Search Results (510)

Somatic cell preservation is an effective strategy for conserving the genetic potential of endangered species. To contribute to the conservation of the Milu deer (Elaphurus davidianus), this study aimed to establish and characterize an immortalized skin fibroblast cell line (ML-iSFC). The cell line is based on fibroblasts from the skin tissue of a male fawn of Milu deer. Optimal culture conditions were determined by supplementing the culture medium with different growth factors, and immortalization was achieved through simian virus 40 large T antigen (SV40T) transduction. Optimal culturing conditions for the cells were determined by adding a range of growth factors. The cellular morphology, growth characteristics, and marker expression of the cells were further evaluated. Cell cycle and proliferation were assessed by flow cytometry and CCK-8 assays, respectively. Chromosomes were determined by karyotype analysis. The highest cell growth rate was observed when the culture medium was supplemented with 3 ng/mL of FGF2. The fibroblast-specific marker vimentin (VIM) was expressed in both ML-SFC and ML-iSFC, while the epithelial marker keratin 18 (KRT18) was weakly expressed in ML-SFC cells. Cell proliferation and cell-cycle analysis revealed that ML-iSFC exhibited a higher growth rate and greater vitality compared to ML-SFC. Karyotype analysis showed that ML-iSFC maintained the same chromosome number and morphology as ML-SFC. In summary, this study reports the successful construction of an immortalized fibroblast cell line from Milu deer, which will serve as a valuable tool for Milu deer conservation. Full article

Background: Carbapenem-resistant Klebsiella pneumoniae (CRKp) is a critical global health threat due to its multidrug resistance, primarily driven by carbapenemase production. Rapid and accurate detection of carbapenemases is essential for effective treatment and infection control. This study evaluates the validity of the NG-Test CARBA 5, a rapid immunochromatographic assay, for detecting five major carbapenemases (KPC, NDM, VIM, IMP, OXA-48-like) in clinical CRKp isolates. Methods: Clinical isolates of CRKp were collected from various clinical specimens at the Military Medical Academy in Belgrade, Serbia, between January 2023 and October 2024. Detection of carbapenemases was performed using NG-Test CARBA 5, while PCR served as the reference method. Diagnostic performance was assessed by calculating sensitivity, specificity, and Cohen’s kappa coefficient. Results: Among 312 isolates, OXA-48-like was the most prevalent carbapenemase. NG-Test CARBA 5 showed high sensitivity (98.7%) and specificity (100%) overall, with excellent agreement for NDM (κ = 0.947), OXA-48-like (κ = 0.957), and KPC (κ = 0.978). However, it failed to detect VIM in five PCR-positive isolates, suggesting potential limitations. Conclusions: NG-Test CARBA 5 is a rapid and reliable tool for detecting major carbapenemases in CRKp, though its performance for VIM detection requires further investigation. This assay has the potential to improve clinical diagnostics and strengthen infection control in settings with high antimicrobial resistance. Full article

Background: Antimicrobial resistance (AMR) is a major global health threat, particularly in surgical site infections (SSIs), where multidrug-resistant (MDR) pathogens complicate treatment. Objective: This study aimed to identify antimicrobial resistance genes and assess their prevalence in bacterial species causing SSIs in Lebanon. Materials and Methods: The present research is a multicenter and prospective study that included patients who developed SSIs after surgery in seven hospitals, within the period of January 2024–September 2024. Bacterial isolates from wound swabs or tissue samples were identified using standard microbiological methods. Antimicrobial susceptibility was tested by disk diffusion, and resistance genes were detected by PCR. Data were analyzed using Statistical Package for the Social Sciences (SPSS). Results: Among 6933 surgical patients, 63 developed SSIs (0.91%; 95% CI [0.70–1.15]). Gram-negative bacteria predominated (73%), mainly Escherichia coli and Pseudomonas aeruginosa, while Gram-positive isolates accounted for 27%, mostly Staphylococcus aureus. MDR was observed in 71% of Gram-positive and 61% of Gram-negative isolates. The most frequent genes were mecA in S. aureus (100%) and coagulase-negative staphylococci (83.3%); bla_CTX-M in E. coli, Klebsiella pneumoniae, and Enterobacter cloacae (100%); and bla_NDM in E. cloacae (100%) and Acinetobacter baumannii (60%). bla_KPC was less common, and no isolates carried Imipenemase (IMP), Verona integron-encoded metallo-β-lactamase (VIM), and Oxacillinase-48-like β-lactamase (OXA-48). Conclusions: This study highlights the high prevalence of antibiotic resistance in agents causing SSIs in Lebanese hospitals. Resistance genes, particularly mecA, bla_CTX-M, and blaNDM, were highly prevalent in SSI pathogens, underscoring the urgent need for surveillance and judicious antibiotic use in Lebanese hospitals. Full article

Cross-view geo-localization is a challenging task due to the significant changes in the appearance of target scenes from variable perspectives. Most existing methods primarily adopt Transformers or ConvNeXt as backbone models but often face high computational costs and accuracy degradation in complex scenarios. Therefore, this paper proposes a visual Mamba framework based on the state-space model (SSM) for cross-view geo-localization. Compared with the existing methods, Vision Mamba is more efficient in modeling and memory usage and achieves more efficient cross-view matching by combining the twin architecture of shared weights with multiple mixed losses. Additionally, this paper introduces Dice Loss to handle scale differences and imbalance issues in cross-view images. Extensive experiments on the public cross-view dataset University-1652 demonstrate that Vision Mamba not only achieves excellent performance in UAV target localization tasks but also attains the highest efficiency with lower memory consumption. This work provides a novel solution for cross-view geo-localization tasks and shows great potential to become the backbone model for the next generation of cross-view geo-localization. Full article

Age-related macular degeneration (AMD) progresses to vision-threatening dry and wet forms, with no effective dry AMD treatments available. The sulfated polysaccharide (GNP, 25.8 kDa) derived from Gelidium crinale exhibits diverse biological activities and represents a potential source of novel therapeutic agents. This study employed a hydrogen peroxide (H₂O₂)-induced oxidative stress and epithelial–mesenchymal transition (EMT) model in retinal pigment epithelial (RPE) cells to investigate GNP’s protective mechanisms against both oxidative damage and EMT. The results demonstrated that GNP effectively suppressed oxidative stress, with the 600 μg/mL dose significantly inhibiting excessive reactive oxygen species (ROS) generation to levels comparable to untreated controls. Concurrently, at concentrations of 200–600 μg/mL, GNP inhibited NF-κB signaling and increased the Bax/Bcl-2 ratio, effectively counteracting H₂O₂-induced oxidative damage and cell apoptosis. Furthermore, in H₂O₂-treated ARPE-19 cells, 600 μg/mL GNP significantly reduced the secretion of N-cadherin (N-cad), Vimentin (Vim), and α-smooth muscle actin (α-SMA), while increasing E-cadherin (E-cad) expression, consequently inhibiting cell migration. Mechanistically, GNP activated the Nrf2/HO-1 pathway, thereby mitigating oxidative stress. These findings suggest that GNP may serve as a potential therapeutic agent for dry AMD. Full article

Background: Class B carbapenemases confer high-level resistance to carbapenems in Klebsiella pneumoniae. In Brazil, data on the national burden and geographic distribution of these genes among clinical K. pneumoniae isolates are sporadic. We performed a systematic review and meta-analysis to estimate the prevalence of MβL genes in Brazilian clinical K. pneumoniae. Methods: We searched SciELO, PubMed, ScienceDirect and LILACS for original studies published between 2006 and 2024 reporting molecular detection of MβL in clinical K. pneumoniae isolates from Brazil. Articles were independently screened, along with the extracted data and appraised study quality using Joanna Briggs Institute checklists. A random-effects meta-analysis estimated the pooled prevalence of MβL producers and assessed heterogeneity and publication bias. Results: Fifteen studies including 3.533 clinical K. pneumoniae isolates met inclusion criteria. Overall, 402 isolates (11.4%) harbored MβL genes, yielding a pooled prevalence of 44.6%. Subgroup analysis demonstrated highest prevalence in the Southeast. bla_NDM was the dominant variant (present in 14/15 studies), with bla_VIM and bla_IMP rarely detected. Meta-regression revealed an inverse association between sample size and reported prevalence, and no significant publication bias was observed. Conclusions: MβLs, particularly NDM, are widespread in Brazilian clinical K. pneumoniae but show marked regional heterogeneity driven by differences in study design, laboratory capacity, and outbreak dynamics. Urgent expansion of standardized and multicenter molecular surveillance, including allele-specific detection, and strengthened laboratory infrastructure are needed and may inform targeted infection-control and antimicrobial-stewardship interventions. Full article

Angiosarcomas are highly aggressive soft tissue tumors with poor prognosis in both humans and dogs. In dogs, visceral hemangiosarcoma offers a relevant spontaneous model for evaluating novel therapies. Surgery alone yields a median survival of 1–3 months, and treatment with doxorubicin (DOX), alone or in combination (e.g., with propranolol), modestly extends median survival time to 5–7 months, with a 1-year survival of around 10%. We developed a conjugate vaccine technology, called immune Boost (iBoost), and hypothesized that combining DOX with an iBoost vaccine targeting extracellular vimentin (eVim) could improve survival without added toxicity. Twenty-three dogs with visceral hemangiosarcoma received six cycles of DOX every two weeks post-splenectomy, alongside four doses every other week of eVim iBoost immunotherapy, followed by maintenance vaccinations every two months. Outcomes were compared to historical controls treated with DOX alone. Compared to the control group the median overall survival time increased from 136 to 235 days (NS), restricted mean survival time at one year increased with 81 days (p = 0.02) and 1-year survival rate was 44% versus 14% (p = 0.0344). The combination was well-tolerated, with no systemic vaccine-related toxicity. Adding dog eVim vaccine to DOX appears to improve survival without added toxicity in dogs with hemangiosarcoma. These results support further clinical development, including evaluation in human angiosarcoma. 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

Small and medium-sized enterprises (SMEs) are critical contributors to economic growth, innovation, and employment. However, they often struggle in securing external financing. This financial gap mainly arises from perceived risks and information asymmetries creating barriers between SMEs and potential investors. To address this issue, our study proposes a machine learning (ML) framework for predicting the investment readiness (IR) of SMEs. All the models involved in this study are trained using data provided by the European Central Bank’s Survey on Access to Finance of Enterprises (SAFE). We train, evaluate, and compare the predictive performance of nine (9) machine learning algorithms and various ensemble methods. The results provide evidence on the ability of ML algorithms in identifying investment-ready SMEs in a heavily imbalanced and noisy dataset. In particular, the Gradient Boosting algorithm achieves a balanced accuracy of 75.4% and the highest ROC AUC score at 0.815. Employing a relevant cost function economically enhances these results. The approach can offer specific inference to policymakers seeking to design targeted interventions and can provide investors with data-driven methods for identifying promising SMEs. Full article

The spread of carbapenemase-producing Gram-negative bacteria poses a significant clinical challenge due to their association with severe Difficult-to-Treat nosocomial infections, as available therapies are drastically reduced. Rapid and accurate detection of carbapenemase-producing Gram-negative bacteria is critical for effective patient management, guiding appropriate antibiotic therapy, and implementing infection control measures to limit their dissemination within healthcare settings. Lateral flow immunoassays that detect the five main carbapenemases have become cornerstones in the fight against carbapenemase-producing Gram-negative bacteria. Carbapenemases evolve in response to antibiotic exposure, and therefore regular evaluation of these lateral flow immunoassays is crucial. Here, we have evaluated a novel assay, the KINVO assay (Medomics Medical Technology) and compared it to the Gold Standard of LFIAs for carbapenemase detection, the NG-TEST CARBA 5 assay (NG-Biotech) on a large panel of carbapenemase variants. The comparison between the two assays highlighted that both share key advantages such as rapidity and simplicity. However, NG-Test CARBA 5 demonstrated superior performance overall, particularly in accurately detecting IMP-type carbapenemases and the OXA-48 variant OXA-505. In contrast, the KINVO assay was more effective at detecting a broader range of KPC variants, including some that have lost carbapenem-hydrolyzing activity but gained resistance to ceftazidime/avibactam. If we consider these variants no longer as carbapenemases, and thus that they should not be detected, the NG-Test CARBA 5 performed better for KPC carbapenemase detection. Full article

Carbapenemase-producing Klebsiella pneumoniae (CRKP) is a critical public health threat, particularly in Greece, where high prevalence limits therapeutic options. This retrospective study analyzed 26 CRKP isolates recovered at the General Hospital of Volos between July 2024 and January 2025, aiming to correlate carbapenemase phenotypes with clinical and epidemiological parameters. Demographic, clinical, and microbiological data were extracted from patient records, and isolates underwent phenotypic carbapenemase detection, antimicrobial susceptibility testing, and molecular characterization using real-time PCR; four isolates were further analyzed using whole-genome sequencing. CRKP was detected across multiple hospital departments, notably in the Emergency Department (n = 5) and Intensive Care Unit (n = 6). KPC producers predominated (n = 9), followed by NDM (n = 6), VIM (n = 1), and OXA-48 (n = 6). All VIM- or NDM + VIM-positive cases were associated with mortality. High-risk clones, including ST15, ST11, and ST307, were identified, with one ST15 isolate harboring bla_NDM-1, bla_VIM-1, and chromosomal colistin resistance; this is the first such report in Greece. Colistin and gentamicin were the most active agents in vitro; three isolates were pan-drug-resistant. The findings highlight significant CRKP circulation outside ICUs, the role of horizontal gene transfer in resistance dissemination, and the need to expand screening and rapid diagnostics to non-ICU settings. Enhanced molecular surveillance targeted at infection control and strengthened antimicrobial stewardship programs are essential for limiting the spread of CRKP. Full article

Background/Objectives: Antimicrobial resistance (AMR) poses a significant threat to public health, food security, and sustainable livestock production. Despite increasing concern, AMR remains poorly studied in cuniculture, particularly in regions where rabbit farming is predominantly small-scale and household-based. This study aimed to assess the prevalence and diversity of antimicrobial resistance genes (ARGs) in rabbit farms in northeastern Ukraine. Methods: A total of 100 fecal samples were collected from clinically healthy rabbits housed in two farms and one vivarium. DNA was extracted and analyzed using real-time PCR targeting 21 ARGs associated with resistance to major antibiotic classes, including tetracyclines, sulfonamides, β-lactams, macrolides, quinolones, carbapenems, and multidrug efflux systems. Results: A diverse and widespread resistome was identified. The most prevalent genes included sul1 (96%), blaTEM (95%), tetM (94%), and ermB (93%). AcrB (78%) and qnrS/oqxB (approximately 67%) were also frequently detected. Carbapenemase genes (blaKPC, blaNDM, blaVIM) were identified in 6% of samples, despite limited veterinary use of carbapenems. Notably, 96% of samples harbored ARGs from three or more antibiotic classes, indicating a high potential for multidrug resistance. Conclusions: The findings demonstrate a high prevalence and diversity of ARGs in rabbit farming systems in northeastern Ukraine. The presence of clinically significant resistance genes, including those conferring carbapenem resistance, underscores the urgent need for targeted AMR surveillance and improved antibiotic stewardship in cuniculture, particularly in regions with minimal regulatory oversight of antimicrobial use. Full article

Background: Infections caused by carbapenemase-producing Pseudomonas aeruginosa (CPPA) isolates have become a worldwide clinical challenge for clinicians due to the limited treatment options. This study provides epidemiological data on CPPA clinical isolates recovered from one of the largest tertiary care hospitals in Athens, Greece, serving a diverse patient population during and after the COVID-19 pandemic. Materials and Methods: The study included all consecutive single-patient CPPA clinical isolates identified from January 2020 to December 2023 in the clinical laboratory. Identification and antimicrobial susceptibility testing were performed using the VITEK-2 automated system. A lateral flow immunoassay and the FilmArray system for blood cultures only were used for the detection of the five most prevalent carbapenemases. Their epidemiological and antimicrobial susceptibility trends were retrospectively analyzed. Results: During the study period, a total of 628 single-patient CPPA were identified among 902 carbapenem-resistant P. aeruginosa clinical isolates. An increasing number of CPPA was revealed during the survey, especially in the post-COVID period (rising from n = 102 in 2020, n = 105 in 2021, and n = 123 in 2022 to n = 298 in 2023; p < 0.05). Regarding the type of carbapenemase, VIM metallo-beta-lactamase was the only carbapenemase identified during the first two years of the study (2020 to 2021). In 2022, VIM- and NDM-producing isolates split almost evenly at proportions of 57% and 43%, respectively. In 2023, NDM-producing isolates seem to have surpassed VIM producers with rates of 54.4% and 45.6%, respectively. As far as antimicrobial resistance profiles, high rates of resistance were observed for most of the antipseudomonal drugs, exceeding 90% across all study years, with little significant variation. However, aztreonam exhibited moderate activity and colistin exhibited excellent activity, remaining the most viable drugs in this setting. Conclusions: Following the COVID-19 pandemic, an increase in CPPA pathogens was identified, while an epidemiological shift was also observed, with the carbapenemase NDM dominating over VIM since 2023. Continuous surveillance is required to track resistance patterns and guide empirical therapy. In this context, new antimicrobials and antimicrobial combinations are also urgently needed. Full article

Infections caused by carbapenem-resistant Enterobacterales (CRE), particularly those harboring metallo-β-lactamases (MBLs) such as VIM, constitute a significant public health threat due to the paucity of effective therapeutic options. Cefiderocol (CFD), a novel siderophore-conjugated cephalosporin, exhibits potent activity against CRE by exploiting bacterial iron uptake systems. Nevertheless, the emergence of CFD resistance has been recently documented. This study aimed to characterize the development of CFD resistance in a VIM-producing Klebsiella pneumoniae isolate during antimicrobial treatment. Antimicrobial susceptibility was assessed by broth microdilution using iron-depleted medium according to EUCAST guidelines. Whole-genome sequencing and comparative genomic analyses focused on mutations in genes related to iron transport and CFD resistance, using Illumina MiSeq. Initial isolates (RS, BA1) were susceptible to CFD (MIC 2 mg/L), whereas the isolate recovered after 9 days of CFD therapy (BA2) was resistant (MIC 8 mg/L). In conclusion, this study illustrates for the first time the rapid emergence of CFD resistance in a VIM-producing ST307 K. pneumoniae isolate linked to a missense variant in envZ gene, arising after a 9-day CFD treatment. Full article

Background: The global impact of the SARS-CoV-2 pandemic on antimicrobial resistance (AMR) patterns has been significant. In northern Brazil, Carbapenem-resistant Klebsiella pneumoniae (CRKP) are a major concern, with an observed shift from Klebsiella pneumoniae carbapenemase (KPC) to New Delhi metallo-β-lactamase (NDM) during the pandemic. Methods: This cross-sectional study analyzed 775 carbapenem-resistant K. pneumoniae isolates collected from 25 hospitals in the Brazilian Amazon Region (states of Pará and Acre) between 2018 and 2021. The isolates were tested for the presence of carbapenemase genes (bla_KPC, bla_NDM, bla_OXA-48, bla_IMP, bla_VIM, bla_AIM, bla_DIM, bla_GIM and bla_SIM). Results: Of the isolates analyzed, n = 653/775 (84%) were carbapenemase producers, with the most prevalent being bla_KPC n = 446/775 (57.5%) and bla_NDM n = 243/775 (31.4%). A significant increase in NDM producers was observed during the pandemic, rising from n = 1/250 (8.4%) pre-pandemic to n = 222/525 (42.3%) during the pandemic, while KPC producers declined from n = 172/250 (68.8%) to n = 274/525 (52.2%) (p < 0.001). Adult intensive care units (ICUs) were the primary source of isolates n = 357/775 (46%), with a notable increase in tracheal secretion and surveillance swab samples during the pandemic. Regression analysis confirmed a strong upward trend in the prevalence of bla_NDM (R² = 0.778). Conclusions: The shift from KPC to NDM producers in northern Brazil highlights an evolving AMR landscape, partly driven by the pandemic. Strengthened infection control measures, antimicrobial stewardship and continuous surveillance are essential to mitigate the spread of NDM-producing K. pneumoniae in settings with limited resources. Full article