[Multiresistant Pathogens in Oncological and Cardiological Rehabilitation - Results of a Surveillance Study in Brandenburg]. / Multiresistente Erreger in der onkologischen und kardiologischen Rehabilitation ­ Ergebnisse einer Surveillancestudie in Brandenburg.

We investigated the prevalence of multidrug resistant pathogens in patients of oncologic and cardiologic rehabilitation units with 155 oncologic and 157 cardiologic patients undergoing microbiologic screening. It was found that 4.5% of oncologic as well as cardiologic patients were colonized with multidrug resistant pathogens. 2-MRGN and ESBL were the most encountered species (2.9%). 3-MRGN were found twice as frequent in oncologic patients (2.6 and 1.3%). Overall oncologic and cardiologic patients exhibit comparatively low prevalence rates for multidrug resistant pathogens.

Viability of Lucilia sericata maggots after exposure to wound antiseptics.

After debridement and before dressing a wound with maggots of calliphorid flies, one frequently performed step is the application of antiseptics to the prepared wound bed. However, the concomitant application of antiseptic agents during maggot therapy is regarded controversial as antiseptics may interfere with maggots' viability. In this experimental in vitro study, the viability of fly maggots was investigated after exposure to various antiseptics frequently used in wound care. Here, we show that Lucilia sericata fly maggots can survive up to an hour's exposure to wound antiseptics such as octenidine, povidone-iodine or polihexanide. Concomitant short-term application of wound antiseptics together with maggots on wound beds is tolerated by larvae and does not impair their viability.

In vitro susceptibility of methicillin-resistant and methicillin-susceptible strains of Staphylococcus aureus to two different cold atmospheric plasma sources.

BACKGROUND: Staphylococcus aureus may be the most important wound pathogen and causative for most of surgical site infections. As many anti-staphylococcal drugs are useless because of resistance, novel antimicrobial strategies are strongly needed and may be provided by cold atmospheric plasma (CP), which is being currently investigated for antiseptic efficacy. METHODS: To test the antimicrobial properties of CP against Staphylococcus aureus, 168 methicillin-susceptible isolates (MSSA) and 50 methicillin-resistant isolates (MRSA) were treated with two technically different plasma sources [an atmospheric pressure plasma jet (APPJ) and a dielectric barrier discharge plasma (DBD)] in vitro. RESULTS: CP treatment allowed a reproducible and significant growth reduction of MRSA and MSSA. However, MRSA was significantly less susceptible to treatment with DBD than was MSSA, while no difference between MRSA and MSSA was found using APPJ. CONCLUSIONS: As the initial physical antiseptic on skin, CP may be suitable for rapid decolonization of microbial pathogens in vivo. Each device must undergo validated efficacy testing prior to clinical application, as device related differences may occur.

Antimicrobial Efficacy of a Historical High-Frequency Plasma Apparatus in Comparison With 2 Modern, Cold Atmospheric Pressure Plasma Devices.

BACKGROUND: Today, highly sophisticated devices deliver cold atmospheric pressure plasma (CAPP) with a multitude of bioactive properties, opening the window to a new medical field: plasma medicine. Different techniques to create the optimal plasma device for different medical indications are currently being explored. However, even a 100 years ago, CAPP was briefly used in the related form of high-frequency therapy. The objective of our study was to compare historic with modern techniques regarding antimicrobial efficacy. METHODS: First, 26 different clinical isolates of relevant wound pathogens were treated in vitro with a historic violet wand (VW) and 2 modern plasma sources (kINPen 09 and dielectric barrier discharge [DBD]) and the obtained inhibition areas (IAs) were compared. Second, a biofilm model was used to compare biofilm inactivation by VW, DBD, ethanol, and polyhexanide treatment. RESULTS: DBD with the largest electrode produced the largest IAs. VW showed results similar to 2 different modes of the kINPen 09. IAs of VW were enlargeable by attaching a larger electrode. Against biofilms, VW was less effective than DBD but more effective than ethanol 70% and polyhexanide. CONCLUSION: The proven antimicrobial efficacy of VW may encourage the development of new, potent plasma devices based on the very simple and inexpensive technique of the historic high-frequency apparatus.

Risk factors for MRSA colonization in dermatologic patients in Germany.

BACKGROUND AND OBJECTIVES: Detection of methicillin-resistant Staphylococcus aureus (MRSA) carriage requires well-defined risk factors (RFs). Except for "chronic wounds", RFs are mostly specified in national recommendations. To avoid ineffective and expensive screening, we divided the entity "wounds" into different categories and calculated further RFs in dermatologic patients. PATIENTS AND METHODS: After a surveillance period with general MRSA screening, we correlated MRSA results with wound categories and dermatologically relevant diagnoses. We analyzed the screening efficacy by adding potential new RFs. RESULTS: Ulcers (pressure, arterial, combined pressure/arterial ulcers, ulcers otherwise unclassified), type 2 diabetes mellitus (DM), and atopic dermatitis (AD) were significantly associated with MRSA carriage. Tumors (subgroup basal and squamous cell carcinoma) were also significantly associated with MRSA carriage but had a protective odds ratio. Differentiation of wound types did not provide added benefit. In all MRSA-positive patients with chronic wounds, other RKI-listed RFs or type 2 DM were found. Screening sensitivity was increased combining classic RFs (except wounds) with type 2 DM and AD. CONCLUSIONS: In dermatologic patients, AD and type 2 DM were identified as new RFs. Distinct wound types were also found to be significant RFs, but differentiated screening offers no benefit. When screening patients according to national recommendations, excluding wounds but including type 2 DM and AD, there is no loss of sensitivity.

Skin and wound decontamination of multidrug-resistant bacteria by cold atmospheric plasma coagulation.

BACKGROUND AND OBJECTIVES: Novel concepts to limit the spread of multidrug-resistant bacteria (MDR) are urgently needed. Since treatment with cold atmospheric plasma (CAP) has shown significant antibacterial properties, the purpose of this study was to evaluate the ability of CAP to eliminate MDR- compared to non-MDR-pathogens in chronic wounds. METHODS: Eleven patients with 18 heavily colonized wounds were treated with a CE-certified commercial argon-based CAP source for 10 s/cm(2) in one session. The antimicrobial efficacy was assessed by calculating the microbial load before and after treatment. RESULTS: A single CAP treatment reduced MDR in all wounds. In 14 treatments (63.6 %) and for 16 pathogens (66.7 %), a 100 % reduction of the bacterial load was observed. For 11 of 17 (64.7 %) MDR-pathogens and for 5 of 7 (71.4 %) other non-MDR-pathogens, complete eradication was achieved. The remaining 8 treatments showed reductions of 77.5 ± 18.6 % and the remaining pathogens a reduction of 74.8 ± 25.7 %. CONCLUSIONS: As proof of principle, argon-based CAP serves as a potent treatment modality that was shown to limit MDR microbial colonization. The possible role of CAP in clinical MDR decontamination must be evaluated in clinical trials with repeated plasma treatment embedded in a comprehensive hygienic decontamination concept.

Comparison between cold plasma, electrochemotherapy and combined therapy in a melanoma mouse model.

The study was undertaken to compare antitumor efficacy of electrochemotherapy (ECT) with cold plasma therapy (CP) in a melanoma mouse model. After melanoma implantation into the flank of C57BL/6N mice, CP by two different plasma sources (APPJ and DBD) was applied directly to the tumor surface. ECT was performed with bleomycin intravenously at a field strength of 1000 V/cm without or combined with CP. Primary endpoints were tumor growth acceleration (TGA), daily volume progression (DVP) and survival after treatment. Both plasma sources as single treatment showed a significant TGA delay, which proved less effective than ECT. CP (APPJ) combined with ECT (ECJ) significantly improved per cent mouse survival, with significant superiority compared with ECT. Plasma therapy alone albeit less effective seems a potential alternative to ECT in patients with melanoma and can be applied manifold in a session without general anaesthesia. Accordingly, CP alone and combined with ECT may serve as new option in palliative skin melanoma therapy.

Triple swabbing allows sensitive MRSA detection in dermatologic patients of a university tertiary care hospital.

BACKGROUND: To optimize preventive measures to control MRSA, we investigated retrospectively the suitability of a multiple site screening model and the optimal sampling technique to detect MRSA in a university-based phlebology and skin cancer center in Germany. PATIENTS AND METHODS: During 4.5 years samples of 3 712 inpatients in a dermatologic department were analyzed for MRSA by conventional microbiologic cultures and in parallel by PCR. Samples were taken from nares, wounds and skin lesions. RESULTS: MRSA was detected in 60 inpatients (1.6%). 268 of 7 269 (3.7%) samples at admission and during hospital stay were found positive ñ 96 (35.8%) of these were swabs of nares, 59 (22.0%) surveillance swabs, 53 (19.8%) wound swabs and 42 (15.7%) from other dermatologic lesions. Twenty-five of 60 patients (41.7%) were found positive only in the nares, 10 (16.7%) patients only in wounds and 4 (6.7%) patients only in lesions. 166 (61.9%) of all positive culture samples became positive 24 hours after cultivation, 86 (32.1%) after 48 hours, and 16 (6.0%) after 72 hours. CONCLUSIONS: Highest sensitivity to detect MRSA can be reached by combining three swabs: nares, wounds and skin lesions (ìtriple-testî). Culture of screening specimens for 72 hours is recommended.

[Hyperspectral imaging demonstrates microcirculatory effects of postoperative exercise therapy in Dupuytren's disease]. / Hyperspektralimaging demonstriert mikrozirkulatorische Effekte postoperativer Ergotherapie bei Patienten mit Morbus Dupuytren.

BACKGROUND: Dupuytren's disease (DD) is a common connective tissue disorder of the hand. To prevent recurrence of contractures, patients usually receive early postoperative ergotherapy (ET). However, it is yet unknown how this measure impacts on local blood flow and hence on the occurrence or prevention of postoperative complications. Hyperspectral imaging (HSI) allows for a quantitative evaluation of tissue perfusion by measuring oxygen saturation and tissue water content. OBJECTIVE: The aim of this work was to evaluate the microcirculatory effects of early ET after partial fasciectomy in DD using HSI for optimised treatment and prevention. PATIENTS AND METHODS: In five patients, the oxygen saturation and tissue water content of the hand were measured before and 20 min after exercise therapy on the first two postoperative days using HSI. RESULTS: HSI demonstrated improved tissue perfusion in terms of quantity and quality following ET in all patients. After ET, all patients showed a relative increase in oxygen saturation of up to 20 % and a reduction in tissue water of up to 17 %. CONCLUSION: HIS allows for a fast and non-invasive evaluation of increased oxygen supply and decreased tissue water content in the surgical site after partial fasciectomy in DD following postoperative exercise therapy. This may improve wound healing and decrease the rate of recurrence in DD.

Hyperspectral imaging as a possible tool for visualization of changes in hemoglobin oxygenation in patients with deficient hemodynamics - proof of concept.

There is a lack of imaging tools for the evaluation of spatial alterations in microcirculation including blood oxygen saturation and hemoglobin distribution but recent innovative developments in hyperspectral technology may offer a solution. We examined different hemodynamic disorders in patients suffering from scleroderma, Dupuytren surgery, chronic foot ulcera and skin infections. Superficial and deeper blood oxygen saturation, hemoglobin distribution and water content were determined using hyperspectral imaging (HSI). In the patient with scleroderma, distinct cutaneous low perfused regions correlated with macroscopic skin aspects and seem to be potential therapy control marker. With HSI accurate clinical evaluation of a macroscopic conspicuous wound after Dupuytren surgery was possible and influenced further surveillance decisions. HSI clearly revealed the spatial geometry and also the clinically related perfusion parameters of abscess formation and chronic ulcer wounds. The hemodynamically relevant parameters like blood oxygen saturation (1 mm to approx. 6 mm subcutaneous), total hemoglobin distribution and tissue water content can be easily determined and visualized with HSI in near real time. Hence, this technique seems to be suitable for routine diagnostics of acute and chronic wounds as well as for the examination of systemic hemodynamic disturbances. Special indications may be transplant surveillance and monitoring of therapeutical interventions.

Hyperspectral imaging: innovative diagnostics to visualize hemodynamic effects of cold plasma in wound therapy.

An important clinical potential of cold atmospheric plasma (CAP) lies in tumor and wound treatment, whereby the last-mentioned is well-referenced already. However, the underlying mechanisms of improved wound healing have not been sufficiently clarified yet, in particular the influence of CAP on microcirculation. Hyperspectral imaging (HSI) enables the visualization of microcirculation of large tissue areas, thus this technique seems to be a candidate to examine CAP effects on perfusion and oxygen saturation in wounds. During clinical wound management, one chronic wound caused by peripheral arterial occlusive disease and one acute wound after surgical removal of cervical lymph nodes were examined using HSI before and after CAP treatment. HSI was able to demonstrate CAP effects on microcirculation showing a relevant increase of superficial and deeper cutaneous oxygen saturation together with elevated hemoglobin concentration in treated and also surrounding wound area. For the first time, it was shown that CAP improves the superficial and deeper oxygenation and hemoglobin perfusion in and around the treated area of acute and chronic wounds. This effect may contribute to healing support by CAP in wounds. HSI seems suitable for evaluating and monitoring CAP effects in clinical settings.

Hyperspectral imaging as a novel diagnostic tool in microcirculation of wounds.

Monitoring of perfusion is a cornerstone in surgery, phlebology and basic science to proof wound healing by interventions. In chronic wound management it is of utmost importance to realize and parametrize wound bed perfusion to verify actual, and plan further treatment by noninvasive diagnostics. Up to now monitoring is based on visual inspection of wounds as conventionally practiced over more than decades. The main problems of visual inspection are the lack of standardization and comparability because of interindividual variations. Therefore technical performance with contact free probes based on standardized perfusion measuring is strongly needed. Hyperspectral imaging (HSI) was investigated to overcome manual and visual wound inspection in monitoring of wound healing. HSI works noninvasive, and imaging of relevant perfusion parameters is possible without the need of contrast enhancing drugs. METHODS: HSI technology uses imaging spectroscopic analysis in visual and near infrared spectrum to get information on imaged tissue in less than 10 s. Tissue is radiated by broad spectrum light and the following parameters are calculated from remitted spectra: the grade of oxygenation and the volume proportion of hemoglobin (in superficial and also deeper (8 mm) tissues. The calculated data comprise the "Tissue hemoglobin oxygen saturation" (StO2) as percental oxygenation index to assess superficial perfusion (VIS-spectrum), the "Near infrared perfusion" (NIR) to assess deeper perfusion (near infrared spectrum) and the "Tissue hemoglobin index" (THI) to measure the percental volume of hemoglobin of surface perfusion (VIS-spectrum). The measurements of these parameters are calculated as false color-coded perfusion results on screen.We investigated different kind of wounds (combustion, infection, ulcer wounds, wounds in immune disorders, trauma wounds) determining superficial and deeper oxygen saturation, hemoglobin distribution and water content using hyperspectral imaging with TIVITA™ Tissue system. RESULTS: Hyperspectral Imaging allowed easy real time determination and visualization of hemodynamically relevant parameters- superficial and deeper oxygen saturation, total hemoglobin and tissue water content. In the patient with scleroderma, acral lesions with decreased perfusion correlated well with necrotic skin aspects.HSI clearly revealed macroscopic conspicuous suture wounds after Dupuytren surgery, infected soft tissue wounds with strong inflammatory hyperemia, edema in burn injuries, spatial geometry of abscess formation and chronic ulcer wounds. All measurements influenced further surveillance decisions. Hyperspectral imaging seems suitable for routine diagnostics and monitoring of skin and soft tissue lesions like acute and chronic wounds. It allows surveillance of postoperative suture wounds and burn wounds. Special indications may be transplant surveillance and monitoring of therapeutical interventions.

Does antibiotic resistance impair plasma susceptibility of multi-drug resistant clinical isolates of enterococci in vitro?

BACKGROUND: Cold atmospheric plasma could constitute an alternative against multi-drug resistant pathogens. Susceptibility of enterococci to cold atmospheric plasma was investigated in vitro. METHODS: 39 clinical isolates of enterococci were grouped dependent on the most important resistance patterns and treated on agar using dielectric barrier discharge plasma. These included enterococci with combined vancomycin- and high-level gentamicin resistance, high-level resistance to gentamicin (HLGR) only, vancomycin resistance alone (VRE), and enterococci susceptible to both. Susceptibility to cold atmospheric plasma was evaluated based on the zones of inhibition and examined in terms of the enterococcal group and the "degree" of drug resistance. RESULTS: Cold atmospheric plasma treatment killed all groups. Comparison of VRE and HLGR strains with non-VRE and non-HLGR isolates concerning zones of inhibition revealed that enterococci with special resistance patterns (VRE and HLGR) showed significantly smaller zones of inhibition than the sensitive ones. The mean of all isolates, irrespective of belonging to groups, showed smaller zones of inhibition with increasing "degree" of drug resistance. CONCLUSIONS: Cold atmospheric plasma treatment killed all isolates of enterococci, but its efficacy depended on the "degree" of drug resistance and on membership in special resistance groups with particular clinical-outbreak importance. However, a possible role of the different genetic lineages, which might be prone to acquiring more or less resistance phenotypes, may also play a role in this context.

On the history of plasma treatment and comparison of microbiostatic efficacy of a historical high-frequency plasma device with two modern devices.

BACKGROUND: Cold atmospheric pressure plasma (CAP) with its many bioactive properties has defined a new medical field: the plasma medicine. However, in the related form of high-frequency therapy, CAP was even used briefly a century ago. The aim of this study was to review historic CAP treatments and to obtain data regarding the antimicrobial efficacy of a historical high-frequency plasma device. METHODS: First, historic literature regarding the history of CAP treatment was evaluated, because in the modern literature no data were available. Second, the susceptibility of 5 different bacterial wound isolates, cultured on agar, to a historic plasma source (violet wand [VW]) and two modern devices (atmospheric pressure plasma jet [APPJ] and Dielectric Barrier Discharge [DBD]) was analyzed . The obtained inhibition areas (IA) were compared. RESULTS: First, the most convenient popular historical electromedical treatments produced a so-called effluvia by using glass electrodes, related to today's CAP. Second, all three tested plasma sources showed complete eradication of all tested microbial strains in the treated area. The "historical" cold VW plasma showed antimicrobial effects similar to those of modern APPJ and DBD regarding the diameter of the IA. CONCLUSION: Some retrograde evidence may be deducted from this, especially for treatment of infectious diseases with historical plasma devices. The underlying technology may serve as model for construction of modern sucessive devices.

Influence of preoperative skin sealing with cyanoacrylate on microbial contamination of surgical wounds following trauma surgery: a prospective, blinded, controlled observational study.

OBJECTIVE: Intraoperative bacterial contamination is a risk factor for surgical site infections (SSIs). This prospective, randomized, blinded, controlled trial (Reg. No. BB08/12) investigated the effect of a cyanoacrylate-based skin sealant (InteguSeal) on intraoperative wound contamination during trauma surgery. METHODS: A total of 128 patients undergoing trauma surgery were assigned randomly to an intervention (n=62) or a control group (n=66). Surgical sites were investigated at three locations: maximum incision depth (base), wound margin prior to wound closure (margin), and the surgical sutures (suture). Colony-forming units (CFU) were counted after 48h of incubation. RESULTS: Overall, significantly lower CFU counts were obtained for samples from the intervention group at all three sample sites compared to the control group. The difference, however, was only significant for the suture site (p=0.040). CONCLUSIONS: Preoperative sealing reduced microbial contamination on sutures during surgery, while the overall wound contamination remained unchanged. Hence, prevention of the clinically more relevant deep SSIs may not be expected. However, this study was not designed to detect differences in the rate of SSI. The role of the reduction in suture contamination with regard to the prevention of SSI remains to be evaluated.