Glucosylceramide is synthesized at the cytosolic surface of various Golgi subfractions

In our attempt to assess the topology of glucosylceramide biosynthesis, we have employed a truncated ceramide analogue that permeates cell membranes and is converted into water soluble sphingolipid analogues both in living and in fractionated cells. Truncated sphingomyelin is synthesized in the lumen of the Golgi, whereas glucosylceramide is synthesized at the cytosolic surface of the Golgi as shown by (a) the insensitivity of truncated sphingomyelin synthesis and the sensitivity of truncated glucosylceramide synthesis in intact Golgi membranes from rabbit liver to treatment with protease or the chemical reagent DIDS; and (b) sensitivity of truncated sphingomyelin export and insensitivity of truncated glucosylceramide export to decreased temperature and the presence of GTP-γ-S in semiintact CHO cells. Moreover, subfractionation of rat liver Golgi demonstrated that the sphingomyelin synthase activity was restricted to fractions containing marker enzymes for the proximal Golgi, whereas the capacity to synthesize truncated glucosylceramide was also found in fractions containing distal Golgi markers. A similar distribution of glucosylceramide synthesizing activity was observed in the Golgi of the human liver derived HepG2 cells. The cytosolic orientation of the reaction in HepG2 cells was confirmed by complete extractability of newly formed NBD-glucosylceramide from isolated Golgi membranes or semiintact cells by serum albumin, whereas NBD-sphingomyelin remained protected against such extraction

Upregulation of virulence genes promotes Vibrio cholerae biofilm hyperinfectivity

Vibrio cholerae remains a major global health threat, disproportionately impacting parts of the world without adequate infrastructure and sanitation resources. In aquatic environments, V. cholerae exists both as planktonic cells and as biofilms, which are held together by an extracellular matrix. V. cholerae biofilms have been shown to be hyperinfective, but the mechanism of hyperinfectivity is unclear. Here we show that biofilm-grown cells, irrespective of the surfaces on which they are formed, are able to markedly outcompete planktonic-grown cells in the infant mouse. Using an imaging technique designed to render intestinal tissue optically transparent and preserve the spatial integrity of infected intestines, we reveal and compare three-dimensional V. cholerae colonization patterns of planktonic-grown and biofilm-grown cells. Quantitative image analyses show that V. cholerae colonizes mainly the medial portion of the small intestine and that both the abundance and localization patterns of biofilm-grown cells differ from that of planktonic-grown cells. In vitro biofilm-grown cells activate expression of the virulence cascade, including the toxin coregulated pilus (TCP), and are able to acquire the cholera toxin-carrying CTXФ phage. Overall, virulence factor gene expression is also higher in vivo when infected with biofilm-grown cells, and modulation of their regulation is sufficient to cause the biofilm hyperinfectivity phenotype. Together, these results indicate that the altered biogeography of biofilm-grown cells and their enhanced production of virulence factors in the intestine underpin the biofilm hyperinfectivity phenotype

Search for Axionlike and Scalar Particles with the NA64 Experiment

We carried out a model-independent search for light scalar (s) and pseudoscalar axionlike (a) particles that couple to two photons by using the high-energy CERN SPS H4 electron beam. The new particles, if they exist, could be produced through the Primakoff effect in interactions of hard bremsstrahlung photons generated by 100 GeV electrons in the NA64 active dump with virtual photons provided by the nuclei of the dump. The a(s) would penetrate the downstream HCAL module, serving as shielding, and would be observed either through their $a(s)\to\gamma \gamma$ decay in the rest of the HCAL detector or as events with large missing energy if the a(s) decays downstream of the HCAL. This method allows for the probing the a(s) parameter space, including those from generic axion models, inaccessible to previous experiments. No evidence of such processes has been found from the analysis of the data corresponding to $2.84\times10^{11}$ electrons on target allowing to set new limits on the $a(s)\gamma\gamma$-coupling strength for a(s) masses below 55 MeV.Comment: This publication is dedicated to the memory of our colleague Danila Tlisov. 7 pages, 5 figures, revised version accepted for publication in Phys. Rev. Let

Search for Axionlike and Scalar Particles with the NA64 Experiment

We carried out a model-independent search for light scalar (s) and pseudoscalar axionlike (a) particles that couple to two photons by using the high-energy CERN SPS H4 electron beam. The new particles, if they exist, could be produced through the Primakoff effect in interactions of hard bremsstrahlung photons generated by 100 GeV electrons in the NA64 active dump with virtual photons provided by the nuclei of the dump. The a(s) would penetrate the downstream HCAL module, serving as a shield, and would be observed either through their a(s)→γγ decay in the rest of the HCAL detector, or as events with a large missing energy if the a(s) decays downstream of the HCAL. This method allows for the probing of the a(s) parameter space, including those from generic axion models, inaccessible to previous experiments. No evidence of such processes has been found from the analysis of the data corresponding to 2.84×10^{11} electrons on target, allowing us to set new limits on the a(s)γγ-coupling strength for a(s) masses below 55 MeV

Improved exclusion limit for light dark matter from e+e- annihilation in NA64

The current most stringent constraints for the existence of sub-GeV dark matter coupling to Standard Model via a massive vector boson A′ were set by the NA64 experiment for the mass region mA′≲250 MeV, by analyzing data from the interaction of 2.84×1011 100-GeV electrons with an active thick target and searching for missing-energy events. In this work, by including A′ production via secondary positron annihilation with atomic electrons, we extend these limits in the 200-300 MeV region by almost an order of magnitude, touching for the first time the dark matter relic density constrained parameter combinations. Our new results demonstrate the power of the resonant annihilation process in missing energy dark-matter searches, paving the road to future dedicated e+ beam efforts

Search for pseudoscalar bosons decaying into e+e- pairs in the NA64 experiment at the CERN SPS

We report the results of a search for a light pseudoscalar particle a that couples to electrons and decays to e+e- performed using the high-energy CERN SPS H4 electron beam. If such light pseudoscalar exists, it could explain the ATOMKI anomaly (an excess of e+e- pairs in the nuclear transitions of Be8 and He4 nuclei at the invariant mass ≃17 MeV observed by the experiment at the 5 MV Van de Graaff accelerator at ATOMKI, Hungary). We used the NA64 data collected in the "visible mode"configuration with a total statistics corresponding to 8.4×1010 electrons on target (EOT) in 2017 and 2018. In order to increase sensitivity to small coupling parameter ϵ we also used the data collected in 2016-2018 in the "invisible mode"configuration of NA64 with a total statistics corresponding to 2.84×1011 EOT. The background and efficiency estimates for these two configurations were retained from our previous analyses searching for light vector bosons and axionlike particles (ALP) (the latter were assumed to couple predominantly to γ). In this work we recalculate the signal yields, which are different due to different cross section and lifetime of a pseudoscalar particle a, and perform a new statistical analysis. As a result, the region of the two dimensional parameter space ma-ϵ in the mass range from 1 to 17.1 MeV is excluded. At the mass of the central value of the ATOMKI anomaly (the first result obtained on the beryllium nucleus, 16.7 MeV) the values of ϵ in the range 2.1×10-4<ϵ<3.2×10-4 are excluded