Ethyl acetate extract from Cistus x incanus L. leaves enriched in myricetin and quercetin derivatives, inhibits inflammatory mediators and activates Nrf2/HO-1 pathway in LPS-stimulated RAW 264.7 macrophages.

Abstract Cistus x incanus L. is a Mediterranean evergreen shrub used in folk medicine for the treatment of inflammatory disorders but the underlying mechanisms are not fully understood. We therefore investigated the anti-inflammatory effects of an ethyl acetate fraction (EAF) from C. x incanus L. leaves on lipopolysaccharide (LPS) activated RAW 264.7 macrophages. HPLC analysis revealed myricetin and quercetin derivatives to be the major compounds in EAF; EAF up to 1 µM of total phenolic content, was not cytotoxic and inhibited the mRNA expression of interleukin-6 (IL-6) and cyclooxygenase-2 (COX-2) (p < 0.05) and the production of prostaglandins E2 (PGE2) (p < 0.05). Meanwhile, EAF triggered the mRNA expression of interleukin-10 (IL-10) and elicited the nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2), as well as the expression of its main target gene, heme oxygenase-1 (HO-1) (p < 0.05). These data indicate that EAF attenuates experimental inflammation via the inhibition of proinflammatory mediators and at least in part, by the activation of Nrf2/HO-1 pathway. These effects are likely due to myricetin and quercetin derivatives but the role of other, less abundant components cannot be excluded. Further studies to confirm the relevance of our findings in animal models and to highlight the relative contribution of each component to the anti-inflammatory activity of EAF should be conducted

Relevance of arginine residues in Cu(II)-induced DNA breakage and Proteinase K resistance of H1 histones

This work analyzes the involvement of arginines in copper/H2O2-induced DNA breakage. Copper is a highly redox active metal which has been demonstrated to form compounds with arginines. For this aim we used mixtures of pGEM3 DNA plasmid and two types of H1 histones which differ only in their arginine content. The sperm H1 histone from the annelid worm Chaetopterus variopedatus (arginine content 12.6 mol% K/R ratio 2) and the somatic H1 histone from calf thymus (arginine content 1.8 mol% and K/R ratio 15). Copper/H2O2-induced DNA breakage was observed only in presence of sperm H1 histones, but it was more relevant for the native molecule than for the deguanidinated derivative (K/R ratio 14), in which 80% of arginine residues were converted to ornithine. Further, copper induced proteinase K resistance and increase of DNA binding affinity on native sperm H1 histones. These observations are consistent with a copper induced reorganization of the side-chains of arginine residues. Copper, instead, did not affect DNA binding affinity of somatic and deguanidinated H1 histones, which show similar K/R ratio and DNA binding mode. These results indicate that arginine residues could affect these H1 histones properties and provide new insights into copper toxicity mechanisms

UV radiation promotes flavonoid biosynthesis, while negatively affecting the biosynthesis and de-epoxidation of xanthophylls: consequence for photoprotection?

There is evidence that UV radiation may detrimentally affect the biosynthesis of carotenoids, particularly de-epoxided xanthophylls, while strongly promoting phenylpropanoid, particularly flavonoid biosynthesis in a range of taxa. Here we tested the hypothesis that mesophyll flavonoids might protect chloroplasts from UV-induced photo-oxidative damage, by partially compensating for the UV-induced depression of xanthophyll biosynthesis. To test this hypothesis we grew two members of the Oleaceae family, Ligustrum vulgare L. and Phillyrea latifolia L., under either partial shading or fully exposed to sunlight, in the presence or in the absence of UV radiation. The examined species, which display very similar flavonoid composition, largely differ in their ability to limit the transmission of UV and visible light through the leaf and, hence, in the accumulation of flavonoids in mesophyll cells. We conducted measurements of photosynthesis, chlorophyll a fluorescence kinetics, the concentrations of individual carotenoids and phenylpropanoids at the level of whole-leaf, as well as the content of epidermal flavonoids. We also performed multispectral fluorescence micro-imaging to unveil the intra-cellular distribution of flavonoids in mesophyll cells. UV radiation decreased the concentration of carotenoids, particularly of xanthophylls, while greatly promoting the accumulation of flavonoids in palisade parenchyma cells. These effects were much greater in L. vulgare than in P. latifolia. UV radiation significantly inhibited the de-epoxidation of xanthophyll cycle pigments, while enhancing the concentration of luteolin, and particularly of quercetin glycosides. Flavonoids accumulated in the vacuole and the chloroplasts in palisade cells proximal to the adaxial epidermis. We hypothesize that flavonoids might complement the photo-protective functions of xanthophylls in the chloroplasts of mesophyll cells exposed to the greatest doses of UV radiation. However, UV radiation might result in adaxial mesophyll cells being less effective in dissipating the excess of radiant energy, e.g., by decreasing their capacity of thermal dissipation of excess visible light in the chloroplast

Dissecting Adaptation Mechanisms to Contrasting Solar Irradiance in the Mediterranean Shrub Cistus incanus

Molecular mechanisms that are the base of the strategies adopted by Mediterranean plants to cope with the challenges imposed by limited or excessive solar radiation during the summer season have received limited attention. In our study, conducted on C. incanus plants growing in the shade or in full sunlight, we performed measurements of relevant physiological traits, such as leaf water potential, gas exchange and PSII photochemistry, RNA-Seq with de-novo assembly, and the analysis of differentially expressed genes. We also identified and quantified photosynthetic pigments, abscisic acid, and flavonoids. Here, we show major mechanisms regulating light perception and signaling which, in turn, sustain the shade avoidance syndrome displayed by the &lsquo;sun loving&rsquo; C. incanus. We offer clear evidence of the detrimental effects of excessive light on both the assembly and the stability of PSII, and the activation of a suite of both repair and effective antioxidant mechanisms in sun-adapted leaves. For instance, our study supports the view of major antioxidant functions of zeaxanthin in sunny plants concomitantly challenged by severe drought stress. Finally, our study confirms the multiple functions served by flavonoids, both flavonols and flavanols, in the adaptive mechanisms of plants to the environmental pressures associated to Mediterranean climate

Laryngeal sensitivity in patients with amyotrophic lateral sclerosis

Recent studies have shown the involvement of the sensory nervous system in patients with amyotrophic lateral sclerosis (ALS). The aim of our study was to investigate the correlation between the laryngeal sensitivity deficit and the type of ALS onset (bulbar or spinal) in a large series of 114 consecutive ALS patients. Participants were subdivided into two groups, bulbar and spinal ALS, according to the clinical onset of disease and submitted to a clinical and instrumental evaluation of swallowing, including a fiber-optic endoscopic evaluation of swallowing with sensory testing. Dysphagia severity was scored using the Penetration–Aspiration Scale (PAS) and the Pooling score (P-score). In addition, three patients with laryngeal sensitivity deficit were submitted to a laryngeal biopsy to assess the status of the sensory innervation. All patients showed a normal glottal closure during phonation and volitional cough. Fifty-six subjects (49%), 14 spinal- and 42 bulbar-onset ALS, showed dysphagia at the first clinical observation (PAS score &gt;1; P-score &gt;5). Dysphagia resulted more frequently in bulbar-onset ALS (P&nbsp;&lt;&nbsp;0.01). Thirty-eight (33%) patients had a sensory deficit of the larynx. The sensory deficit of the larynx was significantly more frequent in bulbar-onset ALS (P&nbsp;&lt;&nbsp;0.01). The sensory deficit of the larynx among dysphagic patients was also significantly more frequent in bulbar-onset ALS (P&nbsp;=&nbsp;0.02). Several abnormalities were found in all three subjects who underwent a laryngeal biopsy: in one patient, no intraepidermal fiber was found; in the other two, the fibers showed morphological changes. Our observations are important to consider for assessment and management of dysphagia in patients with AL

Metabolic plasticity in the hygrophyte Moringa oleifera exposed to water stress

Over the past decades, introduction of many fast-growing hygrophilic, and economically valuable plants into xeric environments has occurred. However, production and even survival of these species may be threatened by harsh climatic conditions unless an effective physiological and metabolic plasticity is available. Moringa oleifera Lam., a multipurpose tree originating from humid sub-tropical regions of India, is widely cultivated in many arid countries because of its multiple uses. We tested whether M. oleifera can adjust primary and secondary metabolism to efficiently cope with increasing water stress. It is shown that M. oleifera possesses an effective isohydric behavior. Water stress induced a quick and strong stomatal closure, driven by abscisic acid (ABA) accumulation, and leading to photosynthesis inhibition with consequent negative effects on biomass production. However, photochemistry was not impaired and maximal fluorescence and saturating photosynthesis remained unaffected in stressed leaves. We report for the first time that M. oleifera produces isoprene, and show that isoprene emission increased three-fold during stress progression. It is proposed that higher isoprene biosynthesis helps leaves cope with water stress through its antioxidant or membrane stabilizing action, and also indicates a general MEP (methylerythritol 4-phosphate) pathway activation that further helps protect photosynthesis under water stress. Increased concentrations of antioxidant flavonoids were also observed in water stressed leaves, and probably cooperate in limiting irreversible effects of the stress in M. oleifera leaves. The observed metabolic and phenotypic plasticity may facilitate the establishment of M. oleifera in xeric environments, sustaining the economic and environmental value of this plant

Laboratory-based surveillance of invasive listeriosis in Northern Italy over a fourteen-year period: epidemiological and clinical results

Introduction Invasive listeriosis is a rare foodborne disease with a large public health impact, because of the severity of its clinical manifestations and high fatality rate. In this study, we provide a snapshot of epidemiology of listeriosis in Lombardy Region, Northern Italy, reviewing enhanced surveillance data collected over fourteen years, after the implementation of a voluntary laboratory-based surveillance system for the referral of clinical isolates of Listeria monocytogenes to a regional reference laboratory, since 2005. Methods Invasive listeriosis cases data from 2005 to 2018 were extracted from the regional laboratory-based surveillance system database and compared with the regional mandatory notification disease system data. Results Over the fourteen period under study, 533 Listeria monocytogenes isolates were detected by the laboratory surveillance system, 55 of which from pregnancy-related cases. The median age of non-pregnancy-associated patients was 71 years, with 64.6% of cases observed in the elderly. Cases with underlying medical risk conditions accounted for 92.1%, and the fatality rate was 26.2%. By integrating data from the two sources, a total of 935 cases were recorded. The collection of data through the laboratory surveillance system allowed to increase the surveillance sensitivity by 18%. Conclusions Our results documented the growing epidemiological relevance of listeriosis through the analysis of two information sources. The data we obtained were consistent with the literature, except for pregnancy-related cases, which are often underdiagnosed. This study highlighted the importance of laboratory-based surveillance system, which led to a significant increase in the sensitivity of the mandatory notification system