Characterisation of a major QTL for sodium accumulation in tomato grown in high salinity.

Soil salinity is a serious concern for tomato culture, affecting both yield and quality parameters. Although some genes involved in tomato salt tolerance have been identified, their genetic diversity has been rarely studied. In the present study, we assessed salt tolerance-related traits at juvenile and adult stages in a large core collection and identified salt tolerance quantitative trait loci (QTLs) by genome-wide association study (GWAS). The results suggested that a major QTL is involved in leaf sodium accumulation at both physiological stages. We were able to identify the underlying candidate gene, coding for a well-known sodium transporter, called SlHKT1.2. We showed that an eQTL for the expression of this gene in roots colocalized with the above ground sodium content QTL. A polymorphism putatively responsible for its variation was identified in the gene promoter. Finally, to extend the applicability of these results, we carried out the same analysis on a test-cross panel composed of the core collection crossed with a distant line. The results indicated that the identified QTL retained its functional impact even in a hybrid genetic context: this paves the way for its use in breeding programs aimed at improving salinity tolerance in tomato cultivars.

Seismic Stereometry Reveals Preparatory Behavior and Source Kinematics of Intermediate-Size Earthquakes.

Although moderate-size earthquakes are poorly studied by lack of near-fault observations, they can provide key information about larger damaging earthquakes. Here we propose a new approach, inspired by double-difference relocation, that uses high-coherency waveforms recorded at neighboring sensors, to study the preparation phase and dynamics of moderate-size earthquakes. We validate this technique by analyzing the 2016, M w 5.2 Borrego Springs earthquake in Southern California and find consistent rupture velocities of 2 km/s highlighting two main rupture asperities. The analysis of the 2019, Ml5.2 Le Teil earthquake in France reveals slow nucleation at depth that migrates to the surface and propagates northward with a velocity of ∼2.8 km/s, highlighting two main rupture events also imaged by InSAR. By providing unprecedented resolution in our observation of the rupture dynamics, this approach will be useful in better understanding the preparation phase and rupture of both tectonic and induced earthquakes.

In vivo selection of KPC-94 and KPC-95 in Klebsiella pneumoniae isolates from patients treated with ceftazidime/avibactam.

Ceftazidime/avibactam (CZA) is used to treat infections caused by Klebsiella pneumoniae carbapenemase-producing K. pneumoniae (KPC-Kp). Resistance to CZA is commonly related to point mutations in the blaKPC gene. Here we describe the in vivo emergence of CZA resistance in clinical isolates of KPC-Kp from four patients treated with this combination therapy. Four pre-therapy and five post-therapy KPC-Kp isolates were examined. Antibiogram (microdilution and gradient strips) and whole-genome sequencing were performed. The role of KPC mutations was validated by cloning blaKPC genes into competent Escherichia coli. All KPC-Kp isolates recovered before treatment with CZA were susceptible to CZA and produced KPC-3. Five KPC-Kp isolates recovered after treatment were resistant to this combination. Three post-therapy isolates from two patients produced KPC-31 (D179Y mutation). Additionally, we identified the novel substitution LN169-170H (KPC-94) in one isolate, and the combination of two independently described mutations, D179Y and A172T (KPC-95), in another isolate. All KPC-Kp isolates belonged to sequence type 512 (ST512). All CZA-resistant isolates with blaKPC variants had restoration of carbapenem susceptibility. In conclusion, resistance to CZA was related to blaKPC mutations, including the new KPC-94 and KPC-95 alleles, which do not cause carbapenem resistance.

Protective proteins are differentially expressed in tomato genotypes differing for their tolerance to low-temperature storage.

When stored at low temperature, tomato fruits exhibit chilling injury symptoms, such as rubbery texture and irregular ripening. To identify proteins related to chilling tolerance, we compared two tomato near isogenic lines differing for their texture phenotype at harvest in a fruit-storage trial including two temperatures (4 and 20 degrees C) along several days of conservation. Fruit evolution was followed by assessing fruit color, ethylene emission and texture parameters. The most contrasted samples were submitted to proteomic analysis including two-dimensional electrophoresis and mass spectrometry of protein spots to identify the proteins, whose expression varied according to the genotype or the storage conditions. Unexpectedly, the most firm genotype at harvest was the most sensitive to cold storage. The other genotype exhibited a delay in fruit firmness loss leading to the texture differences observed after 20 days of 4 degrees C storage. The proteome analysis of these contrasted fruits identified 85 proteins whose quantities varied with temperature or genotype. As expected, cold storage decreased the expression of proteins related to maturation process, such as acidic invertase, possibly controlled post-translational regulation of polygalacturonase and up-regulated proteins related to freezing tolerance. However, the study point out proteins involved in the differential resistance to chilling conditions of the two lines. This includes specific isoforms among the large family of small heat shocked proteins, and a set of proteins involved in the defense against of the reticulum endoplasmic stress.

Tomato fruit ascorbic acid content is linked with monodehydroascorbate reductase activity and tolerance to chilling stress.

Quantitative trait loci (QTL) mapping is a step towards the identification of factors regulating traits such as fruit ascorbic acid content. A previously identified QTL controlling variations in tomato fruit ascorbic acid has been fine mapped and reveals that the QTL has a polygenic and epistatic architecture. A monodehydroascorbate reductase (MDHAR) allele is a candidate for a proportion of the increase in fruit ascorbic acid content. The MDHAR enzyme is active in different stages of fruit ripening, shows increased activity in the introgression lines containing the wild-type (Solanum pennellii) allele, and responds to chilling injury in tomato along with the reduced/oxidized ascorbate ratio. Low temperature storage of different tomato introgression lines with all or part of the QTL for ascorbic acid and with or without the wild MDHAR allele shows that enzyme activity explains 84% of the variation in the reduced ascorbic acid levels of tomato fruit following storage at 4 degrees C, compared with 38% at harvest under non-stress conditions. A role is indicated for MDHAR in the maintenance of ascorbate levels in fruit under stress conditions. Furthermore, an increased fruit MDHAR activity and a lower oxidation level of the fruit ascorbate pool are correlated with decreased loss of firmness because of chilling injury.

Evaluation of new GenoType MTBDRplus for detection of resistance in cultures and direct specimens of Mycobacterium tuberculosis.

SETTINGS: Molecular methods frequently used in laboratories can now give us useful information about low growth bacteria. OBJECTIVE: To evaluate the new GenoType MTBDRplus assay for its ability to detect mutations in the 81-bp hotspot region of the rpoB gene, mutations in codon 315 of the katG gene and alterations in the inhA promoter region. DESIGN: Prospective resistance to rifampicin (RMP) and isoniazid (INH) study using Mycobacterium tuberculosis positive specimens and cultures comparing the results of GenoType MTBDRplus with those obtained phenotypically with the Bactec MGIT (Mycobacterial Growth Indicator Tube) 960. RESULTS: In 59 specimens (18 smear microscopy samples and 41 solid and liquid medium cultures), mutations were detected in all of 36 M. tuberculosis strains phenotypically resistant to RMP (100%), and in 35 of 37 strains phenotypically resistant to INH (94.59%). The new assay prompted a 21.6% increase in the direct detection of INH resistance in the strains studied, due to the incorporation of inhA promoter region probes in the test. CONCLUSIONS: The GenoType MTBDRplus assay is a valid method for detecting the most common mutations in strains resistant to RMP and INH. However, further phenotypic testing is required, as the assay failed to detect 100% of INH and RMP resistance.

[Antimicrobial susceptibility of Enterococcus faecalis isolated from patients in Córdoba (Spain)]. / Sensibilidad a los antimicrobianos de Enterococcus faecalis aislados de pacientes en la provincia de Córdoba (España).

Enterococcus faecalis is a pathogenic microorganism. The aim of this investigation was to study the antibiotic susceptibility of the strains isolated in Cordoba in a 20-month period (January 2004 to August 2005). Susceptibility rates to betalactamics were 98% to ampicillin and 99% to amoxicillin/clavulanic acid; high-dose aminoglycosides (streptomycin 1000 microg and gentamycin 500 microg) obtained 56% and 76%, respectively. We found no strains resistant to glycopeptides (vancomycin and teicoplanin) or to linezolid.

Saturated molecular map of the rice genome based on an interspecific backcross population.

A molecular map has been constructed for the rice genome comprised of 726 markers (mainly restriction fragment length polymorphisms; RFLPs). The mapping population was derived from a backcross between cultivated rice, Oryza sativa, and its wild African relative, Oryza longistaminata. The very high level of polymorphism between these species, combined with the use of polymerase chain reaction-amplified cDNA libraries, contributed to mapping efficiency. A subset of the probes used in this study was previously used to construct an RFLP map derived from an inter subspecific cross, providing a basis for comparison of the two maps and of the relative mapping efficiencies in the two crosses. In addition to the previously described PstI genomic rice library, three cDNA libraries from rice (Oryza), oat (Avena) and barley (Hordeum) were used in this mapping project. Levels of polymorphism detected by each and the frequency of identifying heterologous sequences for use in rice mapping are discussed. Though strong reproductive barriers isolate O. sativa from O. longistaminata, the percentage of markers showing distorted segregation in this backcross population was not significantly different than that observed in an intraspecific F2 population previously used for mapping. The map contains 1491 cM with an average interval size of 4.0 cM on the framework map, and 2.0 cM overall. A total of 238 markers from the previously described PstI genomic rice library, 250 markers from a cDNA library of rice (Oryza), 112 cDNA markers from oat (Avena), and 20 cDNA markers from a barley (Hordeum) library, two genomic clones from maize (Zea), 11 microsatellite markers, three telomere markers, eleven isozymes, 26 cloned genes, six RAPD, and 47 mutant phenotypes were used in this mapping project. Applications of a molecular map for plant improvement are discussed.

[Evolution of methicillin resistance in Staphylococcus aureus in Cordoba (Spain) in the years 2002-2005]. / Evolución de la resistencia a la meticilina de Staphylococcus aureus en la provincia de Córdoba (España) en los años 2002-2005.

In the last few years, methicillin-resistant Staphylococcus aureus (MRSA) has become a very important human pathogen. Our aim was to study the evolution of methicillin resistance of Staphylococcus aureus strains isolated in our hospital over a four-year period and to compare our situation with the rest of Spain and Europe. The rates varied from 39.9% in 2002 to 46.4% in 2005. Units with the highest rate were ICU and surgical wards. We found no glycopeptide-resistant strains.

The neuroergonomic evaluation of human machine interface design in air traffic control using behavioral and EGG/ERP measures.

The Air Traffic Control (ATC) environment is complex and safety-critical. Whilst exchanging information with pilots, controllers must also be alert to visual notifications displayed on the radar screen (e.g., warning which indicates a loss of minimum separation between aircraft). Under the assumption that attentional resources are shared between vision and hearing, the visual interface design may also impact the ability to process these auditory stimuli. Using a simulated ATC task, we compared the behavioral and neural responses to two different visual notification designs--the operational alarm that involves blinking colored "ALRT" displayed around the label of the notified plane ("Color-Blink"), and the more salient alarm involving the same blinking text plus four moving yellow chevrons ("Box-Animation"). Participants performed a concurrent auditory task with the requirement to react to rare pitch tones. P300 from the occurrence of the tones was taken as an indicator of remaining attentional resources. Participants who were presented with the more salient visual design showed better accuracy than the group with the suboptimal operational design. On a physiological level, auditory P300 amplitude in the former group was greater than that observed in the latter group. One potential explanation is that the enhanced visual design freed up attentional resources which, in turn, improved the cerebral processing of the auditory stimuli. These results suggest that P300 amplitude can be used as a valid estimation of the efficiency of interface designs, and of cognitive load more generally.

New Anyplex™ II MTB/MDR/XDR kit for detection of resistance mutations in M. tuberculosis cultures.

SETTINGS: The new Anyplex™ II MTB/MDR/XDR PCR assay enables the joint analysis of mutations conferring resistance to first- and second-line anti-tuberculosis drugs and the detection of Mycobacterium tuberculosis. OBJECTIVES: To evaluate the performance of the new Anyplex assay in detecting mutations that confer resistance to first- and second-line drugs in M. tuberculosis cultures. DESIGN: Results obtained using the new technique were compared with those obtained by phenotypic drug susceptibility testing (DST) and with two GenoType tests for the detection of mutations: GenoType(®) MTBDRplus and GenoType(®) MTBDRsl. RESULTS: For rifampicin resistance mutations, Anyplex displayed 97% sensitivity and 100% specificity compared with 100% and 100% for MTBDRplus. For isoniazid (INH) resistance, Anyplex displayed 61% sensitivity and 98% specificity compared with 62% and 98% for MTBDRplus. For second-line drugs, Anyplex recorded 95% sensitivity and 99% specificity in the detection of resistance to quinolones compared with 100% and 98% for the MTBDRsl. While both techniques displayed 100% specificity for aminoglycoside resistance mutations, sensitivity was 100% for Anyplex and 88% for MTBDRsl. CONCLUSIONS: Results obtained using Anyplex agreed strongly with those obtained using the two GenoType molecular techniques and with phenotypic DST, except in the case of INH, due to the large number of genes involved in resistance to this drug.

Complications associated with Brucella melitensis infection: a study of 530 cases.

We carried out a prospective study of 530 patients older than 14 years of age with brucellosis. We describe the incidence and clinical features of the focal forms of the disease, analyzing some of the possible factors associated with their appearance. One hundred sixty-nine patients (31.9%) had a focal form or complication. Osteoarticular complications were the most frequent, totaling 113 cases (66%), followed by genitourinary with 18 cases (5.1% of males), hepatic (2.5%), neurologic (1.7%), and heart (1.5%). Nine patients (1.7%) had more than 1 complication. In a multivariate analysis, diagnostic delay greater than 30 days (OR 2.0), ESR > 40 mm/hr (OR 1.9), and levels of alpha-2 globulin > 7.5 g/L (OR 6.8) were statistically significant independent variables associated with the presence of focal forms. Twenty-five patients with complications (14.8%) required surgical treatment. The relapse rate was 3.6% for those patients without complications and 4.1% for patients with focal forms (p > 0.05). However, when therapeutic failure, relapses, and mortality were considered together, the risk of an unfavorable evolution was significantly greater in patients with focal forms (10.6% versus 3.6% in patients without complications; OR 1.9, 95% CI 1.4-7.1, p < 0.005). Given the worse prognosis, knowledge and early diagnosis of the focal forms of B. melitensis infection is especially important.

Blood levels of water-soluble vitamins in pediatric patients on total parenteral nutrition using a multiple vitamin preparation.

Although guidelines for the parenteral use of vitamin preparations in pediatric patients have been published, there are very limited data on the efficiency of these preparations and on the exact needs of infants and children on total parenteral nutrition (TPN). We report here an open, prospective, study of the blood levels of water-soluble vitamins in infants and children on TPN before and during supplementation with a new water-soluble multivitamin formula containing per vial unit: B1, 3 mg; B2, 3.6 mg; B6, 4 mg; niacin, 40 mg; pantothenate, 15 mg; ascorbate, 100 mg; biotin, 60 micrograms; folic acid, 400 micrograms; B12, 5 micrograms. Thirteen children, 9 months to 15 yr old, on home TPN for 1.5 months to 7 yr, and 17 hospitalized infants and children, 1 week to 15 yr old, receiving TPN were studied for 2 weeks to 4 months. Daily doses were given according to age: 1/2 vial if less than 18 months; 1 vial if greater than 18 months and less than 10 years; 1.5 vials if greater than 10 years. Assays for B1, B2, biotin, niacin, plasma and red blood cell (RBC) folates were performed by microbiologic methods, B12 was measured by radioimmunoassay. During the study, B1 levels were consistently above the upper limit of the normal range, B2 and B12 remained in the normal range although there was a slight decrease in B12 values. Almost half of the patients had initially low levels of biotin, niacin, and folates. Biotin, after a significant sharp rise during the first month of supplementation returned to normal range. Niacin levels were initially low in infants and rose toward normal values during treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

[Incidence and current clinical spectrum of tuberculosis in a metropolitan area in the south of Spain]. / Incidencia y espectro clínico actual de la tuberculosis en un área sanitaria metropolitana del sur de España.

BACKGROUND: To study the incidence and clinical spectrum of tuberculosis in the metropolitan area of Málaga (Spain). METHODS: Prospective study which includes all patients who had a diagnosis of tuberculosis within the referral area of "Carlos Haya" Málaga Regional Hospital from March 1, 1993 to February 28, 1994. RESULTS: During the study period, there were 138 cases of tuberculosis, with an incidence of 43.7 cases/10(5) inhabitants. Ninety one cases (66%) were male, and the mean age (SD) was 33.2 (18.3), with 88% being less than 55 years old. Thirty six patients (26.1%) were HIV-infected. Extrapulmonary tuberculosis made up 27.5% of the cases, and was more frequent in HIV-infected patients (p < 0.01; odds ratio: 2.9; 95% CI: 1.2-7.1). The mean (SD) time to diagnosis was 54.3 (76) days. The diagnosis was microbiologically confirmed in 106 cases (76.8%), histologically in 14 cases (10.1%), and the remaining 18 cases (13.1%) were clinically diagnosed. The global rate of resistance was 10.8%. The rate of primary resistance was 4.6%, and the rate of multidrug-resistant tuberculosis was 3.1%. Eighty nine patients (77.4%) were cured, six patients (5.2%) stopped the treatment, 3 (2.6%) had relapses and 1 (0.9%) was considered a therapeutic failure; 16.7% of patients were lost for follow-up. Sixteen patients died and in nine of them (6.5%) the death was attributed to tuberculosis. CONCLUSIONS: The incidence of tuberculosis in Málaga urban area is high. It mainly affects young males of unfavored socio-economic classes. HIV-infected patients account for a high percentage of the cases. The high number of productive cases and the long time to diagnosis evidence the shortcomings of our sanitary system. These facts, together with the high rate of non-compliance, of treatment may explain the seriousness of the current situation in our country.

[Comparison of different methods for serum folate assay]. / Comparaison de différentes méthodes de dosage des folates sériques.

Folates were determined in 148 patient sera, using four different methods: a microbiological assay (Reference Method), two radioassays (Magic B12 FOL [NB] and SimulTRAC SNB no boil kits) and a non isotopic competition method (Magic Lite kit). Folate mean varied according to the methods 10.0 nmol.l-1 (reference method), 12.2 nmol.l-1 (Magic Lite), 8.7 nmol.l-1 (Magic B12 FOL [NB]) and 10.8 nmol.l-1 (SimulTRAC SNB NO boil). Poor correlations were also noted (0.83 < r < 0.95, figures 1 and 2). Differences between methods were explained according to the literature.

[Investigation of antimicrobial resistance to Enterococcus faecium]. / Investigación de las resistencias a antimicrobianos en Enterococcus faecium.

We performed a antibiotic resistance study on Enterococcus faecium isolated from intrahospitalary and extrahospitalary samples between 2004 and 2010. Three different samples were studied; urine, blood and wound swabs, considering a strain per patient. We included in the study a global amount of 637 E. faecium isolares. We employed semiautomatic system WIDER I for identification and sensitivity testing. We considered susceptibility and resistance criteria recommended by MENSURA group. We found a susceptibility rate of 48.05% to betalactams, 100% to linezolid, and 99.46% to vancomycin. The resistance to aminoglycosides ranged between 41.41 and 73.55%. We obtained 6 isolates resistant to vancomycin one of them from an extrahospitalary strain and five from intrahospitalary strains. It seems that vancomycin resistance should be controlled.

[Evaluation of the variability in the susceptibility of Acinetobacter baumannii to tigecycline in the same medium with two methods of quantitative diffusion different commercial]. / Evaluación de la variabilidad en la sensibilidad de Acinetobacter baumannii a tigeciclina en un mismo medio de cultivo con dos métodos de difusión cuantitativos comerciales diferentes.

INTRODUCTION: Tigecycline may be a therapeutic alternative for the control of multidrug-resistant Acinetobacter baumannii, although there is no consensus on the cutoffs or susceptibility to the variability of the minimum inhibitory concentration (MIC) according to the culture medium and strips for the antibiogram against this microorganism by quantitative diffusion method. Therefore, the objective was to verify this variability and propose epsilometer test strip that more closely resemble to the standard method. MATERIAL AND METHODS: 38 strains of A. baumannii were selected and evaluated their susceptibility to tigecycline with two different commercial strips (E-TEST and Liofilchem). MICs were compared with those obtained by the standard technique of microdilution broth. RESULTS: MICs obtained by the Liofilchem strip were more similar to standard method than those obtained by E-TEST strips. CONCLUSION: In the two studied strips, higher MICs to those obtained by the standard method were observed leading to false-positive tigecicline resistance in many cases. However, the Liofilchem strip showed the results more closely resemble to the standard method.

Morphologic examination of the temporal bone by cone beam computed tomography: comparison with multislice helical computed tomography.

INTRODUCTION: High-resolution CT imaging is essential to diagnosis and follow-up of temporal bone pathology. Morphologically, CT is the reference examination. The requirement of long-term follow-up thus exposes patients to cumulative radiation doses. Limiting exposure to ionizing radiation is an increasing concern of public health authorities. The principal advantage of Cone Beam CT (CBCT) lies in a significant reduction in radiation dose. The main objective of the present study was to assess the morphologic concordance between CBCT and Multislice Helical Computed Tomography (MSCT) on 20 anatomic landmarks corresponding to regions of interest in clinical practice. The secondary objectives were to compare the two techniques qualitatively in stapes and footplate assessment and measurement of footplate thickness, and quantitatively in terms of dosimetry. MATERIAL AND METHODS: An experimental anatomical study was performed on 12 temporal bones from fresh human cadavers of unknown clinical history. Each underwent CBCT and MSCT. RESULTS: There was no significant difference in morphologic assessment of the temporal bones on the two techniques. Exploration of the stapes, incudostapedial joint, anterior stapediovestibular joint and footplate was qualitatively more precise on CBCT, and footplate thickness showed less overestimation than on MSCT. CBCT delivered 22 times less radiation than MSCT under the present experimental conditions. CONCLUSION: CBCT provides reliable morphologic assessment of temporal bone, thanks to higher spatial resolution than on MSCT, with significantly reduced radiation dose.

The distribution of copia-type retrotransposons and the evolutionary history of tomato and related wild species.

Retrotransposons are mobile genetic elements that amplify throughout the genome and may be important contributors of genetic diversity. Their distribution is influenced by element behaviour and host-driven controls. We analysed the distribution of three copia-type retrotransposons, ToRTL1, T135 and Tnt1 using sequence-specific amplification polymorphism in self-compatible (SC) and incompatible (SI) species of Solanum subsection Lycopersicon, and genetically mapped polymorphic insertions in S. lycopersicum (tomato). The majority of polymorphic insertions (61%) are located in centromeric regions of the tomato genome. A significant positive relationship was detected between insertion polymorphisms and mating system, independent of selection as most insertions were found to be neutral. As insertion patterns successfully inferred interspecific relationships of Solanum subsection Lycopersicon, our results suggest that the distribution of ToRTL1, T135 and Tnt1 may essentially be determined by selection removing strongly deleterious insertions, with genetic drift and mating system, but not recombination rate, playing important roles.