Genetic characterization of a Sinorhizobium meliloti chromosomal region involved in lipopolysaccharide biosynthesis

The genetic characterization of a 5.5-kb chromosomal region of Sinorhizobium meliloti 2011 that contains lpsB, a gene required for the normal development of symbiosis with Medicago spp., is presented. The nucleotide sequence of this DNA fragment revealed the presence of six genes: greA and lpsB, transcribed in the forward direction; and lpsE, lpsD, lpsC, and lrp, transcribed in the reverse direction. Except for lpsB, none of the lps genes were relevant for nodulation and nitrogen fixation. Analysis of the transcriptional organization of lpsB showed that greA and lpsB are part of separate transcriptional units, which is in agreement with the finding of a DNA stretch homologous to a "nonnitrogen" promoter consensus sequence between greA and lpsB. The opposite orientation of lpsB with respect to its first downstream coding sequence, lpsE, indicated that the altered LPS and the defective symbiosis of lpsB mutants are both consequences of a primary nonpolar defect in a single gene. Global sequence comparisons revealed that the greA-lpsB and lrp genes of S. meliloti have a genetic organization similar to that of their homologous loci in R. leguminosarum bv. viciae. In particular, high sequence similarity was found between the translation product of lpsB and a core-related biosynthetic mannosyltransferase of R. leguminosarum bv. viciae encoded by the lpcC gene. The functional relationship between these two genes was demonstrated in genetic complementation experiments in which the S. meliloti lpsB gene restored the wild-type LPS phenotype when introduced into lpcC mutants of R. leguminosarum. These results support the view that S. meliloti lpsB also encodes a mannosyltransferase that participates in the biosynthesis of the LPS core. Evidence is provided for the presence of other lpsB-homologous sequences in several members of the family Rhizobiaceae.Facultad de Ciencias ExactasInstituto de Biotecnologia y Biologia Molecula

Inverse Identification of Cable Forces using its Modal Behavior by Direct and Non-Contact Vibration Measurements

Cables are essential in civil engineering for constructing slender, lightweight structures with large spans. To ensure serviceability and load-bearing capacity, a monitoring of the cable forces is necessary. Conventional, static methods are not suitable for systems with highly pre-stressed cables or large cable diameters, so dynamic measurements using the cable's vibration behavior offer an alternative. This study presents laboratory test results on inverse identification of cable forces using eigenmodes and the corresponding frequencies, comparing contact and non-contact dynamic measurement methods. Two methods for determining the cable force will be investigated within this study: (1) the linear theory of vibrating strings neglects internal sag and bending stiffness, and (2) an inverse identification of the cable force for a cable tensioned on both sides, accounting for bending stiffness. Contact based measurement with accelerometers can identify many eigenmodes and frequencies unambiguously and is suitable for simple systems like single span systems. In the conducted investigations, the non-contact measurement with microwave interferometers could only identify up to 4 natural frequencies. The study also examines the influence of the free vibration length, which, in addition to the bending stiffness of the cable, the fork fitting and utilization, has a significant influence on the determined cable forces. The implications for using different fork fittings and cable cross-sections are discussed. This study offers valuable insights into the challenges and limitations of cable force identification and highlights the importance of choosing the appropriate measurement method based on the design of the cable structure

Virtual Axle Detector Based on Analysis of Bridge Acceleration Measurements by Fully Convolutional Network

In the practical application of the Bridge Weigh-In-Motion (BWIM) methods, the position of the wheels or axles during the passage of a vehicle is a prerequisite in most cases. To avoid the use of conventional axle detectors and bridge type-specific methods, we propose a novel method for axle detection using accelerometers placed arbitrarily on a bridge. In order to develop a model that is as simple and comprehensible as possible, the axle detection task is implemented as a binary classification problem instead of a regression problem. The model is implemented as a Fully Convolutional Network to process signals in the form of Continuous Wavelet Transforms. This allows passages of any length to be processed in a single step with maximum efficiency while utilising multiple scales in a single evaluation. This allows our method to use acceleration signals from any location on the bridge structure and act as Virtual Axle Detectors (VADs) without being limited to specific structural types of bridges. To test the proposed method, we analysed 3787 train passages recorded on a steel trough railway bridge of a long-distance traffic line. Results of the measurement data show that our model detects 95% of the axles, which means that 128,599 out of 134,800 previously unseen axles were correctly detected. In total, 90% of the axles were detected with a maximum spatial error of 20 cm, at a maximum velocity of vmax=56.3m/s. The analysis shows that our developed model can use accelerometers as VADs even under real operating conditions

Long‐term validation of virtual sensing of a railway bridge with ballasted superstructure

Railway bridges have a long lifespan, which is challenged by the constant development of vehicles leading to increased loads that they were not originally designed for. To ensure the longest possible use of existing structures, a sensor‐based structural health monitoring system can make a significant contribution. However, due to economic reasons and the inaccessibility of many points of interest, sensors cannot be installed everywhere. Therefore, in most cases, only a few sensors are available at a few points of interest, and methods that aim to reconstruct structural responses at unmeasured points from these measurements are referred to as virtual sensing. In this paper, we have analyzed 19,075 passages recorded on a steel trough bridge with a ballast superstructure and a span of 16.4 m, together with weather data. Our findings show that the influence of train type and speed has a significantly higher impact on the results than environmental factors. The investigation revealed that the model‐based analysis produced similar results to the data‐driven analysis concerning acceleration signals. However, when analyzing strain signals, the two approaches yielded distinctly different results

Genetic characterization of a Sinorhizobium meliloti chromosomal region involved in lipopolysaccharide biosynthesis

The genetic characterization of a 5.5-kb chromosomal region of Sinorhizobium meliloti 2011 that contains lpsB, a gene required for the normal development of symbiosis with Medicago spp., is presented. The nucleotide sequence of this DNA fragment revealed the presence of six genes: greA and lpsB, transcribed in the forward direction; and lpsE, lpsD, lpsC, and lrp, transcribed in the reverse direction. Except for lpsB, none of the lps genes were relevant for nodulation and nitrogen fixation. Analysis of the transcriptional organization of lpsB showed that greA and lpsB are part of separate transcriptional units, which is in agreement with the finding of a DNA stretch homologous to a "nonnitrogen" promoter consensus sequence between greA and lpsB. The opposite orientation of lpsB with respect to its first downstream coding sequence, lpsE, indicated that the altered LPS and the defective symbiosis of lpsB mutants are both consequences of a primary nonpolar defect in a single gene. Global sequence comparisons revealed that the greA-lpsB and lrp genes of S. meliloti have a genetic organization similar to that of their homologous loci in R. leguminosarum bv. viciae. In particular, high sequence similarity was found between the translation product of lpsB and a core-related biosynthetic mannosyltransferase of R. leguminosarum bv. viciae encoded by the lpcC gene. The functional relationship between these two genes was demonstrated in genetic complementation experiments in which the S. meliloti lpsB gene restored the wild-type LPS phenotype when introduced into lpcC mutants of R. leguminosarum. These results support the view that S. meliloti lpsB also encodes a mannosyltransferase that participates in the biosynthesis of the LPS core. Evidence is provided for the presence of other lpsB-homologous sequences in several members of the family Rhizobiaceae.Facultad de Ciencias ExactasInstituto de Biotecnologia y Biologia Molecula

A broad range quorum sensing inhibitor working through sRNA inhibition

Abstract For the last decade, chemical control of bacterial virulence has received considerable attention. Ajoene, a sulfur-rich molecule from garlic has been shown to reduce expression of key quorum sensing regulated virulence factors in the opportunistic pathogen Pseudomonas aeruginosa. Here we show that the repressing effect of ajoene on quorum sensing occurs by inhibition of small regulatory RNAs (sRNA) in P. aeruginosa as well as in Staphylococcus aureus, another important human pathogen that employs quorum sensing to control virulence gene expression. Using various reporter constructs, we found that ajoene lowered expression of the sRNAs RsmY and RsmZ in P. aeruginosa and the small dual-function regulatory RNA, RNAIII in S. aureus, that controls expression of key virulence factors. We confirmed the modulation of RNAIII by RNA sequencing and found that the expression of many QS regulated genes encoding virulence factors such as hemolysins and proteases were lowered in the presence of ajoene in S. aureus. Importantly, our findings show that sRNAs across bacterial species potentially may qualify as targets of anti-virulence therapy and that ajoene could be a lead structure in search of broad-spectrum compounds transcending the Gram negative-positive borderline

Dihydropyrimidine Dehydrogenase Testing prior to Treatment with 5-Fluorouracil, Capecitabine, and Tegafur: A Consensus Paper

Background: 5-Fluorouracil (FU) is one of the most commonly used cytostatic drugs in the systemic treatment of cancer. Treatment with FU may cause severe or life-threatening side effects and the treatment-related mortality rate is 0.2–1.0%. Summary: Among other risk factors associated with increased toxicity, a genetic deficiency in dihydropyrimidine dehydrogenase (DPD), an enzyme responsible for the metabolism of FU, is well known. This is due to variants in the DPD gene (DPYD). Up to 9% of European patients carry a DPD gene variant that decreases enzyme activity, and DPD is completely lacking in approximately 0.5% of patients. Here we describe the clinical and genetic background and summarize recommendations for the genetic testing and tailoring of treatment with 5-FU derivatives. The statement was developed as a consensus statement organized by the German Society for Hematology and Medical Oncology in cooperation with 13 medical associations from Austria, Germany, and Switzerland. Key Messages: (i) Patients should be tested for the 4 most common genetic DPYD variants before treatment with drugs containing FU. (ii) Testing forms the basis for a differentiated, risk-adapted algorithm with recommendations for treatment with FU-containing drugs. (iii) Testing may optionally be supplemented by therapeutic drug monitorin

Diversity Promotes Temporal Stability across Levels of Ecosystem Organization in Experimental Grasslands

The diversity–stability hypothesis states that current losses of biodiversity can impair the ability of an ecosystem to dampen the effect of environmental perturbations on its functioning. Using data from a long-term and comprehensive biodiversity experiment, we quantified the temporal stability of 42 variables characterizing twelve ecological functions in managed grassland plots varying in plant species richness. We demonstrate that diversity increases stability i) across trophic levels (producer, consumer), ii) at both the system (community, ecosystem) and the component levels (population, functional group, phylogenetic clade), and iii) primarily for aboveground rather than belowground processes. Temporal synchronization across studied variables was mostly unaffected with increasing species richness. This study provides the strongest empirical support so far that diversity promotes stability across different ecological functions and levels of ecosystem organization in grasslands

The number of tree species on Earth

One of the most fundamental questions in ecology is how many species inhabit the Earth. However, due to massive logistical and financial challenges and taxonomic difficulties connected to the species concept definition, the global numbers of species, including those of important and well-studied life forms such as trees, still remain largely unknown. Here, based on global groundsourced data, we estimate the total tree species richness at global, continental, and biome levels. Our results indicate that there are 73,000 tree species globally, among which ∼9,000 tree species are yet to be discovered. Roughly 40% of undiscovered tree species are in South America. Moreover, almost one-third of all tree species to be discovered may be rare, with very low populations and limited spatial distribution (likely in remote tropical lowlands and mountains). These findings highlight the vulnerability of global forest biodiversity to anthropogenic changes in land use and climate, which disproportionately threaten rare species and thus, global tree richness