Carbapenem-resistant Citrobacter spp. isolated in Spain from 2013 to 2015 produced a variety of carbapenemases including VIM-1, OXA-48, KPC-2, NDM-1 and VIM-2

Objectives: There is little information about carbapenemase-producing (CP) Citrobacter spp.We studied the molecular epidemiology and microbiological features of CP Citrobacter spp. isolates collected in Spain (2013-15). Methods: In total, 119 isolates suspected of being CP by the EUCAST screening cut-off values were analysed. Carbapenemases and ESBLs were characterized using PCR and sequencing. The genetic relationship among Citrobacter freundii isolates was studied by PFGE. Results: Of the 119 isolates, 63 (52.9%) produced carbapenemases, of which 37 (58.7%) produced VIM-1, 20 (31.7%) produced OXA-48, 12 (19%) produced KPC-2, 2 (3.2%) produced NDM-1 and 1 (1.6%) produced VIM- 2; 9 C. freundii isolates co-produced VIM-1 plus OXA-48. Fourteen isolates (22.2%) also carried ESBLs: 8 CTX-M-9 plus SHV-12, 2 CTX-M-9, 2 SHV-12 and 2 CTX-M-15. Fifty-seven isolates (90.5%) were C. freundii, 4 (6.3%) were Citrobacter koseri, 1 (1.6%) was Citrobacter amalonaticus and 1 (1.6%) was Citrobacter braakii. By EUCAST breakpoints, eight (12.7%) of the CP isolates were susceptible to the four carbapenems tested. In the 53 CP C. freundii analysed by PFGE, a total of 44 different band patterns were observed. Four PFGE clusters were identified: cluster 1 included eight isolates co-producing VIM-1 and OXA-48; blaVIM-1 was carried in a class 1 integron (intI-blaVIM-1 - aacA4-dfrB1-aadA1-catB2-qacE¿1/sul1) and blaOXA-48 was carried in a Tn1999.2 transposon. Conclusions: We observed the clonal and polyclonal spread of CP Citrobacter spp. across several Spanish geographical areas. Four species of Citrobacter spp. produced up to five carbapenemase types, including coproduction of VIM-1 plus OXA-48. Some CP Citrobacter spp. isolates were susceptible to the four carbapenems tested, a finding with potential clinical implications

Comparación de distintas estrategias para la predicción de muerte a corto plazo en el paciente anciano infectado

Objective. The aim of this study was to determine the utility of a post hoc lactate added to SIRS and qSOFA score to predict 30-day mortality in older non-severely dependent patients attended for infection in the Emergency Department (ED). Methods. We performed an analytical, observational, prospective cohort study including patients of 75 years of age or older, without severe functional dependence, attended for an infectious disease in 69 Spanish ED for 2-day three seasonal periods. Demographic, clinical and analytical data were collected. The primary outcome was 30-day mortality after the index event. Results. We included 739 patients with a mean age of 84.9 (SD 6.0) years; 375 (50.7%) were women. Ninety-one (12.3%) died within 30 days. The AUC was 0.637 (IC 95% 0.587-0.688; p= 2 and 0.698 (IC 95% 0.635- 0.761; p= 2. Comparing receiver operating characteristic (ROC) there was a better accuracy of qSOFA vs SIRS (p=0.041). Both scales improve the prognosis accuracy with lactate inclusion. The AUC was 0.705 (IC95% 0.652-0.758; p<0.001) for SIRS plus lactate and 0.755 (IC95% 0.696-0.814; p<0.001) for qSOFA plus lactate, showing a trend to statistical significance for the second strategy (p=0.0727). Charlson index not added prognosis accuracy to SIRS (p=0.2269) or qSOFA (p=0.2573). Conclusions. Lactate added to SIRS and qSOFA score improve the accuracy of SIRS and qSOFA to predict short-term mortality in older non-severely dependent patients attended for infection. There is not effect in adding Charlson index

The menthol receptor TRPM8 is the principal detector of environmental cold

Sensory nerve fibres can detect changes in temperature over a remarkably wide range, a process that has been proposed to involve direct activation of thermosensitive excitatory transient receptor potential (TRP) ion channels. One such channel--TRP melastatin 8 (TRPM8) or cold and menthol receptor 1 (CMR1)--is activated by chemical cooling agents (such as menthol) or when ambient temperatures drop below approximately 26 degrees C, suggesting that it mediates the detection of cold thermal stimuli by primary afferent sensory neurons. However, some studies have questioned the contribution of TRPM8 to cold detection or proposed that other excitatory or inhibitory channels are more critical to this sensory modality in vivo. Here we show that cultured sensory neurons and intact sensory nerve fibres from TRPM8-deficient mice exhibit profoundly diminished responses to cold. These animals also show clear behavioural deficits in their ability to discriminate between cold and warm surfaces, or to respond to evaporative cooling. At the same time, TRPM8 mutant mice are not completely insensitive to cold as they avoid contact with surfaces below 10 degrees C, albeit with reduced efficiency. Thus, our findings demonstrate an essential and predominant role for TRPM8 in thermosensation over a wide range of cold temperatures, validating the hypothesis that TRP channels are the principal sensors of thermal stimuli in the peripheral nervous system

Detección cualitativa alternativa de lactoperoxidasa en leche

"Introducción: La lactoperoxidasa (LPO) es una enzima que se encuentra dentro de los principales componentes de la leche, tiene una función antibacteriana y es un indicador muy utilizado para el tratamiento térmico de la leche. Objetivo: Por lo anterior el objetivo de este estudio fue determinar la presencia de LPO en cinco marcas de leche comerciales procesadas y una muestra de leche no procesada. Metodología: La metodología usada se basas en la interacción de la leche con el sistema guayacol al 1 % y peróxido de hidrógeno al 12% (H2O2) formando el complejo tetraguayacol que corresponde al desarrollo de una coloración salmón. También se determinó los sólidos totales (ST) por gravimetría dichos valores se correlacionaron con la presencia de LPO, con estos datos se estableció la calidad de la leche. Resultados y Discusión: Se obtuvo LPO en la leche cruda (LC) y en las muestras LGV, LSC y LAl no así en las muestras LA y LT. La determinación de ST mostró no tener una correlación con la LPO por lo que solo se establece esta última como indicador de calidad, esta cuando es negativa, indica que la leche se sometió a procesos de sobrecalentamiento (˃77.8°C) y destrucción de la enzima peroxidasa durante la pasteurización. Perdiendo con esto, características de su estado natural y, por lo tanto, el valor nutricional ha disminuido"

Insights into ROS-dependent signalling underlying transcriptomic plant responses to the herbicide 2,4-D

The synthetic auxin 2,4¿dichlorophenoxyacetic acid (2,4¿D) functions as an agro-nomic weed control herbicide. High concentrations of 2,4¿D induce plant growthdefects, particularly leaf epinasty and stem curvature. Although the 2,4¿D triggeredreactive oxygen species (ROS) production, little is known about its signalling. In thisstudy, by using a null mutant in peroxisomal acyl CoA oxidase 1 (acx1¿2), we iden-tified acyl¿coenzyme A oxidase 1 (ACX1) as one of the main sources of ROS pro-duction and, in part, also causing the epinastic phenotype following 2,4¿Dapplication. Transcriptomic analyses of wild type (WT) plants after treatment with2,4¿D revealed a ROS¿related peroxisomal footprint in early plant responses, whileother organelles, such as mitochondria and chloroplasts, are involved in later re-sponses. Interestingly, a group of 2,4¿D¿responsive ACX1¿dependent transcriptspreviously associated with epinasty is related to auxin biosynthesis, metabolism, andsignalling. We found that the auxin receptor auxin signalling F¿box 3 (AFB3), acomponent of Skp, Cullin, F¿box containing complex (SCF) (ASK¿cullin¿F¿box) E3ubiquitin ligase complexes, which mediates auxin/indole acetic acid (AUX/IAA)degradation by the 26S proteasome, acts downstream of ACX1 and is involved in theepinastic phenotype induced by 2,4¿D. We also found that protein degradationassociated with ubiquitin E3¿RING and E3¿SCF¿FBOX in ACX1¿dependent signallingin plant responses to 2,4¿D is significantly regulated over longer treatment periodsThis study was funded by the Spanish Ministry of Science, Innovation and Universities (MCIU), the State Research Agency (AEI) and FEDER grant PGC2018-098372-B-I00. MAP-V was supported by MCIU Research Personnel Training (FPI) grant BES-2016-076518

4-Hydroxynonenal, an endogenous aldehyde, causes pain and neurogenic inflammation through activation of the irritant receptor TRPA1.

TRPA1 is an excitatory ion channel expressed by a subpopulation of primary afferent somatosensory neurons that contain substance P and calcitonin gene-related peptide. Environmental irritants such as mustard oil, allicin, and acrolein activate TRPA1, causing acute pain, neuropeptide release, and neurogenic inflammation. Genetic studies indicate that TRPA1 is also activated downstream of one or more proalgesic agents that stimulate phospholipase C signaling pathways, thereby implicating this channel in peripheral mechanisms controlling pain hypersensitivity. However, it is not known whether tissue injury also produces endogenous proalgesic factors that activate TRPA1 directly to augment inflammatory pain. Here, we report that recombinant or native TRPA1 channels are activated by 4-hydroxy-2-nonenal (HNE), an endogenous alpha,beta-unsaturated aldehyde that is produced when reactive oxygen species peroxidate membrane phospholipids in response to tissue injury, inflammation, and oxidative stress. HNE provokes release of substance P and calcitonin gene-related peptide from central (spinal cord) and peripheral (esophagus) nerve endings, resulting in neurogenic plasma protein extravasation in peripheral tissues. Moreover, injection of HNE into the rodent hind paw elicits pain-related behaviors that are inhibited by TRPA1 antagonists and absent in animals lacking functional TRPA1 channels. These findings demonstrate that HNE activates TRPA1 on nociceptive neurons to promote acute pain, neuropeptide release, and neurogenic inflammation. Our results also provide a mechanism-based rationale for developing novel analgesic or anti-inflammatory agents that target HNE production or TRPA1 activation