The Assessment of Activity of Antiseptic Agents against Biofilm of Staphylococcus aureus Measured with the Use of Processed Microscopic Images.

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 activity of polyhexanide and povidone-iodine than hypochlorite against staphylococcal biofilm. Taking into account the fact that in vitro results of the efficacy of antiseptic agents against staphylococcal biofilm are frequently applied to back up their use in hospitals and ambulatory units, our work should be considered an important tool; providing reliable, quantitative data in this regard.

Diagnosing pleural effusions using mass spectrometry-based multiplexed targeted proteomics quantitating mid- to high-abundance markers of cancer, infection/inflammation and tuberculosis.

Pleural effusion (PE) is excess fluid in the pleural cavity that stems from lung cancer, other diseases like extra-pulmonary tuberculosis (TB) and pneumonia, or from a variety of benign conditions. Diagnosing its cause is often a clinical challenge and we have applied targeted proteomic methods with the aim of aiding the determination of PE etiology. We developed a mass spectrometry (MS)-based multiple reaction monitoring (MRM)-protein-panel assay to precisely quantitate 53 established cancer-markers, TB-markers, and infection/inflammation-markers currently assessed individually in the clinic, as well as potential biomarkers suggested in the literature for PE classification. Since MS-based proteomic assays are on the cusp of entering clinical use, we assessed the merits of such an approach and this marker panel based on a single-center 209 patient cohort with established etiology. We observed groups of infection/inflammation markers (ADA2, WARS, CXCL10, S100A9, VIM, APCS, LGALS1, CRP, MMP9, and LDHA) that specifically discriminate TB-PEs and other-infectious-PEs, and a number of cancer markers (CDH1, MUC1/CA-15-3, THBS4, MSLN, HPX, SVEP1, SPINT1, CK-18, and CK-8) that discriminate cancerous-PEs. Some previously suggested potential biomarkers did not show any significant difference. Using a Decision Tree/Multiclass classification method, we show a very good discrimination ability for classifying PEs into one of four types: cancerous-PEs (AUC: 0.863), tuberculous-PEs (AUC of 0.859), other-infectious-PEs (AUC of 0.863), and benign-PEs (AUC: 0.842). This type of approach and the indicated markers have the potential to assist in clinical diagnosis in the future, and help with the difficult decision on therapy guidance.

Organizational issues of home parenteral nutrition during COVID-19 pandemic: Results from multicenter, nationwide study.

OBJECTIVES: Patients on home parenteral nutrition (HPN) are prone to severe complications of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. The pandemic requires adaptation of the health care standards, including epidemiologic surveillance, logistics of home supply, and monitoring. Potential lack of medical professionals may worsen the standard of care. The aim of this study was to evaluate the medical staff resources in HPN units. RESULTS: The study was conducted by major Polish scientific societies in clinical nutrition. A questionnaire was distributed among all Polish adult HPN centers concerning statistics from the first 3 mo of the pandemic (March through May 2020). Data on medical staff resources and organizational issues of the units were collected. Modifications of the home procedures, SARS-CoV-2 infection rates of HPN patients and health care workers (HCW) were analyzed. Influence of the pandemic on the rates of new qualifications for home artificial nutrition (HAN) was estimated. Fourteen of 17 adult Polish HPN units took part in the study. The point prevalence of HPN in Poland was 30.75/1 million citizens. Of HCWs, 344 were involved in patient care in Polish HPN units; 18.9% were physicians (49% surgeons, 18.46% internal medicine specialists, 15.38% anesthesiologists, 7.69% pediatricians, 1.54% palliative care specialists), 32.27% nurses, 5.23% dietitians, 9.01% pharmacists, 4.94% pharmacy technicians, 3.2% pharmacy assistants, 5.81% administrative workers, 3.49% physiotherapists. HAN patient-to-HCW ratios for physicians, nurses, pharmacists, dietitians were 49.5, 29.15, 111.6, and 181.6, respectively. Medium ages of physicians and nurses were 45.6 and 44.15 y, respectively. Slightly less than half (53.8%) of physicians and 31.53% of nurses worked parallelly in hospital wards. Thirty-one pharmacists overall were working in all HPN units (2.21 per unit) as were 18 dietitians (1.3 per unit). Nine patients had a confirmed COVID-19 infection (four HPN, five home enteral nutrition). All the units introduced telemedicine solutions in the first months of the pandemic. The number of new qualifications for HPN and home enteral nutrition in the units did not significantly decline from March through May in comparison with a similar period in 2019. CONCLUSIONS: A shortage of HPN medical professionals requires attention when planning health care organization, especially during a pandemic. Severe restrictions in public health systems may not reduce the number of new qualifications for the HPN procedure. There is a need for the continuation of data collection during the evolution of the pandemic as it may have a detrimental effect on HPN including serious issues with access to professional HCWs.

In Vitro Evaluation of Polihexanide, Octenidine and NaClO/HClO-Based Antiseptics against Biofilm Formed by Wound Pathogens.

Chronic wounds complicated with biofilm formed by pathogens remain one of the most significant challenges of contemporary medicine. The application of topical antiseptic solutions against wound biofilm has been gaining increasing interest among clinical practitioners and scientific researchers. This paper compares the activity of polyhexanide-, octenidine- and hypochlorite/hypochlorous acid-based antiseptics against biofilm formed by clinical strains of Candida albicans, Staphylococcus aureus and Pseudomonas aeruginosa. The analyses included both standard techniques utilizing polystyrene plates and self-designed biocellulose-based models in which a biofilm formed by pathogens was formed on an elastic, fibrinous surface covered with a fibroblast layer. The obtained results show high antibiofilm activity of polihexanide- and octenidine-based antiseptics and lack or weak antibiofilm activity of hypochlorite-based antiseptic of total chlorine content equal to 80 parts per million. The data presented in this paper indicate that polihexanide- or octenidine-based antiseptics are highly useful in the treatment of biofilm, while hypochlorite-based antiseptics with low chlorine content may be applied for wound rinsing but not when specific antibiofilm activity is required.

Further Data on Wound Healing Rates After Application of Lucilia sericata.

Maggot therapy has recently received a special medical and public attention, and according to the specialists' opinion, it takes us to the future of a wound care. Simultaneously, as new biomechanisms were discovered, statistical analyses of wound healing rates were conducted usually adopting simple parametric and nonparametric tests. In this study, based on a set of statistical methods, we performed an advanced analysis of wound surface reduction using Lucilia sericata larvae in different clinical aspects: status of diabetes mellitus, maggots' density, and pain intensity. Particularly, we employed these factors because, in our statistical analysis, they are easy to obtain and they proved to be the possible risk factors of wound regeneration. Furthermore, these factors represent different clinical, biological, and neurological spectra of knowledge. In our study, we have found further and statistically significant correlations between the analyzed variables and skin regeneration together with different time periods of the healing rate using maggot therapy in patients with lower limb ulceration.

Potential of Biocellulose Carrier Impregnated with Essential Oils to Fight Against Biofilms Formed on Hydroxyapatite.

In this research, bacterial cellulose (BC), one of the most promising biopolymers of the recent years, was saturated with thyme, eucalyptus and clove essential oils (EOs) and applied against staphylococcal and pseudomonal biofilms formed on hydroxyapatite (HA). BC dressings were thoroughly analyzed with regard to their physical properties. Moreover, the exact composition and ability of particular EO molecules to adhere to HA was assessed. Additionally, cytotoxicity of oil-containing, cellulose-based dressings towards osteoblasts and fibroblasts as well as their impact on reactive oxygen species (ROS) production by macrophages was assessed. The results revealed the high ability of BC dressings to absorb and subsequently release EOs from within their microstructure; the highest number of compounds able to adhere to HA was found in the thyme EO. The eucalyptus EO displayed low, while thyme and clove EOs displayed high cytotoxicity towards fibroblast and osteoblast cell lines. The clove EO displayed the highest eradication ability toward staphylococcal, while the thyme EO against pseudomonal biofilm. Taken together, the results obtained indicate the suitability of EO-saturated BC dressings to eradicate pseudomonal and staphylococcal biofilm on HA surface and moreover, to not trigger reactive oxygen species production by immune system effector cells. However, due to cytotoxic effects of thyme and clove EOs towards cell lines in vitro, the eucalyptus EO-saturated BC dressing is of highest potential to be further applied.

An MRM-Based Cytokeratin Marker Assay as a Tool for Cancer Studies: Application to Lung Cancer Pleural Effusions.

PURPOSE: The goal of this work was to develop an LC-MRM assay for the quantitative analysis of a set of established and diagnostically important cytokeratin (CK) markers used in cancer diagnosis, prognosis, and therapy monitoring. Second, the potential of this assay in lung cancer diagnosis through pleural effusion (PE) analysis was examined. EXPERIMENTAL DESIGN: A multiplexed MRM assay was developed for 17 CKs and their select caspase-cleaved fragments. Isotope-labeled standard peptides were used for high assay specificity and absolute peptide quantitation; with robust standard-flow LC coupled to a latest-generation triple-quadrupole instrument for high sensitivity. The potential clinical applicability was demonstrated by the analysis of 118 PE samples. RESULTS: The MRM assay was evaluated for endogenous detection, linearity, precision, upper and lower limits of quantification, selectivity, reproducibility and peptide stability, and is generally applicable to any epithelial cancer study. A set of 118 patients with known pathologies allowed us to define the range of CK levels in clinical PE samples. Specific CKs were able to differentiate cancer-related PEs from those caused by benign ailments. In addition, they allowed to differentiate between PEs from subjects with small cell lung cancer versus non-small cell lung carcinoma, and to further differentiate the latter into its two subtypes, adenocarcinoma and squamous cell carcinoma. CONCLUSION AND CLINICAL RELEVANCE: An MRM-based CK assay for carcinoma studies can differentiate between the three lung cancer histological types using less-invasive PE sampling providing potential therapy-guiding information on patients that are inoperable.

Metabolic profiles of exudates from chronic leg ulcerations.

Chronic leg ulceration is a disease usually associated with other comorbidities, and significantly reduces patient quality of life. Infected leg ulcers can lead to limb-threatening sequelae or mortality. Leg ulcerations are colonized by a number of microbes that are able to cause life-threating infections in susceptible patients. Wound exudate is a body fluid that collects metabolites from patient eukaryotic cells and from prokaryotic bacterial communities inhabiting the wound. This study aimed at identification of metabolites in exudates collected from chronic leg ulcers, and correlation of this metabolome with patient comorbidities and microbiological status of the wound. By means of NMR spectroscopy we detected 42 metabolites of microbial or patient origin. The metabolites that were in abundance in exudates analyzed were lactate, lysine, and leucine. Metabolites were associated with the presence of neutrophils in wounds and destruction of high quantities of microbes, but also with hypoxia typical for venous insufficiency. The combination of nuclear magnetic resonance spectroscopy technique and partial least squares discriminant analysis allowed us to further discriminate groups of metabolites with regards to potential clinical meaning. For example, to discriminate between S.aureus versus all other isolated microbial species, or between patients suffering from type I or II diabetes versus patients without diabetes. Therefore, wound exudate seems to be highly applicable material for discriminant analysis performed with the use of NMR technique to provide for rapid metabolomics of chronic wound status.

A Multiplexed Cytokeratin Analysis Using Targeted Mass Spectrometry Reveals Specific Profiles in Cancer-Related Pleural Effusions.

Pleural effusion (PE), excess fluid in the pleural space, is often observed in lung cancer patients and also forms due to many benign ailments. Classifying it quickly is critical, but this remains an analytical challenge often lengthening the diagnosis process or exposing patients to unnecessary risky invasive procedures. We tested the analysis of PE using a multiplexed cytokeratin (CK) panel with targeted mass spectrometry-based quantitation for its rapid classification. CK markers are often assessed in pathological examinations for cancer diagnosis and guiding treatment course. We developed methods to simultaneously quantify 33 CKs in PE using peptide standards for increased analytical specificity and a simple CK enrichment method to detect their low amounts. Analyzing 121 PEs associated with a variety of lung cancers and noncancerous causes, we show that abundance levels of 10 CKs can be related to PE etiology. CK-6, CK-7, CK-8, CK-18, and CK-19 were found at significantly higher levels in cancer-related PEs. Additionally, elevated levels of vimentin and actin differentiated PEs associated with bacterial infections. A classifier algorithm effectively grouped PEs into cancer-related or benign PEs with 81% sensitivity and 79% specificity. A set of undiagnosed PEs showed that our method has potential to shorten PE diagnosis time. For the first time, we show that a cancer-relevant panel of simple-epithelial CK markers currently used in clinical assessment can also be quantitated in PEs. Additionally, while requiring less invasive sampling, our methodology demonstrated a significant ability to identify cancer-related PEs in clinical samples and thus could improve patient care in the future.

A comparison of an antibacterial sandwich dressing vs dressing containing silver.

The purpose of this study was to compare the efficacy of dressings containing octenidine vs. dressings containing silver in the wound healing in the course of a chronic venous disease. There were two groups of 40 patients who met the inclusion criteria and who did not meet the exclusion criteria. The patients were randomly assigned into the groups (envelope method). The first, "O group" was treated with octenidine-based dressings. The second, "S group" was treated with silver dressings. The study lasted for 56 days. All patients in the research were treated with medical compression stockings with cotton understockings. Microbiological eradication was observed on the 28th day of the study among 33% of patients in the treatment group vs. 6% in control group. On the 56th day of the treatment, these percentages equalled 72% and 35%. The rate of healing was faster in the 0 group than in the S group. In the wounds <10 cm(2) it was faster by 1.35 cm(2)/week and in wounds >10 cm(2) it equalled 3.44 cm(2). The reduction of pain level was 37.5% higher in the O group, in contrast with the S group. One change of a dressing in the O group led to a 0.06 cm(2) greater wound size reduction and in the case of wounds >10 cm(2) to 0.29 cm(2) reduction compared with the S group. The presented results indicate that the efficacy of dressings containing octenidine is higher compared to silver dressings.

[Etiology of bacterial infections and incidence of comorbidities in patients with tuberculosis, treated in Mazovian Treatment Centre of Tuberculosis and Lung Diseases during years 2012-2014].

INTRODUCTION: Tuberculosis (TB) is an infectious disease caused by the Mycobacterium tuberculosis. Infection occurs mostly via inhalation, while the immune system is weakened. TB can take a pulmonary or extrapulmonary form. Treatment involves an intensive, long-term antimycobacterial multidrug therapy. TB cases are recorded on the worldwide scale. The morbidity in Poland varies territorially. AIM: The analysis of bacterial infections and comorbidities cases in the patients with TB, treated in Mazovian Treatment Centre of Tuberculosis and Lung Diseases (MCLChPiG) during years 2012-2014. MATERIALS AND METHODS: The study includes an analysis of 3093 cases of tuberculosis among MCLChPiG patients in years 2012-2014, taking into account the age and gender of patients, forms of the disease, bacterial superinfection in the course of TB (based on the results of microbiological tests) and concomitant diseases. RESULTS: The study showed that TB was more common in men (64.79%). Most cases have been diagnosed in the 50-65 year age group (31.65%). The most common form of TB among MCLChPiG patients was a respiratory tract tuberculosis (96.61%), especially the pulmonary form (82.67%). Concomitant diseases were diagnosed in 244 patients (7.89%), wherein tumors (4.88%) were the most common ones. Bacterial superinfection in the course of TB was observed in 149 patients (4.82%). The most frequently isolated bacteria were H. influenzae (28.65%) and S. aureus (15.79%). CONCLUSIONS: Long-term antimycobacterial treatment leads to the weakening of the patient's immune system, which is a favorable condition for the development of bacterial infections. Superinfection can be associated with concomitant disease, where weakness of immunoresponsiveness increases the risk of developing TB. Bacteria isolated from superinfections in the course of TB are mostly a typical pathogens of the upper and lower respiratory tract.

Laparoscopic splenectomy complicated by pancreatic cyst in the course of Hodgkin's disease: case report.

This paper describes a case of a patient suffering from Hodgkin's disease and treated by means of minimally invasive surgery: laparoscopic splenectomy. The performance of laparoscopic splenectomy led to a complication in the form of a pancreatic cyst. The cyst was subjected to endoscopic procedures (endoscopic retrograde cholangiopancreatography, with a shaft to the pancreatic tract) and percutaneous drainage. The application of the above methods allowed for a quick introduction of causative treatment. Because in the case in question open surgery methods would impede the introduction of systemic treatment, minimally invasive surgery techniques (laparoscopy and endoscopy) were applied and are discussed below.

The content of elements in rainwater and its relation to the frequency of hospitalization for gastric and duodenal peptic ulcers in Opole Voivodship, Poland, during 2000-2002.

The relationship between chemical elements in rainwater and the frequency of hospitalizations for gastric and duodenal peptic ulcers was studied on the population of the province (Voivodship) of Opole, Poland, during the years 2000-2002. There is a high positive correlation between hospitalized cases of gastric peptic ulcers with chromium in rainwater (r = 0.71), cadmium (r = 0.63), and lead (r = 0.70). Mild positive correlations were found with zinc (r = 0.55), copper (r = 0.56), iron (r = 0.57), chloride (r = 0.60), and sulfate (r = 0.52). These correlations were higher on men, suggesting that there are gender correlations involved. In duodenal peptic ulcers, we observed a high positive correlation between chromium in rainwater and hospitalized cases (r = 0.61) and mild positive correlations with lead (r = 0.57), copper (r = 0.52), and cadmium (r = 0.51). Significant gender differences were not found. These positive correlations may be due to the biological activity of the elements, such as their cytotoxic activity, enhanced local adrenergic stimulation on mucosal vasculature, ion channel (mainly calcium channels) remodeling, and, for example promoting infection by Helicobacter pylori.

The content of elements in rainwater and its relation to the frequency of hospitalization for atrial fibrillation in Opole Voivodship, Poland, during 2000-2002.

The elemental composition of rainwater is a suitable indirect indicator of the presence of chemical elements in airborne dust. As such, rainwater is considered a suitable monitor for environmental or natural pollution. The yearly content of chemical elements in rainwater may be considered a good indicator for determining the influence of these environmental factors on human body. We decided to investigate the relationship between chemical elements in rainwater and the frequency of hospitalizations for atrial fibrillation (I48, ICD10) on the population of the region of Opole Voivodship, Poland during the years 2000-2002. The voivodship or province has a population of one million and is divided into 12 counties that have between 44,000 and 151,000 inhabitants. We calculated the yearly average number of hospitalizations by reasons of atrial fibrillation per 10,000 inhabitants in particular counties. The average content of the chosen chemical elements in kilograms per hectare per year was calculated for each county individually and the data evaluated by means of Spearman's correlation coefficient. There is a high positive correlation between chromium in rainwater and hospital admissions for atrial fibrillation (r = 0.62) and mild positive correlation with cadmium (r = 0.57), lead (r = 0.57), zinc (r = 0.50), and chloride (r = 0.48). There are no significant differences between male and female patients. The biological activities of these elements, which may include enhanced adrenergic stimulation, remodeling of calcium or other ion channels, or the cytotoxic effects of hexavalent chromium may explain the positive correlations here reported.

The content of elements in rainwater and its relation to the frequency of hospitalization for diabetes and obesity in Opole Voivodship, Poland, during 2000-2002.

Environmental factors significantly influence the incidence and course of metabolic syndrome diseases such as diabetes and obesity. The content of elements in rainwater is an indirect indicator of their presence in dust suspended in the air. In this paper we present the relationships between the content of selected elements in rainwater and hospitalization frequencies due to diabetes (E10-E13) and obesity (E66). It was assumed that the hospitalization frequency could be taken as a measure of deterioration of the metabolic process in the course of diabetes and its complications. The observations concerned the population of Opole Voivodeship, Poland (one million inhabitants), distributed in small communities of 44,000 to 151,000 inhabitants during the years 2000-2002. In cases of diabetes E10-E13 for all subjects relevant correlation indicators were found for chromium (r = 0.71), cadmium (r = 0.65), and lead (r = 0.66). Borderline relevance was seen for copper (r = 0.57) and zinc (r = 056). For diabetic men the statistically relevant correlations were chromium (r = 0.79), lead (r = 0.77), cadmium (r = 0.74), copper (r = 0.70), chloride (r = 0.69), zinc (r = 0.68), and iron (r = 0.64). For women the only relevant correlations were chromium (r = 0.62) and cadmium (r = 0.55). No significant correlations were found in obese individuals of both sexes.

Infant mortality--gynaecological and unemployment aspects in Opole Province, Poland.

In the study the results of the relation between infant (neonatal + post-neonatal) mortality and the gynaecological and unemployment factors are presented. The findings are based on Bayesian modelling and indicate the possible influence of the unemployment situation on mortality, whereas no correlation was found as regards access to the availability of gynaecological health care. The main conclusion is that elevated unemployment which could be referred to a poor socioeconomic situation (amongst other factors) may force pregnant women to pay more attention to improving their life conditions, than to sufficient medical controlling.