Phenotypic and genomic characteristics of oxacillin-susceptible mecA-positive Staphylococcus aureus,rapid selection of high-level resistance to beta-lactams

European Journal of Clinical Microbiology & Infectious Diseases - The aim of this study is to describe the phenotypic and genetic properties of oxacillin-susceptible methicillin-resistant...     

Towards clinical data-driven eligibility criteria optimization for interventional COVID-19 clinical trials

ObjectiveThis research aims to evaluate the impact of eligibility criteria on recruitment and observable clinical outcomes of COVID-19 clinical trials using electronic health record (EHR) data.Materials and MethodsOn June 18, 2020, we identified frequently used eligibility criteria from all the interventional COVID-19 trials in ClinicalTrials.gov (n = 288), including age, pregnancy, oxygen saturation, alanine/aspartate aminotransferase, platelets, and estimated glomerular filtration rate. We applied the frequently used criteria to the EHR data of COVID-19 patients in Columbia University Irving Medical Center (CUIMC) (March 2020–June 2020) and evaluated their impact on patient accrual and the occurrence of a composite endpoint of mechanical ventilation, tracheostomy, and in-hospital death.ResultsThere were 3251 patients diagnosed with COVID-19 from the CUIMC EHR included in the analysis. The median follow-up period was 10 days (interquartile range 4–28 days). The composite events occurred in 18.1% (n = 587) of the COVID-19 cohort during the follow-up. In a hypothetical trial with common eligibility criteria, 33.6% (690/2051) were eligible among patients with evaluable data and 22.2% (153/690) had the composite event.DiscussionBy adjusting the thresholds of common eligibility criteria based on the characteristics of COVID-19 patients, we could observe more composite events from fewer patients.ConclusionsThis research demonstrated the potential of using the EHR data of COVID-19 patients to inform the selection of eligibility criteria and their thresholds, supporting data-driven optimization of participant selection towards improved statistical power of COVID-19 trials.     

Protection against mouse and avian influenza A strains via vaccination with a combination of conserved proteins NP, M1 and NS1

Background Experimental data accumulated over more than a decade indicate that cross‐strain protection against influenza may be achieved by immunization with conserved influenza proteins. At the same time, the efficacy of immunization schemes designed along these lines and involving internal influenza proteins, mostly NP and M1, has not been sufficient. Objective To test the immunogenicity and protective efficacy of DNA vaccination with a combination of NP, M1 and NS1 genes of influenza virus. Methods The immunogenicity and protective efficacy of DNA vaccination with NP, M1 and NS1 was tested in mice and chickens. Mice were challenged with mouse‐adapted viral strains H3N2 and H5N2 and chicken challenged with avian H5N3 virus. Results In these settings, wild‐type NS1 did not impede the cellular and humoral response to NP/M1 immunization in vivo. Moreover, addition of NS1‐encoding plasmid to the NP/M1 immunization protocol resulted in a significantly increased protective efficacy in vivo. Conclusions The addition of NS1 to an influenza immunization regimen based on conserved proteins bears promise. It is feasible that upon further genetic modification of these and additional conserved influenza proteins, providing for their higher safety, expression and immunogenicity, a recombinant vaccine based on several structural and non‐structural proteins or their epitopes will offer broad anti‐influenza protection in a wide range of species.     

SHP1 expression in bone marrow biopsies of myelodysplastic syndrome patients: a new prognostic factor

Myelodysplastic syndrome (MDS) progresses into acute leukaemia with a variable time course. We analysed 45 newly diagnosed patients and found that the expression of the Src homology 2 domain-containing tyrosine phosphatase 1 (SHP1) had a significant impact on disease severity, progression and overall prognosis. Global or lineage-specific loss of SHP1 was observed by immunohistochemistry in bone marrow biopsies of MDS patients who progressed rapidly (P = 0.0021) and had shorter survival (P < 0.001). Cox regression analysis demonstrated that SHP1 expression in megakaryocytes had prognostic relevance for time to progression (P = 0.009) and overall survival (P = 0.001).     

Helicobacter pylori susceptibility to fosfomycin,rifampin, and 5 usual antibiotics for H. pylori eradication

The aim of the study was to assess Helicobacter pylori resistance to fosfomycin, rifampin, and 5 other antibiotics. Susceptibility of 50 consecutive H. pylori strains was tested by E test and breakpoint susceptibility testing method. Overall and primary resistance rates were amoxicillin 2.0 and 0%, metronidazole 34.0 and 31.2%, tetracycline 2.0 and 2.1%, levofloxacin 18.0 and 16.7%, rifampin 12.0 and 10.4%, and fosfomycin 8.3 and 6.5%, respectively. Clarithromycin resistance was >20.0% in all patients (22.0%) and in untreated subjects (20.8%). Rifampin resistance was higher than those usually reported. Quinolone resistance rose from 2005–2007 (8.7%) to 2012–2013 (18.0%). High double/multidrug resistance rates (overall 22.0% and 20.0% in untreated adults), including a 5-fold resistance, were found. In conclusion, fosfomycin and rifampin resistance rates were much lower than that of metronidazole and slightly lower than those of clarithromycin and levofloxacin. MICs₉₀ of both fosfomycin and rifampin were lower than those of clarithromycin and metronidazole. Thus, in countries of high/increasing H. pylori resistance and multidrug resistance, both rifamycins and fosfomycin could be helpful in rescue regimens.     

Serum profiles of free and conjugated neuroactive pregnanolone isomers in nonpregnant women of fertile age

BACKGROUND: Pregnanolone isomers (PI) with a hydroxy group in the 3alpha-position are neuroinhibitors operating via positive modulation of GABA(A) receptors. The 3beta-PI and sulfates of PI and pregnenolone exert the opposite effect. In addition to the brain's in situ synthesis, some circulating steroids can penetrate the blood-brain barrier. METHODS: To assess the physiological impact of peripheral endogenous neuroactive pregnanolone isomers and their polar conjugates in women, serum allopregnanolone (P3alpha5alpha), isopregnanolone (P3beta5alpha), pregnanolone (P3alpha5beta), epipregnanolone (P3beta5beta), pregnenolone, estradiol (including their polar conjugates), and additional steroids were measured in 16 women in the follicular and luteal phases of the menstrual cycle using gas chromatography/mass spectrometry and RIA for the analysis. Linear models and Spearman's correlations were used for data evaluation. RESULTS AND DISCUSSION: The levels of conjugated PI were from one to almost three orders of magnitude higher in comparison with the free steroids. The results indicate that a substantial proportion of the progesterone is metabolized in the sequence progesterone-->5beta-dihydroprogesterone-->P3alpha5beta-->conjugated P3alpha5beta. The sulfation of PI and particularly of P3alpha5beta moderates the levels of free PI and restrains estradiol biosynthesis via progesterone degradation. PI including the conjugates reflected changing progesterone formation during the menstrual cycle. In the follicular phase, the positive correlation with conjugated pregnenolone, the independence of progesterone, and the negative age relationships of PI indicate their adrenal origin. The dependence on progesterone and the independence of conjugated pregnenolone suggest a gonadal source of PI in the luteal phase. The neuroactivating PI prevailed over neuroinhibiting PI.     

Neuroactive pregnanolone isomers during pregnancy

The pregnanolone isomers (PI) allopregnanolone (3alpha-hydroxy-5alpha-pregnan-20-one), pregnanolone (3alpha-hydroxy-5beta-pregnan-20-one), isopregnanolone (3beta-hydroxy-5alpha-pregnan-20-one), epipregnanolone (3beta-hydroxy-5beta-pregnan-20-one), progesterone, and estradiol were measured in 138 pregnant women. The sampling was carried out from the first through the 10th month of pregnancy. Gas chromatography-mass spectrometry analysis and RIA were used for the measurement of steroid levels. The ratios of individual PI were similar to those found previously around parturition: about 25:10:7:1 for allopregnanolone, pregnanolone, isopregnanolone, and epipregnanolone, respectively. All the PI showed a significant increase during pregnancy, which was more pronounced in the 3alpha-steroids. The results indicated changing ratios between 3alpha- and 3beta-PI and between 5alpha- and 5beta-PI throughout pregnancy. The constant allopregnanolone/isopregnanolone ratio found through pregnancy weakened the hypothesis of the role of isopregnanolone in the onset of parturition. The ratio of estradiol (stimulating uterine activity) to 5alpha-PI and epipregnanolone exhibited significant changes during pregnancy in favor of estradiol up to the sixth or seventh month, in contrast to the constant estradiol/pregnanolone ratio. A pregnancy-stabilizing role of pregnanolone, counterbalancing the stimulating effect of estradiol on the onset of parturition, was suggested.     

Hsp90 and its co-chaperone, Sgt1, as autoantigens in dilated cardiomyopathy

Recently, it has been suggested that some heat shock proteins such as Hsp70 and Hsp60 are involved in autoimmune diseases including cardiospecific ones. In this work we focused on the involvement of another well known heat shock protein, Hsp90, and its novel co-chaperone, Sgt1, in dilated cardiomyopathy (DCM). We found that the level of autoantibodies against these two proteins was significantly higher in patients with DCM and ischemic heart disease than in sera of healthy donors. We have also analyzed the expression level and subcellular localization of Hsp90 and Sgt1 in diseased myocardia. Using Western blot we found changes in subcellular localization of Hsp90 in the left ventricle of DCM hearts while the total level of this protein remained unchanged. Regarding the Sgt1 protein, we found an increased level in DCM and no changes in subcellular localization. Taken together, our data suggest that Hsp90 and Sgt1 might be involved in the progression of heart failure and might serve as markers for cardiomyopathies of different origin.