeared to increase ICU and in-hospital mortality. Full article

The rising prevalence of diseases linked to factors such as excessive alcohol and tobacco use, sedentary lifestyles, and poor nutrition has led to a greater focus on Personal Health Promotion (PHP) as a preventive measure. PHP emphasizes improving quality of life and well-being, driven by advances in medical technology, including virtual and augmented reality. However, as PHP systems grow in popularity, concerns over personal data security, such as account theft and information breaches, have become increasingly important. To address these concerns, this study proposes a game-based personalized health promotion system that integrates Key-Policy Attribute-Based Encryption (KP-ABE), key insulation, time-bound encryption. These methods ensure data security through hierarchical access control and dynamic time management, protecting personal health records and exercise prescriptions. The system also incorporates key insulation to enhance the security of private keys. This multi-layered security approach provides a robust solution for safeguarding sensitive data within PHP systems while accommodating dynamic subscription needs and legal access requirements. Full article

New SARS-CoV-2 lineages continue to evolve and may exhibit new characteristics regarding host cell entry efficiency and potential for antibody evasion. Here, employing pseudotyped particles, we compared the host cell entry efficiency, ACE2 receptor usage, and sensitivity to antibody-mediated neutralization of four emerging SARS-CoV-2 lineages, KP.2, KP.2.3, KP.3, and LB.1. The XBB.1.5 and JN.1 lineages served as controls. Our findings reveal that KP.2, KP.2.3, KP.3, and LB.1 lineages enter host cells efficiently and in an ACE2-dependent manner, and that KP.3 is more adept at entering Calu-3 lung cells than JN.1. However, the variants differed in their capacity to employ ACE2 orthologues from animal species for entry, suggesting differences in ACE2 interactions. Moreover, we demonstrate that only two out of seven therapeutic monoclonal antibody (mAbs) in preclinical development retain robust neutralizing activity against the emerging JN.1 sublineages tested, while three mAbs displayed strongly reduced neutralizing activity and two mAbs lacked neutralizing activity against any of the lineages tested. Furthermore, our results show that KP.2, KP.2.3, KP.3, and LB.1 lineages evade neutralization by antibodies induced by infection or vaccination with greater efficiency than JN.1, particularly in individuals without hybrid immunity. This study indicates that KP.2, KP.2.3, KP.3, and LB.1 differ in ACE2 interactions and the efficiency of lung cell entry and suggest that evasion of neutralizing antibodies drove the emergence of these variants. Full article

Escherichia coli isolates from intensive poultry production are associated with antimicrobial resistance and worldwide health problems. The aim of the study was to detect and evaluate the phenotypic and genotypic antimicrobial resistance, biofilm formation, phylogenetic typing, and virulence factors in E. coli isolates from the rectal swabs of chickens from two farms and swabs of chicken meat purchased from Slovakian food markets. Interpretative readings of minimal inhibitory concentration (MIC) revealed dominant resistance to ampicillin (>50%) in both groups. We also detected higher resistance to ciprofloxacin (45%), tetracycline, ampicillin + sulbactam, and trimethoprim + sulfonamide (each >30%). Here, 28.57% of the strains studied were multidrug-resistant (MDR). The formation of weak biofilms was confirmed in 8.8% of E. coli, while one of the strains obtained from chicken cloacal swabs was classified as a strong biofilm producer. The most frequently confirmed phylogenetic groups in E. coli were B₁ and A₁ in all groups. PCR detection revealed the presence of genes encoding tetracycline resistance (tetAB) and plasmid-mediated quinolone resistance (qnrABS), and Int1 (52.9%), Tn3 (76.5%), kpsMT II (8.8%), fimA (97.1%), cvaC (38.2%), and iutA (76.5%) genes in the strains studied. Our results demonstrate that chickens and chicken meat were the source of antibiotic-resistant, biofilm-forming, and virulent E. coli, representing a potential risk from the point of view of the One Health concept. Full article

Fungal diseases not only reduce the yield of edible mushrooms but also pose potential threats to the preservation and quality of harvested mushrooms. Cobweb disease, caused primarily by fungal pathogens from the Hypocreaceae family, is one of the most significant diseases affecting edible mushrooms. Deciphering the genomes of these pathogens will help unravel the molecular basis of their evolution and identify genes responsible for pathogenicity. Here, we present high-quality genome sequences of three cobweb disease fungi: Hypomyces aurantius Cb-Fv, Cladobotryum mycophilum CB-Ab, and Cladobotryum protrusum CB-Mi, isolated from Flammulina velutipes, Agaricus bisporus, and Morchella importuna, respectively. The assembled genomes of H. aurantius, C. mycophilum, and C. protrusum are 33.19 Mb, 39.83 Mb, and 38.10 Mb, respectively. This is the first report of the genome of H. aurantius. Phylogenetic analysis revealed that cobweb disease pathogens are closely related and diverged approximately 17.51 million years ago. CAZymes (mainly chitinases, glucan endo-1,3-beta-glucosidases, and secondary metabolite synthases), proteases, KP3 killer proteins, lipases, and hydrophobins were found to be conserved and strongly associated with pathogenicity, virulence, and adaptation in the three cobweb pathogens. This study provides insights into the genome structure, genome organization, and pathogenicity of these three cobweb disease fungi, which will be a valuable resource for comparative genomics studies of cobweb pathogens and will help control this disease, thereby enhancing mushroom quality. Full article

The particle size distribution (PSD) in emulsion polymerization (EP) has been modeled in the past using either the pseudo bulk (PB) or the 0-1/0-1-2 approaches. There is some controversy on the proper type of model to be used to simulate the experimental PSDs, which are apparently broader than the theoretical ones. Additionally, the numerical technique employed to solve the model equations, involving hyperbolic partial differential equations (PDEs) with moving and possibly steep fronts, has to be precise and robust, which is not a trivial matter. A deterministic kinetic model for the PSD evolution of ab initio EP of vinyl monomers was developed to investigate these issues. The model considers three phases, micellar nucleation, and particles that can contain $n\ge 0$ radicals. Finite volume (FV) and weighted-residual methods are used to solve the system of PDEs and compared; their limitations are also identified. The model was validated by comparing predictions with data of monomer conversion and PSD for the batch emulsion homopolymerization of styrene (Sty) and methyl methacrylate (MMA) using sodium dodecyl sulfate (SDS)/potassium persulfate (KPS) at 60 °C, as well as the copolymerization of Sty-MMA (50/50; mol/mol) at 50 and 60 °C. It is concluded that the PB model has a structural problem when attempting to adequately represent PSDs with steep fronts, so its use is discouraged. On the other hand, there is no generalized evidence of the need to add a stochastic term to enhance the PSD prediction of EP deterministic models. Full article

Photosynthesis is a key process in plant physiology. Understanding its mechanisms is crucial for optimizing crop yields and for environmental monitoring across a diverse range of plants. In this study, we employed reflectance, transmittance, and absorbance hyperspectral sensors and utilized multivariate statistical techniques to improve the predictive models for chlorophyll a fluorescence (ChlF) parameters in Hibiscus and Geranium model plants. Our objective was to identify spectral bands within hyperspectral data that correlate with ChlF indicators using high-resolution data spanning the electromagnetic spectrum from ultraviolet to shortwave infrared (UV–VIS–NIR–SWIR). Utilizing the hyperspectral vegetation indices (HVIs) tool to align importance projection for wavelength preselection and select the most responsive wavelength by variable importance projection (VIP), we optimized partial least squares regression (PLSR) models to enhance predictive accuracy. Our findings revealed a strong relationship between hyperspectral sensor data and ChlF parameters. Employing principal component analysis, kappa coefficients (k), and accuracy (Acc) evaluations, we achieved values exceeding 86% of the predicted ChlF parameters for both Hibiscus and Geranium plants. Regression models for parameters such as Ψ(EO), ϕ(PO), ϕ(EO), ϕ(DO), δRo, ρRo, Kn, Kp, SFI(abs), PI(abs), and D.F. demonstrated model accuracies close to 0.84 for R² and approximately 1.96 for RPD. The spectral regions linked with these parameters included blue, green, red, infrared, SWIR1, and SWIR2, emphasizing their relevance for noninvasive evaluations. This research demonstrates the ability of hyperspectral sensors to noninvasively predict chlorophyll a fluorescence (ChlF) parameters, which are essential for assessing photosynthetic efficiency in plants. Notably, hyperspectral absorbance data were more accurate in predicting JIP-test-based chlorophyll a kinetic parameters. In conclusion, this study underscores the potential of hyperspectral sensors for deepening our understanding of plant photosynthesis and monitoring plant health. Full article

In many universities, students’ physical ability data are collected and stored in the cloud through various sensing devices to save computational and storage costs. Therefore, how to effectively access data while ensuring data security has become an urgent issue. Key-policy attribute-based encryption (KP-ABE) not only enables secure one-to-many communication and fine-grained access control but also adapts to data sharing in static scenarios, making it more suitable for the cloud sharing of physical ability data. In this paper, we construct an online/offline multi-authority key-policy attribute-based encryption with a cryptographic reverse firewall for physical ability data. This scheme uses multi-authority to avoid the single point of failure crisis of a single authority, and is combined with a cryptographic reverse firewall to resist backdoor attacks. In addition, the scheme uses outsourcing decryption to save users’ computing costs, and utilizes offline/online technology to move a large amount of computing offline, reducing the online burden. Finally, the experiment shows the feasibility of the scheme. Full article

Background: The high prevalence and rapid emergence of antibiotic resistance in high-risk Klebsiella pneumoniae (KP) ST147 clones is a global health concern and warrants molecular surveillance. Methods: A pangenome analysis was performed using publicly available ST147 complete genomes. The characteristics and evolutionary relationships among ST147 members were investigated through a Bayesian phylogenetic analysis. Results: The large number of accessory genes in the pangenome indicates genome plasticity and openness. Seventy-two antibiotic resistance genes were found to be linked with antibiotic inactivation, efflux, and target alteration. The exclusive detection of the bla_OXA-232 gene within the ColKp3 plasmid of KP_SDL79 suggests its acquisition through horizontal gene transfer. The association of seventy-six virulence genes with the acrAB efflux pump, T6SS system and type I secretion system describes its pathogenicity. The presence of Tn6170, a putative Tn7-like transposon in KP_SDL79 with an insertion at the flanking region of the tnsB gene, establishes its transmission ability. The Bayesian phylogenetic analysis estimates ST147’s initial divergence in 1951 and the most recent common ancestor for the entire KP population in 1621. Conclusions: Present study highlights the genetic diversity and evolutionary dynamics of high-risk clones of K. pneumoniae. Further inter-clonal diversity studies will help us understand its outbreak more precisely and pave the way for therapeutic interventions. Full article

This study aimed to characterize a Klebsiella pneumoniae strain (KP411) recovered from the stool samples of poultry (Gallus gallus) in the Brazilian Amazon Region. The whole-genome sequencing of KP411 revealed the presence of an important arsenal of antimicrobial resistance genes to β-lactams (bla_CTX-M-14, bla_TEM-1B, bla_KPC-2, bla_SVH-11), aminoglycosides [aph(3″)- Ib, aph(6)-Id, aph(3′)-Ia], sulfonamides (sul1, sul2), quinolones (oqxAB), fosfomycin (fosA^KP), and macrolides [mph(A)]. Furthermore, our analyses revealed that the KP411 strain belongs to the ST258 clonal lineage, which is one of the main epidemic clones responsible for the dissemination of KPC-2 worldwide. Our data suggest that food-producing animals may act as reservoirs of multidrug-resistant K. pneumoniae belonging to the ST258 clone, and, consequently, contribute to their dissemination to humans and the environment. Full article

Hypervirulent K. pneumoniae (hvKP) strains possess distinct characteristics such as hypermucoviscosity, unique serotypes, and virulence factors associated with high pathogenicity. To better understand the genomic characteristics and virulence profile of the isolated hvKP strain, genomic data were compared to the genomes of the hypervirulent and typical K. pneumoniae strains. The K. pneumoniae strain was isolated from a patient with a recurrent urinary tract infection, and then the string test was used for the detection of the hypermucoviscosity phenotype. Whole-genome sequencing was conducted using Illumina, and bioinformatics analysis was performed for the prediction of the isolate resistome, virulome, and phylogenetic analysis. The isolate was identified as hypermucoviscous, type 2 (K2) capsular polysaccharide, ST14, and multidrug-resistant (MDR), showing resistance to ciprofloxacin, ceftazidime, cefotaxime, trimethoprim-sulfamethoxazole, cephalexin, and nitrofurantoin. The isolate possessed four antimicrobial resistance plasmids (pKPN3-307_type B, pECW602, pMDR, and p3K157) that carried antimicrobial resistance genes (ARGs) (bla_OXA-1,bla_CTX-M-15, sul2, APH(3″)-Ib, APH(6)-Id, and AAC(6′)-Ib-cr6). Moreover, two chromosomally mediated ARGs (fosA6 and SHV-28) were identified. Virulome prediction revealed the presence of 19 fimbrial proteins, one aerobactin (iutA) and two salmochelin (iroE and iroN). Four secretion systems (T6SS-I (13), T6SS-II (9), T6SS-III (12), and Sci-I T6SS (1)) were identified. Interestingly, the isolate lacked the known hypermucoviscous regulators (rmpA/rmpA2) but showed the presence of other RcsAB capsule regulators (rcsA and rcsB). This study documented the presence of a rare MDR hvKP with hypermucoviscous regulators and lacking the common capsule regulators, which needs more focus to highlight their epidemiological role. Full article

Aim: The aim of the study was to evaluate the in vitro activity of N-acetylcysteine (NAC), alone or in combination with beta-lactams, against carbapenem-resistant Klebsiella pneumoniae (CR-Kp) and Acinetobacter baumannii (CR-Ab). Methods: The antibacterial activity of each compound was tested by broth microdilution and the synergism was evaluated by the checkerboard method. Killing studies of NAC alone and in combination with beta-lactams were performed. Bacterial morphological changes were investigated with scanning electron microscopy (SEM). Results: Overall, 30 strains were included (15 CR-Kp and 15 CR-Ab). The NAC Minimal Inhibitory Concentrations (MIC)50/90 were 5/5 and 2.5/5 mg/mL for CR-Kp and CR-Ab, respectively. For both microorganisms, NAC, in addition to beta-lactams (meropenem for CR-Kp, meropenem and ampicillin/sulbactam for CR-Ab, respectively), was able to enhance their activity. The killing studies showed a rapid and concentration-dependent activity of NAC alone; the addition of NAC to meropenem or ampicillin/sulbactam at subinhibitory concentrations induced a fast and lasting bactericidal activity that persisted over time. The SEM analyses showed evident morphological alterations of the bacterial cells following incubation with NAC, alone and in combination with meropenem. Conclusions: NAC demonstrated a high in vitro activity against CR-Kp and CR-Ab and was able to enhance beta-lactams’ susceptibility in the tested strains. The preliminary data on the SEM analyses confirmed the in vitro results. Full article

Introduction: We studied the trend of antimicrobial resistance and consumption at Saint George Hospital University Medical Center (SGHUMC), a tertiary care center in Beirut, Lebanon, with a focus on the SARS-CoV-2 pandemic. Materials and Methods: We calculated the isolation density/1000 patient-days (PD) of the most isolated organisms from 1 January 2015–31 December 2020 that included: E. coli (Eco), K. pneumoniae (Kp), P. aeruginosa (Pae), A. baumannii (Ab), S. aureus (Sau), and E. faecium (Efm). We considered March–December 2020 a surrogate of COVID-19. We considered one culture/patient for each antimicrobial susceptibility and excluded Staphylococcus epidermidis, Staphylococcus coagulase-negative, and Corynebacterium species. We analyzed the trends of the overall isolates, the antimicrobial susceptibilities of blood isolates (BSI), difficult-to-treat (DTR) BSI, carbapenem-resistant Enterobacteriaceae (CRE) BSI, and restricted antimicrobial consumption as daily-defined-dose/1000 PD. DTR implies resistance to carbapenems, beta-lactams, fluoroquinolones, and additional antimicrobials where applicable. Results and Discussion: After applying exclusion criteria, we analyzed 1614 blood cultures out of 8314 cultures. We isolated 85 species, most commonly Eco, at 52%. The isolation density of total BSI in 2020 decreased by 16%: 82 patients were spared from bacteremia, with 13 being DTR. The isolation density of CRE BSI/1000 PD decreased by 64% from 2019 to 2020, while VREfm BSI decreased by 34%. There was a significant decrease of 80% in Ab isolates (p-value < 0.0001). During COVID-19, restricted antimicrobial consumption decreased to 175 DDD/1000 PD (p-value < 0.0001). Total carbapenem consumption persistently decreased by 71.2% from 108DDD/1000 PD in 2015–2019 to 31 DDD/1000 PD in 2020. At SGHUMC, existing epidemics were not worsened by the pandemic. We attribute this to our unique and dynamic collaboration of antimicrobial stewardship, infection prevention and control, and infectious disease consultation. Full article