Risk Factors for Non-Healing Wounds-A Single-Centre Study.

Background: Chronic wounds present a significant clinical, social, and economic challenge. This study aimed to objectify the risk factors of healing outcomes and the duration of chronic wounds from various etiologies. Methods: Patients treated for non-healing wounds at the surgical outpatient clinic of the Olomouc Military Hospital were involved. Data from patients treated between 8/2021 and 9/2023 were selected. Patients were mostly treated as outpatients, with microbiological follow-up indicated in cases of advanced signs of inflammation. Results: There were 149 patients who met our selection criteria (the mean age was 64.4 years). Predominant causes of wounds involved diabetes (30.9%), post-trauma (25.5%), pressure ulcers (14.8%), surgical site infections (14.8%), and vascular ulcers (14.1%). Patient outcomes included wound resolution in 77.2% of patients (with a mean healing time of 110.9 days), amputation in 14.1%, and wound-related death in 8.7% of patients. Non-healing cases (amputation/death) were predicted by several local factors including an initial depth greater than 1 cm, wound secretion, inflammatory base, and a maximum wound size. Systemic factors included most strongly clinically manifested atherosclerosis and its risk factors. Of the 110 swabs performed, 103 identified at least 1 bacterial genus. The dominant risk factor for a prolonged healing duration was bacterial infection. Wounds contaminated by Proteus or Pseudomonas had prolonged healing times of 87 days (p = 0.02) and 72 days (p = 0.045), respectively. Conclusions: The early identification of local and systemic risk factors contributes to the successful resolution of chronic wounds and a reduced duration of healing.

Nonwoven Releasing Propolis as a Potential New Wound Healing Method-A Review.

Wound healing poses a serious therapeutic problem. Methods which accelerate tissue regeneration and minimize or eliminate complications are constantly being sought. This paper is aimed at evaluation of the potential use of biodegradable polymer nonwovens releasing propolis as wound healing dressings, based on the literature data. Propolis is honeybee product with antioxidant, antibacterial, antifungal, anticancer, anti-inflammatory, analgesic, and regenerative properties. Controlled release of this substance throughout the healing should promote healing process, reduce the risk of wound infection, and improve aesthetic effect. The use of biodegradable aliphatic polyesters and polyester carbonates as a propolis carrier eliminates the problem of local drug administration and dressing changes. Well-known degradation processes and kinetics of the active substance release allows the selection of the material composition appropriate to the therapy. The electrospinning method allows the production of nonwovens that protect the wound against mechanical damage. Moreover, this processing technique enables adjusting product properties by modifying the production parameters. It can be concluded that biodegradable polymer dressings, releasing a propolis, may find potential application in the treatment of complicated wounds, as they may increase the effectiveness of treatment, as well as improve the patient's life quality.

THE EFFECT OF SULFASALAZINE AND 5-AMINOSALICYLIC ACID ON THE SECRETION OF INTERLEUKIN 8 BY HUMAN COLON MYOFIBROBLASTS.

Sulfasalazine (SAS) and its therapeutically active derivative--5-aminosalicylic acid (5-ASA) are used in the treatment of inflammatory bowel disease. 5-ASA mechanism of action on the one hand, involves the inhibition of the cyclooxygenase and lipoxygenase activity, and thus decrease of synthesis of prostaglandins, leukotrienes and free radicals, on the other hand, the suppression of the immune response in the intestinal mucosa. Myofibroblasts, which are located just below the basement membrane, are important element of the mucosa. Due to its secretory activity they may interact with other cells, including epithelial cells. Examining SAS and 5-ASA cytotoxic properties on human normal, colon subepithelial myofibroblasts (CSEMF) it was found that the first of these compounds in a concentration of 1 mM significantly reduced the number of these cells as compared to the control, while the latter exhibited an action at the 5-fold higher concentration (5 mM). Moreover, SAS concentration greater than 0.25 mM reduced IL-8 secretion by CSEMF, and 5-ASA had no effect in the tested range of concentrations, i.e., up to 7.5 mM.

Antiproliferative effect of valproic acid and 5,7-dimethoxycoumarin against A2058 human melanoma cells.

Melanoma is one of the most malignant tumors of a dangerous high incidence and high metastatic potential. It grows quickly and in an advanced stage is resistant to radio-, chemo- and immunotherapy, which makes it difficult to cure. Therefore, research efforts are focused on the development of new therapeutics or chemopreventive strategies. The aim of the study was to investigate whether the valproic acid and 5,7-dimethoxycoumarin have an antiproliferative activity against A2058 human melanoma cell line. Investigated compounds inhibited the proliferation of cells, however, no synergistic effect of their co-administration was observed.

The sugar 3-deoxy-d-manno-oct-2-ulosonic acid (Kdo) as a characteristic component of bacterial endotoxin -- a review of its biosynthesis, function, and placement in the lipopolysaccharide core.

The sugar 3-deoxy-d-manno-oct-2-ulosonic acid (Kdo) is a characteristic component of bacterial lipopolysaccharide (LPS, endotoxin). It connects the carbohydrate part of LPS with C6 of glucosamine or 2,3-diaminoglucose of lipid A by acid-labile α-ketosidic linkage. The number of Kdo units present in LPS, the way they are connected, and the occurrence of other substituents (P, PEtn, PPEtn, Gal, or ß-l-Ara4N) account for structural diversity of the inner core region of endotoxin. In a majority of cases, Kdo is crucial to the viability and growth of bacterial cells. In this paper, the biosynthesis of Kdo and the mechanism of its incorporation into the LPS structure, as well as the location of this unique component in the endotoxin core structures, have been described.

The pyrolytic profile of lyophilized and deep-frozen compact part of the human bone.

Background. Bone grafts are used in the treatment of nonunion of fractures, bone tumors and in arthroplasty. Tissues preserved by lyophilization or deep freezing are used as implants nowadays. Lyophilized grafts are utilized in the therapy of birth defects and bone benign tumors, while deep-frozen ones are applied in orthopedics. The aim of the study was to compare the pyrolytic pattern, as an indirect means of the analysis of organic composition of deep-frozen and lyophilized compact part of the human bone. Methods. Samples of preserved bone tissue were subjected to thermolysis and tetrahydroammonium-hydroxide- (TMAH-) associated thermochemolysis coupled with gas chromatography and mass spectrometry (Py-GC/MS). Results. Derivatives of benzene, pyridine, pyrrole, phenol, sulfur compounds, nitriles, saturated and unsaturated aliphatic hydrocarbons, and fatty acids (C12-C20) were identified in the pyrolytic pattern. The pyrolyzates were the most abundant in derivatives of pyrrole and nitriles originated from proteins. The predominant product in pyrolytic pattern of the investigated bone was pyrrolo[1,2-α]piperazine-3,6-dione derived from collagen. The content of this compound significantly differentiated the lyophilized graft from the deep-frozen one. Oleic and palmitic acid were predominant among fatty acids of the investigated samples. The deep-frozen implants were characterized by higher percentage of long-chain fatty acids than lyophilized grafts.

The chemical composition of endotoxin isolated from intestinal strain of Desulfovibrio desulfuricans.

Desulfovibrio desulfuricans anaerobes are constituents of human alimentary tract microflora. There are suggestions that they take part in the pathogenesis of periodontitis and some gastrointestinal inflammatory disorders, such as ulcerative colitis or Crohn's disease. Endotoxin is one of Gram-negative bacteria cellular components that influence these microorganisms pathogenicity. Endotoxin is a lipid-polisaccharide heteropolymer consisting of three elements: lipid A, core oligosaccharide, and O-specific polysaccharide, also called antigen-O. The biological activity of lipopolysaccharide (LPS) is determined by its structure. In this study, we show that rhamnose, fucose, mannose, glucose, galactose, heptose, and 2-keto-3-deoxyoctulosonic acid (Kdo) are constituents of D. desulfuricans endotoxin oligosaccharide core and O-antigen. Lipid A of these bacteria LPS is composed of glucosamine disaccharide substituted by 3-acyloxyacyl residues: ester-bound 3-(dodecanoyloxy)tetradecanoic, 3-(hexadecanoyloxy)tetradecanoic acid, and amide-bound 3-(tetradecanoyloxy)tetradecanoic acid.

Effects of 300 mT static magnetic field on IL-6 secretion in normal human colon myofibroblasts.

Intestinal subepithelial myofibroblasts play crucial role in the growth and development of the intestine. Colitis, small bowel injury, gastric ulcer disease and inflammatory bowel disease (IBD) accompany the increase of number of activated myofibroblasts. In the last few years, the increasing production of electromagnetic (EMF) and static magnetic fields (SMF), due to the expanding use of electronic devices in everyday life, has led to a number of studies on the effects of these fields on living organisms. EMF therapy, because of its anti-inflammatory properties, may be used in medicine in IBD treatment. This mechanism has not been elucidated yet. In the present work normal human colon myofibroblasts CCD-18Co were exposed to SMF with a flux density of 300 mT. After 24 h incubation TNF-alpha-dependent IL-6 secretion was determined with ELISA kit (RandD Systems).The influence of magnetic field and its effect on cell proliferation were determined with TOX-2 (In Vitro Toxicology Assay Kit XTT Based, TOX-2, Sigma) and CyQUANT NF cell proliferation assay kit (Molecular Probes). It was shown that SMF inhibited TNF-alpha-dependent IL-6 secretion. The observed effects were statistically significant and depended on the time of incubation. Moreover, SMF triggered cell proliferation whereas it did not alter cell viability. IL-6 belongs to pro-inflammatory cytokines family and plays a crucial role in IBD. Inhibition of IL-6 secretion by SMF and lack of its cytotoxic effect seem to be advantageous whilst SMF is implicated in the treatment of inflammatory diseases associated by increase in number of activated myofibroblasts.

Comparison of dissolution profiles of theophylline extended-release dosage forms.

Dissolution testing is a very important tool used to demonstrate the similarity between different formulations. Due to a narrow therapeutic range of theophylline, it is crucial to investigate the differences in the rate of release of this drug between the products. The aim of study was to compare the dissolution profiles of theophylline extended-release dosage forms available on Polish market: Theoplus, Theovent, Theospirex retard, and Euphyllin long. The investigation of theophylline release from tablets was performed by the basket apparatus type DT700 (Erweka). The difference and similarity factor was used to compare the obtained dissolution profiles. The obtained values showed that dissolution profiles of investigated formulations were not equivalent to each other. The tablets differed by the mechanism of drug release also.

Chemical composition of Desulfovibrio desulfuricans lipid A.

Lipopolysaccharides also called endotoxins are an integral component of the outer membrane of Gram-negative bacteria. When released from the bacterial surface, they interact with a host immune system, triggering excessive inflammatory response. Lipid A is the biologically most active part of endotoxin, and its activity is modulated by the quantity, quality and arrangement of its fatty acids. Desulfovibrio desulfuricans is sulfate-reducing, Gram-negative bacterium that is supposed to be opportunistic pathogens of humans and animals. In the present study, chemical composition of lipid A from various strains of D. desulfuricans was analyzed by gas chromatography/mass spectrometry. It was found that the fatty acid component of the lipid A contains dodecanoic, tetradecanoic, 3-hydroxytetradecanoic and hexadecanoic acids, and its carbohydrate core is composed of glucosamine. The analysis of 3-acyloxyacyl residue of the lipid A revealed the presence of amide-bound 3-(dodecanoyloxy)tetradecanoic and 3-(hexadecanoyloxy)tetradecanoic acids and ester-bound 3-(tetradecanoyloxy)tetradecanoic acid. It was concluded that both fatty acid and 3-acyloxyacyl residue profiles of the lipid A from the studied bacteria were similar to those of E. coli and S.enterica.

The role of Desulfovibrio desulfuricans lipopolysaccharides in modulation of periodontal inflammation through stimulation of human gingival fibroblasts.

OBJECTIVE: Periodontitis is a destructive disease which is likely to be the result of the activities of different microbial complexes. Recently, sulphate-reducing bacteria (SRB) have been detected in the oral cavity, and they have been found to be common inhabitants of sites showing periodontal destruction. The aim of study was to evaluate the influence of endotoxins of Desulfovibrio desulfuricans bacteria on human gingival fibroblast HGF-1 line. METHODS: The immunological response of gingival fibroblasts was evaluated by determination of their IL-6 and IL-8 secretion upon treatment with D. desulfuricans intestinal and type strain LPS, sodium butyrate (NaB) and IL-1beta. The amounts of cytokines were estimated by ELISA immunoassay. The influence of LPS and NaB on fibroblast proliferation was determined using the CyQUANT Cell Proliferation Assay Kit. RESULTS: No significant growth inhibition of cells exposed to LPS was observed, except for the culture growing in the presence of intestinal strain endotoxin at the highest concentration (100 microg/ml). The secretion of IL-6 and IL-8 by fibroblasts was increased by D. desulfuricans endotoxins. Cells stimulated with proinflammatory cytokine 1L-1beta showed very high levels of both cytokines secretion. The release of IL-6 and IL-8 by cells in response to LPS and 1L-1beta was modulated by butyric acid. CONCLUSIONS: The observed response of gingival fibroblasts to stimulation by endotoxin suggests that D. desulfuricans can be involved in the pathogenesis of periodontitis. Moreover, butyrate present in the oral cavity seems to have immunoregulatory effect on cytokine production by gingival fibroblasts under physiological conditions and during microbe-induced inflammation.

Interstrain diversity of 2-keto-3-deoxyoctonate content in lipopolysaccharides of Desulfovibrio desulfuricans.

Bacteria of Desulfovibrio desulfuricans species are Gram-negative, anaerobic rods selectively reducing sulphates and colonizing oxygen-free ecosystems. They are ubiquitous in the natural environment and have been also found to reside in the human digestive tract. They are suggested to be involved in the pathogenesis ofulcerative colitis and Crohn's disease. The D. desulfuricans wild strains were isolated from feces and bioptate of patients suffering from various digestive tract disorders. LPSs were isolated from the wild enteric strains and soil type strain La 2226 of D. desulfuricans and analyzed in terms of their 2-keto-3-deoxyoctulosonic acid (Kdo) component content. The obtained spectrophotometric data indicate that Kdo content is characteristic of each of the investigated strains and it ranges from 0.48% to 2.86% (w/w) of the total LPS mass. Statistically significant interstrain differences of Kdo quantity seem to suggest the differences in the O-antigen content. Comparative analysis of Kdo content in LPSs of D. desulfuricans strains in relation to that of the reference endotoxin from Salmonella spp. allows us to suggest that D. desulfuricans bacteria possess O-antigen polysaccharides composed of diverse number of carbohydrate units.

[The structural diversity of lipid A from gram-negative bacteria]. / Heterogennosc strukturalna lipidu A bakterii Gram-ujemnych.

The majority of Gram-negative bacteria are pathogenic to humans and animals. Lipopolysaccharide (LPS) is the most biologically active component of these microorganisms. This compound is also called endotoxin to emphasize its negative impact on a macroorganism. Lipid A, one of the three structural components of the LPS molecule, is responsible for the pathophysiological effects associated with Gram-negative bacteria infections. Although lipid A is considered the conservative component of endotoxin, differences in its structure among species and even strains may occur. These differences concern the type of aminosugars, the degree of substitution of the disaccharide core by fatty acids, phosphate, and/or ethanolamine, and also the type, quantity, and distribution of fatty acids. The lipid A saccharide backbone of the majority of Gram-negative bacteria consists of two glucosamine units in beta (1-->6) glycosidic linkage. Amino groups (at positions 2 and 2') and hydroxy groups (at positions 3 and 3') of glucosamines are commonly substituted by 3-hydroxyfatty acids, most often by 3-hydroxytetradecanoic acid. Other fatty acids (usually saturated, unbranched) are ester-linked to hydroxyacids by their hydroxy group. In lipid A of different microorganisms there is a high diversity of fatty acids, from mirystic (tetradecanoic, 14:0) and lauric (dodecanoic, 12:0) acids and their hydroxylated derivatives to such unique structures as cis-11-octadecenoic acid (Rhodospirillum salinarum 40), 3-hydroxy-5-dodecenoic acid (Phenylobacterium immobile), and iso-2,3-dihydroxytetradecanoic acid (Legionella pneumophila). The saccharide core of some bacterial lipid A may consist of sugars different from glucosamine, e.g., 2,3-diamino-2,3-dideoxy-D-glucose. Other substituents of this part of LPS, besides phosphate groups and ethanolamine, are beta -mannopyranose, 4-aminoarabinose, galacturonic acid, and glycine. Therefore, lipid A, though considered the relatively conservative component of endotoxin, reveals relatively large structural diversity, which influences the variety of LPS biological activity.

[Methods of lipopolysaccharide component derivation in evaluating their structures by chromatographic techniques]. / Metody derywatyzacji komponentów lipopolisacharydów w ocenie ich struktury chemicznej technikami chromatograficznymi.

The pathogenicity of Gram-negative bacteria is mediated mainly by the outer membrane-associated lipid-polysaccharide structure, called endotoxin or lipopolysaccharide (LPS). This structure consists of three parts, O-antigen, core oligosaccharide, and lipid A, which differ in chemical composition and biological properties. Gas-liquid chromatography coupled with mass spectrometry (GLC/MS) is commonly used to determine LPS structure. The carbohydrate profiles of endotoxins are analyzed after derivation to alditol acetates or partially methylated alditol acetates, which enables one to establish the site of glycosidic linkages. The chromatographic analysis of LPS lipid components requires their modification to ester derivatives, most frequently to methyl esters.