Antioxidant and antimicrobial properties of an extract rich in proteins obtained from Trametes versicolor.

Introduction: Bioactive proteins and peptides generated from fruit, vegetables, meat or fish have great potential as functional food or substitutes for antibiotics. In recent years it has also been demonstrated that the fungus kingdom could be a source of these compounds. The study investigated the bioactivity of an extract of the lignicolous fungus Trametes versicolor and its hydrolysate. Material and Methods: The fungus was collected in a mixed forest in October, extracted and hydrolysed. To inspect the protein and peptide profiles before and after hydrolysis, matrix-assisted laser desorption/ionisation-time-of-flight mass spectrometric analysis was performed. To evaluate the antioxidant properties of the preparations, 2,2-diphenyl-1-picrylhydrazyl (DPPH•) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS•+) radical scavenging assays were used. The activity of the fungus extract and hydrolysate against Aeromonas veronii, Bacillus cereus, Enterococcus faecalis, Enterococcus faecium, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella Typhimurium, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus agalactiae, Streptococcus dysgalactiae, and Streptococcus uberis was determined by the minimum inhibitory concentration and minimum bactericidal concentration values. Results: The extract and its hydrolysate showed almost 100% ABTS•+ and DPPH• radical scavenging with a low half maximal inhibitory concentration. The water extract and hydrolysate of T. versicolor exhibited antimicrobial activity against two S. aureus strains, E. coli, P. aeruginosa and Salmonella Typhimurium. Conclusion: These results provide compelling evidence that the analysed fungus extract and its hydrolysate hold promise with their antibacterial and antioxidant properties.

Egg Yolk as a New Source of Peptides with Antioxidant and Antimicrobial Properties.

A significant increase in interest in food-derived peptides obtained mostly through enzymatic reactions has been observed in the past few years. One of the best sources of bioactive peptides are defatted egg yolk proteins, which can potentially find application as high-quality nutritional supplements for infants with cow's milk protein intolerance and as natural preservatives. The aim of this study was to obtain peptides from defatted egg yolk protein, to study their antioxidant and antimicrobial properties, and to identify peptides with bioactive properties To control the course of the process, MALDI-TOF/MS (matrix-assisted laser desorption/ionization time-of flight/mass spectrometry) spectra were also examined. The peptide mixture obtained through enzyme digestion was tested for its antioxidant properties by measuring the scavenging activity in 2,2-diphenyl-1-picrylhydrazyl radical (DPPH•), 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical cation decolorization (ABTS•+), and ferric reducing activity (FRAP) assays. Antimicrobial activity was also studied. The peptide mixture exhibited significant antioxidant activity: DPPH-1776.66 ± 32.99, ABTS-390.43 ± 8.92, and FRAP-16.45 ± 0.19. The inhibition of bacterial growth by two concentrations of the peptide mixture was examined. The best result was obtained in Bacillus cereus, with an inhibition zone of 20.0 ± 1.0 and 10.7 ± 0.6 mm at the concentrations of 50 and 25 mg/mL, respectively. The results of the study suggest that the mixture of egg yolk peptides may exhibit a number of health-promoting properties.

Indole-Acrylonitrile Derivatives as Potential Antitumor and Antimicrobial Agents-Synthesis, In Vitro and In Silico Studies.

A series of 2-(1H-indol-2-yl)-3-acrylonitrile derivatives, 2a-x, 3, 4a-b, 5a-d, 6a-b, and 7, were synthesized as potential antitumor and antimicrobial agents. The structures of the prepared compounds were evaluated based on elemental analysis, IR, 1H- and 13NMR, as well as MS spectra. X-ray crystal analysis of the representative 2-(1H-indol-2-yl)-3-acrylonitrile 2l showed that the acrylonitrile double bond was Z-configured. All compounds were screened at the National Cancer Institute (USA) for their activities against a panel of approximately 60 human tumor cell lines and the relationship between structure and in vitro antitumor activity is discussed. Compounds of interest 2l and 5a-d showed significant growth inhibition potency against various tumor cell lines with the mean midpoint GI50 values of all tests in the range of 0.38-7.91 µM. The prominent compound with remarkable activity (GI50 = 0.0244-5.06 µM) and high potency (TGI = 0.0866-0.938 µM) against some cell lines of leukemia (HL-60(TB)), non-small cell lung cancer (NCI-H522), colon cancer (COLO 205), CNS cancer (SF-539, SNB-75), ovarian cancer ((OVCAR-3), renal cancer (A498, RXF 393), and breast cancer (MDA-MB-468) was 3-[4-(dimethylamino)phenyl]-2-(1-methyl-1H-indol-2-yl)acrylonitrile (5c). Moreover, the selected 2-(1H-indol-2-yl)-3-acrylonitriles 2a-c and 2e-x were evaluated for their antibacterial and antifungal activities against Gram-positive and Gram-negative pathogens as well as Candida albicans. Among them, 2-(1H-indol-2-yl)-3-(1H-pyrrol-2-yl)acrylonitrile (2x) showed the most potent antimicrobial activity and therefore it can be considered as a lead structure for further development of antimicrobial agents. Finally, molecular docking studies as well as drug-likeness and ADME profile prediction were carried out.

Anticancer activity of lactic acid bacteria.

Lactic acid bacteria (LAB), a group of Gram-positive microorganisms naturally occurring in fermented food products and used as probiotics, have been gaining the interest of researchers for years. LAB are potent, albeit still not wholly understood, source of bioactive compounds with various functions and activity. Metabolites of LAB, among others, short-chain fatty acids, exopolysaccharides and bacteriocins have promising anticancer potential. Research on the interactions between the bioactive metabolites of LAB and immune mechanisms demonstrated that these substances could exert a strong immunomodulatory effect, which would explain their vast therapeutic potential. The anticancer activity of LAB was confirmed both in vitro and in animal models against cancer cells from various malignancies. LAB inhibit tumor growth through various mechanisms, including antiproliferative activity, induction of apoptosis, cell cycle arrest, as well as through antimutagenic, antiangiogenic and anti-inflammatory effects. The aim of this review was to summarize the most recent data about the anticancer activity of LAB, with particular emphasis on the most promising bioactive compounds with potential clinical application.

Probiotics: Should All Patients Take Them?

The usefulness of probiotics in the treatment as well as prevention of many infections and disorders has been confirmed by previous clinical studies. They can protect not only against gastrointestinal diseases such as diarrhea or enteritis but they have proven efficacy against pneumonia, urogenital infection, depression/anxiety, cancer metastasis, obesity, and others. However, it should be mentioned that not all clinical trials have shown improvement of health in patients undergoing probiotic treatment, and very rarely have even reported that probiotic strains may be the causative agents of opportunistic infections. Studies have documented cases of sepsis/bacteremia, endocarditis, liver abscess, pneumonia, and fungemia caused by probiotic strains, mainly in high-risk groups. This review summarizes the cases of infections caused by probiotic strains and the potential hazard associated with the supplementation of probiotics in seriously ill and hospitalized patients.

Presence of egc-positive major clones ST 45, 30 and 22 among methicillin-resistant and methicillin-susceptible oral Staphylococcus aureus strains.

The oral cavity may comprise a significant reservoir for Staphylococcus aureus but the data on molecular epidemiology and clonal distribution of oral strains are really scarce. This study aimed to evaluate the clonal relatedness in S. aureus isolated from oral cavity and their relationship with carriage of virulence genes, and antimicrobial resistance profiles. A total of 139 oral S. aureus isolates were obtained from 2327 analysed oral samples of dental patients. Antimicrobial susceptibility testing was performed. Isolates were characterized using protein A gene (spa) typing, spa-CC clonal complexes, toxin genes and SCCmec typing for MRSA. High resistance rates for penicillin, tetracycline and gentamicin were detected, respectively 58.3%, 42.4%, and 35.2%. Twelve (8.6%) S. aureus isolates were identified as MRSA. All of MRSA isolates were mecA-positive and mecC-negative. SCCmec IV was the most common type (66.7%), which was typical for community-acquired MRSA (CA-MRSA). Overall, the enterotoxin gene cluster (egc) was the most frequent detected virulence factor (44.9%), both in MSSA and MRSA isolates. Presence of genes encoding for the enterotoxins (sea, seb, sec, seh, sek), exfoliative toxin A (eta), and toxic shock syndrome toxin-1 (tst) was also observed. Strains carrying lukS-PV/lukF-PV genes belonged to SCCmecV- spa type t437. The most prevalent spa types were t091, t015, t084, t002, t571, and t026 among all 57 identified. Spa types, including 3 new ones, grouped in 6 different spa-CC clonal complexes, with four major dominated; CC45, CC30, CC5, and CC15. This study demonstrated that both methicillin-susceptible and methicillin-resistant major European clones of S. aureus could be isolated from the oral cavity of dental patients, with the emergence of PVL-positive CA-MRSA strains. The oral cavity should be considered as a possible source of toxigenic egc-positive S. aureus strains, in terms of potential risk of cross-infection and dissemination to other body sites.

Genetic diversity and antimicrobial resistance of Staphylococcus aureus from recurrent tonsillitis in children.

The aim of this study was to analyze the prevalence of Staphylococcus aureus in the tonsils of children subjected tonsillectomy due to recurrent tonsilitis and to determine the spa types of the pathogens, carriage of virulence genes and antimicrobial resistance profiles. The study included 73 tonsillectomized children. Bacteria, including S. aureus were isolated from tonsillar surface prior to tonsillectomy, recovered from tonsillar core at the time of the surgery, and from posterior pharynx 2-4 weeks after the procedure. Staphylococcus aureus isolates were compared by spa typing, tested for antimicrobial susceptibility and for the presence of superantigenic toxin genes (sea-seu, eta, etb, tst, lukS/lukF-PV) by multiplex polymerase chain reaction. Seventy-three patients (mean 7.1 ± 4.1 years, 61.6% male) were assessed. The most commonly isolated bacteria were S. aureus. The largest proportion of staphylococcal isolates originated from tonsillar core (63%), followed by tonsillar surface (45.1%) and posterior pharynx in tonsillectomized children (18.2%, p = 0.007). Five (6.3%) isolates were identified as MRSA (mecA-positive). Up to 67.5% of the isolates synthesized penicillinases (blaZ-positive isolates), and 8.8% displayed MLSB resistance. The superantigenic toxin genes were detected in more than half of examined isolates (56.3%). spa types t091, t084, and t002, and clonal complexes (CCs) CC7, CC45, and CC30 turned out to be most common. Staphylococcus aureus associated with RT in children showed pathogenicity potential and considerable genetic diversity, and no clones were found to be specific for this condition although further studies are needed.

Denture stomatitis associated with small-colony variants of Staphylococcus aureus: a case report.

BACKGROUND: The small-colony variants (SCVs) of Staphylococcus aureus were isolated from persistent and recurrent infections, especially after placement of medical devices having direct contact with human tissues. The emergence of SCVs is a survival strategy of S. aureus which enables them to hide inside host's cells and induces a less severe immune response than to wild-type S. aureus. However, contrary to other medical devices, dental prosthesis as a surface potentially colonized by SCVs of S. aureus has not been examined thus far. We reported the first case of SCVs - S. aureus infection in denture wearer. CASE PRESENTATION: A 62-year-old woman with a complete removable acrylic denture presented extensive elevated erythematous lesions on the palate, compatible with denture stomatitis. The patient had a history of arterial hypertension, cigarette smoking and wearing denture at night. The fungal colonies, identified as Candida albicans, were cultured on Sabouraud agar. From three swabs (from hard palate mucosa, denture surface and angular cheilitis lesions) were cultured of pinpoint, clear, non-pigmented, and non-haemolytic colonies on Columbia agar. The small colonies turned out to be Gram-positive cocci, catalase-, Pastorex Staph Plus -, and clumping factor-positive, and oxidase-negative. Suspected phenotypically SCVs forms were definitively identified as S. aureus based on PCR amplification of species specific nuc and coa genes. Methicillin-resistance was verified by mecA gene detection. The isolates turned out to be susceptible to methicillin (MSSA) and resistant to gentamicin. The isolate was identified as menadione-auxotrophic variant. CONCLUSIONS: This case demonstrated that oral cavity in denture wearers may be a reservoir of small-colony variants of S. aureus, besides C. albicans. The prevalence of these bacteria and their role in the pathogenesis of oral diseases are not understood. Due to problems with their detection and identification, the true prevalence of oral SCVs may be underestimated.

N-(2-Arylmethylthio-4-Chloro-5-Methylbenzenesulfonyl)amide Derivatives as Potential Antimicrobial Agents-Synthesis and Biological Studies.

Rising resistance of pathogenic bacteria reduces the options of treating hospital and non-hospital bacterial infections. There is a need to search for newer chemotherapies that will show antimicrobial ability against planktonic cells as well as bacterial biofilms. We have synthesized a series of N-(2-arylmethylthio-4-chloro-5-methylbenzenesulfonyl)amides, namely, molecular hybrids, which include a 2-mercaptobenzenosulfonamide fragment and either cinnamic or cyclohexylpropionic acid residues. The antimicrobial activity of compounds 8‒17 was evaluated on Gram-positive, Gram-negative bacteria and fungal species. Experiments took into account investigation of antibacterial activity against planktonic cells as well as biofilms. Compounds 8‒17 showed high bacteriostatic activity against staphylococci, with the most active molecules 10 and 16 presenting low MIC values of 4-8 µg/mL against reference methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive S. aureus (MSSA) strains as well as clinical isolates. Compounds 10 and 16 also showed an ability to inhibit biofilms formed by MRSA and MSSA. The potential of 10 and 16 as antibiofilm agents was supported by cytotoxicity assays that indicated no cytotoxic effect either on normal cells of human keratinocytes or on human cancer cells, including cervical, colon, and breast cancer lines.

Emergence and spread of worldwide Staphylococcus aureus clones among cystic fibrosis patients.

BACKGROUND: The aim of this study was to assess the relatedness of molecular types of Staphylococcus aureus isolates colonizing cystic fibrosis (CF) patients with their antimicrobial resistance and prevalence of toxin genes. METHODS: A total of 215 isolates from the airways of 107 patients with CF were tested for spa and SCCmec type, antimicrobial resistance and carriage of toxin genes. RESULTS: t015, t084, t091, t700 and t002 were the largest group (approximately 25%) among all 69 identified spa types. Five new spa types, t14286, t14287, t14288, t14289 and t14290, were identified and registered. Isolates from CF patients were clustered into 11 multi-locus sequence typing clonal complexes, with CC30, CC22, CC97, CC45, CC15 and CC5 being the most frequent ones. Twelve (5.6%) methicillin-resistant S. aureus (MRSA) isolates and 102 (47.7%) multidrug-resistant isolates were identified, along with three SCCmec types (I, III and V). All isolates (both MRSA and methicillin-sensitive S. aureus) were Panton-Valentine leucocidin-negative, and 56.7% harbored egc genes. This was the first study documenting the presence of ST398-V-t571 livestock-associated MRSA in a European patient with CF. CONCLUSION: These findings imply that individuals with CF can also be colonized with animal-related ST398 MRSA, and justify constant monitoring of staphylococcal colonization and identification of epidemic S. aureus clones in this group.

Staphylococcus aureus isolated from tonsillectomized adult patients with recurrent tonsillitis.

The aim of this study was to analyze the prevalence and antibiotic resistance of Staphylococcus aureus strains from 118 tonsillectomized adults due to recurrent tonsillitis (RT). The study included strains isolated from the tonsillar surface prior to tonsillectomy, recovered from the tonsillar core at the time of surgery, and from the posterior throat 2-4 weeks after the procedure. Susceptibility of isolates to 19 antibiotics was tested in line with the Clinical and Laboratory Standards Institute recommendations. Irrespective of the stage, the most commonly isolated bacteria were gram-positive cocci, and among them S. aureus. The tonsillar core was the most common site of S. aureus isolation (30.5%), followed by the tonsillar surface (10.8%) and the posterior pharynx (5.9%). This difference turned out to be statistically significant (p < 0.001). Beta-hemolytic streptococci, most often Streptococcus pyogenes (5.1%), were isolated from 2.5% to 10.2% of patients. Staphylococcal isolates were susceptible to most tested antibiotics (except from penicillin and ampicillin) and rarely showed methicillin resistance (n = 1). Staphylococcus aureus seems to be the most common pathogen isolated from patients tonsillectomized due to RT. Staphylococcal isolates associated with RT are present mostly within the tonsillar core and susceptible to most antibiotics. They are typically isolated from patients between 21 and 30 years of age. Tonsillectomy results in less frequent isolation of S. aureus strains.