Distribution of virulence determinants among Escherichia coli ST131 and its H30/H30-Rx subclones in Turkey.

Extraintestinal pathogenic Escherichia coli (ExPEC) is the leading pathogen in urinary tract infection. In recent years multidrug-resistant B2-ST131 E. coli clonal group has disseminated worldwide. The ST131 and its subclones H30 and H30-Rx have been identified only in a few studies from Turkey. The aim of this study is to investigate the presence of ST131 and its subclones and to analyze their adhesin virulence genes and antimicrobial resistance. A total of 250 urinary ExPEC isolates were included in the study. Resistance rates of 16 antimicrobial agents were determined by disk-diffusion. Multidrug-resistance and ESBL production were analyzed. Altogether 8 adhesin genes were investigated namely, papAH, fimH, sfa/focDE, focG, afa/draBC, iha, bmaE and gafD. A total of 39 ST131 isolate were determined and 33 (84.6%) were multidrug-resistant. ESBL production was detected in 34 (87.2%) ST131 and 61 (28.9%) of non-ST131 strains. In our study, we found a strong correlation between ST131 strains and fimH, iha, afa/draBC, papAH virulence determinants. Twenty-nine (85.3%) of 34 ST131-O25b-H30 isolates were identified as H30-Rx. All the papAH gene positive isolates were identified within ST131-O25b-H30-Rx lineage. Non-H30-Rx isolates within H30 isolates were identified as pattern 2. Almost 16% of the isolates were identified as ST131 regardless of clinical syndrome and approximately 34% of the multidrug-resistant isolates were H30-Rx subclone. We report H30-Rx as the dominant subclone of ST131 in our study. Imipenem, fosfomycin and nitrofurantoin proved to be the most effective agents according to antibiotic resistance patterns of both ST131 and non-ST131 E. coli strains.

CXCL10/IP10 as a Biomarker Linking Multisystem Inflammatory Syndrome and Left Ventricular Dysfunction in Children with SARS-CoV-2.

BACKGROUND: To investigate the diagnostic accuracy of CXCL10/IP10 for left ventricular (LV) dysfunction in multisystemic inflammatory syndrome (MIS-C). METHODS: This cross-sectional, longitudinal study included 36 patients with MIS-C. Patients were classified as follows: (1) patients presenting with Kawasaki-like features (group I = 11); (2) patients presenting with LV systolic dysfunction (group II = 9); and (3) other presentations (group III = 3). CXCL10/IP10 levels were measured upon admission and on days 3 and 7 of treatment. RESULTS: Twenty patients were male and 16 were female. The median age of patients at diagnosis was 7.5 (1.5-17) years. All patients had a fever lasting for a median of 4 (2-7) days. Ten patients had LV systolic dysfunction. The duration of hospitalization was longer in group II. Lymphocyte and platelet counts were lower, whereas NT-pro-BNP, troponin-I, D-dimer, and CXCL10/IP10 levels were higher in group II. Baseline levels of CXCL10/IP10 were weakly negatively correlated with ejection fraction (r = -0.387, p = 0.022). Receiver operator characteristic curve analysis yielded a cutoff value of CXCL10/IP10 to discriminate patients with LV dysfunction was 1839 pg/mL with sensitivity 88% and specificity 68% (Area under curve (AUC) = 0.827, 95% CI 0.682-0.972, p = 0.003). CONCLUSION: Having a good correlation with cardiac function, CXCL10/IP10 is a potential biomarker to predict LV dysfunction in MIS-C patients.

Comparison of an inactivated Covid19 vaccine-induced antibody response with concurrent natural Covid19 infection.

OBJECTIVES: The risk of contracting SARS-CoV-2 is high among the health care workers (HCW). The comparison between the antibody response to an inactivated Covid19 vaccine and the antibodies that developed during Covid-19 infection has not been elucidated. In this study, vaccine-induced antibody levels were compared with the antibodies developed in naturally infected HCWs. METHODS: Eighty vaccinated individuals and 80 Covid-19 patients enrolled to the study. Both groups were matched on age, gender and antibody testing time. Anti-SARS-CoV-2 total Ig (Roche) and Anti-SARS-CoV-2 ELISA (IgG) (Euroimmun, Germany) were used to detect antibodies. RESULTS: The anti-S positivity were determined to be 96.2% and 92.5% in vaccinated and patient groups (p=0.303) while the anti-N positivity was 51.2% and 98.8%, respectively (p=<0,0001). The median values for anti-S and anti-N antibodies were statistically significant between both groups. When the vaccinated group was compared with the severe and non-severe patient groups, statistically significant differences were found for both regarding anti-S1 and anti-N antibody titers (p=0,012, p=<0,0001, respectively). For the patient group, there was a positive correlation between the age and anti-S1 antibody titers (r=0.333; p=0.003) and there was also a statistically significant increase in anti-N antibody titers in time (r=0.505; p=0.0001). CONCLUSION: The anti-S seroconversion ratio in vaccinated individuals were higher than what was reported by the vaccine manufacturer. The antibody titers in the vaccinated group were lower than the patients group. The decrease in anti-S1 antibody titers in time were considered to be a disadvantage and an undesired phenomenon.

[Comparison of Antiviral Effect of Olive Leaf Extract and Propolis with Acyclovir on Herpes Simplex Virus Type 1]. / Zeytin Yapragi Ekstresi ve Propolisin Herpes Simpleks Virüsü Tip 1 Üzerine Antiviral Etkisinin Asiklovir ile Karsilastirilmasi.

While acyclovir, a nucleoside analogue, is widely used for herpes simplex virus type 1 (HSV-1), emergence of drug-resistant viruses due to frequent usage of this class of medicines, and their toxic side effects require exploring novel active molecules. Despite the studies on developing synthetic molecules in medical sciences and pharmacology, herbs as a natural source of biologically-active compounds remain popular. In this in vitro study, olive leaf extract (OLE) and propolis alone or in combination with acyclovir were investigated for their antiviral efficacy in HSV-1.Toxic doses of OLE, propolis, and dimethyl sulfoxide, propolis diluent, for Hep-2 (ATCC, CCL-23) cells were determined by conventional cell culture. Using "endpoint" method, the viral dose infecting half of the cell culture (TCID50) was calculated, and viral quantity was determined with Spearman-Karber method. Antiviral effects of OLE and propolis on HSV-1 were investigated by conventional cell culture and real-time cell analysis (RTCA). Combinations of the two extracts with one another and with acyclovir were evaluated by RTCA. Active substances prepared at three different dilutions were added to tubes with HSV-1 of logTCID50: 11.5 in descending order starting from the highest non-toxic concentration, and they were left at room temperature for two different durations (one hour and three hours). The aliquots taken from the tubes were cultured in plates containing Hep-2 cells and evaluated after 72 hours. Combinations of extracts and acyclovir at concentrations at least four times lower than the lowest concentration showing antiviral efficacy against HSV-1 were cultured with Hep-2 cells in the e-plates of the xCELLigence RTCA device, measurements were obtained at 30 minute intervals, and data were recorded in real time. In the test with two different durations and at different concentrations of OLE and propolis, antiviral efficacy was observed both with one-hour and three-hour incubation at a concentration of 10 µg/ ml for propolis and 1.2 mg/ml for OLE with RTCA. The duration and concentration of the greatest decrease in viral quantity were in the first one hour and 10 µg/ml for propolis, and in the first one hour and 1.2 mg/ ml for OLE. Combination of propolis and OLE with acyclovir caused no cytopathic effects, and the combination of extracts led to delayed cytopathic effect. According to these results, propolis and OLE, alone and in combinations with acyclovir, have antiviral efficacy against HSV-1. These agents may reduce the dose and side effects of acyclovir in case of co-administration since they exert their effects through a different mechanism than acyclovir,possibly through direct virucidal activity, inhibition of virus internalization or viral inhibition in early stages of replication (inhibition of adsorption/binding of viral particles to the cell). These extracts that do not require conversion to active form have the potential to reduce infectivity in oral lesions, prevent spread, and be used in the topical treatment of acyclovir-resistant HSV infections, particularly in immunocompromised patients. However, in vivo studies should be conducted to determine their medicinal properties and potential toxicities. These results should be supported by further comprehensive studies and the efficacy against other viruses should also be investigated.

In vitro antibacterial activity of ciprofloxacin loaded chitosan microparticles and their effects on human lung epithelial cells.

Chitosan (CS), due to its inherent mucoadhesive property and biofilm penetration ability, can be considered as very potent vehicle for local drug delivery to the lungs. This study reports on the preparation and in vitro antibacterial activity and cytotoxicity determination of ciprofloxacin loaded chitosan (Cipro-CS) microparticles with size in the range of 0.1-1 µm, which may provide advantages of lower nanotoxicity and lower local clearance. Cipro-CS microparticles were prepared by ionic gelation method and their size, zeta potential and drug release pattern determined. The antibacterial activities of CS and Cipro-CS microparticles against pneumonia causing agents, namely Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus, were evaluated by determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The biocompatibility of the microparticles was tested in the human lung epithelial cell (BEAS-2B) culture, and microparticle association with the bacteria and epithelial cells was evaluated by transmission electron microscopy. Only the Cipro-CS microparticles, but not the CS microparticles, inhibited bacterial growth at concentrations not significantly cytotoxic to BEAS-2B cells. The Cipro-CS microparticles were able to damage the cell wall and membrane of the bacteria, and the ones ≤200 nm in size were internalized by both the BEAS-2B cells and the microorganisms.

Production and mechanical properties of Ti-5Al-2.5Fe-xCu alloys for biomedical applications.

In this study, the mechanical, antibacterial properties and cell toxicity response of Ti-5Al2.5Fe alloy with different copper contents were investigated. The alloys were prepared by high-energy ball milling using elemental Ti, Al, Fe, and Cu powders and consolidated by a uniaxial vacuum hot press. Staphylococcus aureus strain ATCC 29213 and Escherichia coli strain ATCC 25922 were used to determine the antibacterial properties of the sintered alloys. The in vitro cytotoxicity of the samples was evaluated with HeLa (ATTC, CCL-2) cells using thiazolyl blue tetrazolium bromide. The mechanical behavior of the samples was determined as a function of hardness and bending tests and analyzed by scanning electron microscopy, energy dispersive x-ray spectroscopy, optical microscopy and x-ray diffraction (XRD). The results showed that the Cu content significantly improved the antibacterial properties. Cu addition prevented the formation of E. coli and S. aureus colonies on the surface of the samples. All samples exhibited very good cell biocompatibility. The alloys with different copper contents showed different mechanical properties, and the results were correlated by microstructural and XRD analyses in detail. Our results showed that Cu has a great effect on the Ti5Al2.5Fe alloy and the alloy is suitable for biomedical applications with enhanced antibacterial activity.

Comparative Analysis of Proteome Patterns of Francisella tularensis Isolates from Patients and the Environment.

Francisella tularensis is the causative agent of tularemia. Although major contributors and the main mechanism of the virulence are well known, some of the molecular details are still missing. Proteomics studies regarding F. tularensis have provided snapshot pictures of the organism grown under different culture conditions to understand the mechanism of virulence. In general, such studies were carried out with standard strains e.g., LVS and did not involve comparisons of F. tularensis isolates from either clinical or environmental sources. In this study, we performed two-dimensional gel electrophoresis (2DE)-based proteomic analysis and compared the protein profiles of the F. tularensis subsp. holarctica strains isolated from the clinical and the environmental samples. Regulations were detected in 14 spots when twofold regulation criteria were applied. The regulated protein spots were subjected to MALDI-TOF/TOF analysis and identified. Classification of the identified proteins based on metabolic functions revealed that the translation machinery was the most varying metabolic processes among the isolates. Using normalized protein spot intensities, PCA analysis was also performed. The results indicated that the strain isolated from water source was different then the strains isolated from the patients. Most interestingly, the isolates were strikingly distinguishable from the standard NCTC 10857 strain.

Evaluation of Effect of Topical Tacrolimus Treatment on Herpetic Stromal Keratitis in a Rat Model.

OBJECTIVES: To investigate the effectiveness of topical tacrolimus treatment on herpetic stromal keratitis (HSK) in a rat model. METHODS: The development of HSK was monitored for 14 days after the inoculation of rats with herpes simplex type 1 virus. Rats that developed HSK were divided into four groups as follows: (1) topical antiviral treatment (control), (2) topical antiviral and 1% prednisolone acetate, (3) topical antiviral and 0.03% tacrolimus ointment, and (4) topical antiviral plus 0.1% tacrolimus ointment. After 14 days of treatment, the severity levels of HSK were scored and compared with the levels before the treatment. The expression of CD3, CD4, and CD8 was evaluated by flow cytometry. The development of the disease was evaluated clinically and histologically. RESULTS: Significant improvement in vascularization was observed in the groups with the drug treatment in addition to the antiviral agent (P<0.05), but there was no obvious difference within groups 2, 3, and 4 in the vascularization severity. The regression of corneal edema was 8.05%±6% in group 1, 25.17%±14.55% in group 2 (P=0.01), 36.40%±21.69% in group 3 (P=0.03), and 46.39%±14.96% in group 4 (P=0.00). A significant decrease in the number of inflammatory cells in the groups with the drug treatment was evaluated by immunohistochemical staining and confirmed by flow cytometry analysis. CONCLUSIONS: Topical tacrolimus treatment caused a significant decrease in corneal vascularization accompanied by a lower number of inflammatory cells in the experimental HSK corneal edema model. Therefore, topical tacrolimus has the potential to be used in the treatment of HSK.

Relationship between phylogenetic groups, antibiotic resistance and patient characteristics in terms of adhesin genes in cystitis and pyelonephritis isolates of Escherichia coli.

Extraintestinal pathogenic Escherichia coli (E. coli) is considered as the main causative agent of urinary tract infections worldwide. The relationship between antimicrobial resistance, phylogenetic groups, patient characteristics and adhesin virulence genes are complex and not fully understood. In this study, among 146 urinary isolates of E. coli, phylogenetic groups and various adhesin virulence genes were examined with multiplex Polymerase Chain Reaction methods. Patient characteristics divided into sex, cystitis and pyelonephritis; community-acquired and hospital-acquired; complicated and uncomplicated infection. Antimicrobial resistance was also determined. The papAH gene was seen more often in pyelonephritis than cystitis and female than male patients. iha gene was more frequent in hospital-acquired infections than in community-acquired infections. sfa/focDE was more frequent in ampicillin, amikacin, gentamicin, nalidixic acid, norfloxacin, cefuroxime, ceftriaxone, cefazolin, cefotaxime, ciprofloxacin and trimethoprim/sulfamethoxazole susceptible and extended-spectrum ß-lactamase (ESBL) and multi-drug resistance (MDR) negative isolates. focG was seen more often in nalidixic acid, norfloxacin, cefuroxime, ceftriaxone, ciprofloxacin susceptible and MDR negative isolates. fimH and papAH were more commonly observed in amoxicillin/clavulanic acid and cefotaxime susceptible isolates, respectively. iha and afa/draBC genes were more frequent in resistant isolates than the susceptible ones; for iha, in ampicillin, amoxicillin/clavulanic acid, nalidixic acid, cefuroxime, ceftriaxone resistant and ESBL and MDR positive isolates; for afa/draBC, in cefotaxime, cefuroxime, ciprofloxacin, trimethoprim/sulfamethoxazole resistant and ESBL and MDR positive isolates, this trend was observed. ST 131 E. coli virulence gene pattern has a direct effect on resistance profile. Isolates belong to that clonal group has MDR and commonly harbour afa/draBC and iha genes. Our findings may provide new insights into the relationships between pathogenesis, patient characteristics and resistance of E. coli UTI.

Evaluation of immunochromatographic test for the detection of antibodies against Echinococcosis granulosus.

BACKGROUND: Echinococcosis in humans is a disease caused by the larvae of Echinococcus granulosus (E. granulosus) and Echinococcus multilocularis (E. multilocularis). Serological tests are valuable, especially in the clarification of unexplained clinical findings and imaging methods. For this reason, indirect hemagglutination (IHA), latex agglutination, immunoelectrophoresis, immunoblotting, immuno-enzymatic tests, indirect fluorescence antibody test (IFAT), and enzyme-linked immunosorbent assay (ELISA) are used. The purpose of this study was to investigate the value of an immunochromatographic test (ICT) specific for E. granulosus antibodies in the diagnosis of echinococcosis. MATERIAL/METHODS: ICT evaluated 102 cases of cystic echinococcosis, 38 cases of other parasitic diseases, and 50 healthy individuals. ELISA (DRG, Germany) that detects IgG antibodies specific for E. granulosus was used as the reference method. RESULTS: The sensitivity, specificity, and positive and negative predictive values of ICT were 96.8%, 87.5%, 98.9%, and 70%, respectively. Diagnostic value was 96.1%. No significant differences and high degrees of agreement were found between ELISA and immunochromatographic test for cystic echinococcosis. Serum samples included 4 taeniasis, 2 leishmaniasis, and 2 healthy individuals were diagnosed to be positive with immunochromatographic test. CONCLUSIONS: The ability of test to give fast results without need for equipment, devices, and specific storage conditions is an advantage. This test may be used due to its advantages in endemic regions for screening and diagnostic purposes.