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Key Clinical Message: This report presents a rare case of invasive infection caused by Enterococcus cecorum. There are no specific guidelines regarding antibiotic therapy for this infection. Based on this case, it can be concluded that linezolid demonstrates in vivo activity against Enterococcus cecorum and can be successfully used in therapy. Abstract: Enterococcus cecorum is an extremely rare pathogen in humans. Since 1984, when the microorganism was first described, only a dozen cases of invasive infections in humans have been reported in the literature. The diagnostic pathway may involve difficulties in correctly identifying this microorganism. Based on the case described, it can be thought that Enterococcus cecorum is a more challenging bacterium than the much more common Enterococcus faecium or Enterococcus faecalis. The described case underscores the importance of medical vigilance in clinical practice. It seems that due to increasingly advanced techniques in molecular biology, we will more frequently detect pathogens that were previously encountered only sporadically. Since not every center has access to modern and advanced microbiological diagnostic methods, publications that practically combine classical microbiological diagnostic methods with those less accessible but more modern are exceptionally valuable. In the case described, it is also worth noting that classical methods still play a significant and crucial role in conducting microbiological diagnostics. In the era of rapid diagnostic tool development, it is important to emphasize the necessity of combining different methods rather than replacing one with another.
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Background: The difficulties in attaining effective antibiotic therapy arising from the multidrug resistance of Gram-negative bacilli compel the exploration of new possibilities for synergistic interactions among existing antibiotics. Research Design and Methods: An analysis was conducted to assess the efficacy of two antibiotic therapy regimens in the treatment of infections caused by Klebsiella pneumoniae strains producing carbapenemases (MBL). Two patient groups were considered: Group A - individuals in whom the treatment of infection involved the application of ceftazidime-avibactam in combination with aztreonam. Group B comprised patients subjected to an alternative antibiotic therapy regimen. Results: In the group subjected to the treatment regimen involving ceftazidime-avibactam and aztreonam, as compared to alternative antibiotic combinations, a statistically lower mortality rate during the course of treatment and a faster clinical response to the administered therapy were evident. Conclusion: The results obtained may be applicable to routine in vitro assays performed and serve as valuable guidance for the potential utilization of the positive effect of antibiotic therapy through the synergy between ceftazidime-avibactam and aztreonam. The selection of antibiotics employed in the therapy of invasive infections caused by K. pneumoniae influences the ultimate treatment outcome.
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In this study, eighteen new ligands (B1-B18) containing a thiosemicarbazide core were synthesized and characterized in terms of physicochemical properties, molecular docking and in vitro biological activity. The structures of eleven ligands were investigated using X-Ray diffraction and Hirschfeld Surface analysis. To study the structure-activity relationship, the organic ligands contained pyridin-2-ylmethyl, pyridin-3-ylmethyl or pyridin-4-ylmethyl moieties and various substituents. Their pharmakokinetic profiles and molecular docking results suggest high potential as new drug candidates. The complexing ability of the selected organic ligands was also evaluated, yielding five new Cu(II) complexes (Cu(B1)Cl2, Cu(B4)Cl2, Cu(B10)Cl2, Cu(B17)Cl2, Cu(B18)Cl2). The obtained results suggest the formation of the polymeric structures. All organic ligands and Cu(II) complexes were tested for anticancer activity against prostate and melanoma cancer cells (PC-3, DU-145, LNCaP, A375, G-361, SK-MEL-28) and normal fibroblasts (BJ), as well as antimicrobial activity against six selected bateria strains. Among B1-B18 compounds, B3, B5, B9, B10, B12 and B14 exhibited cytotoxic activity. The studied Cu(II) complexes were in general more active, with Cu(B1)Cl2 exhibiting antincancer activity agains all three prostate cancer cells and Cu(B10)Cl2 reaching the IC50 value equal to 88 µM against G-361 melanoma cells. Several compounds also exhibited antimicrobial activity against gram-positive and gram-negative bacteria. It was found that the type of specific substituents, especially the presence of -chloro and -dichloro substituents had a greated impact on the cytotoxicity than the position of the nitrogen atom in the pyridylacetyl moiety.
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In recent years, drug-resistant and multidrug-resistant fungal strains have been more frequently isolated in clinical practice. This phenomenon is responsible for difficulties in the treatment of infections. Therefore, the development of new antifungal drugs is an extremely important challenge. Combinations of selected 1,3,4-thiadiazole derivatives with amphotericin B showing strong synergic antifungal interactions are promising candidates for such formulas. In the study, microbiological, cytochemical, and molecular spectroscopy methods were used to investigate the antifungal synergy mechanisms associated with the aforementioned combinations. The present results indicate that two derivatives, i.e., C1 and NTBD, demonstrate strong synergistic interactions with AmB against some Candida species. The ATR-FTIR analysis showed that yeasts treated with the C1 + AmB and NTBD + AmB compositions, compared with those treated with single compounds, exhibited more pronounced abnormalities in the biomolecular content, suggesting that the main mechanism of the synergistic antifungal activity of the compounds is related to a disturbance in cell wall integrity. The analysis of the electron absorption and fluorescence spectra revealed that the biophysical mechanism underlying the observed synergy is associated with disaggregation of AmB molecules induced by the 1,3,4-thiadiazole derivatives. Such observations suggest the possibility of the successful application of thiadiazole derivatives combined with AmB in the therapy of fungal infections.
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Antifúngicos , Tiadiazoles , Antifúngicos/farmacología , Anfotericina B/farmacología , Antibacterianos , Tiadiazoles/farmacología , Análisis Espectral , Pruebas de Sensibilidad MicrobianaRESUMEN
Proteomic analyses based on mass spectrometry provide a powerful tool for the simultaneous identification of proteins and their signatures. Disorders detection at the molecular level delivers an immense impact for a better understanding of the pathogenesis and etiology of various diseases. Acute coronary syndrome (ACS) refers to a group of heart diseases generally associated with rupture of an atherosclerotic plaque and partial or complete thrombotic obstruction of the blood flow in the infarct-related coronary artery. The essential role in the pathogenesis of ACS is related to the abnormal, pathological activation of blood platelets. The multifactorial and complex character of ACS indicates the need to explain the molecular mechanisms responsible for thrombosis. In our study, we performed screening and comparative analysis of platelet proteome from ACS patients and healthy donors. Two-dimensional fluorescence difference gel electrophoresis and nanoscale liquid chromatography coupled to tandem mass spectrometry showed altered expressions of six proteins (i.e., vinculin, transgelin-2, fibrinogen ß and γ chains, apolipoprotein a1, and tubulin ß), with the overlapping increased expression at the mRNA level for transgelin-2. Dysregulation in protein expression identified in our study may be associated with an increased risk of thrombotic events, correlated with a higher aggregability of blood platelets and induced shape change, thus explaining the phenomenon of the hyperreactivity of blood platelets in ACS.
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Síndrome Coronario Agudo , Trombosis , Síndrome Coronario Agudo/metabolismo , Plaquetas/metabolismo , Humanos , Proteínas de Microfilamentos , Proteínas Musculares , Proteoma/metabolismo , Proteómica/métodos , Espectrometría de Masas en Tándem , Trombosis/metabolismo , TranscriptomaRESUMEN
The pathological conditions caused by blood platelet activation constitute a fundamental core in the pathogenesis of Acute Coronary Syndrome (ACS). The hyperactivity of platelets in ACS is well-documented, but there is still little research into the molecular basis of phenotypic changes in platelet functionality. To expand the knowledge of this phenomenon, we analyzed the disturbances in the expression of several key platelet receptors and the aspect of regulating potential abnormalities. Platelet surface receptors are responsible for maintaining the hemostatic balance, platelet interaction with immune cells, and support of the coagulation cascade leading to occlusion of the vessel lumen. Due to their prominent role, platelet receptors constitute a major target in pharmacological treatment. Our work aimed to identify the molecular alteration of platelet surface receptors, which showed augmented mRNA expression of P2Y12, GP1BB, ITGA2B, and ITGB3 and increased protein concentrations of P2Y12 and GP IIb/IIIa in ACS. The upregulation of the P2Y12 level was also confirmed by confocal and cytometric visualization. Furthermore, we evaluated the expression of two microRNAs: miR-223-3p and miR-126-3p, which were suggested to regulate platelet P2Y12 expression. Results of our study present new insight into the molecular background of ACS.
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PURPOSE: Determination of the association between ica genes and phenotypic biofilm formation in staphylococcal isolates involved in conjunctivitis, their antibiotic resistance as well as detection of selected virulence characteristics: adhesion to epithelial cells and in vitro cytotoxicity. METHODS: The study included 26 Staphylococcus aureus (SA) and 26 Staphylococcus epidermidis (SE) isolates. The presence of icaAD genes and ica operon was determined by the PCR assay. Phenotypic biofilm formation was verified using the microtitre plate assay. Antibiotic resistance was performed using the disc diffusion method. Staphylococcal ability to attach to host cells was assessed by flow cytometry. Cytotoxicity on epithelial cells was evaluated by LDH assay. RESULTS: The ica genes were detected in 26.9% of SE and in 42.3% of SA isolates. Only 15.3% of isolates (SE) were positive for both the icaAD and the ica operon. Phenotypically, 19.2% of SE isolates were strong biofilm producers, among which three were both icaAD- and ica operon-positive. About 26.9% of SA isolates were strong biofilm producers. Methicillin resistance (MR) was detected in 34.6% of SE and 26.9% of SA isolates. About 75% of MR isolates were multidrug resistant. SA isolates adhered to host cells more extensively than SE. SA isolates released higher level of LDH than SE. CONCLUSIONS: Adherence abilities were commonly observed in staphylococci associated with conjunctivitis. However, low prevalence of isolates positive for a complete and functional ica locus and low prevalence of strong biofilm producers was detected. SA adhered to a greater extent to eukaryotic cells than SE and were more cytotoxic.
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Conjuntivitis , Infecciones Estafilocócicas , Antibacterianos/farmacología , Biopelículas , Humanos , Pruebas de Sensibilidad Microbiana , Staphylococcus/genética , Staphylococcus aureus/genética , Staphylococcus epidermidisRESUMEN
Garlic has long been known as the most effective plant species in treatment of bacterial infections. Considering the vast potential of garlic as a source of antimicrobial drugs, this study is aimed to evaluate the antibacterial activity of Allium sativum extracts and their interactions with selected antibiotics against drug-sensitive and multidrug-resistant isolates of emerging bacterial pathogens that are frequently found in healthcare settings. As shown by the in vitro data obtained in this study, the whole Allium sativum extract inhibited the growth of a broad range of bacteria, including multidrug-resistant strains with bactericidal or bacteriostatic effects. Depending on the organism, the susceptibility to fresh garlic extract was comparable to the conventional antibiotic gentamycin. Since the combinations of fresh garlic extract with gentamycin and ciprofloxacin inhibited both the drug sensitive and MDR bacteria, in most cases showing a synergistic or insignificant relationship, the potential use of such combinations may be beneficial, especially in inhibiting drug-resistant pathogens. The study results indicate the possibility of using garlic as e.g. a supplement used during antibiotic therapy, which may increase the effectiveness of gentamicin and ciprofloxacin.
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Antibacterianos/farmacología , Bacterias/efectos de los fármacos , Infecciones Bacterianas/tratamiento farmacológico , Ajo/metabolismo , Extractos Vegetales/farmacología , Infecciones Bacterianas/microbiología , Farmacorresistencia Bacteriana Múltiple/efectos de los fármacos , Ajo/química , Pruebas de Sensibilidad MicrobianaRESUMEN
The immune response to Pseudomonas aeruginosa strains could be influenced by differences in antibiotic resistance and virulence. At the present time, it is unclear which type of immune responses enables uncontrolled invasion of opportunistic pathogens. The conditional pathogenicity of Pseudomonas aeruginosa served as an inspiration to begin a study on this bacterium. The aim of this study was to gain insight into selected parameters describing immune responses with regards to the adaptable agents of this pathogen. For the analysis of the specific immune response, the potential of Pseudomonas aeruginosa to stimulate lymphocytes, including Th17 lymphocytes, dendritic cells and other components of the adaptive immune response, was examined. The highest percentage of CD83+CD1a-HLA-DR++ cells was found after stimulation with lysates of strains isolated from the patients with severe systemic infection. We found statistically significant differences in percentages of HLA-DR+ PBMCs and MFI of HLA-DR between groups of Pseudomonas aeruginosa strains isolated from the patients with different clinical courses of infection. Our results suggest that the clinical course and outcomes of Pseudomonas aeruginosa infections are not associated with impairment of the specific immune response.
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Inmunidad , Pseudomonas aeruginosa/inmunología , Antígenos CD/metabolismo , Citocinas/metabolismo , Células Dendríticas/metabolismo , Femenino , Genes Bacterianos , Antígenos HLA-DR/metabolismo , Humanos , Mediadores de Inflamación/metabolismo , Masculino , Persona de Mediana Edad , Infecciones por Pseudomonas/inmunología , Infecciones por Pseudomonas/microbiología , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/patogenicidad , Células Th17/metabolismo , Virulencia/genéticaRESUMEN
PURPOSE: The aim of the study was to evaluate prostate cell infiltration by CD4(+)IL-17(+) and Treg cells in BPH and PCa patients depending on P. acnes infection in the prostate gland. PATIENTS AND METHODS: Prostate fragments were collected from 54 patients with PCa and 34 patients with BPH. Rapid ID 32 was used to identify the bacteria. Cells were analyzed by flow cytometry BD FACSCanto II. Statistical analysis was performed using Statistica 7 software (TIBCO Software Inc, USA). RESULTS: P. acnes was detected in 35% of patients with PCa and 41% of individuals with BPH. The infiltration of CD4(+)IL-17(+) and Treg cells was statistically significantly higher (P = 0.001) in patients with BPH and positive for P. acnes. A statistically considerably higher (P = 0.001) infiltration of Treg cells in treated for PCa with P.acnes infection was also demonstrated. CONCLUSION: Prostatitis caused by P. acnes may contribute to the development of BPH and PCa.
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Staphylococcus aureus is a widely recognized pathogen responsible for many serious diseases in both humans and animals. It is also one of the major causative agents of bovine mastitis. Methicillin-resistant S. aureus (MRSA), although relatively rare in this pathology, has been increasingly reported in livestock animals, mainly in pigs, but also cattle, sheep, and poultry. The recent emergence of livestock-associated (LA-)MRSA is cause for an immediate public health concern due to the risk of zoonotic transmission to humans, and is of particular concern for people who work in animal husbandry or have prolonged contact with livestock animals. This study reports on the first LA-MRSA outbreak in dairy cattle and the first probable case of MRSA transmission between humans and cows in Poland. A single dairy farm located in Eastern Poland was monitored on a regular basis for the occurrence of mastitis. Over a 1-yr study period, 717 quarter-milk samples from 583 cows were collected and examined microbiologically. A total of 5 MRSA isolates from as many cows with subclinical mastitis were cultured. They all belonged to the same outbreak, given a 2-mo time window in which they were identified. During the outbreak, 24 oral and nasal swabs were voluntarily taken from 6 people: a milker, a veterinarian, and 4 members of the veterinarian's family. Eight swabs from a milker, veterinarian, and 2 family members yielded positive MRSA cultures. All MRSA isolates were genotyped with a combination of multiple-locus variable number tandem repeat analysis, multilocus sequence typing, and staphylococcal protein A gene (spa) typing. Eleven bovine (n = 5; 5 cases) and human (n = 6; 4 cases) isolates showed an identical drug-susceptibility profile and were indistinguishable upon multiple-locus variable number tandem repeat analysis (pattern A), multilocus sequence typing (ST398) and spa (t034) typing. The results of this study provide the evidence of transmission of MRSA between humans and cows, and between humans in the family setting. This work, despite being a preliminary investigation, underscores the risk of intra- and interspecies transmission of LA-MRSA and urges enhancement of the existing biosecurity measures aimed at preventing MRSA (and other milk pathogens) spread at both the farm- and household levels.
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Enfermedades de los Bovinos/epidemiología , Mastitis Bovina/microbiología , Staphylococcus aureus Resistente a Meticilina , Infecciones Estafilocócicas/veterinaria , Crianza de Animales Domésticos , Animales , Bovinos , Enfermedades de los Bovinos/transmisión , Brotes de Enfermedades/veterinaria , Granjas , Femenino , Humanos , Staphylococcus aureus Resistente a Meticilina/genética , Leche/microbiología , Tipificación de Secuencias Multilocus/veterinaria , Polonia , Infecciones Estafilocócicas/epidemiología , Infecciones Estafilocócicas/transmisión , Proteína Estafilocócica A/genéticaRESUMEN
Moraxella catarrhalis is considered an important, exclusively human respiratory tract pathogen which, along with Streptococcus pneumoniae and Haemophilus influenzae, is classified as one of the most frequent bacterial etiological factors causing upper respiratory tract infections. In this manuscript, we report the existence of five tetracycline-resistant M. catarrhalis strains with confirmed presence of tetracycline resistance tetB gene. The strains were isolated from children under the age of three with signs of upper respiratory tract infections. Our research also investigated the occurrence of virulence genes in these strains and involved the analysis of drug resistance to five antibiotic groups. It is the first description of clinical strains with confirmed presence of drug resistance tetB genes isolated in Europe.
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Moraxella catarrhalis , Infecciones del Sistema Respiratorio , Resistencia a la Tetraciclina , Antibacterianos/farmacología , Niño , Europa (Continente) , Humanos , Pruebas de Sensibilidad Microbiana , Moraxella catarrhalis/genética , Polonia , Infecciones del Sistema Respiratorio/epidemiología , Resistencia a la Tetraciclina/efectos de los fármacos , Resistencia a la Tetraciclina/genéticaRESUMEN
Despite its low virulence potential and a commensal lifestyle as a member of the human skin microbiota, Brevibacterium casei has been increasingly reported as an opportunistic pathogen, especially in immunocompromised patients. Here, we present the draft genome sequence of the S51 strain isolated from a bloodstream infection. To the best of the authors' knowledge, this is the first report of the draft genome sequence of the B. casei strain isolated from the clinical infection. The strain was identified using phenotypic and molecular methods and subsequently sequenced using the next-generation sequencing. The draft whole genome was assembled de novo, automatically annotated by Rapid Annotations using Subsystems Technology (RAST) server and scrutinized to predict the presence of virulence, resistance, and stress response proteins. The genome size of the S51 strain was 3,743,532 bp and an average G+C content was 68.3%. The predicted genes included 48 genes involved in resistance to antibiotics (including vancomycin, fluoroquinolones, and beta-lactams) and toxic compounds (heavy metals), 16 genes involved in invasion and intracellular resistance (Mycobacterium virulence operons), and 94 genes involved in stress response (osmotic, oxidative stress, cold and heat shock). ResFinder has indicated the presence of a beta-lactamase, and a phenotypic analysis showed resistance to penicillin. This whole-genome NGS project for the S51strain has been deposited at EMBL/GenBank under the accession no. QNGF00000000.
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Bacteriemia/microbiología , Brevibacterium/genética , Genoma Bacteriano , Infecciones por Bacterias Grampositivas/microbiología , Antibacterianos/farmacología , Composición de Base , Brevibacterium/efectos de los fármacos , Brevibacterium/aislamiento & purificación , Farmacorresistencia Bacteriana Múltiple , Humanos , Análisis de Secuencia de ADN , Virulencia , Secuenciación Completa del GenomaRESUMEN
Unravelling of the interplay between the immune system and non-diphtheria corynebacteria would contribute to understanding their increasing role as medically important microorganisms. We aimed at the analysis of pro- (TNF, IL-1ß, IL-6, IL-8, and IL-12p70) and anti-inflammatory (IL-10) cytokines produced by Jurkat T cells in response to planktonic and biofilm Corynebacterium amycolatum. Two reference strains: C. amycolatum ATCC 700207 (R-CA), Staphylococcus aureus ATCC 25923 (R-SA), and ten clinical strains of C. amycolatum (C-CA) were used in the study. Jurkat T cells were stimulated in vitro by the planktonic-conditioned medium (PCM) and biofilm-conditioned medium (BCM) derived from the relevant cultures of the strains tested. The cytokine concentrations were determined in the cell culture supernatants using the flow cytometry. The levels of the cytokines analyzed were lower after stimulation with the BCM when compared to the PCM derived from the cultures of C-CA; statistical significance (p < 0.05) was observed for IL-1ß, IL-12 p70, and IL-10. Similarly, planktonic R-CA and R-SA stimulated a higher cytokine production than their biofilm counterparts. The highest levels of pro-inflammatory IL-8, IL-1ß, and IL-12p70 were observed after stimulation with planktonic R-SA whereas the strongest stimulation of anti-inflammatory IL-10 was noted for the BCM derived from the mixed culture of both reference species. Our results are indicative of weaker immunostimulatory properties of the biofilm C. amycolatum compared to its planktonic form. It may play a role in the persistence of biofilm-related infections. The extent of the cytokine response can be dependent on the inherent virulence of the infecting microorganism.Unravelling of the interplay between the immune system and non-diphtheria corynebacteria would contribute to understanding their increasing role as medically important microorganisms. We aimed at the analysis of pro- (TNF, IL-1ß, IL-6, IL-8, and IL-12p70) and anti-inflammatory (IL-10) cytokines produced by Jurkat T cells in response to planktonic and biofilm Corynebacterium amycolatum. Two reference strains: C. amycolatum ATCC 700207 (R-CA), Staphylococcus aureus ATCC 25923 (R-SA), and ten clinical strains of C. amycolatum (C-CA) were used in the study. Jurkat T cells were stimulated in vitro by the planktonic-conditioned medium (PCM) and biofilm-conditioned medium (BCM) derived from the relevant cultures of the strains tested. The cytokine concentrations were determined in the cell culture supernatants using the flow cytometry. The levels of the cytokines analyzed were lower after stimulation with the BCM when compared to the PCM derived from the cultures of C-CA; statistical significance (p < 0.05) was observed for IL-1ß, IL-12 p70, and IL-10. Similarly, planktonic R-CA and R-SA stimulated a higher cytokine production than their biofilm counterparts. The highest levels of pro-inflammatory IL-8, IL-1ß, and IL-12p70 were observed after stimulation with planktonic R-SA whereas the strongest stimulation of anti-inflammatory IL-10 was noted for the BCM derived from the mixed culture of both reference species. Our results are indicative of weaker immunostimulatory properties of the biofilm C. amycolatum compared to its planktonic form. It may play a role in the persistence of biofilm-related infections. The extent of the cytokine response can be dependent on the inherent virulence of the infecting microorganism.
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Biopelículas , Infecciones por Corynebacterium/inmunología , Corynebacterium/fisiología , Citocinas/inmunología , Linfocitos T/inmunología , Línea Celular , Corynebacterium/genética , Corynebacterium/inmunología , Infecciones por Corynebacterium/genética , Infecciones por Corynebacterium/microbiología , Citocinas/genética , Humanos , Interleucina-10/genética , Interleucina-10/inmunología , Interleucina-1beta/genética , Interleucina-1beta/inmunología , Interleucina-6/genética , Interleucina-6/inmunología , Interleucina-8/genética , Interleucina-8/inmunología , Plancton/genética , Plancton/fisiologíaRESUMEN
Compounds belonging to the group of 5-substituted 4-(1,3,4-thiadiazol-2-yl) benzene-1,3-diols exhibit a broad spectrum of biological activity, including antibacterial, antifungal, and anticancer properties. The mechanism of the antifungal activity of compounds from this group has not been described to date. Among the large group of 5-substituted 4-(1,3,4-thiadiazol-2-yl) benzene-1,3-diol derivatives, the compound 4-(5-methyl-1,3,4-thiadiazole-2-yl) benzene-1,3-diol, abbreviated as C1, was revealed to be one of the most active agents against pathogenic fungi, simultaneously with the lowest toxicity to human cells. The C1 compound is a potent antifungal agent against different Candida species, including isolates resistant to azoles, and molds, with MIC100 values ranging from 8 to 96 µg/ml. The antifungal activity of the C1 compound involves disruption of the cell wall biogenesis, as evidenced by the inability of cells treated with C1 to maintain their characteristic cell shape, increase in size, form giant cells and flocculate. C1-treated cells were also unable to withstand internal turgor pressure causing protoplast material to leak out, exhibited reduced osmotic resistance and formed buds that were not covered with chitin. Disturbances in the chitin septum in the neck region of budding cells was observed, as well as an uneven distribution of chitin and ß(1â3) glucan, and increased sensitivity to substances interacting with wall polymerization. The ATR-FTIR spectral shifts in cell walls extracted from C. albicans cells treated with the C1 compound suggested weakened interactions between the molecules of ß(1â3) glucans and ß(1â6) glucans, which may be the cause of impaired cell wall integrity. Significant spectral changes in the C1-treated cells were also observed in bands characteristic for chitin. The C1 compound did not affect the ergosterol content in Candida cells. Given the low cytotoxicity of the C1 compound to normal human dermal fibroblasts (NHDF), it is possible to use this compound as a therapeutic agent in the treatment of surface and gastrointestinal tract mycoses.
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Antifúngicos/farmacología , Candida albicans/efectos de los fármacos , Candida glabrata/efectos de los fármacos , Candida parapsilosis/efectos de los fármacos , Tiadiazoles/farmacología , Antifúngicos/síntesis química , Aspergillus niger/química , Aspergillus niger/efectos de los fármacos , Aspergillus niger/aislamiento & purificación , Aspergillus niger/ultraestructura , Candida albicans/química , Candida albicans/aislamiento & purificación , Candida albicans/ultraestructura , Candida glabrata/química , Candida glabrata/aislamiento & purificación , Candida glabrata/ultraestructura , Candida parapsilosis/química , Candida parapsilosis/aislamiento & purificación , Candida parapsilosis/ultraestructura , Candida tropicalis/química , Candida tropicalis/efectos de los fármacos , Candida tropicalis/aislamiento & purificación , Candida tropicalis/ultraestructura , Candidiasis/microbiología , Línea Celular , Supervivencia Celular/efectos de los fármacos , Pared Celular/química , Pared Celular/efectos de los fármacos , Pared Celular/ultraestructura , Quitina/antagonistas & inhibidores , Quitina/química , Quitina/metabolismo , Farmacorresistencia Fúngica/efectos de los fármacos , Fibroblastos/citología , Fibroblastos/efectos de los fármacos , Glucanos/antagonistas & inhibidores , Glucanos/química , Glucanos/metabolismo , Humanos , Pruebas de Sensibilidad Microbiana , Rhodotorula/química , Rhodotorula/efectos de los fármacos , Rhodotorula/aislamiento & purificación , Rhodotorula/ultraestructura , Tiadiazoles/síntesis química , Trichophyton/química , Trichophyton/efectos de los fármacos , Trichophyton/aislamiento & purificación , Trichophyton/ultraestructuraRESUMEN
Amphotericin B (AmB) is a very potent antifungal drug with very rare resistance among clinical isolates. Treatment with the AmB formulations available currently is associated with severe side effects. A promising strategy to minimize the toxicity of AmB is reducing its dose by combination therapy with other antifungals, showing synergistic interactions. Therefore, substances that display synergistic interactions with AmB are still being searched for. Screening tests carried out on several dozen of synthetic 1,3,4-thiadiazole derivatives allowed selection of a compound called 4-(5-methyl-1,3,4-thiadiazole-2-yl) benzene-1,3-diol (abbreviated as C1), which shows strong synergistic interaction with AmB and low toxicity towards human cells. The aim of the present study was to investigate the type of in vitro antifungal interactions of the C1 compound with AmB against fungal clinical isolates differing in susceptibility. The results presented in the present paper indicate that the C1 derivative shows strong synergistic interaction with AmB, which allows the use of a dozen to several dozen times lower AmB concentration necessary for 100% inhibition of the growth of pathogenic fungi in vitro. Synergistic interactions were noted for all tested strains, including strains with reduced sensitivity to AmB and azole-resistant isolates. These observations give hope for the possibility of application of the AmB - C1 combinatory therapy in the treatment of fungal infections.
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Anfotericina B/farmacología , Antifúngicos/farmacología , Sinergismo Farmacológico , Hongos/efectos de los fármacos , Micosis/tratamiento farmacológico , Tiadiazoles/farmacología , Humanos , Micosis/microbiología , Micosis/patología , Tiadiazoles/químicaRESUMEN
Non-diphtherial corynebacteria are Gram-positive rods that cause opportunistic infections, what is supported by their ability to produce biofilm on artificial surfaces. In this study, the characteristic of the biofilm produced on vascular and urological catheters was determined using a confocal microscopy for the most frequently involved in infections diphtheroid species. They were represented by the reference strains of Corynebacterium striatum ATCC 6940 and C. amycolatum ATCC 700207. The effect of ciprofloxacin on the biofilm produced by the antibiotic-susceptible C. striatum strain was evaluated using three concentrations of the antimicrobial agent (2 ×, 4 ×, and 6 × the MIC - the Minimum Inhibitory Concentration). The basis for the interpretation of results was the statistical analysis of maximum points readings from the surface comprising a total of 245 areas of the biofilm image under the confocal microscope. It was observed that ciprofloxacin at a concentration equal to 4 × MIC paradoxically caused an enlargement of areas with live bacteria within the biofilm. Biofilm destruction required the application of ciprofloxacin at a concentration higher than 6 × MIC. This suggests that the use of relatively low doses of antimicrobial agents may increase the number of live bacteria within the biofilm, and further facilitate their detachment from the biofilm's structure thus leading to the spread of bacteria into the bloodstream or to the neighboring tissues. The method of biofilm analysis presented here provides the original and novel approach to the investigation of the diphtheroid biofilms and their interaction with antimicrobial agents.Non-diphtherial corynebacteria are Gram-positive rods that cause opportunistic infections, what is supported by their ability to produce biofilm on artificial surfaces. In this study, the characteristic of the biofilm produced on vascular and urological catheters was determined using a confocal microscopy for the most frequently involved in infections diphtheroid species. They were represented by the reference strains of Corynebacterium striatum ATCC 6940 and C. amycolatum ATCC 700207. The effect of ciprofloxacin on the biofilm produced by the antibiotic-susceptible C. striatum strain was evaluated using three concentrations of the antimicrobial agent (2 ×, 4 ×, and 6 × the MIC the Minimum Inhibitory Concentration). The basis for the interpretation of results was the statistical analysis of maximum points readings from the surface comprising a total of 245 areas of the biofilm image under the confocal microscope. It was observed that ciprofloxacin at a concentration equal to 4 × MIC paradoxically caused an enlargement of areas with live bacteria within the biofilm. Biofilm destruction required the application of ciprofloxacin at a concentration higher than 6 × MIC. This suggests that the use of relatively low doses of antimicrobial agents may increase the number of live bacteria within the biofilm, and further facilitate their detachment from the biofilm's structure thus leading to the spread of bacteria into the bloodstream or to the neighboring tissues. The method of biofilm analysis presented here provides the original and novel approach to the investigation of the diphtheroid biofilms and their interaction with antimicrobial agents.
Asunto(s)
Antibacterianos/farmacología , Biopelículas/efectos de los fármacos , Ciprofloxacina/farmacología , Corynebacterium/efectos de los fármacos , Microscopía Confocal , Pruebas de Sensibilidad MicrobianaRESUMEN
Staphylococcus epidermidis small colony variants can survive inside macrophages and their survival has been proposed as a pivotal process in the pathogenesis of biomaterial associated infections. In the present study the intracellular location of clinical isolates of SCV and parental wild type strains inside macrophages was determined. Furthermore, the effect of IFN-γ and rapamycin on the level of SCV/WT as well as lysosomes colocalisation and iNOS induction in THP-activated macrophages in response to WT and SCV strains of Staphylococcus epidermidis were examined. It was demonstrated that SCV strain of S. epidermidis can survive and persist inside macrophages and its intracellular survival is supported by the induction of phagosomal acidification. The ability to reduce the high proportion of LysoTracker positive SCV containing phagosomes was exclusively found when IFN-γ was used. The findings suggest that IFN-γ mediates SCV killing via two distinct mechanisms, phagosome alkalisation and an increased iNOS synthesis, so the cytokine may control S. epidermidis WT and SCV infection in macrophages. Staphylococcus epidermidis SCV is a less potent stimulus of iNOS than the WT strain and the feature may help SCV to persist in hostile environment of macrophages. Rapamycin treatment did not influence the iNOS synthesis but reduced the percentage of both bacterial strains within acidic organelles. However, the percentage of SCV within LysoTracker positive organelles, even though reduced comparing to non-primed cells, was higher than in the WT strain indicating that Staphylococcus epidermidis possesses unique metabolic features allowing SCV to survive within macrophages.
Asunto(s)
Macrófagos , Viabilidad Microbiana , Fagosomas , Staphylococcus epidermidis/metabolismo , Humanos , Macrófagos/metabolismo , Macrófagos/microbiología , Fagosomas/metabolismo , Fagosomas/microbiología , Células THP-1RESUMEN
PURPOSE: The purpose of this study was to investigate the molecular mechanisms of fluoroquinolone resistance in Moraxella catarrhalis clinical strains isolated in Lublin, Poland. MATERIALS AND METHODS: A total of 150 non-duplicate clinical strains of M. catarrhalis were obtained from individuals with signs of upper respiratory tract infection. Bacterial identification was corroborated on the basis of phenotypic and biochemical characteristics as well as with the use of molecular tests. The antimicrobial susceptibility of M. catarrhalis isolates was determined using the disk diffusion method and Etest. Mutations in the gyrase (gyrA and gyrB) and topoisomerase (parC and parE) genes were determined by polymerase chain reaction and sequencing. RESULTS: It was observed that 16.7% of the studied isolates were drug resistant. Resistance to tetracycline was detected for 12% of the strains. Resistance to nalidixic acid, moxifloxacin, and levofloxacin was exhibited by 2.7% of the strains; 1.3% of the strains were resistant to trimethoprim/sulfamethoxazole and 0.7% to erythromycin. Minimum inhibitory concentration values of the four strains demonstrating fluoroquinolone resistance were: 6-12 mg/L for nalidixic acid, 1-1.5 mg/L for levofloxacin, 1 mg/L for moxifloxacin, and 0.25-0.5 mg/L for ciprofloxacin. The research resulted in the detection of mutations in 4 strains, in gyrase gyrA and gyrB genes. In gyrA gene, there occurred mutation G412C as well as four silent transition mutations. Within gyrB gene, there occurred mutation, substitution A1481G, as well as two identical silent mutations. CONCLUSION: Our findings reveal that resistance to fluoroquinolones in M. catarrhalis is connected with amino acid substitutions in gyrA and gyrB genes. To our knowledge, this work is the first description of fluoroquinolone-resistant clinical strains of M. catarrhalis with described mutations in gyrA and gyrB genes isolated in Poland and in Europe.