Thyme and Savory Essential Oil Efficacy and Induction of Resistance against Botrytis cinerea through Priming of Defense Responses in Apple

The efficacy of thyme and savory essential oils were investigated against Botrytis cinerea on apple fruit. Apples treated with thyme and savory essential oils showed significantly lower gray mold severity and incidence. Thyme essential oil at 1% concentration showed the highest efficacy, with lower disease incidence and smaller lesion diameter. The expression of specific pathogenesis-related (PR) genes PR-8 and PR-5 was characterized in apple tissues in response to thyme oil application and B. cinerea inoculation. After 6 h of pathogen inoculation, thyme essential oil induced a 2.5-fold increase of PR-8 gene expression compared to inoculated fruits. After 24 h of inoculation, PR-8 was highly induced (7-fold) in both thyme oil-treated and untreated apples inoculated with B. cinerea. After 48 h of inoculation, PR-8 expression in thyme-treated and inoculated apples was 4- and 6-fold higher than in inoculated and water-treated apples. Neither thyme oil application nor B. cinerea inoculation markedly affected PR-5 expression. These results suggest that thyme oil induces resistance against B. cinerea through the priming of defense responses in apple fruit, and the PR-8 gene of apple may play a key role in the mechanism by which thyme essential oil effectively inhibits gray mold in apple fruit

Thyme and Savory Essential Oil Vapor Treatments Control Brown Rot and Improve the Storage Quality of Peaches and Nectarines, but Could Favor Gray Mold

The effect of biofumigation, through slow-release diffusors, of thyme and savory essential oils (EO), was evaluated on the control of postharvest diseases and quality of peaches and nectarines. EO fumigation was effective in controlling postharvest rots. Naturally contaminated peaches and nectarines were exposed to EO vapors for 28 days at 0 °C in sealed storage cabinets and then exposed at 20 °C for five days during shelf-life in normal atmosphere, simulating retail conditions. Under low disease pressure, most treatments significantly reduced fruit rot incidence during shelf-life, while, under high disease pressure, only vapors of thyme essential oil at the highest concentration tested (10% v/v in the diffusor) significantly reduced the rots. The application of thyme or savory EO favored a reduction of brown rot incidence, caused by Monilinia fructicola, but increased gray mold, caused by Botrytis cinerea. In vitro tests confirmed that M. fructicola was more sensitive to EO vapors than B. cinerea. Essential oil volatile components were characterized in storage cabinets during postharvest. The antifungal components of the essential oils increased during storage, but they were a low fraction of the volatile organic compounds in storage chambers. EO vapors did not influence the overall quality of the fruit, but showed a positive effect in reducing weight loss and in maintaining ascorbic acid and carotenoid content. The application of thyme and savory essential oil vapors represents a promising tool for reducing postharvest losses and preserving the quality of peaches and nectarines

Oncogenic RET Kinase domain mutations perturb the autophosphorylation trajectory by enhancing substrate presentation in trans

To decipher the molecular basis for RET kinase activation and oncogenic deregulation, we defined the temporal sequence of RET autophosphorylation by label-free quantitative mass spectrometry. Early autophosphorylation sites map to regions flanking the kinase domain core, while sites within the activation loop only form at later time points. Comparison with oncogenic RET kinase revealed that late autophosphorylation sites become phosphorylated much earlier than wild-type RET, which is due to a combination of an enhanced enzymatic activity, increased ATP affinity, and surprisingly, by providing a better intermolecular substrate. Structural analysis of oncogenic M918T and wild-type RET kinase domains reveal a cis-inhibitory mechanism involving tethering contacts between the glycine-rich loop, activation loop, and αC-helix. Tether mutations only affected substrate presentation but perturbed the autophosphorylation trajectory similar to oncogenic mutations. This study reveals an unappreciated role for oncogenic RET kinase mutations in promoting intermolecular autophosphorylation by enhancing substrate presentation

Microcantilver-based DNA hybridization sensors for Salmonella identification

The detection of pathogenic microorganisms in foods remains a challenging since the safety of foodstuffs has to be ensured by the food producing companies. Conventional methods for the detection and identification of bacteria mainly rely on specific microbiological and biochemical identification. Biomolecular methods, are commonly used as a support for traditional techniques, thanks to their high sensitivity, specificity and not excessive costs. However, new methods like biosensors for example, can be an exciting alternative to the more traditional tecniques for the detection of pathogens in food. In this study we report Salmonella enterica serotype Enteritidis DNA detection through a novel class of label-free biosensors: microcantilevers (MCs). In general, MCs can operate as a microbalance and is used to detect the mass of the entities anchored to the cantilever surface using the decrease in the resonant frequency. We use DNA hybridization as model reaction system and for this reason, specific single stranded probe DNA of the pathogen and three different DNA targets (single-stranded complementary DNA, PCR product and serial dilutions of DNA extracted from S. Enteritidis strains) were applied. Two protocols were reported in order to allow the probe immobilization on cantilever surface: i) MC surface was functionalized with 3-aminopropyltriethoxysilane and glutaraldehyde and an amino-modified DNA probe was used; ii) gold-coated sensors and thiolated DNA probes were used in order to generate a covalent bonding (Th-Au). For the first one, measures after hybridization with the PCR product showed related frequency shift 10 times higher than hybridization with complementary probe and detectable signals were obtained at the concentrations of 103 and 106 cfu/mL after hybridization with bacterial DNA. There are currently optimizations of the second protocol, where preliminary results have shown to be more uniform and therefore more precise within each of the three hybridizations

Development of an innovative microcantilever-based biosensor for 17β-estradiol detection in bovine muscles: preliminary results

17β-estradiol is the most powerful substance with estrogenic effect, commonly used as illegal growth promoter in livestock production. To avoid health risks for consumers, sensitive, reliable and low-cost methods for quantification of extremely low concentrations of such carcinogenic residues in food are needed. Antibody-immobilised microcantilever resonators were proposed as innovative biosensors able to quantify an adsorbed target mass thanks to a shift in resonance frequency. Furthermore, the quantification of masses on the order of few picograms has recently shown to be successfully achievable with very high precision. In this study, we analysed the performance of our microcantilever sensors using extracted samples of bovine muscle from experimental animals, containing variable concentrations of 17β-estradiol (HPLC-MS/MS tested). Preliminary data showed that treated animals are correctly revealed, exhibiting large negative frequency shifts. More experiments, though, are needed to obtain a correct quantification of 17β-estradiol concentration

BZLF1, an Epstein–Barr virus immediate–early protein, induces p65 nuclear translocation while inhibiting p65 transcriptional function

AbstractWe have previously demonstrated that the Epstein–Barr virus immediate–early BZLF1 protein interacts with, and is inhibited by, the NF-κB family member p65. However, the effects of BZLF1 on NF-κB activity have not been intensively studied. Here we show that BZLF1 inhibits p65-dependent gene expression. BZLF1 inhibited the ability of IL-1, as well as transfected p65, to activate the expression of two different NF-κB-responsive genes, ICAM-1 and IκB-α. BZLF1 also reduced the constitutive level of IκB-α protein in HeLa and A549 cells, and increased the amount of nuclear NF-κB to a similar extent as tumor necrosis factor-alpha (TNF-α) treatment. In spite of this BZLF1-associated increase in the nuclear form of NF-κB, BZLF1 did not induce binding of NF-κB to NF-κB responsive promoters (as determined by chromatin immunoprecipitation assay) in vivo, although TNF-α treatment induced NF-κB binding as expected. Overexpression of p65 dramatically inhibited the lytic replication cycle of EBV in 293-EBV cells, confirming that NF-κB also inhibits BZLF1 transcriptional function. Our results are consistent with a model in which BZLF1 inhibits the transcriptional function of p65, resulting in decreased transcription of IκB-α, decreased expression of IκB-α protein, and subsequent translocation of NF-κB to the nucleus. This nuclear translocation of NF-κB may promote viral latency by negatively regulating BZLF1 transcriptional activity. In situations where p65 activity is limiting in comparison to BZLF1, the ability of BZLF1 to inhibit p65 transcriptional function may protect the virus from the host immune system during the lytic form of infection

CI and CO in Nearby Spiral Galaxies -- I. Line Ratio and Abundance Variations at ~ 200 pc Scales

We present new neutral atomic carbon [CI](3P1-3P0) mapping observations within the inner ~7 kpc and ~4 kpc of the disks of NGC3627 and NGC4321 at a spatial resolution of 190 pc and 270 pc, respectively, using the ALMA Atacama Compact Array (ACA). We combine these with the CO(2-1) data from PHANGS-ALMA, and literature [CI] and CO data for two other starburst and/or active galactic nucleus (AGN) galaxies (NGC1808, NGC7469), to study: a) the spatial distributions of CI and CO emission; b) the observed line ratio RCICO = I_[CI](1-0)/I_CO(2-1) as a function of various galactic properties; and c) the abundance ratio of [CI/CO]. We find excellent spatial correspondence between CI and CO emission and nearly uniform RCICO ~0.1 across the majority of the star-forming disks of NGC3627 and NGC4321. However, RCICO strongly varies from ~0.05 at the centre of NGC4321 to >0.2-0.5 in NGC1808's starburst centre and NGC7469's centre with an X-ray AGN. Meanwhile, RCICO does not obviously vary with $U$, similar to the prediction of PDR models. We also find a mildly decreasing RCICO with an increasing metallicity over 0.7-0.85 solar metallicity, consistent with the literature. Assuming various typical ISM conditions representing GMCs, active star-forming regions and strong starbursting environments, we calculate the LTE radiative transfer and estimate the [CI/CO] abundance ratio to be ~0.1 across the disks of NGC3627 and NGC4321, similar to previous large-scale findings in Galactic studies. However, this abundance ratio likely has a substantial increase to ~1 and >1-5 in NGC1808's starburst and NGC7469's strong AGN environments, respectively, in line with the expectations for cosmic-ray dominated region (CRDR) and X-ray dominated region (XDR) chemistry. Finally, we do not find a robust evidence for a generally CO-dark, CI-bright gas in the disk areas we probed. (abbreviated)Comment: 23 pages, 13 figures and one table in total (17 pages and 9 figures in main text). Accepted for publication in A&A. For associated data cubes and moment maps, see https://www.canfar.net/storage/vault/list/phangs/RELEASES/DZLIU_etal_202

The semisynthetic flavonoid monoHER sensitises human soft tissue sarcoma cells to doxorubicin-induced apoptosis via inhibition of nuclear factor-κB

Background:Despite therapeutic advances, the prognosis of patients with metastatic soft tissue sarcoma (STS) remains extremely poor. The results of a recent clinical phase II study, evaluating the protective effects of the semisynthetic flavonoid 7-mono-O-(beta-hydroxyethyl)-rutoside (monoHER) on doxorubicin-induced cardiotoxicity, suggest that monoHER enhances the antitumour activity of doxorubicin in STSs.Methods:To molecularly explain this unexpected finding, we investigated the effect of monoHER on the cytotoxicity of doxorubicin, and the potential involvement of glutathione (GSH) depletion and nuclear factor-kappaB (NF-kappaB) inactivation in the chemosensitising effect of monoHER.Results:MonoHER potentiated the antitumour activity of doxorubicin in the human liposarcoma cell line WLS-160. Moreover, the combination of monoHER with doxorubicin induced more apoptosis in WLS-160 cells compared with doxorubicin alone. MonoHER did not reduce intracellular GSH levels. On the other hand, monoHER pretreatment significantly reduced doxorubicin-induced NF-kappaB activation.Conclusion:These results suggest that reduction of doxorubicin-induced NF-kappaB activation by monoHER, which sensitises cancer cells to apoptosis, is involved in the chemosensitising effect of monoHER in human liposarcoma cells

Activation of NF-kB Pathway by Virus Infection Requires Rb Expression

The retinoblastoma protein Rb is a tumor suppressor involved in cell cycle control, differentiation, and inhibition of oncogenic transformation. Besides these roles, additional functions in the control of immune response have been suggested. In the present study we investigated the consequences of loss of Rb in viral infection. Here we show that virus replication is increased by the absence of Rb, and that Rb is required for the activation of the NF-kB pathway in response to virus infection. These results reveal a novel role for tumor suppressor Rb in viral infection surveillance and further extend the concept of a link between tumor suppressors and antiviral activity

Measuring the mixing scale of the ISM within nearby spiral galaxies

The spatial distribution of metals reflects, and can be used to constrain, the processes of chemical enrichment and mixing. Using PHANGS-MUSE optical integral field spectroscopy, we measure the gas phase oxygen abundances (metallicities) across 7,138 HII regions in a sample of eight nearby disc galaxies. In Paper I (Kreckel et al. 2019) we measure and report linear radial gradients in the metallicities of each galaxy, and qualitatively searched for azimuthal abundance variations. Here, we examine the two-dimensional variation in abundances once the radial gradient is subtracted, Delta(O/H), in order to quantify the homogeneity of the metal distribution and to measure the mixing scale over which HII region metallicities are correlated. We observe low (0.03--0.05 dex) scatter in Delta(O/H) globally in all galaxies, with significantly lower (0.02--0.03 dex) scatter on small (<600 pc) spatial scales. This is consistent with the measurement uncertainties, and implies the two-dimensional metallicity distribution is highly correlated on scales of <600 pc. We compute the two point correlation function for metals in the disc in order to quantify the scale lengths associated with the observed homogeneity. This mixing scale is observed to correlate better with the local gas velocity dispersion (of both cold and ionized gas) than with the star formation rate. Selecting only HII regions with enhanced abundances relative to a linear radial gradient, we do not observe increased homogeneity on small scales. This suggests that the observed homogeneity is driven by the mixing introducing material from large scales rather than by pollution from recent and on-going star formation.Comment: 17 pages, 14 figures. Accepted for publication in MNRA