Justyna Glik

In-vivo expression profiling of Pseudomonas aeruginosa infections reveals niche-specific and strain-independent transcriptional programs.

Pseudomonas aeruginosa is a threatening, opportunistic pathogen causing disease in immunocompromised individuals. The hallmark of P. aeruginosa virulence is its multi-factorial and combinatorial nature. It renders such bacteria infectious for many organisms and it is often resistant to antibiotics. To gain insights into the physiology of P. aeruginosa during infection, we assessed the transcriptional programs of three different P. aeruginosa strains directly after isolation from burn wounds of humans. We compared the programs to those of the same strains using two infection models: a plant model, which consisted of the infection of the midrib of lettuce leaves, and a murine tumor model, which was obtained by infection of mice with an induced tumor in the abdomen. All control conditions of P. aeruginosa cells growing in suspension and as a biofilm were added to the analysis. We found that these different P. aeruginosa strains express a pool of distinct genetic traits that are activated under particular infection conditions regardless of their genetic variability. The knowledge herein generated will advance our understanding of P. aeruginosa virulence and provide valuable cues for the definition of prospective targets to develop novel intervention strategies.

The impact of the types of microorganisms isolated from blood and wounds on the results of treatment in burn patients with sepsis.

UNLABELLED Despite development of combustiology, infections continue to be the most important cause of death among patients with burns. Sepsis is the most severe clinical presentation of infection in patients after thermal injuries who require immediate treatment. Early diagnosis and proper treatment of sepsis are important in the clinical management that is often hampered for multiple reasons, e.g. impaired patient immunity, problems with microorganisms with multi-antibacterial drug resistance. The aim of the study was to assess effect of type of a microorganism isolated from blood and wound on results of treatment of sepsis in patients with burns. MATERIAL AND METHODS Effect of type of microorganisms isolated from blood and wound on the result of treatment of sepsis was studied in 338 patients hospitalized immediately after an injury in Centre for Burn Treatment in Siemianowice Śląskie in years 2003 - 2004 (at the age of 18 - 96 years, 66 women and 272 men). Clinical symptoms of generalized infection were found in all study subjects. The study group was divided into two subgroups: cured patients and patients who died of sepsis. The following parameters were assessed in both subgroups: type of microorganism isolated from blood, type of microorganism isolated from wound as well as occurrence of the same and different infections of blood and burn wound. RESULTS positive blood cultures were found in 165 patients (48.8%), 106 (64.2%) were cured, 59 (35.8%) died. The most commonly isolated microorganisms in cured patients were Gram(+) Staphylococcus epidermidis MRSE (19.81%) and Staphylococcus aureus MRSA (18.87%). Gram(-) intestinal rods were least commonly isolated from this group. The most commonly isolated microorganisms from blood of patients who were to die, included non-fermenting Gram(-) rods Acinetobacter baumannii (35.59%) and Pseudomonas aeruginosa (22.03%). Mixed bacterial flora was found in the blood of 22.03% patients. Among patients who were to die, the same microorganisms were found in the blood and in the wound in 32.2% of patients, while this rate was 17.92 in cured patients. The most commonly found bacteria in the blood and burn wound in the cured patients included Staphylococcus aureus MRSA (31.58%) and Staphylococcus aureus (21.05%). In the group of patients who were to die, the most common bacteria isolated simultaneously from the blood and burn wound included Acinetobacter baumannii (47.37%) and Pseudomonas aeruginosa (36.84%). CONCLUSIONS 1. The patients with thermal injuries are at higher risk of death in the event of sepsis caused by Gram(-) bacteria versus Gram(+) bacteria. 2. Infection of blood and burn wound caused by the same bacteria Pseudomonas aeruginosa and Acinetobacter baumanni increases the risk of death due to sepsis in patients with burns following thermal injuries.

Transfer of fibroblast sheets cultured on thermoresponsive dishes with membranes

In cell or tissue engineering, it is essential to develop a support for cell-to-cell adhesion, which leads to the generation of cell sheets connected by extracellular matrix. Such supports must be hydrophobic and should result in a detachable cell sheet. A thermoresponsive support that enables the cultured cell sheet to detach using only a change in temperature could be an interesting alternative in regenerative medicine. The aim of this study was to evaluate plates covered with thermoresponsive polymers as supports for the formation of fibroblast sheets and to develop a damage-free procedure for cell sheet transfer with the use of membranes as transfer tools. Human skin fibroblasts were seeded on supports coated with a thermoresponsive polymer: commercial UpCell™ dishes (NUNC™) coated with thermoresponsive poly(N-isopropylacrylamide) (PNIPAM) and dishes coated with thermoresponsive poly(tri(ethylene glycol) monoethyl ether methacrylate) (P(TEGMA-EE)). Confluent fibroblast sheets were effectively cultured and harvested from both commercial PNIPAM-coated dishes and laboratory P(TEGMA-EE)-coated dishes. To transfer a detached cell sheet, two membranes, Immobilon-P® and SUPRATHEL®, were examined. The use of SUPRATHEL for relocating the cell sheets opens a new possibility for the clinical treatment of wounds. This study established the background for implementing thermoresponsive supports for transplanting in vitro cultured fibroblasts.

The efficiency of burn wounds debridement with use of hydrosurgery--our experiences.

UNLABELLED The aim of the study was to present experience of doctors from the Center for Burns Treatment in Siemianowice Śląskie who use a Versajet system. MATERIAL AND METHODS Debridement with the use of a water jet was applied in 70 patients with IIb° /III° thermal burns in the period between 2009-2013. The corresponding evaluation involved duration of operation, locations, wound surfaces and technique of debridement after operation. Microbiological tests were also performed before and after debridement. RESULTS AND CONCLUSIONS In the paper, its authors draw attention to the short time of debridement. The application of a water jet allows precise debridement, particularly in hardly accessible places, preparation of wounds to be covered with autologous split thickness skin graft, protection of healthy tissues.

Infections in the tissue material and their impact on the loss of transplants in the Laboratory of in vitro Cell and Tissue Culture with Tissue Bank in the years 2011–2015

Radiation sterilization eliminates microbiological infections but causes the degradation of the cell factor. The negative result of microbiological examination for tissue transplants is one of the conditions for approval for distribution in patients. The study attempts to verify impact of the presence of microbes onto material for transplant loss. In the 2011–2015 period, we analyzed 293 donors of skin and amnion. Microbiological sampling was performed. The total of 21 strains of bacteria, molds and fungi was identified in collected tissue. The widest spectrum of strains was found in skin (17), followed by amnia (8). The total number of positive findings was 147 and was again highest in skin (129), while the number of positive findings in amnia was 18 only. The general percentage of fungal infections was very low. The presence of fungal strains was only observed in allogeneic skin (2%). Large number of microorganisms isolated from the skin before sterilization was observed, so it seems impossible to use allogeneic intravital skin. However, the intravital application of allogeneic amnion obtained from cesarean section remains to be considered.

Literature Review Concerning Cell and Skin Substitute Cultures Obtained by Means of Tissue Engineering used in the Treatment of Burns

Center For The Treatment of Burns in Siemianowice Śląskie1 Dyrektor: dr n. med. M. nowak Technical-Humanistic Academy in Bielsko Biała. Faculty of Health Science. Faculty of Emergency Medicine2 Dziekan Wydziału: dr hab. M. Mikulska, prof. ATH Higher School of Applied Sciences in Ruda Śląska. Department of Health3 Dziekan Wydziału: doc. dr n. med. I. Caus-Woźniak Center of Polymer and Carbon Resources of the Polish Academy of Science in Zabrze4 Dyrektor: prof. dr hab. a. dworak Chair and Department of General, Molecular, and Genetic Biology, Silesian Medical University in Katowice5 Kierownik: prof. dr hab. A. L. Sieroń

Allogeneic vs. Autologous Skin Grafts in the Therapy of Patients with Burn Injuries: A Restrospective, Open-label Clinical Study with Pair Matching

BACKGROUND Early application of autologous skin may lead to the loss of split thickness skin graft due to unclarified wound bed. Allogeneic skin grafts are performed on patients with extensive burn injuries after escharotomy, tangential excisions and deep debridement for the purpose of stabilizing the general condition and reducing the scope of local complications. OBJECTIVES The aim of this paper is to determine how the use of allografts improves the conditions for the intake of autografts in burns treatment, and how it accelerates wound healing in comparison to the autografts-only option. MATERIAL AND METHODS In 2012-2013, allogeneic skin was grafted on 46 patients, and in 8 cases grafting was repeated several times. An autologous split-thickness skin graft was applied to 32 patients. The analysis included the relationship between the duration of hospitalization and the number of skin transplantations, the relationship between the time of admission to debridement of the necrotic tissues and the total duration of hospitalization. Statistical analysis encompassed also pain assessment. RESULTS The results suggest that multiple applications of autografts not only do not lead to quicker recovery, but even lengthen the hospitalization time. The dependency is visible also in the patients who underwent the skin grafting procedure in allogeneic and autologous systems twice or more. There was a statistical significant difference between the duration of hospitalization in groups of patients who underwent STSG preceded by allogeneic skin graft transplantation when compared to the group of patients who underwent allogeneic skin application (p < 0.05) and the group of patients who were grafted with autologous skin (p < 0.05). CONCLUSIONS Allogeneic skin grafts are a perfect dressing when wound vascularization is insufficient to take free split-thickness skin graft. In patients with comparable burn surface areas, multiple applications of free autologous split-thickness skin grafts (STSG) extend the hospitalization time in comparison to application of allogeneic skin dressing as the first-line therapy.

A 2000 patient retrospective assessment of a new strategy for burn wound management in view of infection prevention and treatment

Infections in burn patients are still the principal cause of complications in burn injuries. The aim of this study is to assess a new strategy for burn wound management in view of infection prevention and treatment in the experience of the Burn Treatment Center in Siemianowice Śląskie. The applied methodology involved the analysis of patient records describing the hospitals epidemiological situation between 2014 and 2016. The analysis also included the use and cost of antibiotics, silver‐containing dressings, and other antiseptics relative to the number of sepsis cases, including those caused by Pseudomonas aeruginosa, as well as the mortality ratio. The total costs of prevention and treatment of infections were reduced, while the use of silver‐containing dressings and antiseptics increased. The number of patients with sepsis decreased, including cases caused by P. aeruginosa, and the mortality ratio was reduced. Introducing a strategy for burn wound‐oriented infection prevention and treatment in burn patients provides a number of benefits. It is also cost‐effective. Using locally applied active dressings and antiseptics can be a welcome choice for often‐unnecessary antibiotic therapy of a suspected or existing burn wound infection.

Own Experience From The Use Of A Substitute Of An Allogeneic Acellular Dermal Matrix Revitalized With In Vitro Cultured Skin Cells In Clinical Practice.

As a result of the removal of cells from human allogeneic dermis, a collagen scaffold is obtained, which can be populated de novo with autologous/allogeneic skin cells and transplanted onto the area of skin loss. The optimal method for production of acellular dermal matrices (ADM) has been selected. Three female patients (a mean age of 54 years) were subjected to the transplantation of either autologous or allogeneic keratinocytes and fibroblasts into the holes of acellular dermal matrix (ADM) mesh graft. The method for burn wound treatment based on the use of a viable dermal-epidermal skin substitute (based on ADM and in vitro cultured fibroblasts and keratinocytes) may be the optimal method of burn treatment.

The potential of bacteriophages in the treatment of burn wounds.

Bacteriophages were identified in 1915 and have been used since 1919. Despite the prevalence of their application the first users did not understand the nature of bacteriophages, thus, the efficiency of phage therapy proved controversial. Phage therapy was replaced by antibiotics. Fortunately, some centers in Middle and Eastern Europe continued research on bacteriophages. The renaissance of phage therapy was an obvious consequence of the increased percentage of hospital infections caused by multi drug-resistant strains (1). Burn wound infections are a major problem in the recovery process, considering patients with III degree burns, where survival is determined by the severity of the burn and burnrelated infections. Microbial infections which appear during the initial 24 hours are drugresistant and enhance the complexity of the therapeutic problem. The more and more common alternative to antibiotics, widespread by Polish and Georgian centers is the use of phage therapy (2). Based on study results it is estimated that phage therapy is effective in 80% of Enterococcus infections, and even in 90% of Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Klebsiella pneumoniae infections. Bacteriophages may be effectively used in the treatment of burn wounds, especially opportunistic pathogens such as Pseudomonas spp (2). One of the most important areas of phagotherapy interest are gram-negative bacteria, which show multi-drug resistance. Pseudomonas aeruginosa is the flagship member of the above-mentioned and at the same time an etiologic factor of infection with questionable therapeutic success of antibiotics. Simultaneously, Pseudomonas aeruginosa displays a number of properties, which make it a suitable target for phagotherapy, including the creation of a biofilm, which is hydrolized during the phage infection (1). Studies in mice, infected with resistant to imipenem P. aeruginosa, who developed bacteremia and died within 24 hours, is evidence of the high efficacy of phage ØA392, which was represented by 100% of survival of mice. The phage remained in circulation for 48 hours, that is until complete bacterial eradication without resulting in an unspecific immunological response (3). The biggest problem in the treatment of burn wounds is their infection, which often prevents skin grafting. Studies demonstrated that the use of phages, prior to grafting, hinders P. aeruginosa growth, but also has a positive effect on wound healing (1). Based on clinical and laboratory study results with the use of bacteriophages, one may come to the conclusion that the above-mentioned lead to faster recovery from septic com-