Search Results (778)

Background/Objectives: The study addresses the global health challenge posed by multidrug-resistant (MDR) pathogens, highlighting the urgent need for alternative antimicrobial solutions. This study investigated the in vitro antimicrobial and antibiofilm potential of endophytic mediated-synthesized silver nanoparticles (AgNPs). Methods: An endophytic bacterium was isolated from the medicinal plant Commiphora gileadensis in Saudi Arabia and identified as Bacillus subtilis CG1 through 16S rRNA gene sequencing. The bacterium was utilized for the green synthesis of AgNPs, as confirmed by Ultraviolet-visible (UV–Vis) spectroscopy. AgNPs characterization was done using Fourier-transform infrared (FTIR) spectroscopy, Transmission and scanning electron microscopy (TEM and SEM), energy-dispersive X-ray spectroscopy (EDX), and dynamic light scattering (DLS). The antimicrobial efficacy of the fabricated AgNPs was tested against eight clinically relevant pathogens using standard in vitro assays such as the agar disk diffusion method, minimum inhibitory concentration (MIC), minimum bactericidal and fungicidal concentrations (MBC and MFC). Additionally, AgNPs were tested for antibiofilm activity against P. aeruginosa and S. epidermidis. Tested pathogens included Methicillin-Resistant Staphylococcus aureus (MRSA), Staphylococcus epidermidis, Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Candida auris, Candida albicans, and Candida tropicalis. The antibiofilm efficacy was tested using the Crystal violet assay. Results: UV–Vis spectroscopy confirmed AgNP formation with a characteristic absorption peak at 412 nm. FTIR analysis identified the presence of hydroxyl, nitrile, and alkyne functional groups, which are involved in nanoparticle reduction and stabilization. TEM and SEM revealed predominantly spherical AgNPs with sizes ranging from 17 to 72 nm, while EDX confirmed silver as the major elemental component. DLS analysis showed a Z-average particle size of 113.9 ± 67.75 nm and a zeta potential of −24.2 mV. The synthesized AgNPs exhibited concentration-dependent antimicrobial activity, producing inhibition zones of 10–20 mm at 240 µg/mL. MIC values ranged from 6.25 to 25 µg/mL, whereas MBC and MFC values ranged from 6.25 to 50 µg/mL and 25 to 100 µg/mL, respectively. Moreover, bacterial growth kinetics analysis demonstrated a concentration-dependent inhibition of growth by AgNPs at MIC and sub-MIC concentrations. Additionally, AgNPs demonstrated significant antibiofilm activity against P. aeruginosa and S. epidermidis.Conclusions: Overall, B. subtilis CG1-mediated AgNPs exhibited promising physicochemical properties and antimicrobial and antibiofilm activities, suggesting their potential as alternatives for combating resistant and biofilm-associated infections. Full article

The antibiotic and drug resistance of various bacterial and fungal strains poses a significant challenge to medicine and industry. The subject of numerous studies is how to limit the spread of microorganisms and biofilm formation on various surfaces. This research focuses on the antibacterial and antifungal properties of cross-linked methacrylate-based composites for specific applications. These composites were modified using 10 wt.% of compounds with scientifically proven antimicrobial properties. These include nanosilver, copper sulphate, benzethonium chloride, and zinc oxide. The antimicrobial potential against the following bacteria and fungi was determined: Gram-positive bacteria (Staphylococcus aureus); Gram-negative bacteria (Pseudomonas aeruginosa and Escherichia coli); and the pathogenic fungi Candida albicans and Aspergillus niger. Using the modified disc-diffusion method alongside a serial dilution method demonstrated an inhibitory effect on the viability and formation of bacterial and fungal biofilms. It was demonstrated that—in liquid cultures—composites containing benzethonium chloride inhibited the growth of P. aeruginosa by over 75%, more than 50% of E. coli and more than 70% of S. aureus. Growth inhibition of C. albicans exceeded 80% for selected composites (BPA.DM + NVP + CuSO₄, BPA.DM + NVP + ZnO), while all composites inhibited the growth of A. niger by more than 45%, and in some cases (BPA.DM + HEMA + CuSO₄, BPA.DM + HEMA + Ag, BPA.DM + MMA + Ag and BPA.DM + AEH + CuSO₄) by more than 90%. Additionally, these composites significantly reduced biofilm formation on their surfaces. Modification with zinc oxide and benzethonium chloride resulted in materials that were non-toxic to normal human skin fibroblasts. To sum up the obtained results, it can be stated that these multifunctional materials with antibacterial properties could be used in medical devices, coatings, and other specialised applications where microbial contamination is a significant issue. Full article

Background: Surface roughness and microbial adhesion are key determinants of the biological performance of prosthetic base resins. This study evaluated Candida albicans (ATCC 11225) and Streptococcus mutans (DSM 20523) adhesion to a conventional heat-polymerized acrylic resin (Probase Hot) and a 3D-printed resin (V-Print dentbase) fabricated at 0° and 90° build orientations. Methods: Fifteen disc-shaped specimens (10 mm × 2 mm) were polished using a standardized protocol and surface roughness was characterized by contact profilometry (Ra). After UV sterilization, specimens were incubated for 24 h with standardized suspensions of C. albicans or S. mutans. Microbial adhesion was assessed qualitatively by scanning electron microscopy (SEM). Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc test after confirmation of normality (Shapiro–Wilk) and homogeneity of variances (Levene). Statistical significance was set at p < 0.05, and Cohen’s d was calculated to estimate effect size. Results: Surface roughness differed significantly among fabrication conditions (p = 0.017), with the conventional heat-polymerized resin showing the highest Ra values and the 3D-printed resin fabricated at 90° presenting the lowest roughness. SEM observations suggested a similar adhesion pattern for both C. albicans and S. mutans, with greater apparent microbial presence on 90°-printed specimens, followed by the conventional heat-polymerized resin, whereas 0°-printed specimens showed lower apparent adhesion in the analyzed fields. Conclusions: These findings suggest that printing orientation may influence surface-associated microbial adhesion patterns in 3D-printed prosthetic resins; however, further studies with larger sample sizes, orientation-specific surface characterization, and mono- and multispecies biofilm models are warranted to confirm these preliminary findings. Full article

Candida spp. can cause systemic infections with high mortality in immunocompromised patients. Phytotherapy may be an alternative for adjunctive treatment of fungal infections. This study evaluated the phytochemical profile, cytotoxicity, genotoxicity, and antibiofilm activity of the hydroalcoholic extract of Origanum vulgare L. against Candida albicans, Candida tropicalis, and Candida dubliniensis. The extraction and quantification (flavonoids and phenols) were performed, and its antioxidant activity (DPPH) and the presence of bio-active compounds were investigated using high-performance liquid chromatography with Diode Array Detection (HPLC-DAD). Cytotoxicity and genotoxicity tests were conducted using HaCat cell lines. Antifungal activity on planktonic cultures was evaluated using the standard (CLSI M27-S4). The analysis of the extract on biofilms was verified with different exposure times. The extract demonstrated the presence of bioactive molecules, and antioxidant activity. The cytotoxicity test showed viability >70%. Genotoxicity revealed the presence of a few micronuclei in some dilutions. The Minimum Fungicidal Concentration was obtained for C. albicans and C. tropicalis. In the biofilm analysis, there was a reduction of more than 60% in all Candida spp. species at the 24 h exposure time. The findings suggest that the O. vulgare extract exhibits activity against Candida spp. and shows biocompatibility in human keratinocytes. Full article

Candida albicans (C. albicans) is a commensal of the vaginal mucosa and the main etiological agent of acute and recurrent vulvovaginal candidiasis (VVC/RVVC). Disease severity is thought to depend on a dysregulated host inflammatory response to Candida, not necessarily associated with increased fungal burden and/or morphogenesis. The role of strain-specific differences leading to epithelial immune response or tolerance remains undefined. In this study, we compared the virulence profile of vaginal C. albicans isolates from women with acute VVC/RVVC (VVC/RVVC), asymptomatic colonizer (Colonizing), and VVC/RVVC associated with microbial co-infections (Co-infections). Isolates were evaluated for growth and biofilm formation under standard culture conditions and tested in an in vitro vaginal epithelial cell (VEC) infection model to assess fungal shedding, epithelial damage, and cytokine production. Corresponding vaginal samples were analyzed for C. albicans morphology, polymorphonuclear neutrophil presence, microbiota composition, cytokines levels, and anti-C. albicans IgA production. No significant differences in growth or biofilm formation were observed among isolates under culture conditions. However, VEC infection revealed strain-dependent differences: acute VVC/RVVC and Co-infections isolates induced greater fungal shedding, while VVC/RVVC isolates caused increased epithelial damage and showed a trend toward higher cytokine production. Vaginal samples from symptomatic groups displayed increased neutrophils, hyphal morphology, elevated IL-1α, IL-1β, and anti-Candida IgA, but not IL-1Ra, without differences in lactobacilli abundance or Community-State-Type (CST) distribution. These findings suggest that C. albicans pathogenicity in VVC depends on strain-specific interactions with VEC driving differential host responses. Full article

Background: Medical device-associated infections represent a major component of healthcare-associated infections. Biofilm formation promotes microbial persistence on device surfaces, reduces antimicrobial susceptibility, and contributes to multidrug resistance (MDR), complicating diagnosis and treatment. Materials and Method: This study investigated biofilm production and antimicrobial resistance in microorganisms recovered from 100 indwelling and implantable medical devices, including urinary and venous catheters, urethral stents, catheter tips, and orthopedic or prosthetic materials, collected at a tertiary-care hospital (AOU “San Giovanni di Dio e Ruggi d’Aragona”, Salerno, Italy). Microbiological cultures were performed using direct and enrichment methods. Microbial identification was carried out by MALDI-TOF MS, antimicrobial susceptibility testing by VITEK^® (bioMérieux, Marcy-l'Étoile, France) 2 according to EUCAST criteria, and biofilm production was assessed using the crystal violet tissue culture plate assay. MDR status was defined according to international guidelines. Results: Microbial growth was detected in the majority of analized devices, frequently with polymicrobial contamination. Within the study cohort, coagulase-negative staphylococci (CoNS) were the most frequently recovered microorganisms (20%), followed by Klebsiella pneumoniae (10%), Candida albicans (9%), Staphylococcus aureus (9%), Enterococcus faecalis (8%), and Escherichia coli (8%). A significant association was observed between multidrug resistance and biofilm production, with MDR isolates showing a markedly higher likelihood of being biofilm producers compared with non-MDR isolates (OR 9.50; 95% CI 2.72–42.96; p < 0.005). Biofilm formation also differed significantly among device types (p = 0.028). Conclusions: These findings indicate a high prevalence of biofilm-producing MDR microorganisms among isolated recovered from medical devices in our cohort and highlight a significant association between MDR phenotype and biofilm production. These results provide a microbiological characterization of device-associated isolates that may support future studies on infection dynamics and control strategies. Full article

This study describes extracellular vesicles (EVs) isolated from the culture supernatant of a Kluyveromyces marxianus strain deriving from an artisanal sourdough. Previous work had clearly shown the probiotic properties of the yeast isolate and its antagonistic activities against clinical fluconazole-resistant Candida albicans strains. Characterization of the isolated EVs by nanotracking particle analysis showed they had a mean diameter of 157.7 nm. Proteomic characterization of the purified EVs identified a complex array of 100 proteins. Both C. albicans planktonic growth and biofilm formation were inhibited by K. marxianus EVs, as well as adhesion and invasion of Candida cells in the vaginal epithelial A-431 cells. In the same cell model, K. marxianus EVs exerted an immunomodulatory effect affecting the secretion of pro-inflammatory and anti-inflammatory cytokines. Further, the expression of C. albicans SAP2 and SAP6 genes, coding for two aspartyl proteases involved in the invasion and damage of the epithelial mucosa, was affected by the presence of the yeast EVs. Overall, the results of this study show that K. marxianus EVs retain, at least in part, the beneficial features of the live microorganism, representing a postbiotic cell-free alternative preparation potentially useful for the management of C. albicans vaginal infections. Full article

Intestinal fungal overgrowth (IFO) is an increasingly recognized yet underexplored component of gut dysbiosis with potential implications for gastrointestinal and systemic disease. While bacterial microbiota have historically garnered research attention, recent advances in sequencing technologies have highlighted the importance of the gut mycobiome in maintaining intestinal homeostasis. Disruption of fungal–bacterial balance, particularly involving Candida albicans, C. tropicalis, and C. glabrata, may contribute to symptom generation through immune activation, epithelial barrier dysfunction, biofilm formation, and the production of toxic metabolites such as acetaldehyde and candidalysin. Emerging clinical evidence suggests that IFO is associated with persistent gastrointestinal symptoms, including bloating, abdominal discomfort, and altered bowel habits, particularly in patients who do not respond to conventional therapies targeting bacterial overgrowth. Furthermore, fungal dysbiosis involving Malassezia restricta and Saccharomyces cerevisiae has been associated with inflammatory bowel disease, metabolic disorders, and systemic immune dysregulation; however, the nature and directionality of these relationships remain incompletely understood. Despite increasing recognition, the diagnosis of IFO remains challenging due to a lack of standardized criteria and validated non-invasive tools. Therapeutic strategies, including antifungal agents such as fluconazole and nystatin, as well as microbiome-targeted interventions, show promise but require further validation. This review provides a comprehensive synthesis of current evidence regarding the epidemiology, pathophysiology, clinical manifestations, diagnostic challenges, and therapeutic implications of IFO, with particular emphasis on species-specific mechanisms. Recognition of the intestinal mycobiome as a potentially important component of gut health may provide new perspectives for understanding gastrointestinal disorders and inform future precision medicine approaches. Full article

Invasive candidiasis remains a major cause of morbidity and mortality worldwide, with increasing relevance of non-Candida albicans species, particularly Pichia kudriavzevii, which is associated with high mortality and intrinsic resistance to fluconazole. This study evaluated the effect of voriconazole (VRC) on P. kudriavzevii growth, biofilm formation, and metabolic activity under different nutritional conditions. Planktonic growth and biofilm development were analyzed in Sabouraud dextrose broth (SDB), RPMI-1640, and RPMI-1640 supplemented with glucose (20 g·L⁻¹). Antifungal activity was assessed by optical density (OD₅₇₀) and XTT reduction assays, and biofilm morphology was examined by light microscopy. Glucose consumption was also determined during growth. VRC showed dose-dependent inhibition in SDB, reducing growth and biofilm metabolic activity by up to 94% and 98%, respectively. In contrast, in RPMI-1640, inhibition was significantly lower (≤27% growth and ≤77% biofilm reduction). Glucose supplementation partially restored antifungal susceptibility and increased biofilm metabolic activity. Growth kinetics confirmed VRC-induced delays in proliferation and impaired glucose utilization. These results demonstrate that VRC activity against P. kudriavzevii is strongly dependent on environmental nutrient availability, particularly glucose, which modulates fungal metabolism, biofilm development, and antifungal susceptibility, highlighting the importance of standardized antifungal susceptibility testing conditions and the role of metabolic state in azole efficacy. Full article

Azole resistance in Candida albicans is an increasing clinical challenge. Upc2 is a key transcription factor regulating ergosterol biosynthesis, but its additional roles in azole tolerance remain unclear. This study investigated whether Upc2 contributes to azole resistance through pathways beyond ergosterol synthesis. Chemical sensitivity screening, RNA sequencing, flow cytometry, and molecular assays were performed to compare wild-type C. albicans and the upc2Δ/upc2Δ mutant under fluconazole (FLC) treatment. The UPC2 gene deletion affected physiological processes that are dependent on the calcineurin signaling pathway and led to an overall negative enrichment trend in the unfolded protein response (UPR) pathway gene set. Mechanistically, the UPC2 gene deletion impaired unconventional splicing of HAC1 mRNA, leading to accumulation of unfolded proteins and phenotypically its deletion enhanced sensitivity of C. albicans to FLC in planktonic growth, hyphal development, and biofilm formation. Our findings reveal that Upc2 regulates proteostasis in C. albicans, and its absence enhances FLC efficacy by disrupting the UPR pathway. Targeting Upc2-mediated UPR signaling may represent a promising strategy to combat azole resistance. Full article

Background: Fungal ocular infections, including keratitis and endophthalmitis, remain difficult to treat due to limited antifungal efficacy, poor tissue penetration, and biofilm-mediated resistance. This study evaluated the antifungal and host-protective potential of N-(4-methoxycinnamoyl)-anthranilic acid (NMCA) against Candida albicans and the multidrug-resistant Candidozyma auris. Methods: The antifungal activity of NMCA was assessed by analyzing fungal viability over time, ergosterol levels, and its interaction with fluconazole. Its antibiofilm activity was evaluated through biomass and metabolic activity measurements, together with the expression of genes involved in adhesion (ALS3, ALS5, HWP1) and membrane homeostasis (ERG11, OLE1). In addition, infected epithelial models were used to investigate epithelial damage, intracellular fungal burden, oxidative stress, and wound closure. Results: NMCA showed promising antifungal activity (MIC80 75 μg mL⁻¹ against C. albicans and 100 µg mL⁻¹ against C. auris), inducing a time-dependent reduction in fungal viability of about 4-log10 after 24 h. The compound also reduced ergosterol levels and showed synergistic interaction with fluconazole, as indicated by FICI values of 0.203 for C. albicans and 0.375 for C. auris. Moreover, NMCA markedly inhibited biofilm formation by reducing both biomass and metabolic activity up to approximately 80%, while modulating the expression of key adhesion- and membrane-related genes. Beyond its direct antifungal effects, NMCA reduced epithelial damage and intracellular fungal burden, attenuated oxidative stress, and significantly improved wound closure (reaching 76.26% and 90.46% closure in C. albicans- and C. auris-infected cells, respectively) in infected epithelial models. Conclusions: Although limited by the use of in vitro systems, these findings highlight the multifunctional profile of NMCA, which combines antifungal, antibiofilm, and tissue-protective activities. By simultaneously targeting pathogen viability, biofilm formation, and host cell integrity, NMCA appears to be a promising adjunctive candidate for the treatment of ocular fungal infections, where both pathogen eradication and tissue preservation are crucial for clinical outcomes. Full article

Background/Objectives: Wound infections represent a major clinical challenge due to their polymicrobial nature, biofilm formation, and increasing antimicrobial resistance, which compromise conventional treatments. This study aimed to develop and evaluate ligand-stabilized silver nanoparticles (AgNPs) with improved antimicrobial activity and cytocompatibility, and to investigate their incorporation into electrospun nanofibers for wound management. Methods: Four AgNP formulations stabilized with citrate, cysteine, ketorolac, and diclofenac were synthesized via chemical reduction. Physicochemical characterization included surface plasmon resonance and zeta potential measurements. Antimicrobial activity was assessed through minimum inhibitory concentration (MIC) and bactericidal assays against Gram-positive, Gram-negative, and fungal strains. Toxicity was evaluated using the HET-CAM assay, while cytocompatibility was determined in fibroblasts, MG-63 cells, and mesenchymal stem cells. Diclofenac-stabilized AgNPs were incorporated into electrospun PCL/PEO nanofibers to generate a functional nanocomposite system. Results: All AgNPs exhibited a characteristic SPR at ~400 nm and high colloidal stability. Diclofenac-stabilized AgNPs (dc-AgNPs) showed the highest antimicrobial activity, with MIC values of 18.8 mg/L against Staphylococcus aureus and Pseudomonas aeruginosa, and 4.7 mg/L against Candida albicans, along with strong bactericidal effects. HET-CAM assays indicated negligible irritation at concentrations up to 75 mg/L. Cytocompatibility results revealed a dose-dependent response, with fibroblasts being more sensitive. Electrospun nanofibers loaded with dc-AgNPs achieved a 2.6 log reduction against Streptococcus mutans and moderate reductions (0.4–0.7 log) against other pathogens. Conclusions: Ligand engineering critically influences the antimicrobial efficacy and biocompatibility of AgNPs. The incorporation of dc-AgNPs into electrospun nanofibers represents a promising approach for treating biofilm-associated wound infections. Full article

The mitochondrial membrane protein UPS1, a conserved intermembrane space protein in Saccharomyces cerevisiae, possesses phosphatidic acid transfer activity and plays a positive regulatory role in processes such as cardiolipin metabolism and transport. The role of UPS1 protein in pathogenic fungi such as Candida albicans has not been explored, especially in relation to its influence on virulence factors like hyphal growth and biofilm formation, which are crucial for the pathogenicity of C. albicans. The research investigated the function of the UPS1 protein in C. albicans by using gene knockout techniques, analyzing mitochondrial function, and conducting tests for hyphal and biofilm development. The results revealed that deletion of the UPS1 gene leads to altered mitochondrial morphology, increased reactive oxygen species levels, and reduced intracellular ATP content, thereby causing severe growth defects in C. albicans. In addition, transcriptomic analysis indicated that loss of UPS1 significantly represses the expression of genes associated with hyphal growth and biofilm formation. Functional assays further confirmed that UPS1 deficiency markedly impairs cell adhesion capability, hyphal development, and biofilm formation of C. albicans. Notably, deletion of the UPS1 protein markedly reduces the susceptibility of C. albicans to membrane-targeted antifungal drugs. Finally, infection models using Galleria mellonella larvae and a murine vulvovaginal candidiasis model verified that UPS1 gene knockout attenuates the pathogenicity of C. albicans. In summary, our findings demonstrate that UPS1 protein modulates the pathogenicity of C. albicans by regulating mitochondrial function, hyphal growth, and biofilm formation. Full article

Background/Objectives: Catheter-associated urinary tract infections (CAUTIs) are one of the most common healthcare-related morbidities. Also, severe clinical outcomes derived from CAUTIs demand an urgent need for the development of novel antimicrobial catheter materials. Since CAUTIs are primarily driven by a wide range of microorganisms causing biofilm formation on the surfaces of catheters, evaluating the effectiveness of innovative antimicrobial materials against a broad spectrum of known uropathogens is warranted. We aim (1) to demonstrate the ability of incorporated organo-selenium versus that of an FDA-cleared antimicrobial catheter and (2) to show that the results of the study are consistent against the most common microorganisms causing urinary tract infections in humans. Methods: Based on encouraging preliminary studies, three percent of organo-selenium (weight-based), as a novel antimicrobial catheter material, was incorporated into thermoplastic polyurethane (TPU). This was compared in vitro with plain polyurethane catheters and the SILQ ClearTract catheter with a zwitterionic coating (FDA-cleared for its antimicrobial properties in December 2022). The antimicrobial activity was studied against Candida albicans, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, Staphylococcus aureus, MRSA, and Staphylococcus epidermidis by assessment of colony-forming unit counts along with visual confirmation using Scanning Electron Microscopy (SEM). Results: Plain polyurethane catheters (control group) showed 7- to 8-log of in vitro growth for all tested microbes, whereas the antimicrobial zwitterionic SILQ ClearTract catheters still showed 6- to 7-log of microbial growth. In contrast, organo-selenium-incorporated catheters demonstrated no detectable in vitro growth for all tested microbes (C. albicans, E. coli, K. pneumoniae, P. mirabilis, P. aeruginosa, S. aureus, MRSA, and S. epidermidis). SEM analysis also validated the findings. Conclusions: Potentially non-leaching organo-selenium, as a novel urinary catheter material, significantly inhibited microbial attachment, growth, and biofilm formation across a wide spectrum of common uropathogenic organisms compared to a zwitterionic catheter, providing a strong foundation for further detailed in vivo and clinical studies. Full article

Thirteen new decaline polyketides, namely, zosteropenillines T–W (1–4), 8-hydroxypallidopenilline A (5), 13-epi-zosteropenilline P (6), 11-epi-zosteropenilline N (7), 15-hydroxyzosteropenilline M (8), 8-hydroxyzosteropenilline M (9), 11-epi-zosteropenilline M (10), and zosteropenillines X–Z (11–13), along with 17 known related compounds (14–30) were isolated from the ethyl acetate extract of the marine-derived fungus Penicillium yezoense KMM 4679 cultivated on MgCl₂-containing nutrient medium. The structures of the isolated compounds were established based on spectroscopic methods. The absolute configurations of zosteropenillines T (1) and V (3) were determined using time-dependent density functional theory (TD-DFT) calculations of the ECD spectra. X-ray diffraction analysis data were obtained for the known zosteropenilline S (28). A biogenetic pathway for 1–13 was proposed. The effects of the compounds on Staphylococcus aureus and Candida albicans growth and biofilm formation were observed. Zosteropenillines U (2), Y (12) and Z (13) with higher activity against C. albicans biofilms were nontoxic for normal cardiomyocyte H9c2 cells, making them promising anti-candidal agents. Moreover, zosteropenillines U and Y demonstrated cardioprotective effects in acute ischemia/reperfusion and CoCl₂-mimicking hypoxia in vitro models. Full article