Staphylococcal biofilms are major causative factors of non-healing wound infections. Their treatm... more Staphylococcal biofilms are major causative factors of non-healing wound infections. Their treatment algorithms recommend the use of locally applied antiseptic agents to counteract the spread of infection. The efficacy of antiseptics against biofilm is assessed in vitro by a set of standard quantitative and semi-quantitative methods. The development of software for image processing additionally allowed for the obtainment of quantitative data from microscopic images of biofilm dyed with propidium iodine and SYTO-9 reagents, differentiating dead cells from live ones. In this work, the method of assessment of the impact of antiseptic agents on staphylococcal biofilm in vitro, based on biofilms’ processed images, was proposed and scrutinized with regard to clinically relevant antiseptics, polyhexanide, povidone–iodine and hypochlorite. The standard quantitative culturing method was applied to validate the obtained data from processed images. The results indicated significantly higher ac...
The biofilm-associated infections of bones are life-threatening diseases, requiring application o... more The biofilm-associated infections of bones are life-threatening diseases, requiring application of dedicated antibiotics in order to counteract the tissue damage and spread of microorganisms. The in vitro analyses on biofilm formation and susceptibility to antibiotics are frequently carried out using methods that do not reflect conditions at the site of infection. To evaluate the influence of nutrient accessibility on Staphylococcus aureus biofilm development in vitro, a cohesive set of analyses in three different compositional media was performed. Next, the efficacy of four antibiotics used in bone infection treatment, including gentamycin, ciprofloxacin, levofloxacin, and vancomycin, against staphylococcal biofilm, was also assessed. The results show a significant reduction in the ability of biofilm to grow in a medium containing elements occurring in the serum, which also translated into the diversified changes in the efficacy of used antibiotics, compared to the setting in which...
Urinary infections related to the presence of bacterial biofilm on catheters are responsible for ... more Urinary infections related to the presence of bacterial biofilm on catheters are responsible for loss of patients’ health and, due to their high frequency of occurrence, generate a significant economic burden for hospitals. Klebsiella pneumoniae is a pathogen frequently isolated from this type of infection. In this study, using a cohesive set of techniques performed under stationary and flow conditions, we assessed the ability of 120 K. pneumoniae strains to form biofilm on various surfaces, including catheters, and evaluated the usefulness of clinically applied and experimental compounds to remove biofilm. The results of our study indicate the high impact of intraspecies variability with respect to K. pneumoniae biofilm formation and its susceptibility to antimicrobials and revealed the crucial role of mechanical flushing out of the biofilm from the catheter’s surface with use of locally active antimicrobials. Therefore, our work, although of in vitro character, may be considered a...
Staphylococcus aureus is one of the most prevalent pathogens associated with several types of bio... more Staphylococcus aureus is one of the most prevalent pathogens associated with several types of biofilm-based infections, including infections of chronic wounds. Mature staphylococcal biofilm is extremely hard to eradicate from a wound and displays a high tendency to induce recurring infections. Therefore, in the present study, we aimed to investigate in vitro the interaction between S. aureus biofilm and fibroblast cells searching for metabolites that could be considered as potential biomarkers of critical colonization and infection. Utilizing advanced microscopy and microbiological methods to examine biofilm formation and the staphylococcal infection process, we were able to distinguish 4 phases of biofilm development. The analysis of staphylococcal biofilm influence on the viability of fibroblasts allowed us to pinpoint the moment of critical colonization—12 h post contamination. Based on the obtained model we performed a metabolomics analysis by 1H NMR spectroscopy to provide new ...
Noworodki urodzone przed ukończonym 37. tygodniem ciąży klasyfikuje się jako wcześniaki. Wcześnia... more Noworodki urodzone przed ukończonym 37. tygodniem ciąży klasyfikuje się jako wcześniaki. Wcześniactwo często związane jest z niską masą urodzeniową dzieci, tj. wagą poniżej 2500 g, i stanowi najczęstszą przyczynę śmierci niemowląt w pierwszym miesiącu życia oraz drugą najczęstszą przyczynę zgonów dzieci poniżej 5. roku życia. U około 20% noworodków i niemowląt z bardzo niską masą urodzeniową występują infekcje układowe, z czego przyczyną ponad połowy zgonów jest sepsa. Ze względu na czas wystąpienia zakażenia wyróżnia się sepsę wczesną (do 72 godzin po urodzeniu) i późną. W Polsce najczęściej izolowanymi patogenami w zakażeniach krwi u noworodków są: S. agalactiae, E. coli, K. pneumoniae, Enterobacter sp., A. baumannii, gronkowce koagulazo-ujemne i C. albicans; rzadziej – S. aureus, H. influenzae i L. monocytogenes. Skuteczna dezynfekcja skóry za pomocą miejscowo działających środków przeciwdrobnoustrojowych stanowi istotny czynnik prewencyjny w ograniczaniu występowania zakażeń szp...
The in vitro efficacy of locally applied antiseptic molecules against staphylococcal biofilm is f... more The in vitro efficacy of locally applied antiseptic molecules against staphylococcal biofilm is frequently assessed by a set of standard quantitative and semi-quantitative methods. The development of software for parametric image processing allowed to obtain parametric data also from microscopic images of biofilm dyed with a variety of dyes, especially with propidium iodine and SYTO-9, differentiating dead from live cells. In this work, using confocal/epifluorescent microscopy, we analyzed such major properties of staphylococcal biofilm in vitro as its thickness, cellular density and share of Live/Dead cells within its individual parts. We also scrutinized the impact of sample preparation and antiseptic introduction on the outcome obtained. As a result of our analyses we developed a revelatory method of assessment of the impact of antiseptic agents on staphylococcal biofilm in vitro, in which the microscopic images are processed with the use of ABE formula (Antiseptic’s Biofilm Erad...
The ongoing search for effective treatment of Acne vulgaris is concentrated, i.a., on natural pep... more The ongoing search for effective treatment of Acne vulgaris is concentrated, i.a., on natural peptides with antimicrobial properties. The aim of this work was the development of new amino acid derivatives with potential activity on dermal infections against selected microorganisms, including the facultative anaerobe C. acne. The peptides P1–P6 were synthesized via Fmoc solid phase peptide synthesis using Rink amide AM resin, analyzed by RP-HPLC-MS, FTIR, DPPH radical scavenging activity, and evaluated against C. acne and S. aureus, both deposited and non-deposited in BC. Peptides P1–P6 presented a lack of cytotoxicity, antimicrobial activity, or antioxidative properties correlated with selected structural properties. P2 and P4–P6 sorption in BC resulted in variable data, i.a., confirming the prospective topical application of these peptides in a BC carrier.
The problem of hard-to-heal wounds is associated with an increase in the incidence of civilizatio... more The problem of hard-to-heal wounds is associated with an increase in the incidence of civilization diseases. Venous leg ulcers, ischemic ulcers and wounds in diabetic foot syndrome are a particular problem, and the early detection of local wound infections is a serious interdisciplinary challenge in everyday practice. Local wound infections are often undetected or detected too late. They can impair physiological wound healing as well as lead to systemic infection and even sepsis. According to the statements that: “The fate of the wounded rests with the one who applies the first dressing” (Col. Nicholas Senna, 1897), and “The responsibility for the patient rests with the doctor who comes to the patient first” (John Murphy, Medical Society, Chicago, 1889) the success of the treatment of infected wounds depends to a particular extent on the prompt and proper response of those who come in contact with the patient first. The purpose of this document is to briefly present issues related t...
The staphylococcal biofilm-based infections of wounds still pose a significant therapeutical chal... more The staphylococcal biofilm-based infections of wounds still pose a significant therapeutical challenge. Treated improperly, they increase the risk of limb amputation or even death of the patient. The present algorithms of infected wound treatment include, among others, the application of antiseptic substances. In vitro wound biofilm models are applied in order to scrutinize their activity. In the present work, using a spectrum of techniques, we showed how the change of a single variable (medium composition) in the standard in vitro model translates not only to shift in staphylococcal biofilm features but also to the change of efficacy of clinically applied wound antimicrobials such as octenidine, polyhexamethylene biguanide, chlorhexidine, hypochlorite solutions, and locally applied gentamycin. The data presented in this study may be of a pivotal nature, taking into consideration the fact that results of in vitro analyses are frequently used to propagate application of specific anti...
The intestinal barrier plays an extremely important role in maintaining the immune homeostasis of... more The intestinal barrier plays an extremely important role in maintaining the immune homeostasis of the gut and the entire body. It is made up of an intricate system of cells, mucus and intestinal microbiota. A complex system of proteins allows the selective permeability of elements that are safe and necessary for the proper nutrition of the body. Disturbances in the tightness of this barrier result in the penetration of toxins and other harmful antigens into the system. Such events lead to various digestive tract dysfunctions, systemic infections, food intolerances and autoimmune diseases. Pathogenic and probiotic bacteria, and the compounds they secrete, undoubtedly affect the properties of the intestinal barrier. The discovery of zonulin, a protein with tight junction regulatory activity in the epithelia, sheds new light on the understanding of the role of the gut barrier in promoting health, as well as the formation of diseases. Coincidentally, there is an increasing number of rep...
The aim of the article is to analyze the level of viral infections transmitted through sexual con... more The aim of the article is to analyze the level of viral infections transmitted through sexual contact and blood-borne infections in Polish penal institutions and detention centers. The description of the issue presented in the study is primarily aimed at signaling the issue and at trying to indicate possible preventive solutions. It seems inevitable to confront the actual living conditions and situations in the prison environment with the scale of the problem of HIV and HBV/HCV infections. Therefore, the authors consider it necessary to monitor the situation and familiarize the interested specialists in the field of probation with the problem in order to encourage reflection on the possible introduction of preventive measures.
Herein, we present a new test, dubbed AntiBioVol, to be used for the quantitative evaluation of a... more Herein, we present a new test, dubbed AntiBioVol, to be used for the quantitative evaluation of antibiofilm activity of volatile compounds in vitro. AntiBioVol is performed in two 24-well plates using a basic microbiological laboratory equipment. To demonstrate AntiBioVol usability, we have scrutinized the activity of volatilized eucalyptus, tea tree, thyme essential oils, and ethanol (used for method suitability testing) against biofilms of Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans. We have also compared AntiBioVol with the standard disc volatilization method, placing a special stress on evaluating the impact of various technical parameters on the outcomes of the latter method. The obtained results indicate that AntiBioVol allows analyzing the antibiofilm activity of volatile compounds in a high number of repeats and provides semi-quantitative or quantitative results of high repeatability. In comparison to disc volatilization, AntiBioVol is a more space- a...
Introduction: Over the past few decades, Klebsiella pneumoniae strains increased their pathogenic... more Introduction: Over the past few decades, Klebsiella pneumoniae strains increased their pathogenicity and antibiotic resistance, thereby becoming a major therapeutic challenge. One of the few available therapeutic options seems to be intravenous fosfomycin. Unfortunately, the determination of sensitivity to fosfomycin performed in hospital laboratories can pose a significant problem. Therefore, the aim of the present research was to evaluate the activity of fosfomycin against clinical, multidrug-resistant Klebsiella pneumoniae strains isolated from nosocomial infections between 2011 and 2020, as well as to evaluate the methods routinely used in hospital laboratories to assess bacterial susceptibility to this antibiotic. Materials and Methods: 43 multidrug-resistant Klebsiella strains isolates from various infections were tested. All the strains had ESBL enzymes, and 20 also showed the presence of carbapenemases. Susceptibility was determined using the diffusion method (E-test) and th...
Staphylococcal biofilms are major causative factors of non-healing wound infections. Their treatm... more Staphylococcal biofilms are major causative factors of non-healing wound infections. Their treatment algorithms recommend the use of locally applied antiseptic agents to counteract the spread of infection. The efficacy of antiseptics against biofilm is assessed in vitro by a set of standard quantitative and semi-quantitative methods. The development of software for image processing additionally allowed for the obtainment of quantitative data from microscopic images of biofilm dyed with propidium iodine and SYTO-9 reagents, differentiating dead cells from live ones. In this work, the method of assessment of the impact of antiseptic agents on staphylococcal biofilm in vitro, based on biofilms’ processed images, was proposed and scrutinized with regard to clinically relevant antiseptics, polyhexanide, povidone–iodine and hypochlorite. The standard quantitative culturing method was applied to validate the obtained data from processed images. The results indicated significantly higher ac...
The biofilm-associated infections of bones are life-threatening diseases, requiring application o... more The biofilm-associated infections of bones are life-threatening diseases, requiring application of dedicated antibiotics in order to counteract the tissue damage and spread of microorganisms. The in vitro analyses on biofilm formation and susceptibility to antibiotics are frequently carried out using methods that do not reflect conditions at the site of infection. To evaluate the influence of nutrient accessibility on Staphylococcus aureus biofilm development in vitro, a cohesive set of analyses in three different compositional media was performed. Next, the efficacy of four antibiotics used in bone infection treatment, including gentamycin, ciprofloxacin, levofloxacin, and vancomycin, against staphylococcal biofilm, was also assessed. The results show a significant reduction in the ability of biofilm to grow in a medium containing elements occurring in the serum, which also translated into the diversified changes in the efficacy of used antibiotics, compared to the setting in which...
Urinary infections related to the presence of bacterial biofilm on catheters are responsible for ... more Urinary infections related to the presence of bacterial biofilm on catheters are responsible for loss of patients’ health and, due to their high frequency of occurrence, generate a significant economic burden for hospitals. Klebsiella pneumoniae is a pathogen frequently isolated from this type of infection. In this study, using a cohesive set of techniques performed under stationary and flow conditions, we assessed the ability of 120 K. pneumoniae strains to form biofilm on various surfaces, including catheters, and evaluated the usefulness of clinically applied and experimental compounds to remove biofilm. The results of our study indicate the high impact of intraspecies variability with respect to K. pneumoniae biofilm formation and its susceptibility to antimicrobials and revealed the crucial role of mechanical flushing out of the biofilm from the catheter’s surface with use of locally active antimicrobials. Therefore, our work, although of in vitro character, may be considered a...
Staphylococcus aureus is one of the most prevalent pathogens associated with several types of bio... more Staphylococcus aureus is one of the most prevalent pathogens associated with several types of biofilm-based infections, including infections of chronic wounds. Mature staphylococcal biofilm is extremely hard to eradicate from a wound and displays a high tendency to induce recurring infections. Therefore, in the present study, we aimed to investigate in vitro the interaction between S. aureus biofilm and fibroblast cells searching for metabolites that could be considered as potential biomarkers of critical colonization and infection. Utilizing advanced microscopy and microbiological methods to examine biofilm formation and the staphylococcal infection process, we were able to distinguish 4 phases of biofilm development. The analysis of staphylococcal biofilm influence on the viability of fibroblasts allowed us to pinpoint the moment of critical colonization—12 h post contamination. Based on the obtained model we performed a metabolomics analysis by 1H NMR spectroscopy to provide new ...
Noworodki urodzone przed ukończonym 37. tygodniem ciąży klasyfikuje się jako wcześniaki. Wcześnia... more Noworodki urodzone przed ukończonym 37. tygodniem ciąży klasyfikuje się jako wcześniaki. Wcześniactwo często związane jest z niską masą urodzeniową dzieci, tj. wagą poniżej 2500 g, i stanowi najczęstszą przyczynę śmierci niemowląt w pierwszym miesiącu życia oraz drugą najczęstszą przyczynę zgonów dzieci poniżej 5. roku życia. U około 20% noworodków i niemowląt z bardzo niską masą urodzeniową występują infekcje układowe, z czego przyczyną ponad połowy zgonów jest sepsa. Ze względu na czas wystąpienia zakażenia wyróżnia się sepsę wczesną (do 72 godzin po urodzeniu) i późną. W Polsce najczęściej izolowanymi patogenami w zakażeniach krwi u noworodków są: S. agalactiae, E. coli, K. pneumoniae, Enterobacter sp., A. baumannii, gronkowce koagulazo-ujemne i C. albicans; rzadziej – S. aureus, H. influenzae i L. monocytogenes. Skuteczna dezynfekcja skóry za pomocą miejscowo działających środków przeciwdrobnoustrojowych stanowi istotny czynnik prewencyjny w ograniczaniu występowania zakażeń szp...
The in vitro efficacy of locally applied antiseptic molecules against staphylococcal biofilm is f... more The in vitro efficacy of locally applied antiseptic molecules against staphylococcal biofilm is frequently assessed by a set of standard quantitative and semi-quantitative methods. The development of software for parametric image processing allowed to obtain parametric data also from microscopic images of biofilm dyed with a variety of dyes, especially with propidium iodine and SYTO-9, differentiating dead from live cells. In this work, using confocal/epifluorescent microscopy, we analyzed such major properties of staphylococcal biofilm in vitro as its thickness, cellular density and share of Live/Dead cells within its individual parts. We also scrutinized the impact of sample preparation and antiseptic introduction on the outcome obtained. As a result of our analyses we developed a revelatory method of assessment of the impact of antiseptic agents on staphylococcal biofilm in vitro, in which the microscopic images are processed with the use of ABE formula (Antiseptic’s Biofilm Erad...
The ongoing search for effective treatment of Acne vulgaris is concentrated, i.a., on natural pep... more The ongoing search for effective treatment of Acne vulgaris is concentrated, i.a., on natural peptides with antimicrobial properties. The aim of this work was the development of new amino acid derivatives with potential activity on dermal infections against selected microorganisms, including the facultative anaerobe C. acne. The peptides P1–P6 were synthesized via Fmoc solid phase peptide synthesis using Rink amide AM resin, analyzed by RP-HPLC-MS, FTIR, DPPH radical scavenging activity, and evaluated against C. acne and S. aureus, both deposited and non-deposited in BC. Peptides P1–P6 presented a lack of cytotoxicity, antimicrobial activity, or antioxidative properties correlated with selected structural properties. P2 and P4–P6 sorption in BC resulted in variable data, i.a., confirming the prospective topical application of these peptides in a BC carrier.
The problem of hard-to-heal wounds is associated with an increase in the incidence of civilizatio... more The problem of hard-to-heal wounds is associated with an increase in the incidence of civilization diseases. Venous leg ulcers, ischemic ulcers and wounds in diabetic foot syndrome are a particular problem, and the early detection of local wound infections is a serious interdisciplinary challenge in everyday practice. Local wound infections are often undetected or detected too late. They can impair physiological wound healing as well as lead to systemic infection and even sepsis. According to the statements that: “The fate of the wounded rests with the one who applies the first dressing” (Col. Nicholas Senna, 1897), and “The responsibility for the patient rests with the doctor who comes to the patient first” (John Murphy, Medical Society, Chicago, 1889) the success of the treatment of infected wounds depends to a particular extent on the prompt and proper response of those who come in contact with the patient first. The purpose of this document is to briefly present issues related t...
The staphylococcal biofilm-based infections of wounds still pose a significant therapeutical chal... more The staphylococcal biofilm-based infections of wounds still pose a significant therapeutical challenge. Treated improperly, they increase the risk of limb amputation or even death of the patient. The present algorithms of infected wound treatment include, among others, the application of antiseptic substances. In vitro wound biofilm models are applied in order to scrutinize their activity. In the present work, using a spectrum of techniques, we showed how the change of a single variable (medium composition) in the standard in vitro model translates not only to shift in staphylococcal biofilm features but also to the change of efficacy of clinically applied wound antimicrobials such as octenidine, polyhexamethylene biguanide, chlorhexidine, hypochlorite solutions, and locally applied gentamycin. The data presented in this study may be of a pivotal nature, taking into consideration the fact that results of in vitro analyses are frequently used to propagate application of specific anti...
The intestinal barrier plays an extremely important role in maintaining the immune homeostasis of... more The intestinal barrier plays an extremely important role in maintaining the immune homeostasis of the gut and the entire body. It is made up of an intricate system of cells, mucus and intestinal microbiota. A complex system of proteins allows the selective permeability of elements that are safe and necessary for the proper nutrition of the body. Disturbances in the tightness of this barrier result in the penetration of toxins and other harmful antigens into the system. Such events lead to various digestive tract dysfunctions, systemic infections, food intolerances and autoimmune diseases. Pathogenic and probiotic bacteria, and the compounds they secrete, undoubtedly affect the properties of the intestinal barrier. The discovery of zonulin, a protein with tight junction regulatory activity in the epithelia, sheds new light on the understanding of the role of the gut barrier in promoting health, as well as the formation of diseases. Coincidentally, there is an increasing number of rep...
The aim of the article is to analyze the level of viral infections transmitted through sexual con... more The aim of the article is to analyze the level of viral infections transmitted through sexual contact and blood-borne infections in Polish penal institutions and detention centers. The description of the issue presented in the study is primarily aimed at signaling the issue and at trying to indicate possible preventive solutions. It seems inevitable to confront the actual living conditions and situations in the prison environment with the scale of the problem of HIV and HBV/HCV infections. Therefore, the authors consider it necessary to monitor the situation and familiarize the interested specialists in the field of probation with the problem in order to encourage reflection on the possible introduction of preventive measures.
Herein, we present a new test, dubbed AntiBioVol, to be used for the quantitative evaluation of a... more Herein, we present a new test, dubbed AntiBioVol, to be used for the quantitative evaluation of antibiofilm activity of volatile compounds in vitro. AntiBioVol is performed in two 24-well plates using a basic microbiological laboratory equipment. To demonstrate AntiBioVol usability, we have scrutinized the activity of volatilized eucalyptus, tea tree, thyme essential oils, and ethanol (used for method suitability testing) against biofilms of Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans. We have also compared AntiBioVol with the standard disc volatilization method, placing a special stress on evaluating the impact of various technical parameters on the outcomes of the latter method. The obtained results indicate that AntiBioVol allows analyzing the antibiofilm activity of volatile compounds in a high number of repeats and provides semi-quantitative or quantitative results of high repeatability. In comparison to disc volatilization, AntiBioVol is a more space- a...
Introduction: Over the past few decades, Klebsiella pneumoniae strains increased their pathogenic... more Introduction: Over the past few decades, Klebsiella pneumoniae strains increased their pathogenicity and antibiotic resistance, thereby becoming a major therapeutic challenge. One of the few available therapeutic options seems to be intravenous fosfomycin. Unfortunately, the determination of sensitivity to fosfomycin performed in hospital laboratories can pose a significant problem. Therefore, the aim of the present research was to evaluate the activity of fosfomycin against clinical, multidrug-resistant Klebsiella pneumoniae strains isolated from nosocomial infections between 2011 and 2020, as well as to evaluate the methods routinely used in hospital laboratories to assess bacterial susceptibility to this antibiotic. Materials and Methods: 43 multidrug-resistant Klebsiella strains isolates from various infections were tested. All the strains had ESBL enzymes, and 20 also showed the presence of carbapenemases. Susceptibility was determined using the diffusion method (E-test) and th...
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