Antilisterial effect of citrus essential oils and their performance in edible film formulations

[EN] The antimicrobial activity of eight essential oils (EOs) extracted from the fruit peel of Citrus genotypes (orange, mandarin and lemon) was evaluated against 76 strains of Listeria monocytogenes, previously isolated from different food matrices. EOs showing the most (EO L2 and EO L8) and least (EO O3 and EO M7) effective inhibition activities were chemically characterized by gas chromatography coupled with mass spectrometry (GC/MS) to compare their composition. EO L2 and EO L8 were chosen to determine the MIC and to evaluate the cell viability of the most sensitive strains (L. monocytogenes LM35 and LM69) after 1, 2, 4 and 6 h of exposure. The effectiveness of chitosan (CH) and methylcellulose (MC) edible films, alone and in combination with EO L2 and EO L8, was determined against LM35 and LM69 at 37 C for 0, 8 and 24 h and at 8 C for 0, 1, 3 and 7 days. In addition, the analysis of the microstructure of the films were performed by scanning electron microscope (SEM) to evidence the interactions between the polymers and EOs. Thirty-five and twenty-nine strains were clearly inhibited by EO L2 and EO L8, respectively, while the other Citrus EOs showed poor (EO M1, O4, O5, O6) or minimal (EO O3 and M7) antimicrobial activity. A total of 36 chemical volatile substances was identified by GC/MS to detect the compounds that might play an important role in the characterization of the EOs. The chemical characterization points to oxygenated monoterpenes as relevant compounds in inhibiting Listeria strains, since they have been detected in lemon EOs in concentrations four/five folds higher than orange EOs. Generally, CH- and MCbased films containing EO L2 and EO L8 showed antilisterial activities, even though, the best performances were observed in case of CH-films at 8 C, with a major reduction up to 3 log (CFU/cm2) in case of EO L2 incorporation. The microstructures observed by SEM suggested a better incorporation of the EOs in CH matrix, where a higher amount of oil droplets was distinguished. Therefore, lemon EOs incorporated into chitosan films could be an efficient tool to control Listeria monocytogenes, especially in refrigerated applied conditions.WR was supported by the "Student Mobility for Placement - SMP" grant of the EU Life Learning Program. The authors thank the "Azienda Sperimentale Palazzelli C.R.A. - Centro di ricerca per l'agrumicoltura e le colture mediterranee Contrada Palazzelli Scordia" (CT, Italy) for providing some of the fruits used for EOs extractions.Randazzo, W.; Jiménez Belenguer, AI.; Settanni, L.; Perdones Montero, A.; Moschetti, M.; Palazzolo, E.; Guarrasi, V.... (2016). Antilisterial effect of citrus essential oils and their performance in edible film formulations. Food Control. 59:750-758. doi:10.1016/j.foodcont.2015.06.057S7507585

Itraconazole inhibits nuclear delivery of extracellular vesicle cargo by disrupting the entry of late endosomes into the nucleoplasmic reticulum

Extracellular vesicles (EVs) are mediators of intercellular communication under bothhealthy and pathological conditions, including the induction of pro-metastatic traits,but it is not yet known how and where functional cargoes of EVs are delivered to theirtargets in host cell compartments. We have described that after endocytosis, EVsreach Rab+late endosomes and a fraction of these enter the nucleoplasmic reticu-lum and transport EV biomaterials to the host cell nucleoplasm. Their entry thereinand docking to outer nuclear membrane occur through a tripartite complex formedby the proteins VAP-A, ORP and Rab (VOR complex). Here, we report that theantifungal compound itraconazole (ICZ), but not its main metabolite hydroxy-ICZor ketoconazole, disrupts the binding of Rab to ORP–VAP-A complexes, leadingto inhibition of EV-mediated pro-metastatic morphological changes including cellmigration behaviour of colon cancer cells. With novel, smaller chemical drugs, inhi-bition of the VOR complex was maintained, although the ICZ moieties responsiblefor antifungal activity and interference with intracellular cholesterol distributionwere removed. Knowing that cancer cells hijack their microenvironment and thatEVs derived from them determine the pre-metastatic niche, small-sized inhibitors ofnuclear transfer of EV cargo into host cells could nd cancer therapeutic applications,particularly in combination with direct targeting of cancer cell

Effetti di diverse concentrazioni di NaCl applicate su Morus nigra L. attraverso la tecnologia dell’incapsulamento

In questo studio è stata valutata l’influenza di differenti condizioni di stress salino sul genotipo “Fontanarossa nera” (Morus nigra L.), applicando la tecnologia del seme sintetico. Le concentrazioni di NaCl pari a 0, 50, 100, 150 e 200 mM, sono state aggiunte alle tre soluzioni (incapsulante, complessante e di lavaggio) costituenti l’endosperma artificiale, utilizzato per rivestire microtalee uninodali di 4 mm di lunghezza. Dopo 45 giorni dalla semina su substrato privo di ormoni, sono state registrate le percentuali di vitalità, ripresa e conversione ed i parametri morfologici (numero e lunghezza di germogli e di radici, callo). Sono state effettuate inoltre, analisi per la stima di indicatori fisiologici associati allo stress salino: prolina (Pro), catalasi (CAT), malondialdeide (MDA), guaiacolo perossidasi (POD), letture di clorofilla, flavonoidi, antociani e stima della conducibilità elettrica. Dai risultati è emerso che alla concentrazione salina maggiore la percentuale di vitalità è del 76%; mentre il tasso di conversione raggiunto è stato pari al 52%. Nel presente lavoro è stato studiato per la prima volta, in base alle nostre conoscenze, l’effetto di stress salino applicato congiuntamente alla tecnica di incapsulamento per la produzione di semi sintetici di gelso

Insulin Instability in Parenteral Nutrition Admixtures

Background Biosynthetic human recombinant short-acting insulin is added to parenteral nutrition (PN) admixtures to nourish glucose-intolerant patients. Insulin, however, is electrostatically attracted and inactivated by ethyl-vinyl-acetate (EVA) bags and filling system tubes. Our aim was to verify and quantify the presence of insulin in PN with and without intravenous lipid emulsion (ILE), just after addition (T0) until the infusion's end (T24). Methods Four undiluted samples of 12 different PN complete admixtures (6 with ILE and 6 without), each containing 250 g of glucose in a 2000 mL volume, were taken and analyzed at T0 and T24 by an automated electrochemiluminescence immunoassay after the addition of biosynthetic human recombinant short-acting insulin at increasing doses (from 6 to 72 IU/bag) by an automated compounding device. Assay sensitivity was set at 2 μIU/mL. Admixtures with and without ILE were compared in terms of insulin-detected amounts at T0 and T24. Results Regardless of the amount initially provided, insulin was missing in PN without ILE. In admixtures with ILE, the greater the insulin and ILE doses initially included, the higher the insulin availability at T0 and T24, both in absolute terms and as a percentage of the initial amount (from 3 to 81% at T0 and from 2.5 to 72.5% at T24). ILE may prevent insulin attraction to plastic surfaces. Conclusions Insulin is recovered in the presence of ILE in PN even though considerable amounts are untraceable. This aspect needs verification. Until then, insulin should safely be injected in a different manner in uncontrolled situations

Gametic embryogenesis through isolated microspore culture in Corylus avellana L.

Haploid technology is a valuable plant breeding tool for obtaining homozygosity particularly in woody plants. Hazelnut, the world’s sixth ranking nut tree crop is a monoecious, anemophilous species. It is characterized by a sporophytic incompatibility system that prevents production of homozygous plants with conventional methods, involving several self-pollination cycles. In this study, gametic embryogenesis, in particular isolated microspore culture, was tried with five genotypes. Two culture media were tested and four temperature stress treatments were applied to the isolated microspores that were cultivated at the vacuolated developmental stage. To our knowledge, this is the first report of early embryos being recovered from the five hazelnut genotypes via isolated microspore culture. Analysis of the embryo genotypes using SSR markers demonstrated the embryos had a single allele per each locus whereas the parent cultivar was heterozygous, indicating they developed from haploid microspores. The response to the culture treatments, was, however, genotype-dependent, as previously reported for male gamete embryogenesis in other fruit crops

Bovine pericardium patch wrapping intestinal anastomosis improves healing process and prevents leakage in a pig model.

Failure of intestinal anastomosis is a major complication following abdominal surgery. Biological materials have been introduced as reinforcement of abdominal wall hernia in contaminated setting. An innovative application of biological patch is its use as reinforcement of gastrointestinal anastomosis. The aim of study was to verify whether the bovine pericardium patch improves the healing of anastomosis, when in vivo wrapping the suture line of pig intestinal anastomosis, avoiding leakage in the event of deliberately incomplete suture. Forty-three pigs were randomly divided: Group 1 (control, n = 14): hand-sewn ileo-ileal and colo-colic anastomosis; Group 2 (n = 14): standard anastomosis wrapped by pericardium bovine patch; Group 3 (n = 1) and 4 (n = 14): one suture was deliberately incomplete and also wrapped by patch in the last one. Intraoperative evaluation, histological, biochemical, tensiometric and electrophysiological studies of intestinal specimens were performed at 48 h, 7 and 90 days after. In groups 2 and 4, no leak, stenosis, abscess, peritonitis, mesh displacement or shrinkage were found and adhesion rate decreased compared to control. Biochemical studies showed mitochondrial function improvement in colic wrapped anastomosis. Tensiometric evaluations suggested that the patch preserves the colic contractility similar to the controls. Electrophysiological results demonstrated that the patch also improves the mucosal function restoring almost normal transport properties. Use of pericardium bovine patch as reinforcement of intestinal anastomosis is safe and effective, significantly improving the healing process. Data of prevention of acute peritonitis and leakage in cases of iatrogenic perforation of anastomoses, covered with patch, is unpublished

Prolonged lifespan with enhanced exploratory behavior in mice overexpressing the oxidized nucleoside triphosphatase hMTH1

The contribution that oxidative damage to DNA and/or RNA makes to the aging process remains undefined. In this study, we used the hMTH1-Tg mouse model to investigate how oxidative damage to nucleic acids affects aging. hMTH1-Tg mice express high levels of the hMTH1 hydrolase that degrades 8-oxodGTP and 8-oxoGTP and excludes 8-oxoguanine from both DNA and RNA. Compared to wild-type animals, hMTH1-overexpressing mice have significantly lower steady-state levels of 8-oxoguanine in both nuclear and mitochondrial DNA of several organs, including the brain. hMTH1 overexpression prevents the age-dependent accumulation of DNA 8-oxoguanine that occurs in wild-type mice. These lower levels of oxidized guanines are associated with increased longevity and hMTH1-Tg animals live significantly longer than their wild-type littermates. Neither lipid oxidation nor overall antioxidant status is significantly affected by hMTH1 overexpression. At the cellular level, neurospheres derived from adult hMTH1-Tg neural progenitor cells display increased proliferative capacity and primary fibroblasts from hMTH1-Tg embryos do not undergo overt senescence in vitro. The significantly lower levels of oxidized DNA/RNA in transgenic animals are associated with behavioral changes. These mice show reduced anxiety and enhanced investigation of environmental and social cues. Longevity conferred by overexpression of a single nucleotide hydrolase in hMTH1-Tg animals is an example of lifespan extension associated with healthy aging. It provides a link between aging and oxidative damage to nucleic acids. © 2013 John Wiley & Sons Ltd and the Anatomical Society

Role of neuroinflammation in hypertension-induced brain amyloid pathology

Hypertension and sporadic Alzheimer's disease (AD) have been associated but clear pathophysiological links have not yet been demonstrated. Hypertension and AD share inflammation as a pathophysiological trait. Thus, we explored if modulating neuroinflammation could influence hypertension-induced β-amyloid (Aβ) deposition.Possible interactions among hypertension, inflammation and Aβ-deposition were studied in hypertensive mice with transverse aortic coarctation (TAC). Given that brain Aβ deposits are detectable as early as 4 weeks after TAC, brain pathology was analyzed in 3-week TAC mice, before Aβ deposition, and at a later time (8-week TAC mice).Microglial activation and interleukin (IL)-1β upregulation were already found in 3-week TAC mice. At a later time, along with evident Aβ deposition, microglia was still activated. Finally, immune system stimulation (LPS) or inhibition (ibuprofen), strategies described to positively or negatively modulate neuroinflammation, differently affected Aβ deposition.We demonstrate that hypertension per se triggers neuroinflammation before Aβ deposition. The finding that only immune system activation, but not its inhibition, strongly reduced amyloid burden suggests that stimulating inflammation in the appropriate time window may represent a promising strategy to limit vascular-triggered AD-pathology

(A–D) Electrophysiological measurements.

<p>Short circuit current (I_SC) and tissue resistance (R_T) in ileal and colic specimens. Bars represent mean ± SEM in C (before surgery, N = 15), A-ES (after surgery without patch, early stage, N = 15), P-ES (after surgery with patch, early stage, N = 15), A-LS (after surgery without patch, late stage, N = 10), P-LS (after surgery with patch, late stage, N = 10) in ileal and colic specimens, respectively. Significance was determined by Student's t test for unpaired data of follow-up stage <i>vs</i> control (*p≤0.05; **p≤ 0.002; ***p≤0.0001) and for anastomoses with patch <i>vs</i> anastomoses without patch control (#: p≤0.05).</p

Microscopic Findings and Mean Histological Score.

<p>h: hours; pod: post-operative day; mean score ± SEM. *Score 0-4: 0 =  absent; 1 =  minimal; 2 =  moderate; 3 =  distinctive; 4 =  severe.</p