Role of gut-liver axis and glucagon-like peptide-1 receptor agonists in the treatment of metabolic dysfunction-associated fatty liver disease.

Metabolic dysfunction-associated fatty liver disease (MAFLD) is a hepatic manifestation of the metabolic syndrome. It is one of the most common liver diseases worldwide and shows increasing prevalence rates in most countries. MAFLD is a progressive disease with the most severe cases presenting as advanced fibrosis or cirrhosis with an increased risk of hepatocellular carcinoma. Gut microbiota play a significant role in the pathogenesis and progression of MAFLD by disrupting the gut-liver axis. The mechanisms involved in maintaining gut-liver axis homeostasis are complex. One critical aspect involves preserving an appropriate intestinal barrier permeability and levels of intestinal lumen metabolites to ensure gut-liver axis functionality. An increase in intestinal barrier permeability induces metabolic endotoxemia that leads to steatohepatitis. Moreover, alterations in the absorption of various metabolites can affect liver metabolism and induce liver steatosis and fibrosis. Glucagon-like peptide-1 receptor agonists (GLP-1 RAs) are a class of drugs developed for the treatment of type 2 diabetes mellitus. They are also commonly used to combat obesity and have been proven to be effective in reversing hepatic steatosis. The mechanisms reported to be involved in this effect include an improved regulation of glycemia, reduced lipid synthesis, ß-oxidation of free fatty acids, and induction of autophagy in hepatic cells. Recently, multiple peptide receptor agonists have been introduced and are expected to increase the effectiveness of the treatment. A modulation of gut microbiota has also been observed with the use of these drugs that may contribute to the amelioration of MAFLD. This review presents the current understanding of the role of the gut-liver axis in the development of MAFLD and use of members of the GLP-1 RA family as pleiotropic agents in the treatment of MAFLD.

Multilocus-sequence typing reveals clonality of Staphylococcus aureus in atopic dermatitis.

BACKGROUND: Atopic dermatitis (AD) is exacerbated by Staphylococcus aureus, which is capable of displacing not only the physiological microbiota, but also other strains of its own species. Analyses of the molecular characteristics and relationships of S. aureus strains present in different microniches are lacking. OBJECTIVES: To determine, using multilocus sequence typing (MLST), the relationship of S. aureus isolates from the lesional and nonlesional skin and anterior nares of patients with AD, and to review the characteristics of the dominant clones. METHODS: Sixty-three individuals with active AD were enrolled. Ten patients with moderate-to-severe AD (SCoring of Atopic Dermatitis score ≥ 25) colonized by S. aureus in all analysed locations were included in the MLST analysis. RESULTS: The most prevalent sequence types were 7 (10/30 strains; 33.3%), 15 and 97 (both 5/30 strains; 16.7%) all of which were associated with the expression of adhesins and toxins promoting chronic microbial dysbiosis, skin barrier damage and inflammation. Six patients (60%) were carriers of clonal S. aureus strains at all analysed locations, three (30%) carriers in lesional and nonlesional skin, and one (10%) was a carrier in nonlesional skin and the anterior nares. CONCLUSIONS: The results imply that the identified S. aureus lineages are better adapted to dominate the microbiota in AD. Decontaminating the identified reservoirs of S. aureus (i.e. anterior nares and nonlesional skin) could reduce the severity of AD.

The Impact of Harsh Stratospheric Conditions on Survival and Antibiotic Resistance Profile of Non-Spore Forming Multidrug Resistant Human Pathogenic Bacteria Causing Hospital-Associated Infections.

Bacteria are constantly being lifted to the stratosphere due to air movements caused by weather phenomena, volcanic eruptions, or human activity. In the upper parts of the atmosphere, they are exposed to extremely harsh and mutagenic conditions such as UV and space radiation or ozone. Most bacteria cannot withstand that stress, but for a fraction of them, it can act as a trigger for selective pressure and rapid evolution. We assessed the impact of stratospheric conditions on the survival and antibiotic resistance profile of common non-spore-forming human pathogenic bacteria, both sensitive and extremely dangerous multidrug-resistant variants, with plasmid-mediated mechanisms of resistance. Pseudomonas aeruginosa did not survive the exposure. In the case of strains that were recovered alive, the survival was extremely low: From 0.00001% of Klebsiella pneumoniae carrying the ndm-1 gene and methicillin-resistant Staphylococcus aureus mecA-positive with reduced susceptibility to vancomycin (MRSA/VISA), to a maximum of 0.001% of K. pneumoniae sensitive to all common antibiotics and S. aureus sensitive to vancomycin (MRSA/VSSA). We noticed a tendency towards increased antibiotic susceptibility after the stratospheric flight. Antimicrobial resistance is a current real, global, and increasing problem, and our results can inform current understandings of antibiotic resistance mechanisms and development in bacteria.

Enterotoxin Gene Cluster and selX Are Associated with Atopic Dermatitis Severity-A Cross-Sectional Molecular Study of Staphylococcus aureus Superantigens.

Staphylococcus aureus superantigens (SAgs) have been reported to aggravate atopic dermatitis. However, comprehensive analyses of these molecules in multiple microniches are lacking. The present study involved 50 adult patients with active atopic dermatitis. S. aureus was isolated from the lesional skin, nonlesional skin, and anterior nares. Multiplex-PCR was performed to identify genes encoding (1) selX (core genome); (2) seg, selI, selM, selN, selO, selU (enterotoxin gene cluster, EGC); and (3) sea, seb, sec, sed, see, tstH (classic SAgs encoded on other mobile genetic elements). The results were correlated to clinical parameters of the study group. selx and EGC were the most prevalent in all microniches. The number of SAg-encoding genes correlated between the anterior nares and nonlesional skin, and between the nonlesional and lesional skin. On lesional skin, the total number of SAg genes correlated with disease severity (total and objective SCORAD, intensity, erythema, edema/papulation, lichenification and dryness). Linear regression revealed that AD severity was predicted only by selx and EGC. This study revealed that selX and EGC are associated with atopic dermatitis severity. Anterior nares and nonlesional skin could be reservoirs of SAg-positive S. aureus. Restoring the physiological microbiome could reduce the SAg burden and alleviate syndromes of atopic dermatitis.

Genomic Insights of First ermB-Positive ST338-SCCmecVT/CC59 Taiwan Clone of Community-Associated Methicillin-Resistant Staphylococcus aureus in Poland.

We report the first Polish representative of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA), lukS/F-PV-positive, encoding the ermB gene, as a genetic determinant of constitutive resistance to macrolides, lincosamides, and streptogramin B antibiotics, cMLS-B. This is the first detection of the CA-MRSA strain responsible for nosocomial infection in the Warsaw Clinical Hospital. Resistance to ß-lactams associates with a composite genetic element, SCCmec cassette type VT (5C2&5). We assigned the strain to sequence type ST338 (single-locus variant of ST59), clonal complex CC59, spa-type t437, and agr-type I. Genomic-based comparison was designated SO574/12 as an international Taiwan clone, which has been so far described mainly in the Asia-Pacific region. The ermB gene locates on the chromosome within the 14,690 bp mobile element structure, i.e., the MESPM1-like structure, which also encodes aminoglycoside- and streptothricin-resistance genes. The MESPM1-like structure is a composite transposon containing Tn551, flanked by direct repeats of IS1216V insertion sequences, which probably originates from Enterococcus. The ermB is preceded by the 273 bp regulatory region that contains the regulatory 84 bp ermBL ORF, encoding the 27 amino acid leader peptides. The latest research suggests that a new leader peptide, ermBL2, also exists in the ermB regulatory region. Therefore, the detailed function of ermBL2 requires further investigations.

The Propensity to Form Biofilm in vitro by Staphylococcus aureus Strains Isolated from the Anterior Nares of Patients with Atopic Dermatitis: Clinical Associations.

BACKGROUND: Atopic dermatitis is a chronic inflammatory dermatosis with complex pathogenesis. The skin microbiome in atopic dermatitis is dominated by Staphylococcus aureus which shows the ability to produce biofilm. OBJECTIVES: The aim of this work was to assess the influence of S. aureus biofilm on the course of atopic dermatitis. METHODS: Disease severity was evaluated based on the SCORAD index in 56 adult patients with atopic dermatitis. Microtiter plate assay of the propensity to form biofilm was performed on S. aureus strains isolated from the anterior nares, lesional skin, and nonlesional skin. Microbiological results were correlated to the clinical parameters and total IgE concentration. RESULTS: Biofilm-producing strains of S. aureus were identified in 76.3% (29/38) and 79.1% (34/43) of samples from the anterior nares and lesional skin, respectively (p > 0.05), and in 48.5% (16/33) of samples from nonlesional skin (p < 0.03). Patients colonized by biofilm-producing strains of S. aureus within the anterior nares showed statistically higher mean values of total and objective SCORAD and its components (extent, dryness), and of the largest extent of skin lesions during the flares in the last year when compared to patients colonized by non-biofilm-producing strains. Carriage of biofilm-producing S. aureus on lesional skin was associated with higher mean values of the extent of skin lesions during stable periods of the disease. CONCLUSIONS: The results of this study may suggest a relationship between the production of biofilm by S. aureus strains colonizing the anterior nares and the course of atopic dermatitis. Biofilm seems crucial for dispersal and persistent colonization of large areas of the skin by this pathogen. Destruction of S. aureus biofilm could positively affect the course of atopic dermatitis.

Prevalence of methicillin resistant and mupirocin-resistant Staphylococcus aureus strains among medical students of Medical University in Warsaw

INTRODUCTION: Staphylococcus aureus is a microorganism, which is able to colonize the human body without any pathogenic effect, but it also can cause life-threatening infections (opportunistic pathogen). Asymptomatic colonization with both methicillin resistant (MRSA) and methicillin susceptible (MSSA) S.aureus strains state is an important predisposing factor for infections. The risk of infection for carriers of MSSA is even three-times higher than for non-colonized people, and in the case of MRSA it is even four-times higher than in MSSA carriers. Carriers can be also a source of infection for other people, especially those belonging to high-risk groups. The drug of choice used for the local eradication of S.aureus is mupirocin (Mup). In recent years, the failure of decolonization therapy has been observed. The aim of the study was to assess and compare the level of colonization of S.aureus (MRSA or MSSA) among medical students and to evaluate the sensitivity of the strains to mupirocin. For MRSA/MupRSA isolates the molecular mechanism of resistance phenotype was determined. MATERIALS AND METHODS: 955 swabs from 2014-2016 from pre-clinical students of medicine of the Medical University of Warsaw. The strains were identified using Pastorex-Staph-Plus (BioRad) and/or the VITEK-MS system (Biomerieux), according to manufacturer's instructions. Susceptibility to methicillin and mupirocin was determined by disk diffusion and/or broth microdilution method, according to EUCAST. The presence of the mecA/mecC and mupA genes were detected with PCR technique. RESULTS: Asymptomatic colonization with S.aureus strains was found in 245/955 (25,7%) students, in particular years in the range of 21,7-29,9%. 243 isolates expressed the MSSA/MupSSA phenotype, one strain was resistant to mupirocin MSSA/MupRSA (genotype mecA/mecC-negative, mupA-positive) and one showed simultaneous resistance to methicillin and mupirocin (mecA/mupA-positive genotype). The level of MRSA and MupRSA colonization was 0,1% and 0,2%, respectively. SUMMARY AND CONCLUSIONS: The level of S.aureus colonization among surveyed students, didn't differ from the norm for a generally healthy population, but showed an upward trend. The carriage of S.aureus, especially of multi-resistant strains among medical students at the beginning of their clinical activities, consist of a real threat to patients and other people.

The presence of antibiotic resistance genes and bft genes as well as antibiotic susceptibility testing of Bacteroides fragilis strains isolated from inpatients of the Infant Jesus Teaching Hospital, Warsaw during 2007-2012.

The purpose of this study was to assess drug susceptibility of clinical B. fragilis strains and to determine any correlation between drug resistance and the presence of specific genes. Antimicrobial susceptibility was assessed using E-tests. All isolates were analyzed with the PCR technique for the presence of antibiotic resistance genes (cepA, cfxA, cfiA, ermF, ermB, ermG, nim), insertion sequences elements (IS1186, IS1187, IS1188, IS942), and enterotoxin-encoding genes (bft). Susceptibility tests yielded the following rates of resistance to the evaluated antibiotics: penicillin G (100%), clindamycin (22.5%), cefoxitin (6.3%), amoxicillin/clavulanic acid (1.8%). All strain were susceptible to imipenem, and metronidazole. The following antibiotic resistance genes were detected in the evaluated isolates: cepA (in 96.4% of isolates), cfxA (in 12.6%), cfiA (in 1.8%), and ermF (in 25.2%). Genes ermB, ermG, and nim were not found. The presence of the cepA gene showed no correlation with the penicillin G MIC. However, we observed a high correlation between cefoxitin MIC values and the presence of gene cfxA as well as a nearly complete correlation between clindamycin MIC values and the presence of gene ermF. The presence of a bft gene was detected in 14.4% of the analyzed B. fragilis isolates; with the bft-1 allele found in 75%, bft-2 in 25%, and bft-3 in none of the isolates. Antibiotic susceptibility profiles of enterotoxin gene-positive isolates in our study did not differ from those of enterotoxin gene-negative isolates.

Characteristics of glycopeptide-resistant Staphylococcus aureus strains isolated from inpatients of three teaching hospitals in Warsaw, Poland.

Background: Vancomycin is still one of the most commonly used drug for treatment of severe methicillin-resistant Staphylococcus aureus (MRSA) infections. Vancomycin-resistant S. aureus (VRSA) strains are a serious danger for public health. This study aimed to characterize healthcare-associated MRSA (HA-MRSA) strains, resistant to at least one of glycopeptide antibiotics: vancomycin (VRSA) and/or teicoplanin (TRSA), isolated at three Warsaw hospitals over a period of 17-years (1991-2007). Methods: Among 600 HA-MRSA strains, isolated from patients with symptomatic infections, 47 were subjected to detailed analysis. In the study, mechanisms behind VRSA phenotypes were determined (E-tests, GRD-test, agar-dilution method and vanA/B detection). Characteristics of selected isolates on molecular level: i) by detection of resistance genes ermA/ermB/ermC, msrA/msrB, linA/linA', aacA-aphD, aadD, aph(3")-IIIa; ii) SCCmec-typing and iii) MLST-typing was done. Results: In general population of studied strains, 11/47 (23.4%) were VRSA and 36/47 (76.6%) were resistant only to teicoplanin. All isolates exhibited van-independent mechanisms of resistance. Over 80% of isolates belonged to clonal complex CC8, with the following predominant sequence types (STs)/clones: ST247-IA/Iberian, ST241-III/Finland-UK, and ST239-III/Brazilian. Most of the isolated strains harboured ermA and aacA-aphD genes, encoding additional resistance to macrolides, lincosamides, streptogramin B, and majority of aminoglycosides. They occurred also in Polish VRSA/TRSA population over the period, which was subjected for analysis: an increase in MIC values for glycopeptides, evolution in terms of the level and extent of resistance, and genetic re-assortment in epidemic clones. Conclusions: VRSA strains isolated from patients hospitalized at three Warsaw teaching hospitals in Poland, over a period of 17-years do not pose a threat as potential donors of van genes in horizontal-gene transfer processes, but are constantly evolving and represent international epidemic clones.

In vitro effect of clindamycin against Bacteroides and Parabacteroides isolates in Poland.

OBJECTIVES: The aims of this study were (i) to analyse strains of the genera Bacteroides and Parabacteroides isolated from clinical specimens for phenotypic resistance to clindamycin, (ii) to detect erm genes in the isolates and (iii) to determine any correlation between in vitro resistance and the presence of erm genes. METHODS: The Bacteroides and Parabacteroides isolates analysed were obtained from patients hospitalised at teaching hospitals in Poland. Antimicrobial susceptibility testing was performed by Etest and the results were interpreted according to European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines. All isolates were analysed by PCR for the presence of the resistance genes ermF, ermB and ermG. RESULTS: Resistance to clindamycin was detected in 31.0% (62/200) of all evaluated isolates, with the ermF and ermB genes detected in 31.0% (62/200) and 0.5% (1/200) of isolates, respectively. No isolates with ermG were detected among the evaluated strains. Pearson's test showed an almost perfect correlation between clindamycin minimum inhibitory concentrations (MICs) and the presence of ermF in Bacteroides spp. and Parabacteroides distasonis isolates, although the ermF gene was also present in 10 clindamycin-susceptible isolates of Bacteroides spp. CONCLUSIONS: This study demonstrated a substantial proportion of Bacteroides (22.5-100% depending on the species) and 50.0% of Parabacteroides strains exhibiting resistance to clindamycin. The clindamycin MIC for resistant strains in each case was ≥256mg/L. Resistance to clindamycin in Bacteroides and Parabacteroides species is correlated mainly with the presence of the ermF gene.

Production of extracellular polysaccharide and biofilm under different oxygen conditions by clinical isolates of Staphylococcus aureus non-susceptible to glycopeptides

INTRODUCTION: Staphylococcus aureus, which is able to produce an extracellular mucopolysaccharide (MP) and biofilm (SP), is an important etiologic agent in persistent and implant-related infections. This phenotype may be expressed in different levels and character depending on various environmental and/or global intracellular regulatory mechanisms. It may also be induced by sub-inhibitory concentrations of some antibiotics, for example vancomycin. The main aim of the study was to assess the ability to produce MP and SP in different oxygen conditions by clinical isolates of S.aureus nonsusceptible to glycopeptides. MATERIALS AND METHODS: Clinical isolates of health-care associated methicillin resistant S. aureus (HA-MRSA) strains, non-susceptible to glycopeptides (GRSA, 47) and heterogeneous vancomycin intermediate S. aureus isolates (h-VISA, 8). Control group consisted of the following strains: 55 belonging to MRSA, vancomycin susceptible, VSSA and 19 as methicillin susceptible, MSSA/VSSA. The ability to produce MP was investigated according to Freeman method. SP production was tested by means of Christensen procedure. RESULTS: In aerobic conditions MRSA/GRSA and MRSA/h-VISA isolates were the strongest mucopolysaccharide (SMP) producers (12.2% and 28.6% SMP/MP), but MSSA/VSSA were the most frequent MP (100%). In anaerobic atmosphere, all isolates from all groups were MP-positive. MRSA/h-VISA were the strongest MP producers (75% SMP/MP), but MSSA/VSSA were the most susceptible to oxidative stress (the percentage of SMP among MP for MSSA/VSSA increased by 15.8 times). Each evaluated group of clinical S. aureus isolates in aerobic condition had representation in SP positive phenotype: MRSA/GRSA and MRSA/h-VISA, 63.9% and 62.5%; MRSA/VSSA and MSSA/VSSA, respectively 80% and 94.7%. For all mentioned groups of bacteria, SSP variants were present and the amount of values was higher than in similar results obtained in CRA method. The strongest slime producers (60%) were h-VISA strains. The results obtained in Christensen method for anaerobic conditions, were not conclusive due to insufficient optimization of the test parameters. SUMMARY AND CONCLUSIONS: Both methods reveal that MRSA isolates non-susceptible to glycopeptides are the strongest producers of both MP and SP. That is probably due to cell wall alterations and global regulatory system Agr disorders. The Christensen procedure allow to assess both ica- dependent and ica- independent (adhesive) mechanisms of slime production and allow to notice that, as a phenotyping "biofilm booster effect". ica- dependent mechanism, which dominated in MSSA/VSSA strains, demonstrate phenotype with more susceptibility to oxygen stress conditions than adhesive one.

[Comparison of phenotypic methods for the detection of beta-lactamases MBL in strains from the Enterobacteriaceae family and non-fermentative bacilli isolated from clinical specimens]. / Porównanie metod fenotypowych wykrywania beta-laktamaz MBL u szczepów z rodziny Enterobacteriaceae oraz paleczek niefermentujacych izolowanych z próbek materialów klinicznych.

INTRODUCTION: Bacterial resistance is growing because of treatment a broad spectrum antibio- tics. Gram-negative pathogens which producing carbapenemase are a one of major problem in many hospitals. Rapid detection those strains provide an early inhibition of infection and control the expansion of microorganisms. The aim of work was to characterize the frequency of appearance MBLs in specific groups of Gram-negative bacilli which are resistant or intermediate to at least one of carbapenems. METHODS: Bacterial isolates were collected from Baby Jesus Clinical Hospital from 2003 to 2009. Pathogens were isolated from urine, blood, fluids, swab of the wound, pharyngeal swab. They were identified by the ID 32 E (bioMérieux, France) and Vitek2. Antimicrobial resistance was marked by the ATB G-5 and ATB UR (bioMérieux, France). Detection of metalo-beta-lactamases was tested by disk diffusion test recommended by the EUCAST. The DDS test using imipenem, ceftazidime, ethylenediaminetetraacetic acid (EDTA) and 2-mercaptopropionic acid (2-MPA). Positive test was reading as enlargement of inhibition zone about imipenem- or ceftazidime-impregnated disk. RESULTS: Of the 88 isolates, 32 come fromEnterobacteriaceae and 56 from non-fermentative bacilli. All strains were tested of production of MBL by disk diffusion test. This method used two inhibitors: ethylenediaminetetraacetic acid and 2-mercaptopropionic acid. As a result of EDTA there was 45 MBL positive strains. In apply 2-MPA there was 55 MBL positive strains. Both the EDTA and 2-MPA disk test showing the highest percentage of positive result in Enterobacter cloacae, Serratia marcescens, Pseudomonas aeruginosa and Pseudomonas putida. CONCLUSIONS: Resistance to carbapenems in the non-fermentative bacilli occurs more often than in the Enterobacteriaceae. Method with 2-mercaptopropionic acid was more effective to detect metallo-beta-lactamases than EDTA. Concerns especially bacilli from Enterobacteriaceae.

[Occurrence of aminoglycoside transpherase genes in methicillin-resistant strains of Staphylococcus aureus]. / Wystepowanie genów transferaz aminoglikozydowych u metycylino- opornych szczepów Staphylococcus aureus.

Fifty MRSA strains originated from clinical specimens were examined by the PCR method, for the presence of three genes: aacA-aphD, aadD oraz aph(3")-IIIa. The obtained results were correlated with the susceptibility of the strains to gentamicin, tobramicin, kanamicin, neomicin, amikacin and netilmicin. The susceptibility results were interpreted according with CLSI and EUCAST guidelines. aacA-aphD gene was found in 34 strains, aadD in 27 strains and aph(3")-IIIa was present in 22 strains. In 19 strains (38%) was present one of the investigated genes, in 29 (58%) strains two genes and in two strains (4%) all three genes were found. The most frequent variant was combination of aacA-aphD and aadD genes.