Enterococcal bloodstream infection. Design and validation of a mortality prediction rule.

BACKGROUND: To develop a prediction rule to describe the risk of death as a result of enterococcal bloodstream infection. METHODS: A prediction rule was developed by analysing data collected from 122 patients diagnosed with enterococcal BSI admitted to the Clínica Universidad de Navarra (Pamplona, Spain); and validated by confirming its accuracy with the data of an external population (Hospital del Mar, Barcelona). RESULTS: According to this model, independent significant predictors for the risk of death were being diabetic, have received appropriate treatment, severe prognosis of the underlying diseases, have renal failure, received solid organ transplant, malignancy, source of the bloodstream infection and be immunosuppressed. The prediction rule showed a very good calibration (Hosmer-Lemeshow statistic, P = 0.93) and discrimination for both training and testing sets (area under ROC curve = 0.84 and 0.83 respectively). CONCLUSIONS: The predictive rule was able to predict risk of death as a result of enterococcal bloodstream infection as well as to identify patients, who being below the threshold value, will have a low risk of death with a negative predictive value of 96%.

[Personal resources relevant to psychological well-being in nursing]. / Recursos personales relevantes para el bienestar psicológico en el personal de enfermería.

AIMS: To determine differences in social support, resilience, coping, and psychological well-being (PWB) among intensive care nursing and nursing staff of other hospital services, as well as to establish a structural model in these professionals where relevant personal resources to PWB were included. METHOD: Correlational and cross-sectional study. A sample of 208 nursing professionals from University Hospital of Fuenlabrada (Madrid) took part in the study. This sample consisted of nurses (n=133), nursing assistants (n=61), and midwives (n=14), of whom 44 worked in intensive care unit, 50 in other special units, and 114 in wards. INSTRUMENTS: Social Support Subscale, 10-Item CD-RISC (resilience), Brief-Cope (coping), Scales of PWB, and sociodemographic variables. RESULTS: No differences were found in any assessed psychological variables as regards hospital service worked in. A structural model was found and showed that social support, resilience, and coping determined PWB of nursing professionals. The most important personal resource was coping strategies, which determined PWB directly (ß=0.68). Social support influenced PWB directly (ß=0.33), and indirectly (ß=0.32), whereas resilience influenced it indirectly (ß=0.57). CONCLUSIONS: Differences in PWB, coping, social support and resilience are not determined by hospital service. Coping strategies focused on engagement (or adaptive), social support, and resilience, constitute three relevant personal resources that determine the PWB of nursing staff, which can be developed and improved by specific programs. The most important PWB dimensions are self-acceptance and environment mastery.

Comparative Genomics Within the Bacillus Genus Reveal the Singularities of Two Robust Bacillus amyloliquefaciens Biocontrol Strains.

Bacillus amyloliquefaciens CECT 8237 and CECT 8238, formerly known as Bacillus subtilis UMAF6639 and UMAF6614, respectively, contribute to plant health by facing microbial pathogens or inducing the plant's defense mechanisms. We sequenced their genomes and developed a set of ad hoc scripts that allowed us to search for the features implicated in their beneficial interaction with plants. We define a core set of genes that should ideally be found in any beneficial Bacillus strain, including the production of secondary metabolites, volatile compounds, metabolic plasticity, cell-to-cell communication systems, and biofilm formation. We experimentally prove that some of these genetic elements are active, such as i) the production of known secondary metabolites or ii) acetoin and 2-3-butanediol, compounds that stimulate plant growth and host defense responses. A comparison with other Bacillus genomes permits us to find differences in the cell-to-cell communication system and biofilm formation and to hypothesize variations in their persistence and resistance ability in diverse environmental conditions. In addition, the major protection provided by CECT 8237 and CECT 8238, which is different from other Bacillus strains against bacterial and fungal melon diseases, permits us to propose a correlation with their singular genetic background and determine the need to search for additional blind biocontrol-related features.

Cyclo(Pro-Tyr) elicits conserved cellular damage in fungi by targeting the [H+]ATPase Pma1 in plasma membrane domains.

Bioactive metabolites play a crucial role in shaping interactions among diverse organisms. In this study, we identified cyclo(Pro-Tyr), a metabolite produced by Bacillus velezensis, as a potent inhibitor of Botrytis cinerea and Caenorhabditis elegans, two potential cohabitant eukaryotic organisms. Based on our investigation, cyclo(Pro-Tyr) disrupts plasma membrane polarization, induces oxidative stress and increases membrane fluidity, which compromises fungal membrane integrity. These cytological and physiological changes induced by cyclo(Pro-Tyr) may be triggered by the destabilization of membrane microdomains containing the [H+]ATPase Pma1. In response to cyclo(Pro-Tyr) stress, fungal cells activate a transcriptomic and metabolomic response, which primarily involves lipid metabolism and Reactive Oxygen Species (ROS) detoxification, to mitigate membrane damage. This similar response occurs in the nematode C. elegans, indicating that cyclo(Pro-Tyr) targets eukaryotic cellular membranes.

[Is subjective well-being perceived by non-health care workers different from that perceived by nurses? Relation with personality and resilience]. / El bienestar subjetivo percibido por los profesionales no sanitarios ¿es diferente al de enfermería de intensivos? Relación con personalidad y resiliencia.

Subjective well-being (SWB), usually called 'happiness', is influenced directly by psychological factors. Personality and resilience (capacity of recover from adversity) are included among these factors. Empirical evidence has demonstrated that resilience is an essential and inherent characteristic for the nursery staff. This study has aimed to analyze personality factors (including resilience) related with SWB (satisfaction with life, positive and negative affect) in a nursery staff sample (n=59) of intensive care and cardiological units, and a non-health care workers sample (n=50) mainly made up of government employees and teachers. Multiple regression analyses showed that SWB was associated with more resilience and less neuroticism in the nursery staff. Extraversion and conscientiousness (positively related), and neuroticism (negatively related) were the significant predictors of SWB in the non-health care workers group. Finally, mediational analyses revealed that resilience measured the relationships between extraversion (total mediation) and neuroticism (partial mediation) with SWB in the nursery staff group, but not in the group of non-health care workers. The results show the importance of resilience for nursery staff of intensive care units, since they are constantly exposed to human suffering and to a continually adverse occupational environment. Likewise, the discussion stresses that resilience is a means for nursing staff to cope with the occupational stress and that resilient nurses are a crucial element in our health care system.

Bacillus subtilis biofilm matrix components target seed oil bodies to promote growth and anti-fungal resistance in melon.

Beneficial microorganisms are used to stimulate the germination of seeds; however, their growth-promoting mechanisms remain largely unexplored. Bacillus subtilis is commonly found in association with different plant organs, providing protection against pathogens or stimulating plant growth. We report that application of B. subtilis to melon seeds results in genetic and physiological responses in seeds that alter the metabolic and developmental status in 5-d and 1-month-old plants upon germination. We analysed mutants in different components of the extracellular matrix of B. subtilis biofilms in interaction with seeds and found cooperation in bacterial colonization of seed storage tissues and growth promotion. Combining confocal microscopy with fluorogenic probes, we found that two specific components of the extracellular matrix, amyloid protein TasA and fengycin, differentially increased the concentrations of reactive oxygen species inside seeds. Further, using electron and fluorescence microscopy and metabolomics, we showed that both TasA and fengycin targeted the oil bodies in the seed endosperm, resulting in specific changes in lipid metabolism and accumulation of glutathione-related molecules. In turn, this results in two different plant growth developmental programmes: TasA and fengycin stimulate the development of radicles, and fengycin alone stimulate the growth of adult plants and resistance in the phylloplane to the fungus Botrytis cinerea. Understanding mechanisms of bacterial growth promotion will enable the design of bespoke growth promotion strains.

Proteomic analysis of Strongyloides stercoralis L3 larvae.

Strongyloidiasis can be perpetuated by autoinfection with the filariform larvae L3, causing asymptomatic chronic infections and creating a population of carriers, affecting not only developing countries. So far, very little is known about the proteins that interact with the human host, and few proteins from the infective Strongyloides stercoralis L3 have been characterized. Here, we report results obtained from a proteomic analysis of the proteins from S. stercoralis L3 larvae obtained from patients. Since the genome of S. stercoralis is not yet available, we used proteomic analysis to identify 26 different proteins, 13 of them released by short digestion with trypsin, which could represent surface-associated proteins. The present work extends our knowledge of host-parasite interactions by identifying proteins that could be of interest in the development of diagnostic tools, vaccines, or treatments for a neglected disease like strongyloidiasis.

First Occurrence of Cucurbit Powdery Mildew Caused by Race 3-5 of Podosphaera fusca in Spain.

A new race of cucurbit powdery mildew was observed for the first time on melon (Cucumis melo) in three research greenhouses in the Axarquia area of southern Spain during the spring of 2008. Fungal growth appeared as white powdery colonies initially restricted to the upper leaf surfaces. Morphological characteristics of colonies, conidiophores, conidia, germ tubes, and appressoria indicated that the powdery mildew fungus was Podosphaera fusca (also known as P. xanthii) (3), a fungal pathogen extensively reported in the area (1). However, the fungus developed on plants of melon cv. PMR 6, which is resistant to races 1 and 2 of P. fusca, suggesting that the fungus could belong to race 3, a race of P. fusca not yet reported in Spain. Race determination was carried out by inoculating the third true leaf of a set of differential melon genotypes that were maintained in a greenhouse. Symptoms and colonization observed on cvs. Rochet, PMR 45, PMR 6, and Edisto 47 indicated that the isolates belonged to race 3-5 of P. fusca. Fungal strains of races 1, 2, and 5 of P. fusca (all present in Spain) were used as controls. Pathotype designation was determined by inoculating different cucurbit genera and species (2). In addition to melon, the isolates were pathogenic on zucchini (Cucurbita pepo) cv. Diamant F1, but failed to infect cucumber (C. sativus) cv. Marketer and watermelon (Citrullus lanatus) cv. Sugar Baby; therefore, the isolates were pathotype BC (2). Races 1, 2, 4, and 5 of P. fusca have been previously reported in the area (1). The occurrence of race 3-5 represents another challenge in the management of cucurbit powdery mildew in Spain. References: (1) D. del Pino et al. Phytoparasitica 30:459, 2002. (2) E. Krístková et al. Sci. Hortic. 99:257, 2004. (3) A. Pérez-García et al. Mol. Plant Pathol. 10:153, 2009.

First Report of Powdery Mildew Elicited by Podosphaera fusca (Synonym Podosphaera xanthii) on Euryops pectinatus in Spain.

Euryops pectinatus Cass. is an evergreen shrub that is planted extensively in Spain for landscape use. In 2007, powdery mildew outbreaks were observed on E. pectinatus in several nurseries located in the Axarquia area (Malaga, southern Spain). Fungal growth appeared as typical, white, powdery mildew colonies that were restricted to upper leaf surfaces. Initially, individual colonies were small and nearly circular, but later enlarged and coalesced to cover the whole leaf surface. With progress of the disease, all green parts (leaves, stems, and petioles) were covered with a white mycelium. Newly developed leaves especially became rapidly infected. Diseased leaves ultimately dried up and senesced, making nursery plants aesthetically unattractive and unsaleable. Conidiophores were erect, had crenate edge lines, cylindrical foot cells that measured 37.5 to 45.0 × 10.0 to 12.5 µm, and were followed by one to three shorter cells. Conidia were hyaline, ellipsoid to ovoid, borne in chains, and measured 27.5 to 35.0 × 12.5 to 17.5 µm. Conidial length-to-width ratios ranged from 1.6 to 2.4. Conidia possessed conspicuous fibrosin bodies and from their sides produced short germ tubes. No chasmothecia were found. The nuclear rDNA internal transcribed spacer (ITS) regions were amplified by PCR and sequenced (GenBank Accession No. EU424056). On the basis of morphological characteristics of the imperfect state and ITS sequence data, this powdery mildew was identified as Podosphaera fusca (Fr.) U. Braun & N. Shishkoff (1), this isolate belonging to ITS haplotype 15 (group III) (3); this group is considered a separate species, P. xanthii (Castagne) U. Braun & N. Shishkoff by some authors (2). Pathogenicity was confirmed by gently pressing diseased leaves onto leaves of healthy E. pectinatus plants. Plants were incubated in a growth chamber at 25°C, and after 14 days, powdery mildew colonies developed. A similar disease of E. pectinatus was observed in 1999 in California (4). P. fusca parasitizes a large number of asteraceous species including field marigold (Calendula arvensis) and fleabane (Erigeron sp.) weeds, as previously reported in the same area, and ornamentals such as Calendula officinalis, Chrysanthemum spp., and Gerbera spp., which are also grown in the same nurseries and frequently attacked by powdery mildew. References: (1) U. Braun and S. Takamatsu. Schlechtendalia 4:1, 2000. (2) U. Braun et al. Schlechtendalia 7:45, 2001. (3) T. Hirata et al. Can. J. Bot. 78:1521, 2000. (4) G. S. Saenz et al. Plant Dis. 84:1048, 2000.

PAS Kinase deficiency alters the glucokinase function and hepatic metabolism.

The liver controls metabolic homeostasis in response to fasting and refeeding periods. Glucokinase (GCK) adjusts hepatic glucose phosphorylation to blood glucose levels, acting as a glucose sensor. Our objective was to determine whether PAS kinase (PASK), a nutrient sensor, could be affecting the expression or activity of liver GCK and the response to fasting and refeeding states of key hepatic metabolic pathways. PASK-deficient mice have impaired insulin signaling (AKT overactivation). Furthermore, PASK deficiency modified the expression of several transcription factors involved in the adjustment to fasting and refeeding. Foxo1 decreased under fasting conditions, while Ppara and Pparg were overexpressed in PASK-deficient mice. However, PEPCK protein levels were similar or higher, while the expression of Cpt1a decreased in PASK-deficient mice. By contrast, Lxra and Chrebp were overexpressed after refeeding, while the expression of Acc and Fas decreased in PASK-deficient mice. Likewise, with a decreased expression of Gck and increased nuclear location of the complex GCK-GCKR, GCK activity decreased in PASK-deficient mice. Therefore, PASK regulated some of the genes and proteins responsible for glucose sensing, such as glucokinase, and for insulin signalling, affecting glucose and lipid metabolism and consequently certain critical hepatic functions.

Pas Kinase Deficiency Triggers Antioxidant Mechanisms in the Liver.

Metabolic dysfunction in the liver is the cause of numerous pathologies, which are associated with an altered redox state. PASK (PAS Domain Kinase) is a nutrient and bioenergetic sensor. We contend that PASK could act as an oxidative stress sensor in liver and/or control the metabolic balance, playing a role in the mitochondrial homeostasis. Using PASK-deficient mice, we observed that PASK deficiency promotes antioxidant response mechanisms: a lower production of ROS/RNS under non-fasting conditions, overexpression of genes coding to ROS-detoxifying enzymes and mitochondrial fusion proteins (MnSod Gpx, Mfn1 and Opa1), coactivator Ppargc1a, transcription factors (Pparg and FoxO3a) and deacetylase Sirt1. Also, under fasting conditions, PASK deficiency induced the overexpression of Ppargc1a, Ppara, Pparg, FoxO3a and Nrf2 leading to the overexpression of genes coding to antioxidant enzymes such as MnSOD, Cu/ZnSOD, GPx, HO1 and GCLm. Additionally, inducing PINK1 involved in cell survival and mitophagy. These changes kept ROS steady levels and improved the regenerative state. We suggest a new role for PASK as a controller of oxidative stress and mitochondrial dynamics in the liver. In fact, antioxidant response is PASK dependent. PASK-targeting could therefore be a good way of reducing the oxidative stress in order to prevent or treat liver diseases.

Differentiation of fibrotic liver tissue using laser-induced breakdown spectroscopy.

Hepatic cirrhosis is a major cause of morbidity and mortality worldwide due to hepatitis C, alcoholism and fatty liver disease associated with obesity. Assessment of hepatic fibrosis relies in qualitative histological evaluation of biopsy samples. This method is time-consuming and depends on the histopathologists' interpretation. In the last decades, non-invasive techniques were developed to detect and monitor hepatic fibrosis. Laser-induced breakdown spectroscopy (LIBS) is a good candidate for a real-time, independent and fast technique to diagnose hepatic fibrosis. In this work LIBS was employed to characterize rat liver tissues with different stages of fibrosis. Depth profiling measurements were carried out by using a nanosecond Nd:YAG laser operated at the fundamental wavelength and an echelle spectrometer coupled with an ICCD camera. Due to the soft nature of the samples, plasma conditions largely change between consecutives shots. Thus, a theoretically supported procedure to correct the spectral line intensities was implemented. This procedure allows the reduction of the intensities' dispersion from 67% to 12%. After the correction, the LIBS signal shows an enhancement in calcium intensity by a factor of three as the fibrosis progressed. Calcium is known to increase crosslinking of extracellular matrix proteins in the fibrous septa. Therefore, our result singles it out as a key participant in the hepatic fibrosis.

[Epidemiological surveillance for multidrug-resistant microorganisms in a general ICU]. / Vigilancia epidemiológica para microorganismos multirresistentes en una UCI polivalente.

OBJECTIVE: Multidrug resistant (MDR) microorganisms represent a threat for patients admitted in Intensive Care Units (ICUs). The objective of the present study is to analyse the results of epidemiological surveillance cultures for these microorganisms in one of these units. METHODS: General ICU. Retrospective analysis, descriptive statistics. Analysis of epidemiological surveillance cultures for MDR microorganisms in 2015. Studied microorganisms: Methicillin-resistant Staphylococcus aureus (MRSA), ESBL-and/or carbapenemase-producing Klebsiella pneumoniae (CESBL-KP) and MDR Acinetobacter baumannii (MDRAB). RESULTS: One thousand, two hundred and fifty nine patients admitted. A total of 2,234 specimens from 384 patients were analysed (690, 634, 62 and 286 were rectal, throat, nasal and skin swabs respectively). Global APACHE II was 18.3 ± 8 versus 21.7 ± 7.8 in patients colonized/infected on admission. Global mortality was 19.7% versus 22.3% in patients colonized/infected on admission. The higher sensitivities achieved with the different samples for the different microorganism detection were as follows. MRSA: 79% and 90% for nasal and nasal + throat swabs, respectively. MDRAB: 80% and 95% for throat and throat + rectal swabs, respectively. CESBL-KP: 95% and 98% for rectal and rectal + throat swabs, respectively. 94 out of the 384 patients (24.4%) were colonized/infected with MDR at admission. 134 patients (10.6% of the total patients admitted) were colonized/infected with a total of 169 MMR during the hospital stay. MRSA has the earliest colonization/infection (9.2 ± 6.4days) and ESBL-producing Enterobacteriaceae, the latest (18.7± 16.4 days). CONCLUSIONS: 24.4% of patients were colonized/infected by MDR at admission. Nasal, throat and rectal swabs were the most effective specimens for recovering MRSA, MDRAB and CESBL-KP, respectively. The combination of two specimens improves MDR detection except for CESBL-KP. Skin swabs are worthless. The most prevalent MDR at admission were ESBL-producing Enterobacteriaceae while the most frequent hospital acquired MDR was MDRAB..

Single-stage reconstruction of flexor tendons with vascularized tendon transfers.

The reconstruction of finger flexor tendons with vascularized flexor digitorum superficialis (FDS) tendon grafts (flaps) based on the ulnar vessels as a single stage is not a popular technique. We reviewed 40 flexor tendon reconstructions (four flexor pollicis longus and 36 finger flexors) with vascularized FDS tendon grafts in 38 consecutive patients. The donor tendons were transferred based on the ulnar vessels as a single-stage procedure (37 pedicled flaps, three free flaps). Four patients required composite tendon and skin island transfer. Minimum follow-up was 12 months, and functional results were evaluated using a total active range of motion score. Multiple linear regression analysis was performed to evaluate the factors that could be associated with the postoperative total active range of motion. The average postoperative total active range of motion (excluding the thumbs) was 178.05° (SD 50°). The total active range of motion was significantly lower for patients who were reconstructed with free flaps and for those who required composite tendon and skin island flap. Age, right or left hand, donor/motor tendon and pulley reconstruction had no linear effect on total active range of motion. Overall results were comparable with a published series on staged tendon grafting but with a lower complication rate. Vascularized pedicled tendon grafts/flaps are useful in the reconstruction of defects of finger flexor tendons in a single stage, although its role in the reconstructive armamentarium remains to be clearly established.

Predictors of mortality and poor outcome in cancer patients with E. faecium bloodstream infection.

BACKGROUND: To analyze predictors of mortality and poor outcome in cancer patients diagnosed with E. faecium bloodstream infection. METHODS: Demographic, clinical and microbiological data were collected (January 1998-June 2011). RESULTS: After multivariate analysis, presence of a urinary catheter was associated with a worse 7-day prognosis, and higher mortality at discharge. A high Charlson index was also associated with higher 7-day mortality. CONCLUSION: Presence of a urinary catheter was associated with poor 7-day prognosis and higher mortality at discharge in the present series.

Expression of the Avirulence gene Avr9 of the fungal tomato pathogen Cladosporium fulvum is regulated by the global nitrogen response factor NRF1.

Here we describe the role of the Cladosporium fulvum nitrogen response factor 1 (Nrf1) gene in regulation of the expression of avirulence gene Avr9 and virulence on tomato. The Nrf1 gene, which was isolated by a polymerase chain reaction-based strategy, is predicted to encode a protein of 918 amino acid residues. The protein contains a putative zinc finger DNA-binding domain that shares 98% amino acid identity with the zinc finger of the major nitrogen regulatory proteins AREA and NIT2 of Aspergillus nidulans and Neurospora crassa, respectively. Functional equivalence of Nrf1 to areA was demonstrated by complementation of an A. nidulans areA loss-of-function mutant with Nrf1. Nrf1-deficient transformants of C. fulvum obtained by homologous recombination were unable to utilize nitrate and nitrite as a nitrogen source. In contrast to what was observed in the C. fulvum wild-type, the Avr9 gene was no longer induced under nitrogen-starvation conditions in Nrf1-deficient strains. On susceptible tomato plants, the Nrf1-deficient strains were as virulent as wild-type strains of C. fulvum, although the expression of the Avr9 gene was strongly reduced. In addition, Nrf1-deficient strains were still avirulent on tomato plants containing the functional Cf-9 resistance gene, indicating that in planta, apparently sufficient quantities of stable AVR9 elicitor are produced. Our results suggest that the NRF1 protein is a major regulator of the Avr9 gene.

The influence of several factors that intervene in hemodialysis on serum levels of triglycerides and free fatty acids.

The effect of the administration of heparin and glucose, in doses and sequence similar to those of hemodialysis, on serum free fatty acids and triglycerides, was studied in eight patients with chronic renal failure who had not undergone hemodialysis. The results are compared with the action exerted by hemodialysis on the same parameters. During hemodialysis, the values of triglycerides are not significantly different from those obtained at similar intervals after the administration of glucose and/or heparin, with the exception that the values obtained five hours after beginning hemodialysis are lower than those obtained five hours after the administration of glucose (P less than 0.005). The levels of free fatty acids obtained during hemodialysis are higher (P less than 0.005) than those found during the other tests. The possible origin of these high fatty acid values is discussed, and the possibility that they originate from the acetate in the dialysis liquid is suggested.

Bacterial Apical Necrosis of Mango in Southern Spain: A Disease Caused by Pseudomonas syringae pv. syringae.

ABSTRACT A necrotic bacterial disease of mango trees (Mangifera indica) in Spain affecting buds, leaves, and stems is described for the first time. Necrosis of flower and vegetative buds on commercial trees during winter dormancy was the most destructive symptom of the disease. The apical necrosis is caused by Pseudomonas syringae, which was always isolated from mango trees with disease symptoms. Of 95 bacterial strains isolated from symptomatic tissues and characterized from 1992 to 1997, over 90% were identified as P. syringae pv. syringae. Additional strains were isolated from healthy mango trees, and they were identical to the isolates from diseased tissues. Pathogenicity tests on mango plants showed that P. syringae pv. syringae incited the apical necrosis, but that climatic conditions determined the onset of disease development. Populations of total bacteria and of P. syringae and the number of active ice nuclei were monitored over a 3-year period. The largest populations of P. syringae were associated with cool, wet periods that coincided with the highest disease severity, whereas P. syringae was only occasionally detected on healthy trees. The median effective dose was estimated from infectivity titration assays.