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1.
Front Microbiol ; 12: 757215, 2021.
Article in English | MEDLINE | ID: mdl-34733265

ABSTRACT

The cationic proinflammatory cytokine Interleukin 26 (IL-26) shows antibacterial activity and inhibits the replication of cytomegalovirus and hepatitis C virus. This study evaluates the early microbicidal activities of IL-26 against major bacterial species including multi-resistant variants and Candida albicans. Recombinant IL-26 was bacterially expressed and studied for its microbicidal effects in culture. We show that IL-26 has strong 90% bactericidal activities against Enterococcus faecalis, Enterococcus faecium, Staphylococcus aureus, and Acinetobacter baumannii. Similarly, IL-26 sensitivity was also detectable in vancomycin-resistant Enterococcus species, methicillin-resistant S. aureus, and carbapenem-resistant A. baumannii clinical isolates. Additionally, a significant, albeit weak fungicidal effect against Candida albicans was observed. Activities against Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa were not detectable. The proinflammatory cytokine and kinocidin IL-26 shows strong bactericidal activities against A. baumannii and, almost selectively, against Gram-positive bacteria.

2.
APMIS ; 128(6): 451-462, 2020 Jun.
Article in English | MEDLINE | ID: mdl-32358920

ABSTRACT

Bacteria and viruses were analysed in the upper respiratory tract of symptomatic pig farmers and their domestic pigs. Eighty six human nasal and 495 (50 pools) porcine snout swabs were collected in Schleswig-Holstein, Germany. Staphylococcus (S.) aureus (62.8%, 54/86), human rhino- and coronaviruses (HRV, 29.1%, 25/86; HCoV, 16.3%, 14/86) were frequently detected in humans, while Haemophilus parasuis (90.0%, 45/50), Mycoplasma hyorhinis (78.6%, 11/14), Enterovirus G (EV-G, 56.0%, 28/50) and S. aureus (36.0%, 18/50), respectively, were highly prevalent in pigs. The detection of S. aureus in human follow-up samples indicates a carrier status. The methicillin-resistant phenotype (MRSA) was identified in 33.3% (18/54) of nasal swabs and in one of 18 (5.6%) pooled snout swabs that were tested positive for S. aureus. Strains were indicative of the livestock-associated clonal complex CC398, with t011 being the most common staphylococcal protein A type. Enterobacterales and non-fermenters were frequently isolated from swabs. Their detection in follow-up samples suggests a carrier status. All were classified as being non-multiresistant. There was no example for cross-species transmission of viruses. In contrast, transmission of S. aureus through occupational contact to pigs seems possible. The study contributes to the 'One Health' approach.


Subject(s)
Respiratory Tract Infections/microbiology , Respiratory Tract Infections/virology , Staphylococcal Infections/veterinary , Sus scrofa/microbiology , Sus scrofa/virology , Swine Diseases/epidemiology , Animals , Carrier State , Humans , Livestock , Methicillin-Resistant Staphylococcus aureus/genetics , Microbial Sensitivity Tests , Nasal Mucosa/microbiology , Nasal Mucosa/virology , Occupational Diseases/microbiology , Prevalence , Respiratory Tract Infections/epidemiology , Staphylococcal Infections/epidemiology , Staphylococcal Infections/transmission , Swine , Swine Diseases/microbiology , Swine Diseases/transmission , Swine Diseases/virology , Virus Diseases/epidemiology , Virus Diseases/transmission , Virus Diseases/veterinary
3.
Biochim Biophys Acta Biomembr ; 1862(8): 183273, 2020 08 01.
Article in English | MEDLINE | ID: mdl-32171739

ABSTRACT

Antimicrobial peptides (AMPs) play an important role in the host defense against various microbes. One of the most efficient human AMPs is the human beta defensin-3 (hBD-3) which is produced by, e.g. keratinocytes and lung epithelial cells. However, the structure-function relationship for AMPs and in particular for defensins with their typical three disulfide bonds is still poorly understood. In this study the importance of the three disulfide bonds for the activity of the AMPs is investigated with biological assays and with biophysical experiments utilizing different membrane reconstitution systems. The activities of natural hBD-3, hBD-3-c (cyclic variant with one disulfide bond), and hBD-3-l (linear variant without disulfide bonds) and fragments thereof were tested against specific Gram-negative bacteria. Furthermore, hemolytic and cytotoxic activities were analyzed as well as the potency to neutralize immune cell stimulation of lipopolysaccharide (LPS). Experiments using reconstituted lipid matrices composed of phospholipids or LPS purified from the respective Gram-negative bacteria, showed that the membrane activity of all three hBD-3 peptides is decisive for their capability to kill bacteria and to neutralize LPS. In most of the test systems the linear hBD-3-l showed the highest activity. It was also the only peptide significantly active against polymyxin B-resistant Proteus mirabilis R45. However, the stability of hBD-3 against protease activity decreases with decreasing number of disulfide bonds. This study demonstrates that the refining of AMP structures can generate more active compounds against certain strains.


Subject(s)
Antimicrobial Cationic Peptides/chemistry , Bacterial Infections/drug therapy , Gram-Negative Bacteria/drug effects , beta-Defensins/chemistry , Amino Acid Sequence/genetics , Antimicrobial Cationic Peptides/pharmacology , Bacterial Infections/microbiology , Disulfides/chemistry , Drug Resistance, Bacterial/drug effects , Epithelial Cells/drug effects , Epithelial Cells/microbiology , Gram-Negative Bacteria/genetics , Gram-Negative Bacteria/pathogenicity , Humans , Keratinocytes/drug effects , Keratinocytes/microbiology , Lipopolysaccharides/antagonists & inhibitors , Lung/drug effects , Lung/microbiology , Polymyxin B/adverse effects , Polymyxin B/pharmacology , Protein Domains/drug effects , Proteus mirabilis/drug effects , Proteus mirabilis/pathogenicity , Structure-Activity Relationship , beta-Defensins/pharmacology
4.
Microbiome ; 6(1): 37, 2018 02 20.
Article in English | MEDLINE | ID: mdl-29458422

ABSTRACT

BACKGROUND: Multidrug-resistant bacteria represent a substantial global burden for human health, potentially fuelled by migration waves: in 2015, 476,649 refugees applied for asylum in Germany mostly as a result of the Syrian crisis. In Arabic countries, multiresistant bacteria cause significant problems for healthcare systems. Currently, no data exist describing antibiotic resistances in healthy refugees. Here, we assess the microbial landscape and presence of antibiotic resistance genes (ARGs) in refugees and German controls. To achieve this, a systematic study was conducted in 500 consecutive refugees, mainly from Syria, Iraq, and Afghanistan and 100 German controls. Stool samples were subjected to PCR-based quantification of 42 most relevant ARGs, 16S ribosomal RNA gene sequencing-based microbiota analysis, and culture-based validation of multidrug-resistant microorganisms. RESULTS: The fecal microbiota of refugees is substantially different from that of resident Germans. Three categories of resistance profiles were found: (i) ARGs independent of geographic origin of individuals comprising BIL/LAT/CMA, ErmB, and mefE; (ii) vanB with a high prevalence in Germany; and (iii) ARGs showing substantially increased prevalences in refugees comprising CTX-M group 1, SHV, vanC1, OXA-1, and QnrB. The majority of refugees carried five or more ARGs while the majority of German controls carried three or less ARGs, although the observed ARGs occurred independent of signatures of potential pathogens. CONCLUSIONS: Our results, for the first time, assess antibiotic resistance genes in refugees and demonstrate a substantially increased prevalence for most resistances compared to German controls. The antibiotic resistome in refugees may thus require particular attention in the healthcare system of host countries.


Subject(s)
Bacteria , Drug Resistance, Multiple, Bacterial/genetics , Refugees/statistics & numerical data , Afghanistan , Bacteria/drug effects , Bacteria/genetics , Bacteria/isolation & purification , Feces/microbiology , Germany , Humans , Iraq , Microbiota/drug effects , Microbiota/genetics , RNA, Ribosomal, 16S/genetics , Syria
5.
J Craniomaxillofac Surg ; 45(11): 1868-1874, 2017 Nov.
Article in English | MEDLINE | ID: mdl-28927956

ABSTRACT

AIM: The aim of the present study was to demonstrate the possible relation between periodontal health status and infections associated with osteosynthesis materials (OMs) used in the oral and maxillofacial reconstruction. MATERIAL AND METHODS: The study group consisted of 32 individuals which were subdivided into two groups regarding their PSI scores. After the removal of the osteosynthesis plates, microbial colonization was assessed via microbiological cultivation, fluorescence microscopy and scanning electron microscopy. In addition, samples obtained from gingival crevicular fluids were investigated by fluorescence microscopy. RESULTS: A total of 118 osteosynthesis plates were examined. 8.5% (n = 10) of the plates were associated at least one of the clinical signs of infection. There was a positive correlation between periodontal disease and clinical signs of infection (p = 0.022). Patients with infection signs also had a higher number of smoking history (pack years, p = 0.010). Intraorally placed osteosynthesis plates showed wide range of bacterial colonizations compared to extraorally inserted osteosynthesis materials (p = 0.004). CONCLUSION: Patients with poor periodontal health might be potential candidates for OM related infections. Early removal of OMs in patients with poor periodontal health status and/or heavy smokers would have clinical benefits. In addition, preferation of extraoral access to the fracture line might decrease the possibility of plate related infections.


Subject(s)
Bone Plates , Fracture Fixation, Internal/instrumentation , Maxillofacial Injuries/surgery , Oral Health , Periodontitis/microbiology , Prosthesis-Related Infections/microbiology , Skull Fractures/surgery , Adolescent , Adult , Aged , Disease Susceptibility , Female , Gingival Crevicular Fluid/microbiology , Health Status , Humans , Male , Middle Aged , Risk Factors , Smoking/adverse effects , Young Adult
6.
J Infect ; 71(2): 188-99, 2015 Aug.
Article in English | MEDLINE | ID: mdl-25896094

ABSTRACT

Antimicrobial peptides (AP) are important components of the innate immune system. Our previous work revealed a higher mortality rate and up-regulation of proinflammatory gene expression as well as glial cell activation in cathelicidin-related antimicrobial peptide (CRAMP)-deficient mice after bacterial meningitis. However, the influence of CRAMP application on the progression of inflammation and its impact on mortality after bacterial meningitis remains unknown. To assess the effects of continuous CRAMP exposure in the brain, C57BL/6 wildtype mice were given intracerebroventricular infusion of CRAMP to investigate the effects on mortality, glial cell activation and inflammation in a mouse model of pneumococcal meningitis using immunohistochemistry and realtime RT-PCR. Our results revealed a decrease of mortality after CRAMP infusion. The intrathecal CRAMP infusion after pneumococcal meningitis resulted in a decreased mRNA expression of pro-inflammatory cytokines, whereas the immune responses including the expression of pattern recognition receptors and chemokines were increased in bacterial meningitis. Taken together, the results support the important role of CRAMP as part of the innate immune response against pathogens in bacterial CNS infections. The APs may be a promising approach for the development of an adjuvant therapy for bacterial meningitis.


Subject(s)
Antimicrobial Cationic Peptides/therapeutic use , Meningitis, Pneumococcal/drug therapy , Animals , Brain/pathology , Cytokines/biosynthesis , Disease Models, Animal , Immunohistochemistry , Male , Mice, Inbred C57BL , Real-Time Polymerase Chain Reaction , Survival Analysis , Treatment Outcome
8.
Antimicrob Agents Chemother ; 57(7): 2955-66, 2013 Jul.
Article in English | MEDLINE | ID: mdl-23587944

ABSTRACT

Hydramacin-1 (HM1) from the metazoan Hydra exerts antimicrobial activity against a wide range of bacterial strains. Notably, HM1 induces the aggregation of bacterial cells, accompanied by precipitation. To date, the proposed mechanism of peptide-lipid interaction, termed the barnacle model, has not been described on the molecular level. Here, we show by biochemical and biophysical techniques that the lipid-peptide interactions of HM1 are initiated by electrostatic and hydrophobic effects, in particular, by tryptophan and neighboring polar amino acid residues that cause an interfacial localization of the peptide between two self-contained lipid bilayers. The high binding constants of HM1 upon lipid interaction are in the range of other potent antimicrobial peptides, e.g., magainin, and can be reasonably explained by two distinct epitopes on the surface of the peptide's global structure, which both contain SWT(K/R) motifs. The residues of this motif favor localization of the peptide in the head group region of phospholipid bilayers up to a penetration depth of 4 Å and a minor participation of the lipids' hydrocarbon regions. Our results expand the knowledge about the molecular modes of action antimicrobial peptides use to tackle their target cells. Furthermore, the aggregation of living bacteria by HM1 was observed for a broad range of Gram-positive and Gram-negative bacteria. Therefore, the detailed view of peptide-lipid interactions described by the barnacle model consolidates it among the established models.


Subject(s)
Anti-Bacterial Agents/pharmacology , Antimicrobial Cationic Peptides/pharmacology , Bacteria/drug effects , Cell Membrane/metabolism , Lipid Bilayers/metabolism , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/metabolism , Antimicrobial Cationic Peptides/chemistry , Antimicrobial Cationic Peptides/metabolism , Bacteria/metabolism , Burkholderia cepacia/drug effects , Burkholderia cepacia/metabolism , Cell Membrane/drug effects , Escherichia coli/drug effects , Escherichia coli/metabolism , Hydrophobic and Hydrophilic Interactions , Lipid Bilayers/chemistry , Lipids , Protein Binding , Proteus mirabilis/drug effects , Proteus mirabilis/metabolism , Staphylococcus aureus/drug effects , Staphylococcus aureus/metabolism , Staphylococcus haemolyticus/drug effects , Staphylococcus haemolyticus/metabolism , Staphylococcus hominis/drug effects , Staphylococcus hominis/metabolism , Static Electricity , Streptococcus pneumoniae/drug effects , Streptococcus pneumoniae/metabolism
9.
Mater Sci Eng C Mater Biol Appl ; 33(4): 2367-75, 2013 May 01.
Article in English | MEDLINE | ID: mdl-23498271

ABSTRACT

A silver containing coating used in the human body, e.g., on an implant should be both effectively antimicrobial and non-cytotoxic to human cells. It is generally believed that the biologic effect originates from silver ions released from the coating. Nanocomposites with well controlled Ag filling factor were prepared by co-sputtering, and the silver surface concentration and the silver release were determined by XPS and ICP-MS, respectively. Here we show that only a small therapeutic window exists for dissolved silver but the therapeutic window is largely increased at the surface. While the toxicity observed for mammalian cells in contact with the bioactive Ag/TiO2 nanocomposite surface and for silver ions in solution is rather similar the antimicrobial activity is drastically enhanced at the surface. A model is proposed to explain the strong increase of the antimicrobial activity at the surface. The present results not only question well-established tests for antimicrobial activity but they are also important for the design of antimicrobial coatings, e.g., for biomedical devices.


Subject(s)
Coated Materials, Biocompatible/pharmacology , Nanocomposites/chemistry , Silver/pharmacology , Titanium/pharmacology , Anti-Bacterial Agents/pharmacology , Bacterial Adhesion/drug effects , Cell Count , Cell Death/drug effects , Dermis/cytology , Escherichia coli/drug effects , Escherichia coli/growth & development , Fibroblasts/cytology , Fibroblasts/drug effects , Green Fluorescent Proteins/metabolism , Humans , Microbial Sensitivity Tests , Nanocomposites/ultrastructure , Photoelectron Spectroscopy , Silver Nitrate/pharmacology , Solutions , Spectrophotometry, Atomic
10.
Infect Immun ; 81(5): 1788-97, 2013 May.
Article in English | MEDLINE | ID: mdl-23478321

ABSTRACT

The expression and function of psoriasin in the brain have been insufficiently characterized. Here, we show the induction of psoriasin expression in the central nervous system (CNS) after bacterial and viral stimulation. We used a pneumococcal meningitis in vivo model that revealed S100A15 expression in astrocytes and meningeal cells. These results were confirmed by a cell-based in vivo assay using primary rat glial and meningeal cell cultures. We investigated psoriasin expression in glial and meningeal cells using polyinosinic-polycytidylic acid, a synthetic analog of double-stranded RNA that mimics viral infection. Furthermore, previous results showed that antimicrobial peptides have not only bactericidal but also immunomodulatory functions. To test this statement, we used recombinant psoriasin as a stimulus. Glial and meningeal cells were treated with recombinant psoriasin at concentrations from 25 to 500 ng/ml. Treated microglia and meningeal cells showed phosphorylation of the extracellular signal-regulated kinase 1 (ERK1)/ERK2 (ERK1/2) signal transduction pathway. We demonstrated that this activation of ERK depends on RAGE, the receptor for advanced glycation end products. Furthermore, microglia cells treated with recombinant psoriasin change their phenotype to an enlarged shape. In conclusion, our results indicate an occurrence of psoriasin in the brain. An involvement of psoriasin as an antimicrobial protein that modulates the innate immune system after bacterial or viral stimulation is possible.


Subject(s)
Brain/metabolism , Meningitis, Bacterial/metabolism , S100 Proteins/physiology , Analysis of Variance , Animals , Astrocytes/metabolism , Disease Models, Animal , Humans , MAP Kinase Signaling System/physiology , Neuroglia/metabolism , RNA, Messenger/metabolism , Rats , Rats, Wistar , S100 Calcium Binding Protein A7 , S100 Proteins/cerebrospinal fluid , S100 Proteins/metabolism
11.
Clin Oral Investig ; 17(4): 1135-42, 2013 May.
Article in English | MEDLINE | ID: mdl-22961460

ABSTRACT

OBJECTIVE: Information about the spectrum of microorganisms in the intraimplant cavities of two-piece dental implants is scarce. The purpose of this study was to assess the intraimplant microflora of two-piece dental implants by conventional biochemical testing, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), and 16 s rDNA gene sequencing. MATERIALS AND METHODS: Ten patients (six men and four women; average age = 66.7 years; age range = 58-78 years) received 35 two-piece titanium implants carrying ball attachments. Biofilm sampling was performed with sterile microbrushes, and nonadherent microbial samples were obtained by injection and reuptake of predefined volumes of NaCl solution. The samples were cultured and analyzed by conventional biochemical testing, MALDI-TOF MS, and 16 s rDNA gene sequencing. RESULTS: Of the 103 species detected, 27 and 33 were identified only in the biofilm and nonadherent microbial samples, respectively. Forty-three species were identified in both types of samples. CONCLUSIONS: Two-piece dental implants harbored a broad spectrum of gram-positive and gram-negative aerobes and anaerobes, especially rods and cocci. CLINICAL RELEVANCE: These findings confirm bacterial translocation from the oral cavity to intraimplant cavities. Microbiological methods as used in this study are necessary to reveal the complete vital microflora of intraimplant cavities.


Subject(s)
Dental Implant-Abutment Design , Dental Implants/microbiology , Microbiota , Tooth Socket/microbiology , Aged , Bacteria, Aerobic/genetics , Bacteria, Anaerobic/genetics , Biofilms , Colony Count, Microbial , DNA, Bacterial/analysis , DNA, Bacterial/genetics , Female , Gram-Negative Aerobic Rods and Cocci/genetics , Gram-Negative Anaerobic Cocci/genetics , Gram-Negative Anaerobic Straight, Curved, and Helical Rods/genetics , Gram-Positive Cocci/genetics , Gram-Positive Rods/genetics , Humans , Male , Middle Aged , Sequence Analysis, DNA , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
12.
Gut ; 62(4): 520-30, 2013 Apr.
Article in English | MEDLINE | ID: mdl-22442160

ABSTRACT

OBJECTIVE: Cathepsin K is a lysosomal cysteine protease that has pleiotropic roles in bone resorption, arthritis, atherosclerosis, blood pressure regulation, obesity and cancer. Recently, it was demonstrated that cathepsin K-deficient (Ctsk(-/-) ) mice are less susceptible to experimental autoimmune arthritis and encephalomyelitis, which implies a functional role for cathepsin K in chronic inflammatory responses. Here, the authors address the relevance of cathepsin K in the intestinal immune response during chronic intestinal inflammation. DESIGN: Chronic colitis was induced by administration of 2% dextran sodium sulphate (DSS) in distilled water. Mice were assessed for disease severity, histopathology and endoscopic appearance. Furthermore, DSS-exposed Ctsk(-/-) mice were treated by rectal administration of recombinant cathepsin K. Intestinal microflora was assessed by real-time PCR and 16srDNA molecular fingerprinting of ileal and colonic mucosal and faecal samples. RESULTS: Using Ctsk(-/-) mice, the authors demonstrate a protective role of cathepsin K against chronic DSS colitis. Dissecting the underlying mechanisms the authors found cathepsin K to be present in intestinal goblet cells and the mucin layer. Furthermore, a direct cathepsin K-mediated bactericidal activity against intestinal bacteria was demonstrated, which potentially explains the alteration of intestinal microbiota observed in Ctsk(-/-) mice. Rectal administration of recombinant cathepsin K in DSS-treated Ctsk(-/-) mice ameliorates the severity of intestinal inflammation. CONCLUSION: These data identify extracellular cathepsin K as an intestinal antibacterial factor with anti-inflammatory potential and suggest that topical administration of cathepsin K might provide a therapeutic option for patients with inflammatory bowel disease.


Subject(s)
Cathepsin K/pharmacology , Colitis/drug therapy , Colitis/microbiology , Animals , Blotting, Western , Cathepsin K/metabolism , Colitis/chemically induced , Colitis/pathology , Dextran Sulfate , Disease Models, Animal , Endoscopy, Gastrointestinal , Enzyme-Linked Immunosorbent Assay , Immunohistochemistry , Mice , Mice, Inbred C57BL , Real-Time Polymerase Chain Reaction
13.
J Craniomaxillofac Surg ; 41(4): 321-6, 2013 Jun.
Article in English | MEDLINE | ID: mdl-23199627

ABSTRACT

The fight against hospital-acquired infections involving antibiotic-resistant microorganisms has become of critical concern to surgeons worldwide. In addition to the development of new effective antibiotic chemotherapy, exploration of 'forgotten' topical antibacterial agents from the pre-antibiotic era has recently gained new attention. We report the promising efficacy of plant-derived antiseptic oils used in traditional aboriginal and south-east Asian treatments such as Lemongrass, Eucalyptus and Tea Tree Oil in the inhibition of clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus (VRE), multi-resistant Pseudomonas aeruginosa, ESBL-producing Escherichia coli and Klebsiella pneumoniae in the in-vitro setting. Large consistent zones of inhibition were observed for all three plant-derived oils tested in an agar diffusion test. The commonly used antibacterial agents chlorhexidine 0.1%, and ethanol (70%), and standard olive oil consistently demonstrated notably lower or no efficacy in regard to growth inhibition of strains. Notably, Lemongrass oil proved to be particularly active against gram-positive bacteria, while Tea Tree oil showed superior inhibition of gram-negative microorganisms. As proven in vitro, plant-derived antiseptic oils may represent a promising and affordable topical agent to support surgical treatment against multi-resistant and hospital-acquired infections.


Subject(s)
Anti-Infective Agents, Local/pharmacology , Drug Resistance, Multiple, Bacterial/drug effects , Enterococcus/drug effects , Escherichia coli/drug effects , Klebsiella pneumoniae/drug effects , Methicillin-Resistant Staphylococcus aureus/drug effects , Plant Oils/pharmacology , Pseudomonas aeruginosa/drug effects , Chlorhexidine/pharmacology , Cymbopogon , Disinfectants/pharmacology , Ethanol/pharmacology , Eucalyptus , Eucalyptus Oil , Humans , Immunodiffusion , Monoterpenes/pharmacology , Oils, Volatile/pharmacology , Olive Oil , Phytotherapy/methods , Tea Tree Oil/pharmacology , Terpenes/pharmacology , Vancomycin Resistance , beta-Lactamases/drug effects
14.
Arthritis Res Ther ; 14(5): R203, 2012 Oct 17.
Article in English | MEDLINE | ID: mdl-23031229

ABSTRACT

INTRODUCTION: In granulomatosis with polyangiitis (GPA), a complex autoimmune small-vessel vasculitis frequently associated with chronic necrotizing inflammation of the nasal mucosa, elevated nasal Staphylococcus (S.) aureus carrier rates are a risk factor for relapse. As cytokines are primarily involved in the regulation of defense against potentially pathogenic microorganisms, the aim of this study was to compare healthy individuals and GPA patients with respect to their baseline cytokine expression of nasal epithelial cells (NEC), which form the first barrier against such triggers. The ability of S. aureus to influence the nasal microenvironment's cytokine secretion was assessed by exemplary stimulation experiments. METHODS: Baseline expression of 19 cytokines of primary NEC of GPA patients and normal controls (NC) was quantified by a multiplex cytokine assay. Stimulation experiments were performed with supernatants of S. aureus and expression of interleukin-8 was determined by ELISA. RESULTS: In GPA, an altered pattern of baseline cytokine expression with significantly up-regulated G-CSF and reduced interleukin (IL)-8 concentrations was observed. Both NEC of GPA patients and NC responded to stimulation with S. aureus, but GPA patients displayed a significantly lower IL-8 secretion and a diminished dynamic range of response towards the stimulus. CONCLUSIONS: The data presented underline the hypothesis of a disturbed epithelial nasal barrier function in GPA. The dysregulated baseline expression of G-CSF and IL-8 and the reduced response to microbial stimulation may facilitate changes in the composition of the nasal flora and favour an imbalanced inflammatory response, which might be relevant for the disease course.


Subject(s)
Cytokines/metabolism , Granulomatosis with Polyangiitis/metabolism , Nasal Mucosa/metabolism , Nasal Mucosa/microbiology , Adolescent , Adult , Aged , Case-Control Studies , Cells, Cultured , Cellular Microenvironment , Epithelial Cells/metabolism , Epithelial Cells/microbiology , Epithelial Cells/pathology , Female , Granulocyte Colony-Stimulating Factor/metabolism , Granulomatosis with Polyangiitis/microbiology , Granulomatosis with Polyangiitis/pathology , Humans , Interleukin-8/metabolism , Male , Middle Aged , Nasal Mucosa/pathology , Staphylococcus aureus/physiology , Young Adult
15.
Arch Dermatol Res ; 304(8): 633-7, 2012 Oct.
Article in English | MEDLINE | ID: mdl-22955175

ABSTRACT

Staphylococcus aureus is a major human pathogen causing cutaneous infections to life-threatening bacteremia. These infections are often caused by strains derived from the own microflora suggesting that a disturbed epidermal barrier may promote invasion of S. aureus. Antimicrobial peptides and proteins (AMP) such as human beta-defensin-3 and RNase 7 contribute to control the colonization of S. aureus on the skin surface. This leads to the hypothesis that strains with a decreased susceptibility toward skin-derived AMP may better overcome the innate cutaneous defence barrier increasing the possibility of invading into the blood stream. To address this hypothesis we determined whether S. aureus strains from bacteremia patients are less susceptible to various skin-derived AMP than strains from healthy carriers. No differences in the AMP-killing activity against bacteremia-derived S. aureus and control strains were detected suggesting that the onset of S. aureus bacteremia is not based on the varying susceptibilities against skin-derived AMP.


Subject(s)
Antimicrobial Cationic Peptides/immunology , Ribonucleases/immunology , Skin/immunology , Staphylococcal Infections/immunology , Staphylococcus aureus/drug effects , beta-Defensins/immunology , Adolescent , Adult , Aged , Antimicrobial Cationic Peptides/pharmacology , Apoptosis/genetics , Bacteremia/immunology , Bacteremia/microbiology , Drug Resistance, Bacterial , Female , Humans , Male , Microbial Sensitivity Tests , Middle Aged , Ribonucleases/pharmacology , Skin/microbiology , Species Specificity , Staphylococcus aureus/genetics , Staphylococcus aureus/pathogenicity , Young Adult , beta-Defensins/pharmacology
16.
J Biol Chem ; 287(17): 14246-58, 2012 Apr 20.
Article in English | MEDLINE | ID: mdl-22396551

ABSTRACT

The tertiary structures of theromacin and neuromacin confirmed the macin protein family as a self-contained family of antimicrobial proteins within the superfamily of scorpion toxin-like proteins. The macins, which also comprise hydramacin-1, are antimicrobially active against Gram-positive and Gram-negative bacteria. Despite high sequence identity, the three proteins showed distinct differences with respect to their biological activity. Neuromacin exhibited a significantly stronger capacity to permeabilize the cytoplasmic membrane of Bacillus megaterium than theromacin and hydramacin-1. Accordingly, it is the only macin that displays pore-forming activity and that was potently active against Staphylococcus aureus. Moreover, neuromacin and hydramacin-1 led to an aggregation of bacterial cells that was not observed with theromacin. Analysis of the molecular surface properties of macins allowed confirmation of the barnacle model as the mechanistic model for the aggregation effect. Besides being antimicrobially active, neuromacin and theromacin, in contrast to hydramacin-1, were able to enhance the repair of leech nerves ex vivo. Notably, all three macins enhanced the viability of murine neuroblastoma cells, extending their functional characteristics. As neuromacin appears to be both a functional and structural chimera of hydramacin-1 and theromacin, the putative structural correlate responsible for the nerve repair capacity in leech was located to a cluster of six amino acid residues using the sequence similarity of surface-exposed regions.


Subject(s)
Antimicrobial Cationic Peptides/chemistry , Antimicrobial Cationic Peptides/pharmacology , Amino Acid Sequence , Animals , Base Sequence , Cloning, Molecular , Disulfides/chemistry , Humans , Leeches , Magnetic Resonance Spectroscopy/methods , Molecular Sequence Data , Neurons/metabolism , Protein Conformation , Protein Structure, Tertiary , Salts/chemistry , Scattering, Radiation , Sequence Homology, Amino Acid
17.
Platelets ; 23(3): 217-23, 2012.
Article in English | MEDLINE | ID: mdl-21913811

ABSTRACT

Platelet-rich plasma (PRP) is a potent agent that improves soft tissue and bone healing. By the release of growth factors and cytokines, PRP is believed to locally boost physiologic healing processes. Recently, antimicrobial activity of PRP has been demonstrated against S. aureus strains. Major scientific effort is being put into the understanding and prevention of infections i.e. by delivery of antimicrobial substances. In previous studies we showed the ideal antibacterial activity-profile of the human beta-defensin 2 (hBD-2) for orthopaedic infections and therefore hypothesized that hBD-2 may be the effector of antimicrobial platelet action. Platelet concentrates were produced from human platelet phresis obtained from a hospital blood bank. They were screened by immunohistochemistry, Western Blot and ELISA for the human beta defensin-2. In vitro susceptibility to PRP was investigated by a standard disc diffusion test with or without pre-incubation of PRP with anti-hBD-2 antibody. SPSS statistical software was used for statistical analysis. PRP contains hBD-2 470 pg/10(9) platelets or 1786 pg/ml, respectively, (ELISA), which was confirmed by immunohistochemistry and Western Blot. In antimicrobial testing, PRP demonstrates effective inhibition of E. coli, B. megaterium, P. aeruginosa, E. faecalis and P. mirabilis. With this study we confirm the previously reported antimicrobial action of platelet concentrates i.e. PRP. In opposition to previously reported effects against gram positive bacteria our study focuses on gram negative and less common gram positive bacteria that do frequently cause clinical complications. We provide a possible molecular mechanism at least for E. coli and P. mirabilis for this effect by the detection of an antimicrobial peptide (hBD-2). This study may advocate the clinical use of PRP by highlighting a new aspect of platelet action.


Subject(s)
Anti-Infective Agents/metabolism , Blood Platelets/metabolism , Gram-Negative Bacteria/growth & development , Gram-Positive Bacteria/growth & development , beta-Defensins/metabolism , Blood Platelets/cytology , Blood Platelets/microbiology , Female , Humans , Immunohistochemistry , Male
18.
Injury ; 43(2): 137-42, 2012 Feb.
Article in English | MEDLINE | ID: mdl-21561617

ABSTRACT

Today multiple trauma still is associated with a high mortality. Although severe open fractures and wounds can give rise to local infections and sepsis, the overall infection rate of multiply injured patients is surprisingly low. We have investigated serum of multiply injured patients with respect to antibacterial properties and screened for host defence peptides (HDP) that constitute a class of fast acting and rapidly available molecules preventing bacterial infection. Serum specimens were obtained from multiply injured patients. Radial diffusion assays were performed to investigate antimicrobial properties. Ultrafiltration and heat-inactivation were used to rule out antimicrobial activity of large proteins i.e. complement factors. ELISA was performed to analyse serum concentrations of the human beta-defensins 2 and 3 (hBD-2 and hBD-3), LL-37 and the proinflammatory cytokines interferon-gamma (IFN-γ) and interleukin-6 (IL-6). Serum of multiply injured patients showed greater zones of inhibition in antimicrobial testing against Gram negative und positive bacteria. This effect was mediated by proteins smaller than 10 kDa, inactivation of the complement system does not significantly reduce antibacterial action. hBD-2, hBD-3 and LL-37 concentrations were significantly elevated after trauma and followed different characteristic concentration curves. Similar patterns of concentration profiles were recorded for hBD-2/IL-6 and hBD-3/IFN-γ suggesting a stimulatory influence within their induction process. With this study we provide evidence, that serum of multiply injured patients has by far higher antibacterial capacity than that of healthy donors. As possible mediators we have detected the HDP hBD-2, hBD-3 and LL-37 and their inducers in serum of multiply injured patients.


Subject(s)
Antimicrobial Cationic Peptides/blood , Interferon-gamma/blood , Interleukin-6/blood , Multiple Trauma/immunology , Sepsis/prevention & control , beta-Defensins/blood , Adolescent , Adult , Aged , Blood Donors , Enzyme-Linked Immunosorbent Assay , Female , Humans , Male , Middle Aged , Sepsis/immunology , Young Adult , Cathelicidins
19.
Rhinology ; 49(5): 554-61, 2011 12.
Article in English | MEDLINE | ID: mdl-22125786

ABSTRACT

OBJECTIVE: Nasal carriage of Staphylococcus aureus in patients with chronic rhinosinusitis with nasal polyps (NP) is hypothesized to have pathophysiological impact on the disease. Antimicrobial peptides (AMP), especially human beta-defensin-3 (hBD-3) and LL-37, are an important part of the multifactorial defence against microorganisms in barrier organs like the nasal mucosa. The interaction of S. aureus colonization and AMP in nasal secretions and mucosa of NP were investigated in this study. PATIENTS AND METHODS: AMP were quantified in nasal secretions of 13 normal controls (NC) and 12 NP patients, each with and without S. aureus colonization, by ELISA. Immunohistochemistry was used to investigate the cellular sources of AMP in the nasal mucosa. To explore the AMP response of primary nasal epithelial cell cultures (NEC) towards S. aureus stimulation, a functional assay was established. RESULTS: AMP could be demonstrated in nasal secretions of all groups without differences in hBD-3 concentrations comparing S. aureus carriers vs. non-carriers. In NC, higher LL-37 concentrations were observed in S. aureus colonized as compared to non-colonized patients. This effect was not detectable in NP patients. Epithelial cells, submucosal glands and cells of the connective tissue could be identified as sources of AMP by immunohistochemistry. An AMP response of NEC towards S. aureus stimulation was detected in all groups. CONCLUSION: In NP patients, LL-37 response towards S. aureus colonization is disturbed while the ability of NEC to respond on S. aureus challenge is preserved. This deregulation of the nasal barrier could be involved in the multifactorial pathophysiology of NP.


Subject(s)
Anti-Bacterial Agents/metabolism , Antimicrobial Cationic Peptides/metabolism , Cathelicidins/metabolism , Nasal Mucosa/metabolism , Nasal Polyps/metabolism , Rhinitis/epidemiology , Sinusitis/epidemiology , beta-Defensins/metabolism , Adult , Chronic Disease , Comorbidity , Enzyme-Linked Immunosorbent Assay , Female , Humans , Immunohistochemistry , Male , Nasal Mucosa/microbiology , Nasal Polyps/epidemiology , Nasal Polyps/microbiology , Nasal Polyps/physiopathology , Staphylococcus aureus/isolation & purification
20.
Lab Anim ; 45(3): 179-83, 2011 Jul.
Article in English | MEDLINE | ID: mdl-21540281

ABSTRACT

As peri-implantitis is an emerging problem, the development of validated animal models is mandatory. The aim of this pilot study was to provide a first step in describing the normal oral flora of minipigs. In five minipigs, samples of the oral flora were collected with sterile cotton swabs from the buccal gingiva of the lower jaw. Two swabs per animal were collected, followed by bacterial isolation under both aerobe and anaerobe conditions. Microbiological analyses included biochemical tests, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and 16S rDNA gene sequence analysis. A total of 61 taxa were detected, 14-21 different bacterial taxa from each minipig. Among the Gram-positive cocci, mainly staphylococcal and streptococcal species were identified. Different Actinomyces species were the most abundant taxa in the group of Gram-positive rods. Among the anaerobic bacteria, the Gram-negative genera Fusobacterium, Bacteroides and Prevotella were the most often observed taxa. This is the first study which begins to describe the normal oral flora in minipigs in cultures to allow for the detection of a broad spectrum. Several bacterial species identified are different from human ones. No specific species for peri-implantitis could be detected in that healthy sample.


Subject(s)
Bacteria, Anaerobic/isolation & purification , Disease Models, Animal , Gingiva/microbiology , Gram-Positive Cocci/isolation & purification , Gram-Positive Rods/isolation & purification , Peri-Implantitis/microbiology , Animals , Bacteria, Anaerobic/classification , Bacteria, Anaerobic/genetics , Gram-Positive Cocci/classification , Gram-Positive Cocci/genetics , Gram-Positive Rods/classification , Gram-Positive Rods/genetics , Male , Pilot Projects , RNA, Ribosomal, 16S/genetics , Reference Values , Sequence Analysis, DNA , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Swine , Swine, Miniature
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