Patógenos y depredadores naturales del trips del trigo, Haplothrips tritici (Kurdjumov), en Castilla-La Mancha

[ESP] En un estudio de dos años sobre los hongos entomopatógenos que atacan a las larvas del trips del trigo, Haplothrips tritici (Kurdjumov), durante el período de diapusa en el suelo, se encontró que Beauveria bassiana (Balsamo) Vuillemin representa un factor de mortalidad importante, aunque con variaciones acusadas entre años. En los bioensayos de patogenicidad realizados se detectaron diferencias de patogenicidad entre aislados. Se detectaron varios depredadores potenciales de larvas diapausantes, dos especies de carábidos, Paradromius linearis Olivier y Microlestes sp. y varias especies de arañas. Sobre las espigas de trigo se encontraron larvas y adultos de Aeolothrips, que depredan sobre los huevos y las larvas I de H. tritici. Las larvas II no son presas de Aeolothrips ya que se defienden con éxito, llegando a invertirse los papeles, depredando H. triltritici sobre Aeolothrips. En las espigas se recogieron larvas de Chrysopa y sírfidos con síntomas de haberse alimentado sobre larvas de H. tritici, según se pudo comprobar en laboratorio. En cambio, las mariquitas (Coccinella septempunctata L.) no se alimentaron de larvas de H. tritici en cautividad. [ENG] A two year sampling of pathogens and predators of Haplothrips tritici was carried out in Santa Olalla (Toledo) during the years 1994-96. Among the winter mortality factors was found the attack of the fungus Beauveria bassiana, there being differences of pathogenicity detected between isolates. Its incidence was higher in a dry year than in a wet year. Several potential predators of overwintering larvae were detected, two species of Carabidae, Paradromius linearis and Microlestes sp., and several species of spiders. On the spikes larvae and adults of Aeolothrips were found which, according to laboratory observations, prey upon eggs and larvae I of H. tritici. The larvae II are not Aeolothrips preys since they defended themselves with success, the predator-prey role being inverted, H. tritici depredating upon Aeolothrips larvae. In the spikes larvae of Chrysopa and Syrphidae were collected showing symptoms of feeding upon larvae of H. tritici, according to laboratory observations. Coccinella septempunctata did not feed upon larvae of H. tritici

Actividad invernal de Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae) en las Vegas del Guadiana (Extremadura)

[ESP] El trips F. occidentalis es capaz de reproducirse a lo largo de todo el año, sin periodo de diapausa invernal, en las regiones mediterráneas de la geografía peninsular, caracterizadas por inviernos temperados (cf. costas de Levante), precisamente donde más problemas ocasiona, por si mismo o en conjunción con el TSWV. Al contrario, en las regiones de clima continental, con inviernos rigurosos, son sólo los adultos diapausantes los que perpetúan la especie (cf. centro peninsular). Para clarificar esta situación en las Vegas del Guadiana (Extremadura), con un clima transicional entre ambos extremos, se estudió la actividad invernal de F. occidentalis durante los inviernos de 1995/1996 y 1996/1997. La dinámica poblacional se siguió mediante el muestreo sistemático de 30 brotes de alfalfa cada 1-2 semanas y extracción en embudo de Berlese- Tullgren. Paralelamente, se efectuó un muestreo de la vegetación adventicia y un seguimiento del vuelo de los adultos con trampas cromáticas. Se observó una situación antagónica entre ambos inviernos. Durante el invierno de 1995/1996, aunque F. occidentalis disminuyó sus efectivos, tuvo lugar una presencia continuada de larvas, evidenciando que el trips no entró en diapausa y fue capaz de reproducirse. Al contrario, durante el invierno de 1996/1997, F. occidentalis entró en diapausa y detuvo su reproducción, detectándose larvas tan sólo muy esporádicamente. La confrontación de las temperaturas de ambos inviernos con las del año medio, indica que la situación más usual en las Vegas del Guadiana debe corresponderse con la observada en 1996/1997, aunque la gran variabilidad térmica invernal interanual que caracteriza a la zona, especialmente patente en los últimos años, también sugiere que la reproducción invernal de F. occidentalis en las Vegas del Guadiana no debe ser infrecuente. El trips se asoció a 10 de las 13 especies de malas hierbas invernales muestreadas, que confirmaron su papel como reservorio potencial de insectación. El empleo de trampas cromáticas no fue efectivo para el seguimiento de las poblaciones de adultos durante el invierno. Se sugiere que las temperaturas invernales pueden ser un factor determinante de la irregular incidencia del complejo F. occidentalis-JSWW constatada en las Vegas del Guadiana. [ENG] The western flower thrips F. occidentalis does not enter in diapause and maintains its reproduction throughout the winter in temperate areas of Spain, precisely where more damage produces itself or together with the TSWV. In the contrary, in cold winter continental areas the thrips overwinters as diapausing adult. To clarify this situation in the Vegas del Guadiana, Extremadura, southwestern Spain, with a transitional climate between both extremes, the winter activity of F. occidentalis was studied during 1995/1996 and 1996/1997. Thrips population dynamic was monitored sampling systematically 30 lucerne shoots every 1-2 weeks and extracting thrips with a Berlese-Tullgren funnel. Additionally, population dynamic was monitored on winter weeds and with chromatic sticky traps. An opposed situation was observed between both winters. During 1995/1996 winter, even though F. occidentalis population was limited, larvae occurred throughout the winter, pointing out that F. occidentalis did not enter in diapause and maintained reproduction. In the contrary, during 1996/1997 winter, larvae were detected only very sporadically, which suggests that thrips reproduction was stopped and adult diapause induced. The comparison of temperatures of both winters with mean-year temperatures, indicates that the usual situation in the Vegas del Guadiana could be the observed during 1996/1997, even though the high winter thermic inter-annual variation that characterizes the region, specially in the last years, also suggests that thrips winter reproduction may be frequent. F. occidentalis occurred on 10 of 13 species of weeds sampled, which confirms their role as potential reservoir to infest crops. Sticky traps were no effective for monitoring adult populations during the winter. It is suggested that winter temperature may be an important factor determining the observed irregular incidence of the F. occidentalis-TSWV complex in the Vegas del Guadiana

Long-term mortality and trajectory of potassium easurements following an episode of acute severe hyperkalemia

Background: Hyperkalaemia is a common condition in patients with comorbidities such as chronic kidney disease (CKD) or congestive heart failure (HF). Moreover, severe hyperkalaemia is a potentially life-threatening condition that is associated with a higher risk of adverse clinical events such as ventricular arrhythmias and sudden cardiac death. Currently, data regarding the prognostic implications of chronic hyperkalaemia are available; however, information about the long-term clinical consequences after an episode of severe hyperkalaemia remains scarce. The objective of this study was to evaluate the association between the trajectory of potassium measurements in patients with acute hyperkalaemia and long-term all-cause mortality. Methods: This is a retrospective observational study that included patients with acute severe hyperkalaemia [potassium (K) >6 mEq/L] without haemolysis in the emergency room of Dr Peset University Hospital in Valencia, Spain searching the lab database from January 2016 to March 2017. The multivariable-adjusted association of serum potassium with mortality was assessed by using comprehensive state-of-the-art regression methods that can accommodate time-dependent exposure modelling. Results: We found 172 episodes of acute hyperkalaemia in 160 patients in the emergency room. The mean ± standard deviation age of the sample was 77 ± 12 years and 60.5% were males. The most frequent comorbidities were CKD (71.2%), HF (35%) and diabetes mellitus (56.9%). Only 11.9% of the patients were on chronic dialysis. A quarter of the patients did not have previous CKD, making hyperkalaemia an unpredictable life-threatening complication. During the acute episode, mean potassium and estimated glomerular filtration rate (eGFR) were 6.6 ± 0.6 (range 6.1-9.2) mEq/L and 23 ± 16 (range 2-84) mL/min/1.73 m2, respectively. After a median (interquartile range) follow-up of 17.3 (2.2-23.7) months, 68 patients died (42.5%). Recurrences of hyperkalaemia (K >5.5 mEq/L) were detected in 39.5% of the patients who were monitored during follow-up. We found that previous potassium levels during an acute severe hyperkalaemia episode were not predictors of mortality. Conversely, the post-discharge longitudinal trajectories of potassium were able to predict all-cause mortality (overall P = 0.0015). The effect of transitioning from hyperkalaemia to normokalaemia (K >5.5 mEq/L to K ≤5.5 mEq/L) after the acute episode was significant, and inversely associated with the risk of mortality. Conclusions: Potassium levels prior to a severe hyperkalaemic event do not predict mortality. Conversely, following an episode of acute severe hyperkalaemia, serial kinetics of potassium trajectories predict the risk of death. Further evidence is needed to confirm these findings and clarify the optimal long-term management of these patients

Phenotypic characterization of small colony variants of Staphylococcus aureus: a new challenge

En infecciones crónicas y recurrentes por Staphylococcus aureus se han descripto subpoblaciones de colonias pequeñas (VCPSa). El objetivo de este trabajo fue reconocer las características fenotípicas de VCPSa para optimizar su detección y caracterización a partir de materiales clínicos provenientes de infecciones crónicas. Se analizaron n=3 VCPSa de pacientes adultos con infecciones crónicas de tejidos blandos. Las muestras se inocularon en agar nutritivo, agar sangre; agar chocolate y agar Schaedler suplementado. Se realizaron tinción de Gram, catalasa, coagulasa libre, pruebas de dependencia para hemina, menadiona y timidina y, desarrollo/ataque del manitol en agar manitol salado. La sensibilidad antibiótica se efectuó en agar Mueller Hinton suplementado, según las pruebas de dependencia. Se investigó la presencia de proteína ligadora de penicilina anómala (PBP2´) por aglutinación con látex. Las VCPSa se detectaron en los medios de cultivo enriquecidos. Estas bacterias dieron positivas las pruebas de catalasa y coagulasa, y eran dependientes de menadiona y hemina. En los tres aislamientos se observó resistencia a cefoxitina y se detectó la PBP2´.In chronic and recurrent infections, small colonies of Staphylococcus aureus subpopulations (SCVSa) have been observed. The objective of the present study was to recognize the phenotypic characteristics of SCVSa isolated from patients with chronic infections to optimize their detection. SCVSa of adult patients n=3 with chronic soft tissue infections were analyzed. Samples were inoculated on nutritive agar, blood-agar, chocolate agar and supplemented Schaedler agar. Subsequently, Gram stain, catalase, free coagulase, dependence tests for hemin, menadione and thymidine, and growth/fermentation of mannitol on salt mannitol agar were performed. Antibiotic susceptibility tests were performed by the agar difSanfusion method on supplemented Mueller Hinton agar, according to dependence assays results. Anomalous penicillin binding protein (PBP2’) was investigated by latex agglutination. SCVSa were detected in all enriched culture media. They showed catalase and coagulase activities, and menadione and hemin dependence. By the agar diffusion test, cefoxitin resistance was found in all isolates; PBP2’ was detected as well.Facultad de Ciencias Médica

Fatty acids homeostasis during fasting predicts protection from chemotherapy toxicity.

Fasting exerts beneficial effects in mice and humans, including protection from chemotherapy toxicity. To explore the involved mechanisms, we collect blood from humans and mice before and after 36 or 24 hours of fasting, respectively, and measure lipid composition of erythrocyte membranes, circulating micro RNAs (miRNAs), and RNA expression at peripheral blood mononuclear cells (PBMCs). Fasting coordinately affects the proportion of polyunsaturated versus saturated and monounsaturated fatty acids at the erythrocyte membrane; and reduces the expression of insulin signaling-related genes in PBMCs. When fasted for 24 hours before and 24 hours after administration of oxaliplatin or doxorubicin, mice show a strong protection from toxicity in several tissues. Erythrocyte membrane lipids and PBMC gene expression define two separate groups of individuals that accurately predict a differential protection from chemotherapy toxicity, with important clinical implications. Our results reveal a mechanism of fasting associated with lipid homeostasis, and provide biomarkers of fasting to predict fasting-mediated protection from chemotherapy toxicity.General: We thank Prof. Jose Maria. Ordovas for his kind suggestions; nutritionists Helena Marcos-Pasero, Elena Aguilar-Aguilar and Isabel Espinosa-Salinas for their help with volunteers management; Rosa Serrano for her help with animal experiments; Susana Molina for her advice with PBMC isolation; Luisa Mariscal, Domingo Fernandez, Lola Martinez, Diego Megias, Patricia Gonzalez, Fernando Pelaez, Anabel Sanz, Carolina Pola, Celia de la Calle, Ana Ortega, Ana Sagrera, Jose Miguel Frade, Elena Lopez-Guadamillas, Maribel Munoz, Susana Llanos, Andres Fernandez, Aranzazu Sierra, Andres Lopez, Noemi Haro and Ildefonso Rodriguez for their excellent technical and scientific support. Work at the laboratory of P.J.F.M. is funded by the Ramon Areces Foundation, (CIVP18A3891), Asociacion Espanola contra el Cancer-AECC (SIRTBIO-LABAE18008FERN), a Ramon y Cajal Award from the Spanish Ministry of Science, Innovation and Universities (MICINN) (RYC-2017-22335), RETOS projects Program of MICINN (SAF2017-85766-R) and the Portuguese Foundation for Science and Technology (FCT-MCTES, SFRH/BD/124022/2016). Work at the laboratory of ARM was funded by the MICINN (PID2019-110183RB-C21), Regional Government of Community of Madrid (P2018/BAA-4343-ALIBIRD2020-CM) and the Ramon Areces Foundation. Work at the laboratory of A.D.R. Funded by the Comunidad de Madrid-Talento Grant 2018-T1/BMD-11966 and the MICINN PID-2019-106893RA-100. Work at the laboratory of L.D. is funded by projects from the Health Research Fund (ISCIII FIS PI14/01374 and FISPI17/00508) and from a Manuel de Oya research fellowship from the Beer and Health Foundation. Work at the laboratory of A.E. is funded by a Ramon y Cajal Award from MICINN (RYC-2013-13546) and RETOS projects Program of the MICINN, co-funded by the European Regional Development Fund (ERDF) (SAF2015-67538-R). Work in the laboratory of M.S. was funded by the IRB and by grants from the Spanish Ministry of Economy co-funded by the European Regional Development Fund (ERDF) (SAF2013-48256-R), the European Research Council (ERC-2014-AdG/669622), and the "laCaixa" Foundation.S

What is the optimal level of vitamin D in non-dialysis chronic kidney disease population?

AIM: To evaluate thresholds for serum 25(OH)D concentrations in relation to death, kidney progression and hospitalization in non-dialysis chronic kidney disease (CKD) population. METHODS: Four hundred and seventy non-dialysis 3-5 stage CKD patients participating in OSERCE-2 study, a prospective, multicenter, cohort study, were prospectively evaluated and categorized into 3 groups according to 25(OH)D levels at enrollment (less than 20 ng/mL, between 20 and 29 ng/mL, and at or above 30 ng/mL), considering 25(OH)D between 20 and 29 ng/mL as reference group. Association between 25(OH)D levels and death (primary outcome), and time to first hospitalization and renal progression (secondary outcomes) over a 3-year follow-up, were assessed by Kaplan-Meier survival curves and Cox-proportional hazard models. To identify 25(OH)D levels at highest risk for outcomes, receiver operating characteristic (ROC) curves were performed. RESULTS: Over 29 ± 12 mo of follow-up, 46 (10%) patients dead, 156 (33%) showed kidney progression, and 126 (27%) were hospitalized. After multivariate adjustment, 25(OH)D < 20 ng/mL was an independent predictor of all-cause mortality (HR = 2.33; 95%CI: 1.10-4.91; P = 0.027) and kidney progression (HR = 2.46; 95%CI: 1.63-3.71; P < 0.001), whereas the group with 25(OH)D at or above 30 ng/mL did not have a different hazard for outcomes from the reference group. Hospitalization outcomes were predicted by 25(OH) levels (HR = 0.98; 95%CI: 0.96-1.00; P = 0.027) in the unadjusted Cox proportional hazards model, but not after multivariate adjusting. ROC curves identified 25(OH)D levels at highest risk for death, kidney progression, and hospitalization, at 17.4 ng/mL [area under the curve (AUC) = 0.60; 95%CI: 0.52-0.69; P = 0.027], 18.6 ng/mL (AUC = 0.65; 95%CI: 0.60-0.71; P < 0.001), and 19.0 ng/mL (AUC = 0.56; 95%CI: 0.50-0.62; P = 0.048), respectively. CONCLUSION: 25(OH)D < 20 ng/mL was an independent predictor of death and progression in patients with stage 3-5 CKD, with no additional benefits when patients reached the levels at or above 30 ng/mL suggested as optimal by CKD guidelines.Abbott and the Spanish Society of Nephrolog

A novel biocompatible polymer derived from D-mannitol used as a vector in the field of genetic engineering of eukaryotic cells

The design and preparation of new vectors to transport genetic material and increase the transfection efficiency continue being an important research line. Here, a novel biocompatible sugar-based polymer derived from D-mannitol has been synthesized to be used as a gene material nanocarrier in human (gene transfection) and microalga cells (transformation process). Its low toxicity allows its use in processes with both medical and industrial applications. A multidisciplinary study about the formation of polymer/p-DNA polyplexes has been carried out using techniques such as gel electrophoresis, zeta potential, dynamic light scattering, atomic force microscopy, and circular dichroism spectroscopy. The nucleic acids used were the eukaryotic expression plasmid pEGFP-C1 and the microalgal expression plasmid Phyco69, which showed different behaviors. The importance of DNA supercoiling in both transfection and transformation processes was demonstrated. Better results were obtained in microalga cells nuclear transformation than in human cells gene transfection. This was related to the plasmid's conformational changes, in particular to their superhelical structure. It is noteworthy that the same nanocarrier has been used with eukaryotic cells from both human and microalga.This work was supported by the Consejería de Conocimiento, Innovación y Universidades de la Junta de Andalucía (FQM-206, FQM-274, FQM-135 and P20–01234); VI Plan Propio Universidad de Sevilla (PP2019/00000748), and the European Union (Feder Funds).Peer reviewe

Supplementary material. A novel biocompatible polymer derived from D-mannitol used as a vector in the field of genetic engineering of eukaryotic cells

S1. Experimental Section: S1.1. Synthesis and Characterization Data. S1.2. Fluorescence Measurements. S1.3. Zeta Potential Measurements. S1.4. Dynamic Light Scattering Measurements. S1.5. Circular Dichroism Spectra. S1.6. Agarose Gel Electrophoresis. S1.7. Atomic Force Microscopy (AFM). S1.8. In Vitro Assays S1.9. Transfection Assays. S1.10. Chlamydomonas reinhardtii Nuclear Transformation. S2. Results and Discussion: S2.1. Characterization of Monomers and Polyurethanes. S2.2. Formation of the polyplexes PUMan/ctDNA. Figures: Fig. S1. FTIR spectra of PUMan and (MBocCis)DTDI. Fig. S2. SEC chromatogram of (MAL)DTDI. Fig. S3. SEC chromatogram of (MBocCis)DTDI. Fig. S4. SEC chromatogram of PUMan. Fig. S5. 1H NMR of (MAL)DTDI. Fig. S6. 1H NMR of (MBocCis)DTDI. Fig. S7. 1H NMR of PUMan. Fig. S8. TGA curve of PUMan. Fig. S9. Plot of EB emission intensities at different N/P values, circular dichroism spectra, zeta potential and hydrodynamic diameters of the PUMan-based polyplexes. Fig.S10. Electrophoresis of polyplexes PUMan/digested pEGFP-C1 and PUMan/digested Phyco69 at different N/P ratios. Fig.S11. Percentage of GFP positive cells after transfection with 3 μg of the plasmid pEGFP-C1 with the indicated reagents. The molar ratio PUMan:FuGENE and PUMan:DOPE was 1:1. Tables: Table S1. Thermal properties of PUMan and its precursors.Peer reviewe

Truncating FLNC Mutations Are Associated With High-Risk Dilated and Arrhythmogenic Cardiomyopathies

BACKGROUND: Filamin C (encoded by the FLNC gene) is essential for sarcomere attachment to the plasmatic membrane. FLNC mutations have been associated with myofibrillar myopathies, and cardiac involvement has been reported in some carriers. Accordingly, since 2012, the authors have included FLNC in the genetic screening of patients with inherited cardiomyopathies and sudden death. OBJECTIVES: The aim of this study was to demonstrate the association between truncating mutations in FLNC and the development of high-risk dilated and arrhythmogenic cardiomyopathies. METHODS: FLNC was studied using next-generation sequencing in 2,877 patients with inherited cardiovascular diseases. A characteristic phenotype was identified in probands with truncating mutations in FLNC. Clinical and genetic evaluation of 28 affected families was performed. Localization of filamin C in cardiac tissue was analyzed in patients with truncating FLNC mutations using immunohistochemistry. RESULTS: Twenty-three truncating mutations were identified in 28 probands previously diagnosed with dilated, arrhythmogenic, or restrictive cardiomyopathies. Truncating FLNC mutations were absent in patients with other phenotypes, including 1,078 patients with hypertrophic cardiomyopathy. Fifty-four mutation carriers were identified among 121 screened relatives. The phenotype consisted of left ventricular dilation (68%), systolic dysfunction (46%), and myocardial fibrosis (67%); inferolateral negative T waves and low QRS voltages on electrocardiography (33%); ventricular arrhythmias (82%); and frequent sudden cardiac death (40 cases in 21 of 28 families). Clinical skeletal myopathy was not observed. Penetrance was >97% in carriers older than 40 years. Truncating mutations in FLNC cosegregated with this phenotype with a dominant inheritance pattern (combined logarithm of the odds score: 9.5). Immunohistochemical staining of myocardial tissue showed no abnormal filamin C aggregates in patients with truncating FLNC mutations. CONCLUSIONS: Truncating mutations in FLNC caused an overlapping phenotype of dilated and left-dominant arrhythmogenic cardiomyopathies complicated by frequent premature sudden death. Prompt implantation of a cardiac defibrillator should be considered in affected patients harboring truncating mutations in FLNC.Instituto de Salud Carlos III [PI11/0699, PI14/0967, PI14/01477, RD012/0042/0029, RD012/0042/0049, RD012/0042/0066, RD12/0042/0069]; Spanish Ministry of Economy and Competitiveness [SAF2015-71863-REDT]; Plan Nacional de I+D+I; Plan Estatalde I+D+I, European Regional Development Fund; Health in Code SLS