Search

The description of the muon capture system units with different targets (gas and solid-state) is presented. Using the muon-event selection system and high-purity germanium (HPGe) detectors, precise measurements of the time-energy distributions following ordinary muon capture in the isotopically-enriched 130Xe, 82Kr, and 24Mg nuclei were performed. The optimal parameters of the muon capture measuring system to ensure maximum muon stops in the targets under study and to collect effective statistics during measurements are reported. The system makes it possible to separate muonic X-ray events from gamma radiation with high accuracy, which is critical in such studies, and it is also suitable for future measurements with other targets.

International audience; AbstractBacteria of the genus Dickeya are phytopathogens characterised by a wide host range and large geographical distribution. The disease is named soft rot because the plant material is liquefied by bacterial enzymes. The massive production of pectate lyases is typical of Dickeya. Secretion by a specialised system named Out enables the export of pectinases that cause soft rot by cleaving pectin present in plant cell walls. The 10 species of the genus Dickeya were isolated from diseased plants or water. They attack a variety of crops and ornamental plants in tropical, subtropical and temperate climates. Dickeya solani and Dickeya dianthicola are the two main species responsible for potato diseases, leading to significant economic losses. Recent diseases on fruit trees are also due to Dickeya. These bacteria have developed different mechanisms to cope with adverse conditions and compete with other microorganisms. In addition, they use a complex regulatory network to adjust the production of pectate lyases during the successive phases of plant infection.Key Concepts Dickeya members are plant pathogens characterised by a broad‐host range and a large geographical distribution. The genus Dickeya is subdivided into ten species, seven acting as phytopathogens and three found in water. Dickeya species are ‘brute force’ pathogens as they use a set of plant cell wall degrading enzymes as their primary weapon. Massive production of pectate lyases and accessory pectinases is the main virulence signature of the Dickeya species; they cause the symptom of rotting of the plant tissues. Since antibiotics are not allowed in agriculture, there is currently no effective chemical control for Dickeya. Dickeya has developed mechanisms to compete with other microorganisms and to cope with stressful conditions found in the environment or during plant colonisation. The infectious process begins with an asymptomatic phase, corresponding to the asymptomatic presence of Dickeya in plant tissues, followed by the symptomatic phase corresponding to soft‐rot appearance. The disease appears when environmental parameters, mainly temperatures and humidity, become favourable. During the plant infection, the expression of pectate lyase genes is finely controlled by a complex regulatory network. Besides pectate lyase secretion, many other functions favour the bacterial virulence, such as protein secretion systems, motility, chemotaxis, iron capture and envelope components.

SummaryThe phytopathogenic proteobacterium Dickeya dadantii secretes an array of plant cell wall degrading enzymes and other virulence factors via the type 2 secretion system (T2SS). T2SSs are widespread among important plant, animal and human bacterial pathogens. This multi-protein complex spans the double membrane cell envelope and secretes fully folded proteins through a large outer membrane pore formed by 15 subunits of the secretin GspD. Secretins are also found in the type 3 secretion system and the type 4 pili. Usually, specialized lipoproteins termed as pilotins assist the targeting and assembly of secretins into the outer membrane. Here, we show that in Dickeya, the pilotin acts in concert with the scaffolding protein GspB. Deletion of gspB profoundly impacts secretin assembly, pectinase secretion, and virulence. Structural studies reveal that GspB possesses a conserved periplasmic Homology Region domain that interacts directly with the N-terminal secretin domain. Site-specific photo cross-linking unravels molecular details of the GspB-GspD complex in vivo. We show that GspB facilitates outer membrane targeting and assembly of the secretin pores and anchors them to the inner membrane while the C-terminal extension of GspB scaffolds the secretin channel in the peptidoglycan cell wall. Phylogenetic analysis shows that in other bacteria, GspB homologs vary in length and domain composition and act in concert with either a cognate ATPase GspA or a pilotin GspS.ImportanceGram-negative bacteria have two cell membranes sandwiching a peptidoglycan net that form together a robust protective cell envelope. To translocate effector proteins across this multilayer envelope, bacteria have evolved several specialized secretion systems. In the type 2 secretion system and some other bacterial machineries, secretins form large multimeric pores that allow transport of effector proteins or filaments across the outer membrane. The secretins are essential for nutrient acquisition and pathogenicity and constitute a target for development of new antibacterials. Targeting of secretin subunits into the outer membrane is often facilitated by a special class of lipoproteins called pilotins. Here, we show that in Dickeya and some other bacteria, the scaffolding protein GspB acts in concert with pilotin, facilitating the assembly of the secretin pore and its anchoring to both the inner membrane and the bacterial cell wall. GspB homologs of varied domain composition are present in many other T2SSs.

The GERmanium Detector Array (GERDA) collaboration searched for neutrinoless double-$\beta$ decay in $^{76}$Ge with an array of about 40 high-purity isotopically-enriched germanium detectors. The experimental signature of the decay is a monoenergetic signal at Q$_{\beta\beta}$ = 2039.061(7)keV in the measured summed energy spectrum of the two emitted electrons. Both the energy reconstruction and resolution of the germanium detectors are crucial to separate a potential signal from various backgrounds, such as neutrino-accompanied double-$\beta$ decays allowed by the Standard Model. The energy resolution and stability were determined and monitored as a function of time using data from regular $^{228}$Th calibrations. In this work, we describe the calibration process and associated data analysis of the full GERDA dataset, tailored to preserve the excellent resolution of the individual germanium detectors when combining data over several years., Comment: 12 pages, 7 figures

The type II secretion system (T2SS) transports fully folded proteins of various functions and structures through the outer membrane of Gram-negative bacteria. The molecular mechanisms of substrate recruitment by T2SS remain elusive but a prevailing view is that the secretion determinants could be of a structural nature. The phytopathogenic γ-proteobacteria, Pectobacterium carotovorum and Dickeya dadantii, secrete similar sets of homologous plant cell wall degrading enzymes, mainly pectinases, by similar T2SSs, called Out. However, the orthologous pectate lyases Pel3 and PelI from these bacteria, which share 67% of sequence identity, are not secreted by the counterpart T2SS of each bacterium, indicating a fine-tuned control of protein recruitment. To identify the related secretion determinants, we first performed a structural characterization and comparison of Pel3 with PelI using X-ray crystallography. Then, to assess the biological relevance of the observed structural variations, we conducted a loop-substitution analysis of Pel3 combined with secretion assays. We showed that there is not one element with a definite secondary structure but several distant and structurally flexible loop regions that are essential for the secretion of Pel3 and that these loop regions act together as a composite secretion signal. Interestingly, depending on the crystal contacts, one of these key secretion determinants undergoes disorder-to-order transitions that could reflect its transient structuration upon the contact with the appropriate T2SS components. We hypothesize that such T2SS-induced structuration of some intrinsically disordered zones of secretion substrates could be part of the recruitment mechanism used by T2SS.

The GERmanium Detector Array (GERDA) experiment searched for the lepton-number-violating neutrinoless double-$\beta$ ($0\nu\beta\beta$) decay of $^{76}$Ge, whose discovery would have far-reaching implications in cosmology and particle physics. By operating bare germanium diodes, enriched in $^{76}$Ge, in an active liquid argon shield, GERDA achieved an unprecedently low background index of $5.2\times10^{-4}$ counts/(keV$\cdot$kg$\cdot$yr) in the signal region and met the design goal to collect an exposure of 100 kg$\cdot$yr in a background-free regime. When combined with the result of Phase I, no signal is observed after 127.2 kg$\cdot$yr of total exposure. A limit on the half-life of $0\nu\beta\beta$ decay in $^{76}$Ge is set at $T_{1/2}>1.8\times10^{26}$ yr at 90% C.L., which coincides with the sensitivity assuming no signal., Comment: 7 pages, 3 figures, submitted to Physical Review Letters

We present the first search for bosonic superweakly interacting massive particles (super-WIMPs) as keV-scale dark matter candidates performed with the GERDA experiment. GERDA is a neutrinoless double-β decay experiment which operates high-purity germanium detectors enriched in Ge76 in an ultralow background environment at the Laboratori Nazionali del Gran Sasso (LNGS) of INFN in Italy. Searches were performed for pseudoscalar and vector particles in the mass region from 60 keV/c2 to 1 MeV/c2. No evidence for a dark matter signal was observed, and the most stringent constraints on the couplings of super-WIMPs with masses above 120 keV/c2 have been set. As an example, at a mass of 150 keV/c2 the most stringent direct limits on the dimensionless couplings of axionlike particles and dark photons to electrons of gae<3×10−12 and α′/α<6.5×10−24 at 90% credible interval, respectively, were obtained.

We present the first search for bosonic superweakly interacting massive particles (super-WIMPs) as keV-scale dark matter candidates performed with the GERDA experiment. GERDA is a neutrinoless double-s decay experiment which operates high-purity germanium detectors enriched in ^{76}Ge in an ultralow background environment at the Laboratori Nazionali del Gran Sasso (LNGS) of INFN in Italy. Searches were performed for pseudoscalar and vector particles in the mass region from 60 keV/c^{2} to 1 MeV/c^{2}. No evidence for a dark matter signal was observed, and the most stringent constraints on the couplings of super-WIMPs with masses above 120 keV/c^{2} have been set. As an example, at a mass of 150 keV/c^{2} the most stringent direct limits on the dimensionless couplings of axionlike particles and dark photons to electrons of g_{ae}

The GERmanium Detector Array (Gerda) experiment at the Gran Sasso underground laboratory (LNGS) of INFN is searching for neutrinoless double-beta (0νββ) decay of 76Ge. The technological challenge of Gerda is to operate in a “background-free” regime in the region of interest (ROI) after analysis cuts for the full 100 kg·yr target exposure of the experiment. A careful modeling and decomposition of the full-range energy spectrum is essential to predict the shape and composition of events in the ROI around Qββ for the 0νββ search, to extract a precise measurement of the half-life of the double-beta decay mode with neutrinos (2νββ) and in order to identify the location of residual impurities. The latter will permit future experiments to build strategies in order to further lower the background and achieve even better sensitivities. In this article the background decomposition prior to analysis cuts is presented for Gerda Phase II. The background model fit yields a flat spectrum in the ROI with a background index (BI) of $$ {16.04}_{-0.85}^{+0.78}\cdotp {10}^{-3} $$ 16.04 − 0.85 + 0.78 · 10 − 3 cts/(keV·kg·yr) for the enriched BEGe data set and $$ {14.68}_{-0.52}^{+0.47}\cdotp {10}^{-3} $$ 14.68 − 0.52 + 0.47 · 10 − 3 cts/(keV·kg·yr) for the enriched coaxial data set. These values are similar to the one of Phase I despite a much larger number of detectors and hence radioactive hardware components.

Apart from monitoring nuclear reactor parameters, the DANSS neutrino experiment is aimed at searching for sterile neutrinos through a detailed analysis of the ratio of reactor antineutrino spectra measured at different distances from the reactor core. The light collection system of the detector is dual, comprising both the vacuum photomultiplier tubes (PMTs) and silicon photomultipliers (SiPMs). In this paper, the techniques developed to calibrate the responses of these photodetectors are discussed in detail. The long-term stability of the key parameters of the detector and their dependences on the ambient temperature are investigated. The results of detector light yield measurements, performed independently with PMTs and SiPMs are reported.

The GERmanium Detector Array (GERDA) experiment located at the INFN Gran Sasso Laboratory (Italy), is looking for the neutrinoless double beta decay of Ge76, by using high-purity germanium detectors made from isotopically enriched material. The combination of the novel experimental design, the careful material selection for radio-purity and the active/passive shielding techniques result in a very low residual background at the Q-value of the decay, about 1e-3 counts/(keV kg yr). This makes GERDA the first experiment in the field to be background-free for the complete design exposure of 100 kg yr. A search for neutrinoless double beta decay was performed with a total exposure of 47.7 kg yr: 23.2 kg yr come from the second phase (Phase II) of the experiment, in which the background is reduced by about a factor of ten with respect to the previous phase. The analysis presented in this paper includes 12.4 kg yr of new Phase II data. No evidence for a possible signal is found: the lower limit for the half-life of Ge76 is 8.0e25 yr at 90% CL. The experimental median sensitivity is 5.8e25 yr. The experiment is currently taking data. As it is running in a background-free regime, its sensitivity grows linearly with exposure and it is expected to surpass 1e26 yr within 2018., 8 pages, to appear in the proceedings of TAUP2017

The GERDA experiment searches for the lepton-number-violating neutrinoless double-β decay of ^{76}Ge (^{76}Ge→^{76}Se+2e^{-}) operating bare Ge diodes with an enriched ^{76}Ge fraction in liquid argon. The exposure for broad-energy germanium type (BEGe) detectors is increased threefold with respect to our previous data release. The BEGe detectors feature an excellent background suppression from the analysis of the time profile of the detector signals. In the analysis window a background level of 1.0_{-0.4}^{+0.6}×10^{-3} counts/(keV kg yr) has been achieved; if normalized to the energy resolution this is the lowest ever achieved in any 0νββ experiment. No signal is observed and a new 90% C.L. lower limit for the half-life of 8.0×10^{25} yr is placed when combining with our previous data. The expected median sensitivity assuming no signal is 5.8×10^{25} yr.

An observation of neutrinoless double beta ([Formula: see text]) decay would allow to shed light onto the nature of neutrinos. Gerda (GERmanium Detector Array) aims to discover this process in a background-free search using [Formula: see text]Ge. The experiment is located at the Laboratori Nazionali del Gran Sasso (LNGS) of the Istituto Nazionale di Fisica Nucleare (INFN) in Italy. Bare, isotopically enriched, high purity germanium detectors are operated in liquid argon. Gerda follows a staged approach. In Phase II 35.6 kg of enriched germanium detectors are operated since December 2015. The application of active background rejection methods, such as a liquid argon scintillation light read-out and pulse shape discrimination of germanium detector signals, allows to reduce the background index to the intended level of [Formula: see text] cts/(keV⋅kg⋅yr). No evidence for the [Formula: see text] decay has been found in 23.2 kg⋅yr of Phase II data, and together with data from Phase I the up-to-date most stringent half-life limit for this process in [Formula: see text]Ge has been established, at a median sensitivity of 5.8⋅10[Formula: see text][Formula: see text]yr the 90[Formula: see text]% C.L. lower limit is 8.0⋅10[Formula: see text][Formula: see text]yr.

International audience; Soft-rot diseases of plants attributed to Dickeya dadantii result from lysis of the plant cell wall caused by pectic enzymes released by the bacterial cell by a type II secretion system (T2SS). Arabidopsis thaliana can express several lines of defence against this bacterium. We employed bacterial mutants with defective envelope structures or secreted proteins to examine early plant defence reactions. We focused on the production of AtrbohD-dependent reactive oxygen species (ROS), callose deposition and cell death as indicators of these reactions. We observed a significant reduction in ROS and callose formation with a bacterial mutant in which genes encoding five pectate lyases (Pels) were disrupted. Treatment of plant leaves with bacterial culture filtrates containing Pels resulted in ROS and callose production, and both reactions were dependent on a functional AtrbohD gene. ROS and callose were produced in response to treatment with a cellular fraction of a T2SS-negative mutant grown in a Pels-inducing medium. Finally, ROS and callose were produced in leaves treated with purified Pels that had also been shown to induce the expression of jasmonic acid-dependent defence genes. Pel catalytic activity is required for the induction of ROS accumulation. In contrast, cell death observed in leaves infected with the wild-type strain appeared to be independent of a functional AtrbohD gene. It was also independent of the bacterial production of pectic enzymes and the type III secretion system (T3SS). In conclusion, the work presented here shows that D. dadantii is recognized by the A. thaliana innate immune system through the action of pectic enzymes secreted by bacteria at the site of infection. This recognition leads to AtrbohD-dependent ROS and callose accumulation, but not cell death.

The GERDA collaboration is performing a sensitive search for neutrinoless double beta decay of $^{76}$Ge at the INFN Laboratori Nazionali del Gran Sasso, Italy. The upgrade of the GERDA experiment from Phase I to Phase II has been concluded in December 2015. The first Phase II data release shows that the goal to suppress the background by one order of magnitude compared to Phase I has been achieved. GERDA is thus the first experiment that will remain background-free up to its design exposure (100 kg yr). It will reach thereby a half-life sensitivity of more than 10$^{26}$ yr within 3 years of data collection. This paper describes in detail the modifications and improvements of the experimental setup for Phase II and discusses the performance of individual detector components., 31 pages, 34 figures

The objective of the present study was to elucidate the specific features of the clinical course of malignant externa otitis in the context of the present-day concepts. A total of 5 patients presenting with the confirmed diagnosis of malignant external otitis were available for the examination. The analysis of the clinical observations provided the basis for the characteristic of pathogenesis of this condition, diagnostic principles, and treatment modalities for the management of the pathology in question. It is concluded that the patients suffering from malignant externa otitis must remain under medical care and observation during a long period and the strategy for the treatment of each concrete patient should be chosen on an individual basis taking into consideration the presence of concomitant pathologies and their adequate therapy.Цель настоящего исследования - изучение особенностей течения злокачественного наружного отита на современном этапе. Под наблюдением авторов находились 5 пациентов с подтвержденным диагнозом: злокачественный наружный отит. На основании клинического материала даны основные представления о патогенезе, принципах диагностики и лечения данной нозологии. Больные злокачественным наружным отитом требуют длительного наблюдения и тщательного выбора способа лечения в каждом конкретном клиническом случае, включая адекватную терапию сопутствующих заболеваний.Цель настоящего исследования — изучение особенностей течения злокачественного наружного отита на современном этапе. Под наблюдением авторов находились 5 пациентов с подтвержденным диагнозом: злокачественный наружный отит. На основании клинического материала даны основные представления о патогенезе, принципах диагностики и лечения данной нозологии. Больные злокачественным наружным отитом требуют длительного наблюдения и тщательного выбора способа лечения в каждом конкретном клиническом случае, включая адекватную терапию сопутствующих заболеваний.

The GERmanium Detector Array (GERDA) experiment, located at the Gran Sasso underground laboratory in Italy, is built for the search of $0\nu\beta\beta$ decay in $^{76}$Ge. GERDA operates bare high purity germanium detectors submersed in liquid Argon (LAr). Phase I of the experiment was completed reaching an exposure of about 21 kg$\cdot$yr with a background level of $10^{-2}$ cts/(ke V $\cdot$ kg $\cdot$ yr). GERDA Phase I set a limit on the $0\nu\beta\beta$ decay of $^{76}$Ge of $T_{1/2}^{0\nu} > 2.1 \cdot 10^{25}$ yr. In Phase II 35 kg of germanium detectors enriched in $^{76}$Ge are operated to reach an exposure of 100 kg$\cdot$yr. The design goal is to reduce the background by one order of magnitude to reach the sensitivity for $T_{1/2}^{0\nu} = \mathcal{O} (10^{26} )$ yr. The Phase II setup comprises thirty newly produced Broad Energy Germanium (BEGe) detectors. They contribute to the background reduction with better energy resolution and enhanced pulse shape discrimination. To achieve the necessary background reduction, the setup was complemented with LAr veto. The hardware upgrade for Phase II was finished and all detectors were deployed in December 2015. We present the first results of Phase II with 10.8 kg$\cdot$yr exposure reached in June 2016.

Internal contaminations of $^{238}$U, $^{235}$U and $^{232}$Th in the bulk of high purity germanium detectors are potential backgrounds for experiments searching for neutrinoless double beta decay of $^{76}$Ge. The data from GERDA Phase~I have been analyzed for alpha events from the decay chain of these contaminations by looking for full decay chains and for time correlations between successive decays in the same detector. No candidate events for a full chain have been found. Upper limits on the activities in the range of a few nBq/kg for $^{226}$Ra, $^{227}$Ac and $^{228}$Th, the long-lived daughter nuclides of $^{238}$U, $^{235}$U and $^{232}$Th, respectively, have been derived. With these upper limits a background index in the energy region of interest from $^{226}$Ra and $^{228}$Th contamination is estimated which satisfies the prerequisites of a future ton scale germanium double beta decay experiment., Comment: 2 figures, 7 pages

International audience; Many Gram-negative commensal and pathogenic bacteria use a type II secretion system (T2SS) to transport proteins out of the cell. These exported proteins or substrates play a major role in toxin delivery, maintaining biofilms, replication in the host and subversion of host immune responses to infection. We review the current structural and functional work on this system and argue that intrinsically disordered regions and protein dynamics are central for assembly, exo-protein recognition, and secretion competence of the T2SS. The central role of intrinsic disorder-order transitions in these processes may be a particular feature of type secretion.

Type II secretion system (T2SS) is a multiprotein trans-envelope complex that translocates fully folded proteins through the outer membrane of Gram-negative bacteria. Although T2SS is extensively studied in several bacteria pathogenic for humans, animals and plants, the molecular basis for exoprotein recruitment by this secretion machine as well as the underlying targeting motifs remain unknown. To address this question, we used bacterial two-hybrid, surface plasmon resonance, in vivo site-specific photo-cross-linking approaches and functional analyses. We showed that the fibronectin-like Fn3 domain of exoprotein PelI from Dickeya dadantii interacts with four periplasmic domains of the T2SS components GspD and GspC. The interaction between exoprotein and the GspC PDZ domain is positively modulated by the GspD N1 domain, suggesting that exoprotein secretion is driven by a succession of synergistic interactions. We found that an exposed 9-residue-long loop region of PelI interacts with the GspC PDZ domain. This loop acts as a specific secretion signal that controls exoprotein recruitment by the T2SS. Concerted in silico and in vivo approaches reveal the occurrence of equivalent secretion motifs in other exoproteins, suggesting a plausible general mechanism of exoprotein recruitment by the T2SS.

Pectate lyases are enzymes involved in plant cell wall degradation. They cleave pectin using a β-elimination mechanism, specific for acidic polysaccharides. They are mainly produced by plant pathogens and plant-associated organisms, and only rarely by animals. Pectate lyases are also commonly produced in the bacterial world, either by bacteria living in close proximity with plants or by gut bacteria that find plant material in the digestive tract of their hosts. The role of pectate lyases is essential for plant pathogens, such as Dickeya dadantii, that use a set of pectate lyases as their main virulence factor. Symbiotic bacteria produce their own pectate lyases, but they also induce plant pectate lyases to initiate the symbiosis. Pectin degradation products may act as signals affecting the plant–bacteria interactions. Bacterial pectate lyases are also essential for using the pectin of dead or living plants as a carbon source for growth. In the animal gut, Bacteroides pectate lyases degrade the pectin of ingested food, and this is particularly important for herbivores that depend on their microflora for the digestion of pectin. Some human pathogens, such as Yersinia enterocolitica, produce a few intracellular pectate lyases that can facilitate their growth in the presence of highly pectinolytic bacteria, at the plant surface, in the soil or in the animal gut.

In recent years, much attention has been focused on neutrino research because it can shed new light on greatest mysteries in physics. A new experiment devoted to detection and investigation of reactor antineutrinos is being performed at the Institute of Experimental and Applied Physics, Czech Technical University in Prague using a highly segmented scintillating detector S3. This detector is polystyrene based and does not contain any dangerous or flammable materials and is absolutely safe to be placed in the close vicinity of the reactor. The most important feature of plastic scintillators for this experiment is the amount of photoelectrons per deposited energy, which defines the final energy resolution of the detector and a lot of effort has been put into improvement of detector element overall quality. The detector has been constructed as a result of these optimization experiments.

The result of the neutrino magnetic moment (NMM) measurement at the Kalinin Nuclear Power Plant (KNPP) with GEMMA spectrometer is presented. The antineutrino-electron scattering is investigated. A high-purity germanium (HPGe) detector with a mass of 1.5 kg placed at a distance of 13.9 m from the 3 GWth reactor core is exposed to the antineutrino flux of 2.7 × 1013 cm−2s−1. The recoil electron spectra taken in 18134 and 4487 h for the reactor ON and OFF periods are compared. The upper limit for the NMM μν < 2.9 × 10−11 μB at 90% C.L. is derived from the data processing.

Low background experiments need a suppression of cosmogenically induced events. The GERDA experiment located at LNGS is searching for the neutrinless double beta decay of $^{76}$Ge. It is equipped with an active muon veto the main part of which is a water Cherenkov veto with 66 PMTs in the watertank surrounding the GERDA cryostat. With this system 806 live days have been recorded, 491 days were combined muon-germanium data. A muon detection efficiency of $\varepsilon_{\mu d}=(99.935\pm0.015)$ \% was found in a Monte Carlo simulation for the muons depositing energy in the germanium detectors. By examining coincident muon-germanium events a rejection efficiency of $\varepsilon_{\mu r}=(99.2_{-0.4}^{+0.3})$ \% was found. Without veto condition the muons by themselves would cause a background index of $\textrm{BI}_{\mu}=(3.16 \pm 0.85)\times10^{-3}$ cts/(keV$\cdot$kg$\cdot$yr) at $Q_{\beta\beta}$., Comment: 11pages, 14 figures

The GERmanium Detector Array (GERDA) experiment is searching for the neutrinoless double beta decay ( 0νββ ) of the isotope 76 Ge. High-purity germanium crystals enriched in 76 Ge, simultaneously used as source and detector, are directly deployed into ultra-pure, cryogenic liquid argon, which acts both as cooling medium and shield against the external radiation. The second phase of the experiment is taking data since end of 2015 with 20 additional kg of custom-made BEGe-type Germanium detectors and an active LAr veto. In this paper we will summarize the results of the last data release of June 2017. No evidence for a possible signal is found: the lower limit for the half-life of 76 Ge is 8.0·1025 yr at 90% CL. The very low residual background found at the Q -value of the decay, about 10-3 cts/(keV · kg · yr), makes Gerda the first experiment in the field to be background-free for the complete design exposure of 100 kg · yr.

The GERmanium Detector Array (GERDA) is a low background experiment at the Laboratori Nazionali del Gran Sasso (LNGS) of INFN designed to search for the rare neutrinoless double beta decay ([Formula: see text]) of [Formula: see text]Ge. In the first phase (Phase I) of the experiment, high purity germanium diodes were operated in a “bare” mode and immersed in liquid argon. The overall background level of [Formula: see text] was a factor of ten better than those of its predecessors. No signal was found and a lower limit was set on the half-life for the [Formula: see text] decay of [Formula: see text]Ge [Formula: see text] yr (90% CL), while the corresponding median sensitivity was [Formula: see text] yr (90% CL). A second phase (Phase II) started at the end of 2015 after a major upgrade. Thanks to the increased detector mass and performance of the enriched germanium diodes and due to the introduction of liquid argon instrumentation techniques, it was possible to reduce the background down to [Formula: see text]. After analyzing 23.2 kg[Formula: see text]⋅[Formula: see text]yr of these new data no signal was seen. Combining these with the data from Phase I a stronger half-life limit of the [Formula: see text]Ge [Formula: see text] decay was obtained: [Formula: see text] yr (90% CL), reaching a sensitivity of [Formula: see text] yr (90% CL). Phase II will continue for the collection of an exposure of 100 kg[Formula: see text]yr. If no signal is found by then the GERDA sensitivity will have reached [Formula: see text] yr for setting a 90% CL. limit. After the end of GERDA Phase II, the flagship experiment for the search of [Formula: see text] decay of [Formula: see text]Ge will be LEGEND. LEGEND experiment is foreseen to deploy up to 1-ton of [Formula: see text]Ge. After ten years of data taking, it will reach a sensitivity beyond 10[Formula: see text] yr, and hence fully cover the inverted hierarchy region.

Summary The type II secretion system (T2SS) is widely exploited by proteobacteria to secrete enzymes and toxins involved in bacterial survival and pathogenesis. The outer membrane pore formed by the secretin OutD and the inner membrane protein OutC are two key components of the secretion complex, involved in secretion specificity. Here, we show that the periplasmic regions of OutC and OutD interact directly and map the interaction site of OutC to a 20-residue peptide named OutCsip (secretin interacting peptide, residues 139–158). This peptide interacts in vitro with two distinct sites of the periplasmic region of OutD, one located on the N0 subdomain and another overlapping the N2-N3′ subdomains. The two interaction sites of OutD have different modes of binding to OutCsip. A single substitution, V143S, located within OutCsip prevents its interaction with one of the two binding sites of OutD and fully inactivates the T2SS. We show that the N0 subdomain of OutD interacts also with a second binding site within OutC located in the region proximal to the transmembrane segment. We suggest that successive interactions between these distinct regions of OutC and OutD may have functional importance in switching the secretion machine.

International audience; The crystallographic structure of the family 3 polysaccharide lyase (PL-3) PelI from Erwinia chrysanthemi has been solved to 1.45 A resolution. It consists of an N-terminal domain harboring a fibronectin type III fold linked to a catalytic domain displaying a parallel beta-helix topology. The N-terminal domain is located away from the active site and is not involved in the catalytic process. After secretion in planta, the two domains are separated by E. chrysanthemi proteases. This event turns on the hypersensitive response of the host. The structure of the single catalytic domain determined to 2.1 A resolution shows that the domain separation unveils a "Velcro"-like motif of asparagines, which might be recognized by a plant receptor. The structure of PelI in complex with its substrate, a tetragalacturonate, has been solved to 2.3 A resolution. The sugar binds from subsites -2 to +2 in one monomer of the asymmetric unit, although it lies on subsites -1 to +3 in the other. These two "Michaelis complexes" have never been observed simultaneously before and are consistent with the dual mode of bond cleavage in this substrate. The bound sugar adopts a mixed 2(1) and 3(1) helical conformation similar to that reported in inactive mutants from families PL-1 and PL-10. However, our study suggests that the catalytic base in PelI is not a conventional arginine but a lysine as proposed in family PL-9.The crystallographic structure of the family 3 polysaccharide lyase (PL-3) PelI from Erwinia chrysanthemi has been solved to 1.45 A resolution. It consists of an N-terminal domain harboring a fibronectin type III fold linked to a catalytic domain displaying a parallel beta-helix topology. The N-terminal domain is located away from the active site and is not involved in the catalytic process. After secretion in planta, the two domains are separated by E. chrysanthemi proteases. This event turns on the hypersensitive response of the host. The structure of the single catalytic domain determined to 2.1 A resolution shows that the domain separation unveils a "Velcro"-like motif of asparagines, which might be recognized by a plant receptor. The structure of PelI in complex with its substrate, a tetragalacturonate, has been solved to 2.3 A resolution. The sugar binds from subsites -2 to +2 in one monomer of the asymmetric unit, although it lies on subsites -1 to +3 in the other. These two "Michaelis complexes" have never been observed simultaneously before and are consistent with the dual mode of bond cleavage in this substrate. The bound sugar adopts a mixed 2(1) and 3(1) helical conformation similar to that reported in inactive mutants from families PL-1 and PL-10. However, our study suggests that the catalytic base in PelI is not a conventional arginine but a lysine as proposed in family PL-9.

Pectate lyases are enzymes involved in plant cell wall degradation. They cleave pectin using a β-elimination mechanism, specific for acidic polysaccharides. They are mainly produced by plant pathogens and plant-associated organisms, and only rarely by animals. Pectate lyases are also commonly produced in the bacterial world, either by bacteria living in close proximity with plants or by gut bacteria that find plant material in the digestive tract of their hosts. The role of pectate lyases is essential for plant pathogens, such as Dickeya dadantii, that use a set of pectate lyases as their main virulence factor. Symbiotic bacteria produce their own pectate lyases, but they also induce plant pectate lyases to initiate the symbiosis. Pectin degradation products may act as signals affecting the plant–bacteria interactions. Bacterial pectate lyases are also essential for using the pectin of dead or living plants as a carbon source for growth. In the animal gut, Bacteroides pectate lyases degrade the pectin of ingested food, and this is particularly important for herbivores that depend on their microflora for the digestion of pectin. Some human pathogens, such as Yersinia enterocolitica, produce a few intracellular pectate lyases that can facilitate their growth in the presence of highly pectinolytic bacteria, at the plant surface, in the soil or in the animal gut.

The GERmanium Detector Array (GERDA) at the Gran Sasso Underground Laboratory (LNGS) searches for the neutrinoless double beta decay (0{\nu}{\beta}{\beta}) of $^{76}$Ge. Germanium detectors made of material with an enriched $^{76}$Ge fraction act simultaneously as sources and detectors for this decay. During Phase I of the experiment mainly refurbished semi-coaxial Ge detectors from former experiments were used. For the upcoming Phase II, 30 new $^{76}$Ge enriched detectors of broad energy germanium (BEGe)-type were produced. A subgroup of these detectors has already been deployed in GERDA during Phase I. The present paper reviews the complete production chain of these BEGe detectors including isotopic enrichment, purification, crystal growth and diode production. The efforts in optimizing the mass yield and in minimizing the exposure of the $^{76}$Ge enriched germanium to cosmic radiation during processing are described. Furthermore, characterization measurements in vacuum cryostats of the first subgroup of seven BEGe detectors and their long-term behavior in liquid argon are discussed. The detector performance fulfills the requirements needed for the physics goals of GERDA Phase~II.

The Gerda experiment at the Laboratori Nazionali del Gran Sasso in Italy uses germanium detectors made from material with an enriched 76Ge isotope fraction to search for neutrinoless double beta decay of this nucleus. Applying a blind analysis we find no signal after an exposure of 21.6 kg·yr and a background of about 0.01 cts/(keV·kg·yr). A half-life limit of Tov1/2> 2.1 · 1025 yr (90% C.L.) is extracted. The previous claim of a signal for 76Ge is excluded with 99% probability in a model independent way.

Type II secretion system (T2SS) is a multiprotein trans-envelope complex that translocates fully folded proteins through the outer membrane of Gram-negative bacteria. Although T2SS is extensively studied in several bacteria pathogenic for humans, animals and plants, the molecular basis for exoprotein recruitment by this secretion machine as well as the underlying targeting motifs remain unknown. To address this question, we used bacterial two-hybrid, surface plasmon resonance, in vivo site-specific photo-cross-linking approaches and functional analyses. We showed that the fibronectin-like Fn3 domain of exoprotein PelI from Dickeya dadantii interacts with four periplasmic domains of the T2SS components GspD and GspC. The interaction between exoprotein and the GspC PDZ domain is positively modulated by the GspD N1 domain, suggesting that exoprotein secretion is driven by a succession of synergistic interactions. We found that an exposed 9-residue-long loop region of PelI interacts with the GspC PDZ domain. This loop acts as a specific secretion signal that controls exoprotein recruitment by the T2SS. Concerted in silico and in vivo approaches reveal the occurrence of equivalent secretion motifs in other exoproteins, suggesting a plausible general mechanism of exoprotein recruitment by the T2SS.

Erwinia chrysanthemi 3937 secretes an arsenal of pectinolytic enzymes including several pectate lyases encoded by the pel genes. We characterized a novel cluster of pectinolytic genes consisting of the three adjacent genes pehV, pehW and pehX, whose products have polygalacturonase activity. The high similarity between the three genes suggests that they result from duplication of an ancestral gene. The transcription of pehV, pehW and pehX is dependent on several environmental conditions. They are induced by pectin catabolic products and this induction results from inactivation of the KdgR repressor which controls almost all the steps of pectin catabolism. The presence of calcium ions strongly reduced the transcription of the three peh genes. Their expression was also affected by growth phase, osmolarity, oxygen limitation and nitrogen starvation. In addition, the pehX transcription is affected by catabolite repression and controlled by the activator protein CRP. PecS, which was initially isolated as a repressor of virulence factors, acts as an activator of the peh transcription. We showed that the three regulators KdgR, PecS and CRP act by direct interaction with the promoter regions of the peh genes. Analysis of simultaneous binding of KdgR, PecS, CRP and RNA polymerase indicated that the activator effect of PecS results from a competition between PecS and KdgR for the occupation of overlapping binding sites. Thus, to activate peh transcription, PecS behaves as an anti-repressor against KdgR.

Erwinia chrysanthemi 3937 secretes several pectinolytic enzymes, among which eight isoenzymes of pectate lyases with an endo-cleaving mode (PelA, PelB, PelC, PelD, PelE, PelI, PelL, and PelZ) have been identified. Two exo-cleaving enzymes, the exopolygalacturonate lyase, PelX, and an exo-poly-α- d -galacturonosidase, PehX, have been previously identified in other E. chrysanthemi strains. Using a genomic bank of a 3937 mutant with the major pel genes deleted, we cloned a pectinase gene identified as pelX , encoding the exopolygalacturonate lyase. The deduced amino acid sequence of the 3937 PelX is very similar to the PelX of another E. chrysanthemi strain, EC16, except in the 43 C-terminal amino acids. PelX also has homology to the endo-pectate lyase PelL of E. chrysanthemi but has a N-terminal extension of 324 residues. The transcription of pelX , analyzed by gene fusions, is dependent on several environmental conditions. It is induced by pectic catabolic products and affected by growth phase, oxygen limitation, nitrogen starvation, and catabolite repression. Regulation of pelX expression is dependent on the KdgR repressor, which controls almost all the steps of pectin catabolism, and on the global activator of sugar catabolism, cyclic AMP receptor protein. In contrast, PecS and PecT, two repressors of the transcription of most pectate lyase genes, are not involved in pelX expression. The pelX mutant displayed reduced pathogenicity on chicory leaves, but its virulence on potato tubers or Saintpaulia ionantha plants did not appear to be affected. The purified PelX protein has no maceration activity on plant tissues. Tetragalacturonate is the best substrate of PelX, but PelX also has good activity on longer oligomers. Therefore, the estimated number of binding subsites for PelX is 4, extending from subsites −2 to +2. PelX and PehX were shown to be localized in the periplasm of E. chrysanthemi 3937. PelX catalyzed the formation of unsaturated digalacturonates by attack from the reducing end of the substrate, while PehX released digalacturonates by attack from the nonreducing end of the substrate. Thus, the two types of exo-degrading enzymes appeared complementary in the degradation of pectic polymers, since they act on both extremities of the polymeric chain.

Erwinia chrysanthemi causes soft rot on various plants. The maceration of plant tissues is mainly due to the action of endopectate lyases. The E. chrysanthemi strain 3937 produces eight endopectate lyases (PelA, PelB, PelC, PelD, PelE, PelI, PelL and PelZ) that are secreted by the Out pathway. The necrotic response elicited by the wild-type E. chrysanthemi strain on tobacco leaves is due to an extracellular protein secreted by the Out machinery. Purification of the active factor revealed that it corresponds to a pectate lyase presenting immunological cross-reaction with PelI. Analysis of pelI and out mutants indicated that the necrosis-inducing pectate lyase results from a post-translational modification of PelI occurring extracellularly both in culture media and in planta. This modification consists of the cleavage of 97 N-terminal amino acids by the extracellular proteases of E. chrysanthemi. The enzymatic properties of the maturated form, PelI-3, are not, or only weakly, modified. However, this maturation gives rise to a small size and basic form that is active as a defence elicitor in plants.

Erwinia chrysanthemi 3937 secretes five major isoenzymes of pectate lyases encoded by the pel4, pelB, pelC, pelD, and pelE genes and a set of secondary pectate lyases, two of which, pelL and pelZ, have been already identified. We cloned the pelI gene, encoding a ninth pectate lyase of E. chrysanthemi 3937. The pelI reading frame is 1,035 bases long, corresponding to a protein of 344 amino acids including a typical amino-terminal signal sequence of 19 amino acids. The purified mature PelI protein has an isoelectric point of about 9 and an apparent molecular mass of 34 kDa. PelI has a preference for partially methyl esterified pectin and presents an endo-cleaving activity with an alkaline pH optimum and an absolute requirement for Ca2+ ions. PelI is an extracellular protein secreted by the Out secretory pathway of E. chrysanthemi. The PelI protein is very active in the maceration of plant tissues. A pelI mutant displayed reduced pathogenicity on chicory leaves, but its virulence did not appear to be affected on potato tubers or Saintpaulia ionantha plants. The pelI gene constitutes an independent transcriptional unit. As shown for the other pel genes, the transcription of pelI is dependent on various environmental conditions. It is induced by pectic catabolic products and affected by growth phase, oxygen limitation, temperature, nitrogen starvation, and catabolite repression. Regulation of pelI expression appeared to be dependent on the three repressors of pectinase synthesis, KdgR, PecS, and PecT, and on the global activator of sugar catabolism, cyclic AMP receptor protein. A functional KdgR binding site was identified close to the putative pelI promoter. Analysis of the amino acid sequence of PelI revealed high homology with a pectate lyase from Erwinia carotovora subsp. carotovora (65% identity) and low homology with pectate lyases of the phytopathogenic fungus Nectria haematococca (Fusarium solani). This finding indicates that PelI belongs to pectate lyase class III. Using immunoblotting experiments, we detected PelI homologs in various strains of E. chrysanthemi and E. carotovora subsp. carotovora but not in E. carotovora subsp. atroseptica.

The GERDA experiment located at the LNGS searches for neutrinoless double beta (0\nu\beta\beta) decay of ^{76}Ge using germanium diodes as source and detector. In Phase I of the experiment eight semi-coaxial and five BEGe type detectors have been deployed. The latter type is used in this field of research for the first time. All detectors are made from material with enriched ^{76}Ge fraction. The experimental sensitivity can be improved by analyzing the pulse shape of the detector signals with the aim to reject background events. This paper documents the algorithms developed before the data of Phase I were unblinded. The double escape peak (DEP) and Compton edge events of 2.615 MeV \gamma\ rays from ^{208}Tl decays as well as 2\nu\beta\beta\ decays of ^{76}Ge are used as proxies for 0\nu\beta\beta\ decay. For BEGe detectors the chosen selection is based on a single pulse shape parameter. It accepts 0.92$\pm$0.02 of signal-like events while about 80% of the background events at Q_{\beta\beta}=2039 keV are rejected. For semi-coaxial detectors three analyses are developed. The one based on an artificial neural network is used for the search of 0\nu\beta\beta\ decay. It retains 90% of DEP events and rejects about half of the events around Q_{\beta\beta}. The 2\nu\beta\beta\ events have an efficiency of 0.85\pm0.02 and the one for 0\nu\beta\beta\ decays is estimated to be 0.90^{+0.05}_{-0.09}. A second analysis uses a likelihood approach trained on Compton edge events. The third approach uses two pulse shape parameters. The latter two methods confirm the classification of the neural network since about 90% of the data events rejected by the neural network are also removed by both of them. In general, the selection efficiency extracted from DEP events agrees well with those determined from Compton edge events or from 2\nu\beta\beta\ decays., Comment: 18 pages, 27 figures