Single-cell phenotypic profiling and backtracing exposes and predicts clinically relevant subpopulations in isogenic Staphylococcus aureus communities.

Isogenic bacterial cell populations are phenotypically heterogenous and may include subpopulations of antibiotic tolerant or heteroresistant cells. The reversibility of these phenotypes and lack of biomarkers to differentiate functionally different, but morphologically identical cells is a challenge for research and clinical detection. To overcome this, we present ´Cellular Phenotypic Profiling and backTracing (CPPT)´, a fluorescence-activated cell sorting platform that uses fluorescent probes to visualize and quantify cellular traits and connects this phenotypic profile with a cell´s experimentally determined fate in single cell-derived growth and antibiotic susceptibility analysis.

Development of an inducer-free, virulence gene promoter-controlled, and fluorescent reporter-labeled CRISPR interference system in .

The dCas9-based Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) interference (CRISPRi) gene regulation technique requires two components: a catalytically inactive Cas9 protein (dCas9) and a single-guide RNA that targets the gene of interest. This system is commonly activated by expressing dCas9 through an inducible gene promoter, but these inducers may affect cellular physiology, and accessibility and permeability of the inducer are limited in relevant model systems. Here, we have developed an alternative approach for CRISPRi activation in the clinical isolate USA300 LAC, where dCas9 was expressed through endogenous virulence gene promoters (vgp); coagulase, autolysin, or fibronectin-binding protein A.

Chitosan and liposomal delivery systems for epicatechin or propyl gallate targeting localized treatment of vulvovaginal candidiasis.

Natural polyphenols are promising alternatives to antifungals for novel treatments of vulvovaginal candidiasis (VVC) in an era of antimicrobial resistance. However, polyphenols are poorly soluble and prone to degradation. To overcome their limitations, we propose incorporation in liposomes.

Deciphering host dynamics using dual activity-based protein profiling of ATP-interacting proteins.

Unlabelled: The utilization of ATP within cells plays a fundamental role in cellular processes that are essential for the regulation of host-pathogen dynamics and the subsequent immune response. This study focuses on ATP-binding proteins to dissect the complex interplay between and human cells, particularly macrophages (THP-1) and keratinocytes (HaCaT), during an intracellular infection. A snapshot of the various protein activity and function is provided using a desthiobiotin-ATP probe, which targets ATP-interacting proteins.

Antimicrobial liposomes-in-nanofiber wound dressings prepared by a green and sustainable wire-electrospinning set-up.

Increasing prevalence of infected and chronic wounds demands improved therapy options. In this work an electrospun nanofiber dressing with liposomes is suggested, focusing on the dressing's ability to support tissue regeneration and infection control. Chloramphenicol (CAM) was the chosen antibiotic, added to the nanofibers after first embedded in liposomes to maintain a sustained drug release.

Interactions between commensal and and clinical isolates of .

() is a versatile pathogen, responsible for multidrug-resistant infections, especially in hospitalized immunocompromised patients. Its population structure has been characterized by diverse clades (A1, A2, and B (reclassified as ()), adapted to different environments, and distinguished by their resistomes and virulomes. These features only partially explain the predominance of clade A1 strains in nosocomial infections.

Comprehensive virulence profiling and evolutionary analysis of specificity determinants in two-component systems.

In the genome, a set of highly conserved two-component systems (TCSs) composed of histidine kinases (HKs) and their cognate response regulators (RRs) sense and respond to environmental stimuli, which drive the adaptation of the bacteria. This study investigates the complex interplay between TCSs in USA300, a predominant methicillin-resistant strain, revealing shared and unique virulence regulatory pathways and genetic variations mediating signal specificity within TCSs. Using TCS-related mutants from the Nebraska Transposon Mutant Library, we analyzed the effects of inactivated TCS HKs and RRs on the production of various virulence factors, infection abilities, and adhesion assays.

Co-culturing with alters transcriptome when exposed to tonsillar cells.

Introduction: Improved understanding of throat colonization in the presence of other co-existing microbes is important for mapping adaptation to the human throat, and recurrence of infection. Here, we explore the responses triggered by the encounter between two common throat bacteria, and , to identify genes in that are important for colonization in the presence of human tonsillar epithelial cells and , and further compare this transcriptome with the genes expressed in as only bacterium.

Methods: We performed an co-culture experiment followed by RNA sequencing to identify interaction-induced transcriptional alterations and differentially expressed genes (DEGs), followed by gene enrichment analysis.

Enterococcal Membrane Vesicles as Vaccine Candidates.

is a leading cause of nosocomial infections, particularly in immunocompromised patients. The rise of multidrug-resistant , including Vancomycin-Resistant Enterococci (VRE), is a major concern. Vaccines are promising alternatives to antibiotics, but there is currently no vaccine available against enterococci.

Age-related differences in the occurrence, severity, and distress of symptoms in older patients at the initiation of chemotherapy.

Background: Evaluate for differences in occurrence, severity, and distress ratings for 32 symptoms between younger older adults (YOA, < 70 years) and older adults (OA, ≥ 70 years) at initiation of chemotherapy.

Methods: Patients (n = 125) were recruited prior to the initiation of chemotherapy and completed the Memorial Symptom Assessment Scale. Differences in occurrence, severity, and distress ratings were evaluated using Independent sample t-tests and Chi-square or Fisher's exact tests.

Polymyxin B stabilized DNA micelles for sustained antibacterial and antibiofilm activity against .

Nucleic acid-based materials showcase an increasing potential for antimicrobial drug delivery. Although numerous reports on drug-loaded DNA nanoparticles outline their pivotal antibacterial activities, their potential as drug delivery systems against bacterial biofilms awaits further studies. Among different oligonucleotide structures, micellar nanocarriers derived from amphiphilic DNA strands are of particular interest due to their spontaneous self-assembly and high biocompatibility.

Exploring differentially expressed genes of Staphylococcus aureus exposed to human tonsillar cells using RNA sequencing.

Background: The nose and the throat are the most predominant colonizing sites of Staphylococcus aureus, and colonization is a risk factor for infection. Nasal colonization is well described; however, we have limited knowledge about S. aureus throat colonization.

eHealth policy framework in Low and Lower Middle-Income Countries; a PRISMA systematic review and analysis.

Background: Low and lower middle-income countries suffer lack of healthcare providers and proper workforce education programs, a greater spread of illnesses, poor surveillance, efficient management, etc., which are addressable by a central policy framework implementation. Accordingly, an eHealth policy framework is required specifically for these countries to successfully implement eHealth solutions.

The validation of the Norwegian Basic Psychological Need Satisfaction and Frustration Scale: A stratified sampling procedure.

This study aimed to validate the Norwegian version of the Basic Psychological Need Satisfaction and Frustration Scale (BPNSFS) and to examine its relations with indicators of well-being and ill-being. Additionally, despite the vast number of studies employing the BPNSFS, norms related to the BPNSFS are currently lacking. Therefore, we also aimed to provide normative data for this scale.

Chitosan-based delivery system enhances antimicrobial activity of chlorhexidine.

Infected chronic skin wounds and other skin infections are increasingly putting pressure on the health care providers and patients. The pressure is especially concerning due to the rise of antimicrobial resistance and biofilm-producing bacteria that further impair treatment success. Therefore, innovative strategies for wound healing and bacterial eradication are urgently needed; utilization of materials with inherent biological properties could offer a potential solution.

Behavioral and neuropsychological profile of a male patient with mosaic PCDH19 mutation.

Mutations in the protocadherin 19 gene ( are associated with a female-restricted form of epilepsy. As the disorder has an X-reversed inheritance pattern, men are usually healthy carriers. PCDH19 epilepsy among men due to mosaicism is rare but probably underdiagnosed.

Biofilm Responsive Zwitterionic Antimicrobial Nanoparticles to Treat Cutaneous Infection.

To avert the poor bioavailability of antibiotics during biofilm infections, a series of zwitterionic nanoparticles containing nucleic acid nanostructures were fabricated for the delivery of vancomycin. The nanoparticles were prepared with three main lipids: (i) neutral (soy phosphatidylcholine; P), (ii) positively charged ionizable (1,2-dioleyloxy-3-dimethylaminopropane; D), and (iii) anionic (1,2-dipalmitoyl--glycero-3-phospho((ethyl-1',2',3'-triazole) triethylene glycolmannose; M) or (cholesteryl hemisuccinate; C) lipids. The ratio of the anionic lipid was tuned between 0 and 10 mol %, and its impact on surface charge, size, stability, toxicity, and biofilm sensitivity was evaluated.

Bacterial Membrane-Derived Vesicles Attenuate Vancomycin Activity against Methicillin-Resistant .

Methicillin-resistant (MRSA) has evolved numerous antimicrobial resistance mechanisms and is identified as a serious public health threat by the World Health Organization and U.S. Centers for Disease Control and Prevention.

Multifunctional Nanofibrous Dressing with Antimicrobial and Anti-Inflammatory Properties Prepared by Needle-Free Electrospinning.

An active wound dressing should address the main goals in wound treatment, which are improved wound healing and reduced infection rates. We developed novel multifunctional nanofibrous wound dressings with three active ingredients: chloramphenicol (CAM), beta-glucan (βG) and chitosan (CHI), of which βG and CHI are active nanofiber-forming biopolymers isolated from the cell walls of and from shrimp shells, respectively. To evaluate the effect of each active ingredient on the nanofibers' morphological features and bioactivity, nanofibers with both βG and CHI, only βG, only CHI and only copolymers, polyethylene oxide (PEO) and hydroxypropylmethylcellulose (HPMC) were fabricated.

Chitosomes-In-Chitosan Hydrogel for Acute Skin Injuries: Prevention and Infection Control.

Burns and other skin injuries are growing concerns as well as challenges in an era of antimicrobial resistance. Novel treatment options to improve the prevention and eradication of infectious skin biofilm-producing pathogens, while enhancing wound healing, are urgently needed for the timely treatment of infection-prone injuries. Treatment of acute skin injuries requires tailoring of formulation to assure both proper skin retention and the appropriate release of incorporated antimicrobials.

Nurses' experiences of delirium and how to identify delirium-A qualitative study.

Aim: Delirium is a serious, acute medical condition which places a heavy burden on the patient, his or her family and healthcare professionals. There have been only a limited number of studies to explore nurses' experiences of delirium and how delirium is identified in community care. The research questions of the study are as follows: "How do community care nurses' experience delirium?" and "How is delirium identified?".

Transcriptome Profiling of Associated Extracellular Vesicles Reveals Presence of Small RNA-Cargo.

Bacterial extracellular vesicles (EVs) have a vital role in bacterial pathogenesis. However, to date, the small RNA-cargo of EVs released by the opportunistic pathogen has not been characterized. Here, we shed light on the association of small RNAs with EVs secreted by MSSA476 cultured in iron-depleted bacteriologic media supplemented with a subinhibitory dosage of vancomycin to mimic infection condition.

Liposomes-In-Hydrogel Delivery System Enhances the Potential of Resveratrol in Combating Vaginal Chlamydia Infection.

is the most common cause of bacterial sexually transmitted infections and causes serious reproductive tract complications among women. The limitations of existing oral antibiotics and treatment of antimicrobial resistance require alternative treatment options. We are proposing, for the first time, the natural polyphenol resveratrol (RES) in an advanced delivery system comprising liposomes incorporated in chitosan hydrogel, for the localized treatment of infection.

Shotgun-Metagenomics on Positive Blood Culture Bottles Inoculated With Prosthetic Joint Tissue: A Proof of Concept Study.

Clinical metagenomics is actively moving from research to clinical laboratories. It has the potential to change the microbial diagnosis of infectious diseases, especially when detection and identification of pathogens can be challenging, such as in prosthetic joint infection (PJI). The application of metagenomic sequencing to periprosthetic joint tissue (PJT) specimens is often challenged by low bacterial load in addition to high level of inhibitor and contaminant host DNA, limiting pathogen recovery.

Liposomal delivery of antibiotic loaded nucleic acid nanogels with enhanced drug loading and synergistic anti-inflammatory activity against S. aureus intracellular infections.

The persistence of Staphylococcus aureus has been accredited to its ability to escape immune response via host cell invasion. Despite the efficacy of many antibiotics against S. aureus, the high extracellular concentrations of conventional antibiotics required for bactericidal activity is limited by their low cellular accumulation and poor intracellular retention.