Co-occurrence of the cephalosporinase cepA and carbapenemase cfiA genes in a Bacteroides fragilis division II strain, an unexpected finding.

BACKGROUND: Bacteroides fragilis, an anaerobic gut bacterium and opportunistic pathogen, comprises two genetically divergent groups (or divisions) at the species level. Differences exist both in the core and accessory genomes and the beta-lactamase genes, with the cephalosporinase gene cepA represented only in division I and the carbapenemase gene cfiA only in division II. METHODS: Multidrug resistance in a clinical B. fragilis strain was examined by whole-genome sequencing. RESULTS: Strain CNM20200260 carried the antimicrobial resistance genes cepA, cfiA2, ant(6'), erm(F), mef(En2), est(T), tet(Q) and cat(A), along with 82-Phe mutation in gyrA (together with 47 amino acid changes in gyrA/B and parC/parE). bexA/B and other efflux pump genes were also observed. None of the detected insertion sequences was located upstream of cfiA2. The genome-based taxonomy coefficients (average nucleotide identity, DNA-DNA hybridization similarity and difference in genomic G + C%) with respect to genomes of the strains of B. fragilis division II and the novel species Bacteroides hominis (both cfiA-positive) met the criteria for CNM20200260 to belong to either species (>95%, >70% and <1%, respectively). No such similarity was seen with type strain NCTC 9343 or the representative genome FDAARGOS 1225 of B. fragilis (division I, cfiA-negative). Strain CNM20200260 harboured four out of nine Kyoto Encyclopedia of Genes and Genomes orthologues defined for division I and one of two defined for division II. CONCLUSIONS: This is the first description of the co-occurrence of cepA and cfiA in a Bacteroides strain, confirming the complexity of the taxonomy of this species.

Multiresistance to Nonazole Fungicides in Aspergillus fumigatus TR34/L98H Azole-Resistant Isolates.

Drug resistance is a worldwide problem affecting all pathogens. The human fungal pathogen Aspergillus fumigatus coexists in the environment with other fungi targeted by crop protection compounds, being unintentionally exposed to the selective pressure of multiple antifungal classes and leading to the selection of resistant strains. A. fumigatus azole-resistant isolates are emerging in both clinical and environmental settings. Since their approval, azole drugs have dominated clinical treatment for aspergillosis infections and the agriculture fungicide market. However, other antifungal classes are used for crop protection, including benzimidazoles (methyl benzimidazole carbamates [MBCs]), strobilurins (quinolone oxidation inhibitors [QoIs]), and succinate dehydrogenase inhibitors (SDHIs). Mutations responsible for resistance to these fungicides have been widely researched in plant pathogens, but resistance has not been explored in A. fumigatus. In this work, the genetic basis underlying resistance to MBCs, QoIs, and SDHIs was studied in azole-susceptible and -resistant A. fumigatus strains. E198A/Q and F200Y mutations in ß-tubulin conferred resistance to MBCs, G143A and F129L substitutions in cytochrome b conferred resistance to QoIs, and H270R/Y mutations in SdhB conferred resistance to SDHIs. Characterization of susceptibility to azoles showed a correlation between strains resistant to these fungicides and the ones with tandem-repeat (TR)-based azole resistance mechanisms. Whole-genome sequencing analysis showed a genetic relationship among fungicide multiresistant strains, which grouped into subclusters that included only strains carrying the TR-based azole resistance mechanisms, indicating a common ancestor/evolution pattern and confirming the environmental origin of this type of azole-resistant A. fumigatus.

Potential models for the simulation of methane adsorption on graphene: development and CCSD(T) benchmarks.

Different force fields for the graphene-CH4 system are proposed including pseudo-atom and full atomistic models. Furthermore, different charge schemes are tested to evaluate the electrostatic interaction for the CH4 dimer. The interaction parameters are optimized by fitting to interaction energies at the DFT level, which were themselves benchmarked against CCSD(T) calculations. The potentials obtained with both the pseudo-atom and full atomistic approaches describe accurately enough the average interaction in the methane dimer as well as in the graphene-methane system. Moreover, the atom-atom potentials also correctly provide the energies associated with different orientations of the molecules. In the atomistic models, charge schemes including small charges allow for the adequate representation of the stability sequence of significant conformations of the methane dimer. Additionally, an intermediate charge of -0.63e on the carbon atom in methane leads to bond energies with errors of ca. 0.07 kcal mol-1 with respect to the CCSD(T) values for the methane dimer. For the graphene-methane interaction, the atom-atom potential model predicts an average interaction energy of 2.89 kcal mol-1, comparable to the experimental interaction energy of 3.00 kcal mol-1. Finally, the presented force fields were used to obtain self-diffusion coefficients that were checked against the experimental value found in the literature. The no-charge and Hirshfeld charge atom-atom models perform extremely well in this respect, while the cheapest potential considered, a pseudo-atom model without charges, still performs reasonably well.

MP2 Study of Physisorption of Molecular Hydrogen onto Defective Nanotubes: Cooperative Effect in Stone-Wales Defects.

We use large-scale MP2 calculations to investigate the physisorption of molecular hydrogen on (9,0) defective carbon nanotubes (CNTs) of C72H18. These large (supra)molecular systems are typically studied using conventional DFT methods, which do not describe well the van der Waals interactions responsible for this process. Here we use CCSD(T)-calibrated MP2 calculations to estimate binding energies by considering four defective structures (hydrogenated divacancy, octagon-pentagon, and two Stone-Wales defects). The largest physisorption energies for the nondefective CNT are for configurations in which H2 points toward the center of one ring. The computed interaction energies for defect-free CNT are in the range 5.7 to 5.9 kJ/mol, in good agreement with the experimental value of 5.98 kJ/mol. The defects introduced in the (9,0)-CNT increase the surface area of the nanotube, such that the largest surface in found in the 55-77 Stone-Wales defective CNT that furthermore is the most aromatic. Only that defect enlarges the physisorption binding energy, which can become >25% larger. Moreover, a cooperative effect in the adsorption of H2 not appearing in the regular structure is found.

Y155H amino acid substitution in influenza A(H1N1)pdm09 viruses does not confer a phenotype of reduced susceptibility to neuraminidase inhibitors.

The Y155H amino acid substitution in the neuraminidase gene (NA) has previously been associated with highly reduced inhibition by neuraminidase inhibitors in the seasonal H1N1 influenza A virus which circulated in humans before the 2009 pandemic. During the 2012/13 epidemic season in Spain, two A(H1N1) pdm09 viruses bearing the specific Y155H substitution in the NA were detected and isolated from two patients diagnosed with severe respiratory syndrome and pneumonia requiring admission to the intensive care unit. Contrary to what was observed in the seasonal A(H1N1) viruses, neither of the Y155H A(H1N1) pdm09 viruses described here showed a phenotype of reduced inhibition by NAIs as determined by the neuraminidase enzyme inhibition assay (MUNANA). High-throughput sequencing of the NA of both Y155H viruses showed that they were composed to >99% of H155 variants. We believe that this report can contribute to a better understanding of the biological significance of amino acid substitutions in the neuraminidase protein with regard to susceptibility of influenza viruses to neuraminidase inhibitors. This is of critical importance for optimal management of influenza disease patients.

Candida tropicalis antifungal cross-resistance is related to different azole target (Erg11p) modifications.

Candida tropicalis ranks between third and fourth among Candida species most commonly isolated from clinical specimens. Invasive candidiasis and candidemia are treated with amphotericin B or echinocandins as first-line therapy, with extended-spectrum triazoles as acceptable alternatives. Candida tropicalis is usually susceptible to all antifungal agents, although several azole drug-resistant clinical isolates are being reported. However, C. tropicalis resistant to amphotericin B is uncommon, and only a few strains have reliably demonstrated a high level of resistance to this agent. The resistance mechanisms operating in C. tropicalis strains isolated from clinical samples showing resistance to azole drugs alone or with amphotericin B cross-resistance were elucidated. Antifungal drug resistance was related to mutations of the azole target (Erg11p) with or without alterations of the ergosterol biosynthesis pathway. The antifungal drug resistance shown in vitro correlated very well with the results obtained in vivo using the model host Galleria mellonella. Using this panel of strains, the G. mellonella model system was validated as a simple, nonmammalian minihost model that can be used to study in vitro-in vivo correlation of antifungals in C. tropicalis. The development in C. tropicalis of antifungal drug resistance with different mechanisms during antifungal treatment has potential clinical impact and deserves specific prospective studies.

[Burnout syndrome in health professionals at a university hospital in Spain]. / Síndrome de Burnout en profesionales sanitarios en un hospital universitario de España.

OBJECTIVE: To evaluate the risk of Burnout in the health professionals of the Huesca University Hospital, 50 years after its inauguration, to investigate the variables related to the work motivation of the hospital health personnel and to assess the predisposing and protective factors of the risk of Burnout. MATERIAL AND METHODS: An observational, analytical, prospective and unicentric study was conducted from September 2017 to April 2019, evaluating all the health professionals who worked at the Hospital San Jorge de Huesca (n=209). RESULTS: The mean age was 42.86 years. 72.2% were women. 12.4% had moderate risk of burnout. There was 12.4% of high emotional exhaustion, 36.8% of high depersonalization and 44.5% of low personal accomplishment. Burnout was statistically significant associated with the professional category (P=.010), work experience (P=.026), hours of work per week (P=.036), choice of the same profession (P=.001) and recommendation to the offspring (P<.001). CONCLUSIONS: One tenth of the sample had a moderate risk of burnout. Almost half of the health workers confirmed a high degree of satisfaction with the work environment and the majority expressed an adequate use of well-being strategies and a high degree of autonomy, recognition and satisfaction at work.

Frequency of voriconazole resistance in vitro among Spanish clinical isolates of Candida spp. According to breakpoints established by the Antifungal Subcommittee of the European Committee on Antimicrobial Susceptibility Testing.

A total of 4,226 Spanish clinical isolates of Candida spp. were analyzed to assess resistance to voriconazole according to breakpoints established by the European Committee for Antimicrobial Susceptibility Testing (where susceptibility [S] to voriconazole corresponds to a MIC of ≤ 0.12 mg/liter). Resistance was uncommon among Candida albicans (5%), C. parapsilosis (1.2%), and C. tropicalis (11%) isolates. Voriconazole MICs of >0.12 mg/liter were more frequent among Candida glabrata and C. krusei isolates. A significant percentage of voriconazole-resistant strains came from oropharyngeal infections and exhibited high MICs of other azoles.

Assessment of the CTOCD-DZ method in a hierarchy of coupled cluster methods.

Gauge origin independent calculations of nuclear magnetic shielding tensors are carried out inside the formalism of the continuous transformation of the origin of the current density leading to formal annihilation of its diamagnetic contribution (CTOCD-DZ). We employ the unrelaxed linear response approach with a hierarchy of different coupled cluster methods in order to assess the importance of the level of approximation in the coupled cluster expansion. The basis set dependence of the computed nuclear magnetic shielding constants is also analyzed in the series of correlation consistent basis sets, with the aim of designing optimized basis sets of relatively small size.

Glucolytic fingerprinting reveals metabolic groups within the genus Bifidobacterium: an exploratory study.

Microorganisms of the genus Bifidobacterium are inhabitants of diverse niches including the digestive tract of humans and animals. The species Bifidobacterium adolescentis, Bifidobacterium animalis, Bifidobacterium bifidum, Bifidobacterium breve and Bifidobacterium longum have qualified presumption of safety status granted by EFSA and several strains are considered probiotic, and are being included in functional dairy fermented products. In the present work we carried out a preliminary exploration of general metabolic characteristics and organic acid production profiles of a reduced number of strains selected from these and other species of the genus Bifidobacterium. The use of resting cells allowed obtaining metabolic fingerprints without interference of metabolites accumulated during growth in culture media. Acetic acid was the most abundant organic acid formed per mol of glucose consumed (from 1.07 ± 0.03 to 1.71 ± 0.22 mol) followed by lactic acid (from 0.34 ± 0.06 to 0.90 ± 0.12 mol), with moderate differences in production among strains; pyruvic, succinic and formic acids were also produced at considerably lower proportions, with variability among strains. The acetic to lactic acid ratio showed lower values in stationary phase as regard to the exponential phase for most, but not all, the microorganisms; this was due to a decrease in acetic acid molar proportions together with increases of lactic acid proportions in stationary phase. A linear discriminant analysis allowed to cluster strains into species with 51-100% probability, evidencing different metabolic profiles, according to the relative production of organic acids from glucose by resting cells, of microorganisms collected at the exponential phase of growth. Looking for a single metabolic marker that could adequately discriminate metabolic groups, we found that groups established by the acetic to lactic acid ratio fit well with differences previously evidenced by the discriminant analysis. The proper establishment of metabolic groups within the genus Bifidobacterium could help to select the best suited probiotic strains for specific applications.

[Implementation of an assisted electronic prescription system applied to parenteral nutrition in a general hospital]. / Implantación de un sistema de prescripción electrónica asistida aplicada a la nutrición parenteral en un hospital general.

Integrated and shared information systems allow obtaining a high degree of information about processes, costs and outcomes, and considerably reducing prescription errors. Assisted electronic prescription, in the setting of total parenteral nutrition, integrated with other hospital databases and with the hospital drugs management system, is a tool that allows increasing patient' safety (by reducing prescription errors), improving quality assistance, improving information systems and information management and the efficiency of used resources. In this work, implementation of an assisted electronic prescription system applied to parenteral nutrition in a hospital and processes reengineering performed in the nutrition setting are described. This implementation was performed by medical staff from the Nutrition and Diet Department and pharmacists from the Pharmacy Department of Ramón y Cajal Hospital using "Nutriwin" computer software. For two months prior and after its implementation, a follow-up of time consumed in the circuit prescription-validation-elaboration-dispensation of parenteral nutrition formulas has been performed. After implementation, treatment orders reach on average 1 h and 15 minutes sooner the Pharmacy Department; by avoiding transcription, a saving of 3 min per nutrition formula calculations is achieved, besides reducing potential errors; elaboration of nutrition formulas can be started on average 1 h and 20 minutes sooner as compared to manual prescription. Besides, the staff that writes down the prescription may know in real time the nutritional profile for each patient in the current episode and the patient's historic. Electronic prescription of treatment orders in this area has represented for our hospital an optimization of the employed resources, a reduction of potential errors that may occur, an improvement in consumption management, and an increase in the whole process quality.

Subspecialty referrals for pauciarticular juvenile rheumatoid arthritis.

OBJECTIVES: To examine referral patterns from primary care physicians for children with pauciarticular juvenile rheumatoid arthritis (JRA) and to determine whether children with pauciarticular JRA referred to pediatric rheumatologists differ in clinical presentation from children referred to other specialists. DESIGN: A retrospective records review of 49 patients with pauciarticular jRA was performed. Records were reviewed to determine the specialty of the referring physician and whether the children referred had symptoms and signs compatible with a synovitis at the time primary care was sought. SETTING: Inner-city tertiary pediatric rheumatology referral center. PARTICIPANTS: Children with pauciarticular JRA. MAIN OUTCOME MEASURES: Identification of referral patterns of primary care physicians. Associated morbidity owing to JRA was ascertained at the time of referral. RESULTS: Most children with pauciarticular JRA (62%) were referred to orthopedic surgeons prior to referral for pediatric rheumatology care. No differences in clinical symptoms were seen between children referred to pediatric rheumatologists and those referred to orthopedic surgeons. Children referred initially to orthopedic surgeons were younger than those referred to pediatric rheumatologists. CONCLUSION: A notable number of children with pauciarticular JRA are referred to orthopedic surgeons prior to the establishment of that diagnosis, even when such children present with unequivocal signs of synovitis. This may be owing to the misconception that arthritis is rare in preschool-aged children or to the difficulty of ascertaining the presence of synovitis in younger children.

Blau syndrome (familial granulomatous arthritis, iritis, and rash) in an african-american family.

Blau syndrome (familial granulomatous arthritis, iritis, and rash) was originally described in 1985, in 11 members of a family of Dutch ancestry. Inheritance is autosomal dominant. Several more Caucasian families have been described since. Skin and synovial biopsy specimens show noncaseating sarcoid like granulomas, but the lung is not involved as in classic sarcoidosis. This report describes 3 members of an African American family with Blau syndrome. It is important to differentiate this genetic disorder from other childhood arthritides, such as, juvenile rheumatoid arthritis, juvenile spondyloarthropathies, and early-onset sarcoidosis, because of the need for genetic counseling, treatment and differing potential for selective involvement of other organs (eye, skin, and tendons/joints). All children of an affected individual have a 50% chance of inheriting the disease. Unaffected children do not have to be concerned about subsequent generations being affected. The response to conventional treatments used in juvenile rheumatoid arthritis and to etanercept in our patients has not been satisfactory. Joint disease responds to corticosteroids, but these agents are not suitable for a disease that is lifelong. The eye involvement is aggressive and can lead to blindness. These patients need close follow-up by an ophthalmologist.

Phosphorylation of the human respiratory syncytial virus N protein provokes a decrease in viral RNA synthesis.

When HEp-2 cells are infected by human respiratory syncytial virus (HRSV) its N protein becomes phosphorylated at tyrosine (Y) Y38, in a strictly regulated way. To determine how this phosphorylation affects nucleocapsid (NC) template activity during viral RNA synthesis, N protein variants were analysed in which Y38 and nearby Y residues were substituted by phenylalanine (F; Y23F, Y38F and Y69F) or aspartic acid (D; Y23D and Y38D). While the capacity of these proteins to form the NC and to interact with the P protein was maintained, their NC template activity was altered affecting distinctly viral transcription and replication of HRSV based minigenomes. Thus, Y38 phosphorylation of the HRSV N protein modulates NC template activity probably by altering the interactions of the monomeric components of the NC.

In vitro activity of antifungals against Zygomycetes.

To date, no reference standard for therapy for zygomycosis has been established because there are insufficient clinical data with which to make such a judgement. Knowledge of the species responsible for the infection and its antifungal susceptibility profile has become increasingly important in the management of patients. Amphotericin B is the most active drug against all the species involved, followed by posaconazole, whereas voriconazole has no activity. Echinocandins are completely inactive in vitro, but may be an interesting option when used in combination with other drugs.

Diatropicity of tetraazanaphthalenes.

Tetraazanaphthalenes are diatropic molecules, whose magnetic response to a magnetic field perpendicular to the molecular plane closely resembles that of naphthalene. The out-of-plane component of the magnetic susceptibility tensor and its strong anisotropy can be used as quantifiers of magnetic aromaticity. Maps showing streamlines and modulus of the current density field provide clear evidence for diatropicity of these systems. They also explain the strong anisotropy of carbon and nitrogen magnetic shielding, which is determined by the big out-of-plane component of the nuclear shielding tensor. The electronic ring currents observed in the map deshield the nuclei of ring hydrogens by enforcing the local magnetic field and diminishing the out-of-plane component of proton shielding.

Trot asymmetry in relation to physical performance and metabolism in equine endurance rides.

REASON FOR PERFORMING STUDY: Trot is a symmetric gait and asymmetry might appear or increase during endurance rides due to lameness, pain or excessive fatigue. HYPOTHESIS: To assess whether trot asymmetry increases during endurance competitions, whether it is possible to discriminate between horses with different performance and also its possible relationship with metabolism. METHODS: Fifty-eight horses were filmed at trot during the lameness examinations in the vet-gates, before the competition (BCO), and after phases 1 (at 29 km), 2 (at 59 km) and 3 (at the end of the ride, at 80 km) and stride duration (SD) measured in all 4 limbs. A locomotion symmetry index (LSI) was calculated by comparing SD in the diagonal pairs of limbs. In all the cases, the longer diagonal pair SD was compared to the shorter. Horses were classified as symmetric (SyH) when LSI was lower than mean + 2s.d. and asymmetric (ASyH) when LSI was higher than mean + 2s.d. of the data obtained in BCO. Venous samples were withdrawn in the vet-gates and PCV, WBC, creatinine, TPP, uric acid, CK, AST, LDH, Na, K and Cl were measured. Horses were divided into different performance groups: successful (SH), lame (LH) and metabolic (MH), and according to ride velocity, the SH group was also divided into faster finishers (FF) and slower finishers (SF). RESULTS: The percentages of SyH were high (>80%) and very similar between the different performance groups. LSI became more asymmetrical in the vet-gates, especially in the LH and MH groups. LSI presented negative correlations with the velocities during the rides and in the vet-gates and CK. The ASyH had higher velocities during the rides and plasma CK and lower velocities during the vet-gates and plasma uric acid concentrations. CONCLUSIONS: Although trot asymmetry increases during endurance events, LSI calculated comparing SD in the two diagonal pair of limbs did not allow the differentiation of horses with different performance.