Search Results (165)

Background/Objectives: Ofatumumab is a fully human anti-CD20 monoclonal antibody approved for the treatment of relapsing forms of multiple sclerosis (MS). While its efficacy and safety have been demonstrated in clinical trials, real-world data focusing on laboratory changes and detailed immunophenotyping during treatment remain limited. The objective of this study was to assess routine laboratory parameters and immunophenotyping profiles in ofatumumab-treated patients in a real-world setting. Methods: We conducted a retrospective, single-center real-world study including 59 patients with relapsing–remitting MS treated with ofatumumab. Routine laboratory parameters were analyzed at the baseline and 6–12 months after treatment initiation. Immunophenotyping by flow cytometry was available for a subset of 29 patients. Infections were assessed during a follow-up period of at least six months. Paired comparisons were performed using the Wilcoxon signed-rank test. Results: Ofatumumab induced a profound and sustained depletion of CD19+ B cells (p < 0.001). Total T cells, CD4+ and CD8+ T-cell counts, the CD4/CD8 ratio, and natural killer (NK) cells remained largely stable over time. NK cells and helper T cells showed a numerical increase without statistical significance. IgM levels and relative lymphocyte percentages showed a statistically significant decrease compared with baseline (p = 0.047 and p = 0.016, respectively), while remaining within reference ranges. Other routine laboratory parameters remained stable. Reported infections were infrequent and predominantly mild. Conclusions: In this real-world cohort, ofatumumab demonstrated a favorable immunological and laboratory profile consistent with its known mechanism of action. These findings suggest that routine laboratory monitoring is sufficient for most patients, while immunophenotyping may be reserved for selected clinical scenarios. Further prospective studies integrating clinical and radiological outcomes are needed to better define the clinical relevance of these immunological findings. Full article

Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE) are high-burden healthcare-associated pathogens that increase mortality, prolong hospitalisation, and drive substantial healthcare costs worldwide. These infections are associated with high morbidity, increased mortality, prolonged hospital stays, and significant costs, particularly among immunocompromised patients or those with extended hospitalizations. This systematic review was conducted and reported in accordance with PRISMA 2020, aiming to synthesise existing data on the epidemiology, resistance mechanisms, clinical manifestations, and strategies for the diagnosis, treatment, and prevention of MRSA and VRE infections. Data were qualitatively synthesised. A total of 113 records published between 2020 and 2025 met the inclusion criteria and were identified through searches in multiple bibliographic databases and publisher platforms (e.g., PubMed, Scopus, Web of Science). MRSA and VRE are implicated in numerous severe infections, including ventilator-associated pneumonia, catheter-associated urinary tract infections, endocarditis, and bacteraemia. Antimicrobial resistance is driven by the mecA, vanA, and vanB genes, while biofilm formation further complicates therapeutic efforts. Biofilm formation can promote antibiotic tolerance (slower killing without an increase in MIC) and persistence (survival of ‘persister’ cells), distinct from genetic resistance, and may complicate therapy in selected infections. Effective strategies include appropriate anti-MRSA/anti-VRE agents (e.g., ceftaroline for MRSA; linezolid or daptomycin for VRE), active screening, stringent infection prevention and control measures, and antimicrobial stewardship programmes. Implementation is often hindered by institutional barriers, limited resources, and insufficient staff training. A multidisciplinary, evidence-based approach is essential for the effective management of these infections. Reducing this burden requires coordinated implementation of rapid diagnostics, stringent infection prevention and control, and antimicrobial stewardship, supported by sustained institutional and public health investment. Full article

We report a rare case of polymicrobial late-onset knee prosthetic joint infection (PJI) caused by Parvimonas micra and Staphylococcus aureus. An 80-year-old woman with multiple comorbidities presented, five years after total knee arthroplasty, with progressive pain and radiographic signs of prosthetic loosening. Synovial fluid analysis revealed marked neutrophilic inflammation, and intraoperative tissue cultures, including sonication fluid yielded both pathogens. Identification was confirmed by MALDI-TOF MS and whole-genome sequencing (WGS). The P. micra strain showed 97.2% identity to reference strain SAMN29629855, and carried virulence genes such as groEL, tufA, clpP, ctrD, srtC4, and gaIE, associated with oxidative stress response, adhesion, immune evasion, and biofilm formation. Resistance genes vanW, vanT, and vanY from the van operon were also detected, though vanA and vanB were absent. The patient underwent a two-stage revision surgery and a 12-week course of pathogen-targeted antimicrobial therapy, with complete resolution of symptoms and no recurrence after 12 months. This case highlights the overlooked pathogenicity of P. micra in chronic PJIs, especially in polymicrobial biofilm-related infections. The integration of WGS provided valuable insights into possible genetic characteristics of virulence determinants of this rare cause of PJI. Full article

Background/Objectives: Streptococcus pyogenes (Group A beta-hemolytic streptococcus, GAS) is the most common cause of bacterial pharyngitis and a major contributor to morbidity and mortality worldwide. There has been an increase in invasive GAS infections and related deaths in several European countries post-COVID-19 pandemic. We aimed to assess parental knowledge, attitudes, and practices regarding GAS pharyngotonsillitis, with a focus on antibiotic use and misuse. Methods: A cross-sectional questionnaire-based study was conducted on a convenience sample of parents of children admitted to the Pediatric Ward or visiting the Pediatric Emergency Department of the University General Hospital of Patras, Greece (September 2024–February 2025). For knowledge assessment, the questionnaire consisted of 10 True/False questions, based on which a total knowledge score was calculated. For attitude and practice assessment, the questionnaire consisted of 10 Likert scale questions. A Negative Practice Score was calculated as a sum of the answers in five practices with a negative perspective, with higher scores indicating worse practices and lower scores indicating better practices. Results: The study enrolled 378 parents, 79% of them were aware that not all children with a sore throat need antibiotics, and 61% believed asymptomatic children with a positive strep antigen test should receive antibiotics. Concerns about GAS transmissibility were high (76%), while attitudes about severity were mixed. A median GAS total knowledge score of 6 (IQR: 4–7) indicated moderate knowledge. Multivariable analysis revealed that male parents, non-immigrants, those previously hospitalized for GAS infection, and those informed by pediatricians or reliable websites had significantly higher knowledge scores. Regarding practices, most parents (72%) disagreed with requesting antibiotics from pediatricians, and 93.9% did not administer leftover antibiotics. Additionally, 58% expressed more concerns in recent years due to the increase in invasive infections. The median GAS Negative Practice Score was 10.5 (IQR: 7.0–13.0), indicating generally good practices, as lower scores correspond to fewer negative practices. Older parents and those with higher knowledge scores were also linked to fewer negative practices in multivariable analysis. Conclusions: These findings highlight the importance of targeted education on GAS pharyngotonsillitis and the need to focus on specific population groups to reduce antibiotic misuse. Full article

Vancomycin-resistant Enterococcus faecium (VREfm) is an emerging pathogen responsible for healthcare-associated infections. For this reason, 44 VREfm isolates collected during 2021–2023 were characterized using phenotypic and genomic approaches. VREfm isolates were identified by MALDI-TOF and antimicrobial susceptibility tests were performed with Vitek 2, Sensititre, or E-test. Sequence type (ST), antibiotic resistance genes, virulence factors and genetic relatedness were determined using Next Generation Sequencing. Forty-three isolates had a VanA phenotype and vanHAX genotype and one had a VanB phenotype and vanHBX genotype. Isolates showed high antibiotic resistance to various antibiotics, but generally remained susceptible to quinupristin/dalfopristin, tigecycline and eravacycline. Two isolates were resistant to linezolid, showing the chromosomal mutation G2576T in domain V of the 23S rRNA gene in one isolate, and the transferable linezolid resistance genes cfr(D) and optrA in the other. Thirty-eight isolates belonged to ST80, one to ST17 (ST80 and ST17 are included in CC17) and one to ST697. Genomic analysis of the ST80 isolates showed that nearly all of them belonged to a single cluster. To prevent further spread of VREfm in the nosocomial environment, in addition to the application of up-to-date infection control strategies and antibiotic stewardship programs, the implementation of genomic surveillance is recommended. Full article

People who inject drugs (PWIDs) remain underserved in HIV care. Evidence on rapid antiretroviral therapy (ART) for PWID is limited. We evaluated feasibility, effectiveness, safety, and patient-reported outcomes (PROs) for rapid initiation of bictegravir/emtricitabine/tenofovir alafenamide (BIC/FTC/TAF) supported by a peer navigation in Greece. This is a single-arm, multicenter pilot study including PWIDs (≥18 years) newly diagnosed or relinking after >3 months off ART. Participants started BIC/FTC/TAF on the same day or within 7 days and received peer navigation for 48 weeks. Co-primary endpoints were Week-24 virologic suppression (HIV-1 RNA < 50 copies/mL; FDA Snapshot) and grade 3–4 adverse events (AEs). Secondary endpoints included complete-case suppression at Weeks 24/48, CD4 recovery, retention, and PROs. Outcomes were compared with historical controls from the same centers. Thirty-seven participants were enrolled (83.8% male; median age 33.3 years). Median time to ART was 0 days (vs 78 in controls, p < 0.001). Retention was 67.6% at Week 24 and 54.1% at Week 48. In the primary (FDA Snapshot) analysis, suppression was 62.2% and 54.1% at Weeks 24 and 48; in complete-case analyses, results were 92.0% and 100%, respectively. Mean CD4 count increased by 208 cells/μL (95% CI 141–275) at Week 48. Quality of life improved and symptom burden decreased. No grade 3–4 AEs occurred. Rapid BIC/FTC/TAF with peer navigation eliminated delays to ART and achieved favorable virologic, immunologic, and PROs among those retained, with good tolerability. Despite retention challenges, this model appears feasible for PWID and may help close HIV care gaps toward UNAIDS 95–95–95 targets. Full article

Teacher identity emerges from the intertwined relationship of past, present, and future experiences, shaped through active reflection within individual, social, and cultural contexts. This study investigates how initial teacher education programs for future primary school teachers and future mathematics teachers influence their perceptions of mathematics education. Questionnaires with predominantly open-ended questions were distributed to 369 students: 195 enrolled in mathematics teacher education programs and 174 in primary school teacher education programs. The survey examined their preferences for school subjects in prior education, motivations for choosing teaching as a profession, important teacher qualities, and attitudes toward mathematics. The results reveal significant differences between the two groups: future primary school teachers emphasize the focus on children and relational aspects of teaching, while future mathematics teachers highlight mathematics as a scientific discipline. Both groups’ responses reflect their respective educational programs and experiences. The findings suggest that adopting a holistic approach in teacher education, integrating content knowledge with pedagogical practice, and supporting reflective processes can foster deeper self-understanding and the development of a professional teacher identity. Full article

Maize, one of the most cultivated crops worldwide, has multiple uses, one of which is human food. Maize flour intended for human consumption is preferably produced from var. indurata. This maize variety, although it has some desirable traits, generally has a lower yield capacity. In order to obtain high-yielding hybrids that would have some traits necessary to obtain flour for human consumption, fourteen lines with dent or semi-dent grains were crossed with four inbred lines with flint grain in a cyclic system. The 56 resulting hybrids were tested in two experimental years for yield, the percentage of unlodged plants, grain dry matter at harvest, as well as other traits, such as ASI (anthesis-to-silking interval), the interval from sowing to the appearance of stigmas and to physiological maturity, and plant senescence. The maternal lines A478 and A480 were noted for transmitting higher yields. Three hybrids were identified with higher yields, good silking–flowering coincidence, stay-green, and a high unlodged plants percentage: A478 × D328, A480 × B330, and A480 × D328. Full article

Background/Objectives: This study was aimed to investigate the safety profile of traditional Montenegrin Njeguški cheese by quantifying genes associated with resistance to clinically important antibiotics. Methods: Samples of Njeguški cheese were sourced from three artisan producers in Montenegro, identified as A, B, and C, with three individual batches selected per producer. Quantitative PCR (qPCR) was performed on bacterial DNA extracted directly from samples to detect genes encoding resistance to macrolide–lincosamide–streptogramin B (MLS_B) [erm(A), erm(B), erm(C)], vancomycin (vanA, vanB), tetracyclines [tet(M), tet(O), tet(S), tet(K), tet(W)], β-lactams (mecA, blaZ), aminoglycosides [aac (6′)-Ie aph (2″)-Ia], and carbapenems (bla_KPC, bla_OXA-48, bla_NDM-1, bla_GES, and bla_VIM). Results: Among the MLS_B resistance genes, erm(B) was detected in all samples, erm(C) was present only in those from producer B, while erm(A) was found exclusively in batch 3 from producer C. Tetracycline resistance genes were widely distributed, except for tet(O), which was absent in batch 3 from producers A and B. Regarding β-lactam resistance, both blaZ and mecA were consistently detected across all samples, with statistically significant differences observed between producers. None of the samples tested positive for vancomycin resistance genes or the aminoglycoside resistance gene, regardless of producer. Among the carbapenemase genes analyzed, bla_NDM-1 was the only one detected, found in most samples from producers B and C. Conclusions: This research provides the first risk assessment of artisanal and commercial Njeguški cheese regarding antimicrobial resistance genes. The findings offer valuable insights to enhance the microbiological safety of traditional Montenegrin cheeses, supporting consumer confidence in local and international markets. Full article

Background: Vancomycin-resistant Enterococcus faecium (VREfm), particularly vanA-positive strains, represents a growing threat in hospital settings worldwide. These bacteria are able to survive under severe environmental conditions, including high temperatures and saline concentrations. High genome plasticity and advanced ability of inheriting antimicrobial resistance determinants defined the success of E. faecium as a hospital pathogen. Methods: This study presents the whole genomic characterization of vanA-positive VREfm isolates, analyzing 10 clinical isolates collected from three tertiary care hospitals in Moscow, Russia. Several typing approaches, including two MLST schemes and cgMLST profiles, were used to elucidate the relationship between the isolates. Phylogenetic analysis placed the isolates in context with global VREfm populations, demonstrating both local clonal expansion and possible international connections. Phenotypic and genomic antimicrobial resistance profiles were obtained, as well as data regarding the repertoire of virulence factors and plasmid content. Results: Whole genome sequencing revealed that all isolates belonged to the clinically significant CC17 lineage, specifically sequence types ST80 and ST552. Notably, two isolates possessed truncated Tn1546-type transposons lacking vanY and vanZ genes, representing a potentially emerging variant of the vanA operon in Russian clinical settings. A plasmid carrying a truncated vanA operon was reconstructed using long-read sequencing. Conclusions: The study highlights the utility of genomic investigation for tracking resistance mechanisms and strain dissemination, providing crucial baseline data for epidemiological surveillance of infections caused by VREfm in Russia. These findings emphasize the need for continued genomic monitoring to understand the evolution and spread of antimicrobial resistance in clinically important enterococcal lineages. Full article

Methicillin-resistant Staphylococcus aureus (MRSA), characterized by high-level β-lactam resistance and increasing multi-drug resistance, poses a severe and growing global threat to human health and public safety. This review examines MRSA’s complex resistance mechanisms, including mecA/mecC-mediated expression of low-affinity PBP2a, regulatory roles of auxiliary genes like fem and vanA, enzymatic inactivation by β-lactamases and modifying enzymes, efflux pump activity, and biofilm formation. We also systematically review novel therapeutic strategies, such as combination therapies, phage-derived biofilm disruptors, membrane-targeting silver nanoparticles, cell-penetrating antimicrobial peptides, colonization-competitive probiotics, and antibiotic-synergizing phytochemicals. These advances provide critical insights for developing effective countermeasures against MRSA, while highlighting the urgent need for global collaboration, antibiotic stewardship, and innovative drug development to combat antimicrobial resistance. Full article

Introduction: Staphylococcus aureus is a high-priority foodborne pathogen contributing to several food poisoning outbreaks. Methicillin- and vancomycin-resistant S. aureus (MRSA and VRSA), pose significant public health concerns due to their potential for serious illness, antibiotic resistance, and transmission within both healthcare and community settings. These bacteria can cause numerous infections, ranging from skin and soft tissue infections to life-threatening conditions like bloodstream infections, pneumonia, and endocarditis. Although several publications are concerned with Staphylococcus aureus contamination in ready-to-eat (RTE) food products, little published data is available about its prevalence in pizza, which is widely distributed and consumed worldwide. Methods: The current study is intended to determine the prevalence, virulence genes, and antimicrobial resistance profiles of S. aureus in three hundred ready-to-eat pizza samples (100 each of meat, chicken, and canned tuna pizzas) collected from different restaurants in Mansoura City, Egypt. The typical colonies on Baird–Parker selective agar supplemented with egg yolk tellurite emulsion were counted and further confirmed based on Gram staining, coagulase testing, catalase testing, carbohydrate fermentation, and thermostable nuclease production. The genomic DNA of the confirmed coagulase-positive isolates was prepared and subjected to PCR analyses for detecting the nuc gene, mecA (methicillin resistance gene), and vancomycin resistance gene (vanA), as well as six selected S. aureus virulence genes: sea, seb, sec, sed, hla, and tsst. The antimicrobial resistance profile of the S. aureus isolates was determined against 16 antimicrobial agents belonging to six classes using the agar disc diffusion method according to the Clinical and Laboratory Standards Institute guidelines (CLSI), except for oxacillin and vancomycin, which were assessed using the MIC test. Results: The results revealed that 56% (56/100), 56% (56/100), and 40% (40/100) of chicken, meat, and canned tuna pizzas were positive for S. aureus, with an overall prevalence of 50.7% (152/300). All 560 isolates (100%) were verified as S. aureus based on molecular confirmation of the nuc gene. Interestingly, 48.6% (272/560) and 8.6% (48/560) of the isolates tested were identified as methicillin- and vancomycin-resistant S. aureus (MRSA and VRSA) through detection of mecA and vanA genes, respectively. Among the S. aureus isolates tested, the hla gene was detected in 87.1% (488/560), while the enterotoxin genes sea, seb, sec, and sed were identified in 50% (280/560), 78.6% (440/560), 9.8% (55/560), and 24.5% (137/560) of isolates, respectively. All recovered isolates (n = 560) were classified as multidrug-resistant and were resistant to penicillin, oxacillin, and ampicillin. Moreover, 77% (431/560), 24% (134/560), 8% (45/560), and 8.6% (48/560) of isolates were resistant to cefotaxime, ciprofloxacin, azithromycin, and vancomycin, respectively. Conclusions: The current study emphasizes that ready-to-eat pizza is highly contaminated with multidrug-resistant S. aureus, highlighting the urgent need for rationalizing antibiotic use in both veterinary and human medicine to prevent the transmission of resistant bacteria through the food chain. Additionally, strict adherence to good hygienic practices throughout all stages of the food chain is essential to minimize overall contamination and enhance food safety. Full article

Background/Objectives: Vancomycin-resistant enterococci (VRE) are significant nosocomial pathogens worldwide, potentially transmitted by food-producing animals and related products. This study investigates the epidemiological role of bovine raw milk in the transmission of VRE to humans. Methods: Bulk milk samples were screened for van gene presence using a multiplex PCR. Mastitogenic enterococci isolated from individual milk samples were tested for antimicrobial susceptibility using the broth microdilution method. Strains not susceptible to vancomycin were whole genome sequenced. Results: Overall, vanC genes were detected in 299/1026 (29.14%) bulk milk samples. Specifically, vanC1 was found in 204 samples (19.88%) and vanC2/3 in 57 samples (5.56%), with both detected simultaneously in 38 samples (3.70%). Clinically significant vanA and vanB genes were not identified. A total of 163 mastitogenic Enterococcus strains were isolated from individual milk samples. Eight different Enterococcus species were detected, with E. faecium (104/163, 63.80%) and E. faecalis (34/163, 20.86%) being the most common. Multidrug resistance was observed in 106/163 (65.03%) isolates. The most common resistance frequencies were to ciprofloxacin and erythromycin (102/163, 62.58% both), followed by quinupristin/dalfopristin (93/163, 57.06%), linezolid (65/163, 39.88%), tetracycline (58/163, 35.58%), daptomycin (46/163, 28.22%), chloramphenicol (33/163, 20.25%), ampicillin, tigecycline, and high-dosage gentamycin (8/163, 4.91% all). Resistance to teicoplanin was not observed. Two vancomycin non-susceptible strains were identified: one vanC2/3 E. casseliflavus and one vanC1 E. gallinarum. Whole genome sequencing confirmed the presence of the complete vanC gene cluster and several virulence genes in both strains. Conclusions: Our findings suggest that while raw milk is unlikely to be a source of vancomycin resistance genes of highest clinical importance (vanA or vanB), it may contribute to the spread of vanC enterococci, which are increasingly associated with human infections. Full article

Background/Objectives: Enterococci, particularly Enterococcus faecalis and Enterococcus faecium, are Gram-positive cocci that can cause severe infections in hospitalized patients. The rise of vancomycin-resistant enterococci (VRE) and vancomycin-variable enterococci (VVE) poses significant challenges in healthcare settings due to their resistance to multiple antibiotics. Methods: We conducted a point prevalence survey (PPS) to assess the prevalence of VRE and VVE colonization in hospitalized patients. Rectal swabs were collected from 160 patients and analyzed using molecular assays (MAs) and culture. Whole-genome sequencing (WGS) and core-genome multilocus sequence typing (cgMLST) were performed to identify the genetic diversity. Results: Of the 160 rectal swabs collected, 54 (33.7%) tested positive for the vanA and/or vanB genes. Culture-based methods identified 47 positive samples (29.3%); of these, 44 isolates were identified as E. faecium and 3 as E. faecalis. Based on the resistance profiles, 35 isolates (74.5%) were classified as VRE, while 12 (25.5%) were classified as VVE. WGS and cgMLST analyses identified seven clusters of E. faecium, with sequence type (ST) 80 being the most prevalent. Various resistance genes and virulence factors were identified, and this study also highlighted intra- and inter-ward transmission of VRE strains. Conclusions: Our findings underscore the potential for virulence and resistance of both the VRE and VVE strains, and they highlight the importance of effective infection control measures to prevent their spread. VVE in particular should be carefully monitored as they often escape detection. Integrating molecular data with clinical information will hopefully enhance our ability to predict and prevent future VRE infections. Full article