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Anti-sigma factors play a critical role in regulating the expression of sigma factors in response to environmental stress signals. PG1659 is cotranscribed with an upstream gene PG1660 (rpoE), which encodes for a sigma factor that plays an important role in oxidative stress resistance and the virulence regulatory network of P. gingivalis. PG1659, which is annotated as a hypothetical gene, is evaluated in this study. PG1659, composed of 130 amino acids, is predicted to be transmembrane protein with a single calcium (Ca(2+)) binding site. In P. gingivalis FLL358 (ΔPG1659::ermF), the rpoE gene was highly upregulated compared to the wild-type W83 strain. RpoE induced genes were also upregulated in the PG1659-defective isogenic mutant. Both protein-protein pull-down and bacterial two-hybrid assays revealed that the PG1659 protein could interact with/bind RpoE. The N-terminal domain of PG1659, representing the cytoplasmic fragment of the protein, is critical for interaction with RpoE. In the presence of PG1659, the initiation of transcription by the RpoE sigma factor was inhibited. Taken together, our data suggest that PG1659 is an anti-sigma factor which plays an important regulatory role in the modulation of the sigma factor RpoE in P. gingivalis.

As a non-covalent interaction, halogen bonding is now acknowledged to be useful in all fields where the control of intermolecular recognition plays a pivotal role. Halogen-bond basicity scales allow quantification of the halogen bonding of referential donors with organic functional groups from a thermodynamic point of view. Herein we present the pKBAtI basicity scale to provide the community an overview of halogen-bond acceptor strength towards astatine, the most potent halogen-bond donor element. This experimental scale is erected on the basis of complexation constants measured between astatine monoiodide (AtI) and sixteen selected Lewis bases. It spans over 6 log units and culminates with a value of 5.69 ± 0.32 for N,N,N′,N′-tetramethylthiourea. On this scale, the carbon π-bases are the weakest acceptors, the oxygen derivatives cover almost two-thirds of the scale, and sulphur bases exhibit the highest AtI basicity. Regarding the applications of 211At in targeted radionuclide therapy, stronger labelling of carrier agents could be envisaged on the basis of the pKBAtI scale., Based on the halogen bonding between astatine monoiodide (AtI) and 16 Lewis bases, the newly established pKBAtI scale indicates that the halogen bond basicity of AtI follows the order C ≤ O ≤ S ≈ Se for the acceptor atomic site.

International audience; Sugar-enriched media are used to produce extracellular substances (ECS) by Lactobacillus plantarum WCSF1, with a focus on growing stages and carbon source substrates. Combination of size exclusion chromatography and ATR-FTIR spectroscopy provides physicochemical patterns of bulk ECS produced along culture growing time. Secreted biopolymers present polydisperse and high molecular weight distributions, with significant amounts of carbohydrates and proteins. Results, supported by a multivariate statistical analysis, enable to differentiate the macromolecular content of bacterial ECS along the growing stages regardless of the growing media, highlighting a higher production of proteinaceous materials compared to polysaccharides. At the end of the exponential phase, common exoproteins were present in all the tested sugar-enriched media such as transglycosylases between 20 and 35 kDa, a muropeptidase at 36.9 kDa and a cell wall hydrolase. Additionally, L. plantarum WCFS1 secretes ECS with a greater diversity of proteins, when growing in the sucrose-enriched media. Graphical abstract.

Rice is one of the most important crops but its productivity is severely threatened by both biotic and abiotic stresses. Jasmonic acid and its derivatives (referred to JA) are the lipid-based plant hormones that were commonly known as regulators of plant growth, development and defense responses. In this study, we compare the physiological and molecular responses of two rice accessions, G38 (Nep_nuong) and G11 (Tam-tron) to JA treatment. G38 plants under JA treatment experienced a reduction in shoot length, root weight, shoot weight and total plant weight which suggested its sensitiveness to JA, whereas G11 plants showed a less reduction in these traits. The expression levels of 12 JA-related genes were investigated in order to better understand how the JA biosynthesis and responses differ in these two contrasting rice accessions. A significantly higher expression level of a set of genes related to JA biosynthesis, signaling and response in G11 compare to G38 was observed. Furthermore, the inorganic phosphorus starvation (Pi) response was also examined in the two varieties G11 and G38. In low Pi condition (40 µM), G11 plants showed more roots, longer root length and shoot length, higher weight compared to the G38 plants which suggest that G11 did not suffer much effect of Pi deficiency. This study highlights the differences in JA growth response in 2 contrasting rice genotypes and also suggests the link between JA developmental response and the tolerance to the Pi starvation condition in rice.

International audience; The affinity of AtO+ for around 20 model ligands (L), carrying functionalized oxygen, sulfur, and nitrogen atoms, has been assessed through a combined experimental and theoretical methodology. Significant equilibrium constants (KL ∼ 104) have been measured for sulfur-containing compounds, in agreement with the previously highlighted, relatively stable radiolabeling of SH-containing proteins with 211At. Conversely, no interaction occurs in the aqueous phase for their oxygenated counterparts, but higher affinities (KL > 106) have been determined for nitrogen-based ligands, including aromatic nitrogen heterocycles. The quantum mechanical calculations definitively ruled out any rationale based on either the metallic character of astatine or its guessed softness; the favored interactions all involve specifically the oxygen atom of AtO+, leading to the formation of covalent O–S or O–C single bonds.

International audience; The halogen bond is a powerful tool for the molecular design and pushing the limits of its strength is of major interest. Bearing the most potent halogen‐bond donor atom, astatine monoiodide (AtI) was recently successfully probed [Nat. Chem. 2018, 10, 428–434]. In this work, we continue the exploration of adducts between AtI and Lewis bases with the tributylphosphine oxide (Bu3PO) ligand, revealing the unexpected experimental occurrence of two distinct chemical species with 1:1 and 2:1 stoichiometries. The 1:1 Bu3PO⋅⋅⋅AtI complex is found to exhibit the strongest astatine‐mediated halogen bond so far (with a formation constant of 10(4.24±0.35)). Quantum chemical calculations unveil the intriguing nature of the 2:1 2Bu3PO⋅⋅⋅AtI adduct, involving a halogen bond between AtI and one Bu3PO molecular unit plus CH⋅⋅⋅O hydrogen bonds chelating the second Bu3PO unit.

Deoxyfluorination is a primary method for the formation of C-F bonds. Bespoke reagents are commonly used because of issues associated with the low reactivity of metal fluorides. Reported here is the development of a simple strategy for deoxyfluorination, using first-row transition-metal fluorides, and it overcomes these limitations. Using CuF2 as an exemplar, activation of an O-alkylisourea adduct, formed in situ, allows effective nucleophilic fluoride transfer to a range of primary and secondary alcohols. Spectroscopic investigations have been used to probe the origin of the enhanced reactivity of CuF2 . The utility of the process in enabling 18 F-radiolabeling is also presented.

Contact lenses are biomaterials worn on the eye to correct refractive errors. Bacterial adhesion and colonization of these lenses results in adverse events, such as microbial keratitis. The adsorption of tear proteins to contact lens materials enhances bacterial adhesion. Glycoprotein 340 (Gp340), a tear component, is known to promote microbial colonization in the oral cavity; however, it has not been investigated in any contact lens-related adverse event. Therefore, this study examined the adsorption of Gp340 and its recombinantly expressed scavenger receptor cysteine-rich (iSRCR(1)(Gp340)) domain on two common contact lens materials, etafilcon A and lotrafilcon B, and the concomitant effects on the adherence of clinical isolates of microbial keratitis causative agents, Pseudomonas aeruginosa (PA6206; PA6294), and Staphylococcus aureus (SA38; USA300). Across all strains and materials, iSRCR(1)(Gp340) enhanced adherence of bacteria in a dose-dependent manner. However, (i)SRCR(1)(Gp340) did not modulate the lysozyme’s or lactoferrin’s effects on bacterial adhesion to the contact lens. The Gp340 binding serine-rich surface protein (SraP) significantly enhanced the binding of USA300 to iSRCR(1)(Gp340)-coated lenses. In addition, iSRCR(1)(Gp340)-coated surfaces had significantly diminished biofilms with the SraP mutant (ΔSraP), and there was a further reduction in biofilms with the sortase A mutant (ΔSrtA), indicating the likely involvement of additional surface proteins. Finally, the binding affinities between iSRCR(1)(Gp340) and SraP were determined using surface plasmon resonance (SPR), where the complete SraP binding region displayed nanomolar affinity, whereas its smaller fragments adhered with micromolar affinities. This study concludes that Gp340 and its SRCR domains play an important role in bacterial adhesion to the contact lens.

The pathogenicity of de novo donor-specific antibodies (dnDSA) varies according to their characteristics. While their MFI, complement-fixing ability, and IgG3 subclass are associated with ABMR occurrence and graft loss, they are not fully predictive of outcomes. We investigated the role of the Fc glycosylation of IgG3 dnDSA in ABMR occurrence using mass spectrometry after isolation by single HLA antigen beads. Between 2014 and 2018, we enrolled 54 patients who developed dnDSA (ABMR- n = 24; ABMR+ n = 30) in two French transplant centers. Fucosylation, galactosylation, GlcNAc bisection, and sialylation of IgG3 dnDSA were compared between ABMR+ and ABMR- patients. IgG3 dnDSA from ABMR+ patients exhibited significantly lower sialylation (7.5% vs. 10.5%, p .001) and higher GlcNAc bisection (20.6% vs. 17.4%, p = .008). Fucosylation and galactosylation were similar in both groups. DSA glycosylation was not correlated with DSA MFI. In a multivariate analysis, low IgG3 sialylation, high IgG3%, time from transplantation to kidney biopsy, and tacrolimus-free regimen were independent predictive factors of ABMR. We conclude that a proinflammatory glycosylation profile of IgG3 dnDSA is associated with a risk of ABMR occurrence. Further studies are needed to confirm the clinical interest of DSA glycosylation and to clarify its role in determining the risk of ABMR and graft survival.

Small proteins of up to ∼50 amino acids are an abundant class of biomolecules across all domains of life. Yet, due to the challenges inherent in their size, they are often missed in genome annotations, and are difficult to identify and characterize using standard experimental approaches. Consequently, we still know few small proteins even in well-studied prokaryotic model organisms. Mass spectrometry (MS) has great potential for the discovery, validation, and functional characterization of small proteins. However, standard MS approaches are poorly suited to the identification of both known and novel small proteins due to limitations at each step of a typical proteomics workflow, i.e., sample preparation, protease digestion, liquid chromatography, MS data acquisition, and data analysis. Here, we outline the major MS-based workflows and bioinformatic pipelines used for small protein discovery and validation. Special emphasis is placed on highlighting the adjustments required to improve detection and data quality for small proteins. We discuss both the unbiased detection of small proteins and the targeted analysis of small proteins of interest. Finally, we provide guidelines to prioritize novel small proteins, and an outlook on methods with particular potential to further improve comprehensive discovery and characterization of small proteins. IMPORTANCE Small proteins of up to ∼50 amino acids play important physiological roles across all domains of life. Mass spectrometry is an ideal approach to detect and characterize small proteins, but many aspects of standard mass spectrometry workflows are biased against small proteins due to their size. Here, we highlight applications of mass spectrometry to study small proteins, emphasizing modifications to standard workflows to optimize the detection of small proteins.

In 2018, the International Organization for Standardization (ISO) 19867–1 “Harmonized laboratory test protocols” were released for establishing improved quality and comparability for data on cookstove air pollutant emissions, efficiency, safety, and durability. This is the first study that compares emissions [carbon dioxide, carbon monoxide, total hydrocarbons, methane, nitrogen oxides, fine particulate matter (PM(2.5)), organic carbon, elemental carbon, and ultrafine particles] and efficiency data between the ISO protocol and the Water Boiling Test (WBT). The study examines six stove/fuel combinations [liquefied petroleum gas (LPG), pellet, wood fan, wood rocket, three stone fire, and charcoal] tested in the same US EPA laboratory. Evaluation of the ISO protocol shows improvements over previous test protocols and that results are relatively consistent with former WBT data in terms of tier ratings for emissions and efficiency, as defined by the ISO 19867–3 “Voluntary Performance Targets.” Most stove types remain similarly ranked using ISO and WBT protocols, except charcoal and LPG are in higher PM(2.5) tiers with the ISO protocol. Additionally, emissions data including polycyclic aromatic hydrocarbons are utilized to compare between the ISO and Firepower Sweep Test (FST) protocols. Compared to the FST, the ISO protocol results in generally higher PM(2.5) tier ratings.

Complete proteolytic digestion in the preparation of proteins for bottom-up proteomic analysis is substantially improved by the use of detergents for complete denaturation. This however is incompatible with many proteases, and highly detrimental to LC-MS/MS data collection. Recently, filter-based methods such as FASP (Filter-Aided Sample Prep) have seen wide use due to their ability to remove detergents and other harmful reagents prior to digestion and mass spectrometric analysis. Unfortunately, these techniques can be variable and time consuming. Suspension trapping (S-Trapping) is a newer method that utilizes a depth-filter to trap flocculated proteins, and has proven to be a faster approach for proteomic analysis. Sample preparation by these methods requires careful control of protein concentrations in order to flocculate the sample for collection, and the cost of commercial solutions can be high. We hypothesized that protein suspensions also retain on silica-based filters due to ionic interactions mediated by the presence of sodium (Na+), SO42- and PO43-. As such, we sought to investigate if very low-cost DNA purification spin-filters, so called ‘minipreps’ could efficiently and reproducibly trap proteins for digest and LC-MS/MS analysis. Using model proteins and whole-cell lysates we compared digestion efficiencies, capacities, recovery and identification rates from samples prepared using DNA-minipreps and FASP-based protocols. Samples were analyzed using nano uHPLC MS-MS/MS and Label-Free-Quantitative (LFQ) proteomics. DNA-filters show low variability, excellent recovery, sensitivity, and proteome depth from a commercially obtainable device which costs < $0.25 (US) per sample.

The analysis of the vacuum spark spectrum of erbium in the wavelength region 705 - 2460 A has been extended. A total of 1022 spectral lines are newly identified as transitions between the low lying configurations 4 f 11 , 4 f 10 5 d , 4 f 10 6 s and 4 f 10 6 p of the Er 3 + ion (Er IV), in addition to the confirmed previously identified 582 ones. The extended analysis has led to the determination of 168 new energy levels in Er 3 + , in addition to the 120 previously known ones. Energy values have been optimised by a least-squares procedure based on an increased number of measured wavelengths, resulting in a reduction of their uncertainties. Parametric calculations have been performed using the Cowan codes, in which radial integrals are parameters fitted by least-squares minimisation of the differences calculated - experimental energies. The increased number of experimentally known levels leads to better defined parameters and improved predictions of unknown energy levels and electric dipole transition probabilities. Fitted parameters are compared with ab initio Hartree-Fock integrals including relativistic corrections (HFR). The mean errors of the fits are respectively 51 cm − 1 for 65 known levels of the odd configurations 4 f 11 + 4 f 10 6 p with 9 free parameters, and 53 cm − 1 for 223 known levels of the even configurations 4 f 10 5 d + 4 f 10 6 s with 17 free parameters.

Malignant struma ovarii (MSO) is a rare malignant ovarian tumor, histologically identical to differentiated thyroid cancers. Given the rarity of this disease, there are no treatment guidelines, and the place of imaging for response assessment remains controversial. We report a metabolic response assessed by F-FDG PET/CT in a 71-year-old woman with radioiodine-refractory metastatic MSO treated by targeted therapies (first line with lenvatinib and second line with pazopanib). This case of exceptional response also highlights the usefulness of F-FDG PET/CT for therapeutic assessment of targeted drugs in such a rare clinical entity of malignant MSO.

(+)-Oxo-tomaymycin, a naturally occurring substance of the pyrrolo-1,4-benzodiazepine family, was synthesized using a short and efficient route. The key construction of the seven-membered ring by a...

Background The dietary management of methylmalonic acidaemia (MMA) is a low-protein diet providing sufficient energy to avoid catabolism and to limit production of methylmalonic acid. The goal is to achieve normal growth, good nutritional status and the maintenance of metabolic stability. Aim To describe the dietary management of patients with MMA across Europe. Methods A cross-sectional questionnaire was sent to European colleagues managing inherited metabolic disorders (IMDs) (n=53) with 27 questions about the nutritional management of organic acidaemias. Data were analysed by different age ranges (0–6 months; 7–12 months; 1–10 years; 11–16 years; >16 years). Results Questionnaires were returned from 53 centres. Twenty-five centres cared for 80 patients with MMA vitamin B12 responsive (MMAB12r) and 43 centres managed 215 patients with MMA vitamin B12 non-responsive (MMAB12nr). For MMAB12r patients, 44% of centres (n=11/25) prescribed natural protein below the World Health Organization/Food and Agriculture Organization/United Nations University (WHO/FAO/UNU) 2007 safe levels of protein intake in at least one age range. Precursor-free amino acids (PFAA) were prescribed by 40% of centres (10/25) caring for 36% (29/80) of all the patients. For MMAB12nr patients, 72% of centres (n=31/43) prescribed natural protein below the safe levels of protein intake (WHO/FAO/UNU 2007) in at least one age range. PFAA were prescribed by 77% of centres (n=33/43) managing 81% (n=174/215) of patients. In MMAB12nr patients, 90 (42%) required tube feeding: 25 via a nasogastric tube and 65 via a gastrostomy. Conclusions A high percentage of centres used PFAA in MMA patients together with a protein prescription that provided less than the safe levels of natural protein intake. However, there was inconsistent practices across Europe. Long-term efficacy studies are needed to study patient outcome when using PFAA with different severities of natural protein restrictions in patients with MMA to guide future practice.

Objective: This study examined the secondary mental health outcomes of two contrasting alcohol prevention approaches, whereby one intervention targets common underlying personality risk for alcohol use and mental health problems ( Preventure) and the other targets alcohol- and drug-related behaviours and cognitions ( Climate Schools). Methods: A 2 × 2 cluster randomised controlled factorial design trial was conducted in 26 Australian schools randomised to the following 4 conditions: Climate Schools ( n = 6), Preventure ( n = 7), combined Climate Schools and Preventure (CAP; n = 6) or treatment as usual (TAU; n = 7). Participants completed questionnaires at baseline, 6, 12, 24 and 36 months post-baseline including the Brief Symptom Inventory anxiety and depression scales and hyperactivity and conduct scales of the Strengths and Difficulties Questionnaire. Analyses focused on students who were at high-risk based on personality traits ( n = 947; Mage = 13.3). The effectiveness of each approach in reducing symptoms of internalising and externalising problems was assessed using multi-level mixed effects analysis. Results: Main effects for each intervention relative to not receiving that intervention revealed significant main effects of Preventure in reducing anxiety symptoms ( d = −0.27, 95% confidence interval [CI] = [−0.53, −0.01], p Conclusion: This study is the first to report the effectiveness of personality-targeted alcohol prevention in reducing internalising and externalising symptoms relative to an active control, providing evidence in favour of its specificity in preventing concurrent substance use and mental health problems among high-risk youth.

Protein-rich coacervates are liquid phases separate from the aqueous bulk phase that are used by nature for compartmentalization and more recently have been exploited by engineers for delivery and formulation applications. They also serve as an intermediate phase in an assembly path to more complex structures, such as vesicles. Recombinant fusion protein complexes made from a globular protein fused with a glutamic acid-rich leucine zipper (globule-ZE) and an arginine-rich leucine zipper fused with an elastin-like polypeptide (ZR-ELP) show different phases from soluble, through an intermediate coacervate phase, and finally to vesicles with increasing temperature of the aqueous solution. We investigated the phase transition kinetics of the fusion protein complexes at different temperatures using dynamic light scattering and microscopy, along with mathematical modeling. We controlled coacervate growth by aging the solution at an intermediate temperature that supports coacervation and confirmed that the size of the coacervate droplets dictates the size of vesicles formed upon further heating. With this understanding of the phase transition, we developed strategies to induce heterogeneity in the organization of globular proteins in the vesicle membrane through simple mixing of coacervates containing two different globular fusion proteins prior to the vesicle transition. This study gives fundamental insights and practical strategies for development of globular protein-rich coacervates and vesicles for drug delivery, microreactors, and protocell applications.

Secondary organic aerosol (SOA) is a complex mixture of largely unspeciated compounds. The volatility and viscosity of the bulk organic aerosol influence new particle formation, processing, and lif...

Transport And Golgi Organization protein 2 (TANGO2) deficiency has recently been identified as a rare metabolic disorder with a distinct clinical and biochemical phenotype of recurrent metabolic crises, hypoglycemia, lactic acidosis, rhabdomyolysis, arrhythmias, and encephalopathy with cognitive decline. We report nine subjects from seven independent families, and we studied muscle histology, respiratory chain enzyme activities in skeletal muscle and proteomic signature of fibroblasts. All nine subjects carried autosomal recessive TANGO2 mutations. Two carried the reported deletion of exons 3 to 9, one homozygous, one heterozygous with a 22q11.21 microdeletion inherited in trans. The other subjects carried three novel homozygous (c.262C>T/p.Arg88*; c.220A>C/p.Thr74Pro; c.380+1G>A), and two further novel heterozygous (c.6_9del/p.Phe6del); c.11‐13delTCT/p.Phe5del mutations. Immunoblot analysis detected a significant decrease of TANGO2 protein. Muscle histology showed mild variation of fiber diameter, no ragged‐red/cytochrome c oxidase‐negative fibers and a defect of multiple respiratory chain enzymes and coenzyme Q10 (CoQ10) in two cases, suggesting a possible secondary defect of oxidative phosphorylation. Proteomic analysis in fibroblasts revealed significant changes in components of the mitochondrial fatty acid oxidation, plasma membrane, endoplasmic reticulum‐Golgi network and secretory pathways. Clinical presentation of TANGO2 mutations is homogeneous and clinically recognizable. The hemizygous mutations in two patients suggest that some mutations leading to allele loss are difficult to detect. A combined defect of the respiratory chain enzymes and CoQ10 with altered levels of several membrane proteins provides molecular insights into the underlying pathophysiology and may guide rational new therapeutic interventions., Agència de Gestió d'Ajuts Universitaris i de Recerca (AGAUR), Grant/Award Number: 2014: SGR 393; Association Française contre les Myopathies (FR), Grant/Award Number: 21644; CERCA Programme/ Generalitat de Catalunya, the Hesperia Foundation, the Secretariat for Universities and Research of the Ministry of Business and Knowledge of the Government of Catalonia, Grant/Award Number: [2017SGR1206]; European Research Council, Grant/Award Number: 309548; FEuropean Union Seventh Framework Programme, Grant/Award Number: (FP7/2007‐2013); Instituto de la Marató de TV3, Grant/Award Number: 345/C/2014rm Care (CA); Instituto de Salud Carlos III, Grant/Award Number: grants PI17‐01286, PI17/00109, PI16/00579, PI16/01048, PI14/00581 and CP09/00011; Medical Research Council, Grant/Award Number: MR/N025431/1; Mitochondrial Disease Patient Cohort (UK) , Grant/Award Number: (G0800674); National Institute for Health Research (NIHR) doctoral fellowship, Grant/Award Number: NIHR‐HCS‐D12‐03‐04; Newton Fund, Grant/Award Number: MR/N027302/1; Wellcome Centre for Mitochondrial Research, Grant/Award Number: (203105/Z/16/Z); Wellcome Investigator fund, Grant/Award Number: 109915/Z/15/Z; Wellcome Trust Pathfinder Scheme, Grant/Award Number: 201064/Z/16/Z

Although vaccines have been the primary defense against widespread infectious disease for decades, there is a critical need for improvement to combat complex and variable diseases. More control and specificity over the immune response can be achieved by using only subunit components in vaccines. However, these often lack sufficient immunogenicity to fully protect, and conjugation or carrier materials are required. A variety of protein and peptide biomaterials have improved effectiveness and delivery of subunit vaccines for infectious, cancer, and autoimmune diseases. They are biodegradable and have control over both material structure and immune function. Many of these materials are built from naturally occurring self-assembling proteins, which have been engineered for incorporation of vaccine components. In contrast, others are de novo designs of structures with immune function. In this review, protein biomaterial design, engineering, and immune functionality as vaccines or immunotherapies are discussed.

This paper focuses on the study of thermomechanical properties of polyester acrylate (PEA) resins based on poly(butylene succinate) (PBS) and isosorbide with incorporation of 1/ diacid chains ranging from succinic acid (C4) to azelaic acid (C9) and 2/ diols with various structures such as butanediol and poly(ethyleneglycol) (PEG) with the aim to affect the stiffness of photopolymerized coatings. Therefore, statistic polyester acrylates of Mn 600 g/mol with low polydispersity and Tg were successfully synthetized and characterized by SEC, DSC and DMA. The relative acid value is between 60 and 80 mgKOH/g and the biorenewable carbon content can reach 71 to 90%. Incorporation of the different diols in the backbone was confirmed by NMR. The mixtures of mono and difunctionnal oligomers were crosslinked under LED irradiation and benefits from flexible chain extenders was enlighetened first from improved rate of polymerization and with higher conversion.

Background The intravenous, rapidly acting P2Y12 inhibitor cangrelor reduces the rate of ischemic events during PCI with no significant increase in severe bleeding. However, the efficacy and safety of cangrelor compared with clopidogrel in patients treated with single vessel (SV)-percutaneous coronary intervention (PCI) or multivessel (MV)-PCI remains unexplored. Methods We studied the modified intention-to-treat population of patients from the CHAMPION PHOENIX trial who were randomized to either cangrelor or clopidogrel. We used logistic regression and propensity score matching to evaluate the effect of cangrelor compared with clopidogrel on the primary efficacy outcome (composite of death, myocardial infarction, ischemia-driven revascularization, or stent thrombosis) at 48 hours. The safety outcome was moderate or severe Global Utilization of Streptokinase and tPA for Occluded Arteries bleeding at 48 hours. Hypothesis Cangrelor is as efficacious and safe as clopidogrel in both SV and MV PCI. Results Among 10 854 patients, 9204 (85%) underwent SV- and 1650 (15%) MV-PCI. After adjustment, cangrelor was associated with similar reductions vs clopidogrel in the primary efficacy outcome in patients undergoing SV-PCI (4.5% vs 5.2%; odds ratio [OR] 0.81 [0.66-0.98]) or MV-PCI (6.1% vs 9.8%, OR 0.59 [0.41-0.85]; Pint 0.14). Similar results were observed after propensity score matching (SV-PCI: 5.5% vs 5.9%, OR 0.93 [0.74-1.18]; MV-PCI: 6.2% vs 8.9%, OR 0.67 [0.44-1.01]; Pint 0.17). There was no evidence of heterogeneity in the treatment effect of cangrelor compared with clopidogrel for the safety outcome. Conclusions In patients undergoing SV- or MV-PCI, cangrelor was associated with similar relative risk reductions in ischemic complications and no increased risk of significant bleeding compared with clopidogrel, which highlights the expanding repertoire of options for use in complex PCI.

2D heterostructures are more than a sum of the parent 2D materials, but are also a product of the interlayer coupling, which can induce new properties. In this paper we present a method to tune the interlayer coupling in Bi2Se3/MoS2 2D heterostructures by regulating the oxygen presence in the atmosphere, while applying laser or thermal energy. Our data suggests the interlayer coupling is tuned through the diffusive intercalation and de-intercalation of oxygen molecules. When one layer of Bi2Se3 is grown on monolayer MoS2, an influential interlayer coupling is formed that quenches the signature photoluminescence (PL) peaks. However, thermally annealing in the presence of oxygen disrupts the interlayer coupling, facilitating the emergence of the MoS2 PL peak. DFT calculations predict intercalated oxygen increases the interlayer separation ~17%, disrupting the interlayer coupling and inducing the layers to behave more electronically independent. The interlayer coupling can then be restored by thermally annealing in N2 or Ar, where the peaks will re-quench. Hence, this is an interesting oxygen-induced switching between "non-radiative" and "radiative" exciton recombination. This switching can also be accomplished locally, controllably, and reversibly using a low-power focused laser, while changing the environment from pure N2 to air. This allows for the interlayer coupling to be precisely manipulated with submicron spatial resolution, facilitating site-programmable 2D light-emitting pixels whose emission intensity could be precisely varied by a factor exceeding 200x. Our results show that these atomically-thin 2D heterostructures may be excellent candidates for oxygen sensing.

The physical state and mechanical properties of extruded cereal based products were studied as a function of sucrose content and relative humidity (RH) to evaluate how the presence of sucrose affects glass transition temperature ( T g ), sorption isotherm, and texture parameters. Extrudates were prepared with different sucrose content (0–20 %wt). Sorption isotherm showed the water content of extrudates decreased when product contains high sucrose at low aw range and the inverse effect was observed at high aw. T g s were determined using differential scanning calorimetry (DSC) and two transitions were detected on the heat flow first derivative curve. Addition of sucrose or water decreased both T g s in extrudates. Young's modulus showed water acts as anti-plasticizer at low aw, while shows plasticizing effect at high aw. A stability map can explain the brittle-ductile transition occurred while it was below T g .

The purpose of this study was to characterise the production of exopolysaccharide (EPS) by indigenous lactic acid bacteria (LAB) isolated from traditional Algerian dairy products and to evaluate their possible use in agri-foods. Among the collection of isolated strains, the strain Weissella confusa (W4) was selected for its ability to produce EPS once exposed to a sucrose culture medium. EPS produced were first isolated with a standardised method and further characterised in terms of molecular size, antioxidant activity, and rheological properties. Its direct implication in the texture and syneresis of acid milk gel was evaluated offering interesting industrial applications for its use during processes dealing with dairy products.

Streptococcus gordonii is a member of the viridans streptococci and is an early colonizer of the tooth surface. Adherence to the tooth surface is enabled by proteins present on the S. gordonii cell surface, among which SspB belongs to one of the most well studied cell-wall-anchored adhesin families: the antigen I/II (AgI/II) family. The C-terminal region of SspB consists of three tandemly connected individual domains that display the DEv-IgG fold. These C-terminal domains contain a conserved Ca2+-binding site and isopeptide bonds, and they adhere to glycoprotein 340 (Gp340; also known as salivary agglutinin, SAG). Here, the structural and functional characterization of the C123 SspB domain at 2.7 Å resolution is reported. Although the individual C-terminal domains of Streptococcus mutans AgI/II and S. gordonii SspB show a high degree of both sequence and structural homology, superposition of these structures highlights substantial differences in their electrostatic surface plots, and this can be attributed to the relative orientation of the individual domains (C1, C2 and C3) with respect to each other and could reflect their specificity in binding to extracellular matrix molecules. Studies further confirmed that affinity for Gp340 or its scavenger receptor cysteine-rich (SRCR) domains requires two of the three domains of C123 SspB, namely C12 or C23, which is different from AgI/II. Using protein–protein docking studies, models for this observed functional difference between C123 SspB and C123 AgI/II in their binding to SRCR1 are presented.

BACKGROUND: Non-melanoma skin cancer (NMSC) incidence has been dramatically increasing worldwide. Psoralen, a known photocarcinogen, is naturally abundant in citrus products, leading to the hypothesis that high citrus consumption may increase NMSC risk. METHODS: We fitted age- and multivariable-adjusted logistic regression models to evaluate the association between citrus consumption and NMSC risk among 197,372 UKBB participants. A total of 9,613 NMSC cases were identified using international Classification of Disease 10 codes. Citrus consumption data were collected via five rounds of 24-hour recall questionnaires. RESULTS: We found no association between high total citrus consumption and NMSC risk, although a slightly elevated NMSC risk was observed among participants who consumed >0 to half a serving of total citrus per day (OR [95% CI] = 1.08 [1.01-1.16]). There was no association between individual citrus products and NMSC risk. CONCLUSION: High citrus consumption was not associated with an increased risk of NMSC in our UKBB sample. Further studies are needed to clarify these associations.

Dostarlimab is an immune checkpoint inhibitor (ICI) targeting the Programmed-Death-1 (PD-1) co-receptor, recently approved by the European Medicines Agency (EMA) and the Food and Drug Administration (FDA) as a novel therapy for recurrent or advanced endometrial cancer. We report the case of a 64-year-old woman, experiencing vaginal recurrence with microsatellite instability high/hypermutated of a FIGO stage IA grade 2 endometrial endometrioid adenocarcinoma. She received preoperative chemotherapy with four cycles of carboplatin plus paclitaxel, with stable disease on pelvic magnetic resonance imaging (MRI) and fluorine-18 fluorodeoxyglucose positron emission tomography (18F-FDG PET/CT). Dostarlimab (500 mg intravenously every 3 weeks) was then introduced. The subsequent evaluation after three perfusions demonstrated a complete metabolic response on 18F-FDG PET/CT according to immunotherapy-modified PET response criteria in solid tumors (imPERCIST) criteria, then confirmed by MRI according to immune response evaluation criteria in solid tumors (iRECIST). This clinical description suggests that 18F-FDG PET/CT might take place among available tools for guiding the preoperative management for recurrent endometrial cancer patients receiving dostarlimab immunotherapy that should be further explored through clinical trials.

Implants made of ceramics, and more particularly of calcium phosphates (hydroxyapatite: HA, mainly), promoting intimate contact with natural bone are nowadays merging. Addition of copper ions in bio-ceramics is expected to increase the biological compatibilities of bone graft substitutes. Previous works have shown that copper-doped hydroxyapatite (Cu-doped HA) ceramics can be prepared by solid-state sintering between HA and CuO powder mixtures at about 1100 °C; but, copper-substituted HA was found to be metastable leading to apatitic grains and Cu-rich grain boundaries during the sintering process. Ultra-rapid sintering is so needed. Selective laser sintering (SLS) is an additive manufacturing process that possesses the advantage to be based on ultra-fast sintering process under laser irradiation. SLS being used in literature for the application of laser on polymer-ceramic or poymer-metal composites the proper term all along the paper is Ceramic Powder Bed Laser Sintering (CPBLS). To achieve densification of Cu-doped HA ceramics from CPBLS process, one should control the composition/morphology/structure of the powder bed as well as three other important CPBLS parameters: (i) the applied energy from the laser beam, (ii) the laser power and the laser scanning speed, (iii) the distance between two successive lased lines. In this paper, the impact of all the main CPBLS parameters controlling the sintering of dip-coated Cu-doped HA layers on glass substrates is carefully investigated. Possibility of the creation of thin consolidated Cu-doped HA ceramic patterns, using the ultra-fast CPBLS process, is finally shown.

The occurrence of ochratoxin A (OTA) and OTA-producing fungi in cocoa beans has been previously reported to be linked to harvest and post-harvest treatment conditions. The aim of this study was to identify OTA-producing fungi during harvest, fermentation, drying and storage of cocoa beans from two farms located in the most important cocoa-producing region in Mexico, and their relationship with OTA levels of contamination. A high contamination of ochratoxigenic fungi was observed in drying (78.4 % ± 13.6) and storage (70.3 % ± 3.2) steps of cocoa samples from farm A. In farm B, the wash step of cocoa beans after fermentation reduced pulp residues and consequently the OTA-producing fungi infection during drying (12.7 % ± 5.8) and storage (37 % ± 10.4) steps were diminished. The main OTA-producing strain isolated from Malt Extract Agar (MEA) growth medium was Aspergillus carbonarius (14307.5 µg/kg). The presence of ochratoxigenic species was correlated to high levels of OTA measured in cocoa beans during storage in both farm A and B (304.6 ± 15.3 and 11.3 ± 0.5 µg/kg, respectively), and in fresh cocoa beans from farm B (22.2 ± 3.5 µg/kg). This study reports for the first time OTA content and ochratoxigenic species during cocoa process in Mexico. This allowed the identification and evaluation of the sanitary risks linked to Mexican cocoa production.

Bone marrow failure is characterized by a disruption of hematopoietic stem cell (HSC) homeostasis and function, which causes decreased blood counts. Germline and somatic mutations within HSCs and immune dysregulation contribute to the pathogenesis of marrow failure. Allogeneic HSC transplant is a potentially curative therapy for marrow failure, although not all patients are candidates for this procedure. Immune suppressive therapy (IST) is an effective treatment for patients with aplastic anemia (AA) and select patients with myelodysplastic syndromes, but some patients fail to respond or relapse after IST. Over the past decade, the oral thrombopoietin receptor agonist eltrombopag has become a therapeutic option for AA in combination with frontline IST, and as a single agent for relapsed and refractory patients after IST. In this review, we highlight current knowledge of thrombopoietin receptor agonist mechanisms of action, and clinical indications and toxicities in patients with marrow failure, including the risk of clonal evolution.

Alcohol use is a leading cause of burden of disease among young people. Prevention strategies can be effective in the short-term; however little is known about their longer-term effectiveness. The aim of this study was to examine the sustainability of universal, selective, and combined alcohol use prevention across the critical transition period from adolescence into early adulthood.In 2012, a total of 2190 students (mean age, 13.3 years) from 26 Australian high schools participated in a cluster randomized controlled trial and were followed up for 3 years post baseline. Schools were randomly assigned to deliver the following: (1) universal Web-based prevention for all students (Climate Schools); (2) selective prevention for high-risk students (Preventure); (3) combined universal and selective prevention (Climate Schools and Preventure [CAP]); or (4) health education as usual (control). This study extends the follow-up period to 7-years post baseline. Primary outcomes were self-reported frequency of alcohol consumption and binge drinking, alcohol-related harms, and hazardous alcohol use, at the 7-year follow-up.At 7-year follow-up, students in all 3 intervention groups reported reduced odds of alcohol-related harms compared to the control group (odds ratios [ORs] = 0.13-0.33), and the Climate (OR = 0.04) and Preventure (OR = 0.17) groups reported lower odds of hazardous alcohol use. The Preventure group also reported lower odds of weekly alcohol use compared to the control group (OR = 0.17), and the Climate group reported lower odds of binge drinking (OR = 0.12), holding mean baseline levels constant.This study demonstrated that both universal and selective preventive interventions delivered in schools can have long-lasting effects and reduce risky drinking and related harms into adulthood. No added benefit was observed by delivering the combined interventions.The CAP Study: Evaluating a Comprehensive Universal and Targeted Intervention Designed to Prevent Substance Use and Related Harms in Australian Adolescents; https://www.anzctr.org.au/; ACTRN12612000026820.

International audience; The applicability of an alloy as a hydrogen storage media mostly relies on its pressurecomposition-temperature (PCT) diagram. Since the PCT diagram is compositiondependent, the vast compositional field of high entropy alloys, complex concentrated alloys or multicomponent alloys can be explored to design alloys with optimized properties for each application. In this work, we present a thermodynamic model to calculate PCT diagrams of body-centered (BCC) multicomponent alloys. The entropy of the phases is described using the ideal configurational entropy for interstitial solid solutions with site blocking effect. As a first approximation, it is assumed that the hydrogen partial molar enthalpy of a phase is constant, so the enthalpy of hydrogen mixing varies linearly with the hydrogen concentration. Moreover, the hydrogen partial enthalpy of a phase for a multicomponent alloy was approximated by a simple ideal mixture law of this quantity for the alloy's components with the same structure. Experimental data and DFT calculations were used for parametrization of the enthalpy terms of eight elements (Ti, V, Cr, Ni, Zr, Nb, Hf, and Ta), which are the components of the alloys tested in this work. Experimental PCTs of six BCC multicomponent alloys of four different systems were compared against the calculated ones and the agreement was remarkable. The model and parameters presented here can be regarded as a basis for developing powerful alloy design tools for different hydrogen storage applications.

In recent years, protein engineering efforts have yielded a diverse set of binding proteins that hold promise for various therapeutic applications. Despite this, their inability to reach intracellular targets limits their applications to cell surface or soluble targets. To address this challenge, we previously reported a protein carrier that binds antibodies and delivers them to therapeutic targets inside cancer cells. This carrier, known as the Hex carrier, is comprised of a self-assembling coiled coil hexamer at the core, with each alpha helix fused to a linker, an antibody binding domain, and a six Histidine-tag (His-tag). In this work, we designed different versions of the carrier to determine the role of each building block in cytosolic protein delivery. We found that increasing exposure of the Hex coiled coil on the carriers, through molecular design or removing antibodies, increased internalization, pointing to a role of the coiled coil in promoting endocytosis. We observed a clear increase in endosomal disruption events when His-tags were present on the carrier relative to when they were removed, due to an endosomal buffering effect. Finally, we found that the antibody binding domains of the Hex carrier could be replaced with monomeric ultra-stable GFP for intracellular delivery and endosomal escape. Our results demonstrate that the Hex coiled coil, in conjunction with His-tags, could be a generalizable vehicle for delivering small and large proteins to intracellular targets. This work also highlights new biological applications for oligomeric coiled coils and shows the direct and quantifiable impact of histidine residues on endosomal disruption. These findings could inform the design of future drug delivery vehicles in applications beyond intracellular protein delivery.

Phytohormones play a crucial role in regulating plant developmental processes. Among them, ethylene and jasmonate are known to be involved in plant defense responses to a wide range of biotic stresses as their levels increase with pathogen infection. In addition, these two phytohormones have been shown to inhibit plant nodulation in legumes. Here, exogenous salicylic acid (SA), jasmonate acid (JA), and ethephon (ET) were applied to the root system of Casuarina glauca plants before Frankia inoculation, in order to analyze their effects on the establishment of actinorhizal symbiosis. This protocol further describes how to identify putative ortholog genes involved in ethylene and jasmonate biosynthesis and/or signaling pathways in plant, using the Arabidopsis Information Resource (TAIR), Legume Information System (LIS), and Genevestigator databases. The expression of these genes in response to the bacterium Frankia was analyzed using the gene atlas for Casuarina-Frankia symbiosis (SESAM web site).

Mechanical wounding of plant tissues triggers many different responses (Savatin DV, Gramegna G, Modesti V, Front Plant Sci 5:470, 2014). These are primarily mediated by the plant hormone Jasmonic Acid Isoleucine (JA-Ile). Recently, a fluorescent biosensor for JA-Ile showed that sample preparation (i.e., handling of samples) for fluorescent microscopy very often triggers wound response, even without apparent damage to the seedling, affecting downstream analyses (Larrieu A, Champion A, Legrand J, Nat Commun 6:6043, 2015). In this chapter, we describe how to overcome this technical limitation to monitor any fluorescent reporter or dye in response to wounding, using any type of fluorescent or confocal (inverted or upright, laser scanning or spinning disc) microscopes. Pharmacological or wound treatments can easily be performed and responses monitored over long periods of time. We further describe a simple method to extract and analyse quantitative data from confocal images using the open source software Fiji (Fiji Is Just ImageJ (Schindelin J, Arganda-Carreras I, Frise E, Nat Methods 9:676-682, 2012)) and OpenOffice.

Many small proteins move across cellular compartments through narrow pores. In order to thread a protein through a constriction, free energy must be overcome to either deform or completely unfold the protein. In principle, the diameter of the pore, along with the effective driving force for unfolding the protein, as well as its barrier to translocation, should be critical factors that govern whether the process proceeds via squeezing, unfolding/threading, or both. To probe this for a well-established protein system, we studied the electric-field-driven translocation behavior of cytochrome c (cyt c) through ultrathin silicon nitride (SiNx) solid-state nanopores of diameters ranging from 1.5 to 5.5 nm. For a 2.5 nm diameter pore we find that, in a threshold electric field regime of ∼30-100 MV/m, cyt c is able to squeeze through the pore. As electric fields inside the pore are increased, the unfolded state of cyt c is thermodynamically stabilized, facilitating its translocation. In contrast, for 1.5 nm and 2.0 nm diameter pores, translocation occurs only by threading of the fully unfolded protein after it transitions through a higher energy unfolding intermediate state at the mouth of the pore. The relative energies between the metastable, intermediate, and unfolded protein states are extracted using a simple thermodynamic model that is dictated by the relatively slow (∼ms) protein translocation times for passing through the nanopore. These experiments map the various modes of protein translocation through a constriction, which opens new avenues for exploring protein folding structures, internal contacts, and electric field-induced deformability.Significance StatementCan localized electric fields drive the complete unfolding of a protein molecule? Protein unfolding prior to its translocation through a nanopore constriction is an important step in protein transport across biological membranes and also an important step in nanopore-based protein sequencing. We studied here the electric-field-driven translocation behavior of a model protein (cyt c) through nanopores of diameters ranging from 1.5 to 5.5 nm. These single molecule measurements show that electric fields at the nanopore constriction can select both partially and fully unfolded protein conformations. Zero-field free energy gaps between these conformations, found using a simple thermodynamic model, are in remarkable agreement with previously reported studies of cyt c unfolding energetics.

Macrodomains are proteins that recognize and hydrolyze ADP ribose (ADPR) modifications of intracellular proteins. Macrodomains are implicated in viral genome replication and interference with host cell immune responses. They are important to the infectious cycle of Coronaviridae and Togaviridae viruses. We describe crystal structures of the conserved macrodomain from the bat coronavirus (CoV) HKU4 in complex with ligands. The structures reveal a binding cavity that accommodates ADPR and analogs via local structural changes within the pocket. Using a radioactive assay, we present evidence of mono-ADPR (MAR) hydrolase activity. In silico analysis presents further evidence on recognition of the ADPR modification for hydrolysis. Mutational analysis of residues within the binding pocket resulted in diminished enzymatic activity and binding affinity. We conclude that the common structural features observed in the macrodomain in a bat CoV contribute to a conserved function that can be extended to other known macrodomains.

International audience; Addition of copper in biomaterials is currently investigated because it is expected to enhance the biological properties of bone graft substitutes. Copper-doped hydroxyapatite (CuHA) ceramics were prepared by high temperature solid-state reaction sintering between HA and CuO powder mixtures. The reaction occurred from 950 °C and copper-doped apatites were obtained up to 5.3 wt% of copper. For higher copper content, the presence of secondary phases of CuO and Cu2O remained in the material. Structural analyses (XRD, FTIR) showed the substitution of hydrogen by copper into the hydroxyapatite hexagonal channels in agreement with the following chemical composition Ca10(PO4)6CuIIzCuIyO2H2-2z-y with x = y + z and 0 ≤ x ≤ 0.7. Dense single phased apatitic HA ceramics containing up to 5.3 wt% of copper could be produced after natural sintering in air at 1100 °C. But, copper-substituted HA was found to be metastable leading to apatitic grains and Cu-rich grain boundaries during cooling to room temperature, which resulted in the formation of CuO grains at the material surface after annealing at 500 °C. Quenching from the sintering temperature was carried out to prevent this phenomenon and obtain ceramics made of single Cu-HA phase with a homogeneous fine grain microstructure. In vitro biological assays using MC3T3-E1 cells indicated that the sintered CuHA ceramics were biocompatible, neither cell adhesion nor proliferation being affected by copper addition. A negative effect on cell differentiation appears only from 5 wt% of copper in HA.

Recombinant proteins have emerged as promising building blocks for vesicle self-assembly because of their versatility through genetic manipulation and biocompatibility. Vesicles composed of thermally responsive elastin-like polypeptide (ELP) fusion proteins encapsulate cargo during assembly. However, vesicle stability in physiological environments remains a significant challenge for biofunctional applications. Here, incorporation of an unnatural amino acid, para-azido phenylalanine, into the ELP domain is reported to enable photocrosslinking of protein vesicles and tuning of vesicle size and swelling. The size of the vesicles can be tuned by changing ELP hydrophobicity and ionic strength. Protein vesicles are assessed for their ability to encapsulate doxorubicin and dually deliver doxorubicin and fluorescent protein in vitro as a proof of concept. The resulting photocrosslinkable vesicles made from full-sized, functional proteins show high potential in drug delivery applications, especially for small molecule/protein combination therapies or targeted therapies.

International audience; Mn2+-doped Zn3(PO4)2 and Mn2+-doped ZnAl2·2O4+δ compounds were prepared from solid state route and Pechini process, respectively. The luminescent properties evolution of both as-prepared powder compounds versus the annealing temperature (for 10 min annealing treatments) was studied, in the range 500–1400 °C. In parallel, Doctor Blade was used to prepare few micron-thick films for laser irradiation with the 1 μm wavelength laser of our SLS process (Selective Laser Sintering process). The ultra-rapid heating produces by laser irradiation induces on the films very different optical properties evolution in comparison to what was observed for the powder samples annealed into furnaces for the Mn-doped zinc phosphate. On the opposite, for the spinel compound, whatever the heating process (laser on film or furnace for powder samples), the modification of the luminescence properties is directly linked in a same way to the maximum temperature gained during the thermal history. The idea is to use thermal sensor to extract the temperature map of a film while it is subjected to laser irradiation as during SLS process. The study shows that thermal sensors must rely on mechanisms which remain effective even for ultra-rapid heating: a cationic redistribution versus temperature as occurring into spinel matrix is one solution proposed. But changes in the chromaticity resulting from phase transition between different allotropic forms as occurring into the zinc phosphate compound cannot be directly exploited.

Digestion of proteins separated via sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) remains a popular method for protein identification using mass-spectrometry based proteomics. Although robust and routine, the in-gel digestion procedure is laborious and time-consuming. Electroblotting to a capture membrane prior to digestion reduces preparation steps but requires on-membrane digestion that yields fewer peptides than in-gel digestion. This paper develops direct electroblotting through a trypsin-containing membrane to a capture membrane to simplify extraction and digestion of proteins separated by SDS-PAGE. Subsequent liquid chromatography-tandem mass spectrometry (LC-MS/MS) identifies the extracted peptides. Analysis of peptides from different capture membrane pieces shows that electrodigestion does not greatly disturb the spatial resolution of a standard protein mixture separated by SDS-PAGE. Electrodigestion of an Escherichia coli (E. coli) cell lysate requires four hours of total sample preparation and results in only 13% fewer protein identifications than in-gel digestion, which can take 24 h. Compared to simple electroblotting and protein digestion on a poly(vinylidene difluoride) (PVDF) capture membrane, adding a trypsin membrane to the electroblot increases the number of protein identifications by 22%. Additionally, electrodigestion experiments using capture membranes coated with polyelectrolyte layers identifies a higher fraction of small proteolytic peptides than capture on PVDF or in-gel digestion.

N-containing aromatic compounds (NACs) are an important group of light-absorbing molecules in the atmosphere. They are often observed in combustion emissions, but their chemical formulas and structural characteristics remain uncertain. In this study, red oakwood and charcoal fuels were burned in cookstoves using the standard water-boiling test (WBT) procedure. Submicron aerosol particles in the cookstove emissions were collected using quartz (Qf) and polytetrafluoroethylene (PTFE) filter membranes positioned in parallel. A backup quartz filter (Qb) was also installed downstream of the PTFE filter to evaluate the effect of sampling artifacts on NAC measurements. Liquid chromatography–mass spectroscopy (LC–MS) techniques identified 17 NAC chemical formulas in the cookstove emissions. The average concentrations of total NACs in Qb samples (0.37±0.31–1.79±0.77 µg m−3) were greater than 50 % of those observed in the Qf samples (0.51±0.43–3.91±2.06 µg m−3), and the Qb-to-Qf mass ratios of individual NACs had a range of 0.02–2.71, indicating that the identified NACs might have substantial fractions remaining in the gas phase. In comparison to other sources, cookstove emissions from red oak or charcoal fuels did not exhibit unique NAC structural features but had distinct NAC composition. However, before identifying NAC sources by combining their structural and compositional information, the gas-particle partitioning behaviors of NACs should be further investigated. The average contributions of total NACs to the light absorption of organic matter at λ=365 nm (1.10 %–2.57 %) in Qf and Qb samples (10.7 %–21.0 %) are up to 10 times larger than their mass contributions (Qf: 0.31 %–1.01 %; Qb: 1.08 %–3.31 %), so the identified NACs are mostly strong light absorbers. To explain more sample extract absorption, future research is needed to understand the chemical and optical properties of high-molecular-weight (e.g., molecular weight, MW>500 Da) entities in particulate matter.

The formation of persister cells is one mechanism by which bacteria can survive exposure to environmental stresses. We show that Campylobacter jejuni 11168H forms persister cells at a frequency of 10−3 after exposure to 100 × MIC of penicillin G for 24 h. Staining the cell population with a redox sensitive fluorescent dye revealed that penicillin G treatment resulted in the appearance of a population of cells with increased fluorescence. We present evidence, to show this could be a consequence of increased redox protein activity in, or associated with, the electron transport chain. These data suggest that a population of penicillin G treated C. jejuni cells could undergo a remodeling of the electron transport chain in order to moderate membrane hyperpolarization and intracellular alkalization; thus reducing the antibiotic efficacy and potentially assisting in persister cell formation.