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Enniatins (Enns) are mycotoxins produced by Fusarium spp. which are a fungus widely spread throughout cereals and cereal-based products. Among all the identified enniatins, Enn B1 stands as one of the most prevalent analogues in cereals in Europe. Hence, the aim of this study was to evaluate for the first time the presence of Enn B1 and its phase I metabolites in 300 human urine samples using an ultrahigh-performance liquid chromatography high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) methodology. Enn B1 was detected in 94.3% of samples ranging from 0.007 to 0.429 ng/mL (mean value: 0.065 ng/mL). In accordance with previous in vitro and in vivo analysis, hydroxylated metabolites (78.0% samples) and carbonylated metabolites (66.0% samples) were tentatively identified as the major products. Results from this biomonitoring study point to a frequent intake of Enn B1 in the studied population, suggesting that in-depth toxicological studies are needed in order to understand the potential effects in humans.

Bile secretion by hepatocytes is an osmotic process. The output of bile salts and other organic anions (e.g. glutathione), through the bile salt transporter BSEP/ABCB11 and the organic anion transporter MRP2/ABCC2, respectively, are considered to be the major osmotic driving forces for water secretion into bile canaliculi mainly via aquaporin-8 (AQP8) channels. The down-regulated canalicular expression of these key solute transporters and AQP8 would be a primary event in the establishment of hepatocellular cholestasis. Recent studies in animal models of hepatocellular cholestasis show that the hepatic delivery of AdhAQP1, an adenovector encoding for the archetypical water channel human aquaporin-1 (hAQP1), improves bile secretion and restores to normal the elevated serum bile salt levels. AdhAQP1-transduced hepatocytes show that the canalicularly-expressed hAQP1 not only enhances osmotic membrane water permeability but also induces the transport activities of BSEP/ABCB11 and MRP2/ABCC2 by redistribution in canalicular cholesterol-rich microdomains likely through interactions with the cholesterol-binding protein caveolin-1. Thus, the hepatic gene transfer of hAQP1 improves the bile secretory failure in hepatocellular cholestasis by increasing both biliary output and choleretic efficiency of key osmotic solutes, such as, bile salts and glutathione. The study of hepatocyte aquaporins has provided new insights into the mechanisms of bile formation and cholestasis, and may lead to innovative treatments for cholestatic liver diseases.

Using the right start-up strategy can be vital for successful hydrogen production from thermophilic dark fermentation (55 °C), but it needs to be affordable. Hence, three start-up strategies modifying only influent concentration and temperature were assessed in a reactor fed with cheese whey: (i) high temperature (55 °C) and a high organic loading rate (OLRA - 15 kgCOD m−3 d−1) right at the beginning of the operation; (ii) slowly increasing temperature up to 55 °C using a high OLRA and (iii) slowly increasing temperature and OLRA up to the desired condition. Strategy (iii) increased hydrogen productivity in 39% compared to the others. The combination of high temperature and low pH thermodynamically favored H2 producing routes. Synergy between Thermoanaerobacterium and Clostridium might have boosted hydrogen production. Three reactors of 41 m³ each would be needed to treat 3.4 × 103 m3 year−1 of whey (small-size dairy industry) and the energy produced could reach 14 MWh month−1.

Several species of the Myrcia genus have been used in folk medicine to treat diabetes. Therefore, the aim of this work was to investigate the inhibitory activity of α-glucosidase and pancreatic lipase in the crude extract (EBF) and in the ethyl acetate fraction (FFA) of Myrcia hatschbachii, as well as to identify isolated phenolic compounds and to evaluate the antioxidant property and preliminary in vitro toxicity against Artemia salina. EBF (IC50: 3.21 μg/mL) and FFA (IC50: 1.14 μg/mL) showed inhibitory activity superior to acarbose (IC50: 193.65 μg/mL). In addition, they showed inhibitory effects of pancreatic lipase (IC50: 556.58 μg/mL for EBF and 532.68 μg/mL for FFA), antioxidant potential, absence of preliminary toxicity and presence of gallic andellagic acids in FFA. The relevant results in the inhibition of α-glucosidase and pancreatic lipase motivate new studies for the development of herbal medicines that assist in the treatment of diabetic patients.

The Ocotea genus exhibits a variety of pharmacological, antibacterial and antioxidant effects. The aim of the present study was to evaluate the antioxidant potential and antibacterial properties of the ethyl acetate fraction of Ocotea nutans leaves. Isolation and identification of the phenolic compounds from the fraction was also carried out. The isolated compounds were characterised by one- and two-dimensional nuclear magnetic resonance analysis and identified as vitexin (1) and isovitexin (2). The ethyl acetate fraction of Ocotea nutans leaves demonstrated considerable antioxidant potential. The observed minimum inhibitory concentration of 250 µg.mL−1 was classified as a moderate antibacterial activity against Enterococcus faecalis. Findings from this study demonstrate the utility of this plant as a potential source of antioxidant and antibacterial agents.

Diabetes mellitus, which is characterized by hyperglycemia, leads to poor wound healing. Azadirachta indica has antidiabetic properties and further promotes the healing of epidermal wounds. Here, we investigated the effects of topical application of A. indica extract on excisional lesions in hyperglycemic rats. The animals were divided into four groups (n=15): non-hyperglycemic and treated with carbopol gel (NH), non-hyperglycemic and treated with A. indica + carbopol gel (NHT), hyperglycemic and treated with carbopol gel (H), and hyperglycemic and treated with A. indica + carbopol gel (HT). Lesions were made on the animals’ back, with a punch of 1.5-cm diameter. Samples of the skin/lesion were collected on the 2nd, 7th, and 14th days post-injury for histological examination (inflammatory infiltrate, blood vessels, and fibroblasts), biochemical analysis (MPO [myeloperoxidase], neutrophilic infiltrate and NAG [N-acetylglycosaminidase], and macrophagic infiltrate), oxidative stress (oxidants such as NO [nitric oxide] and DCF [2′,7′-dichlorodihydrofluorescein diacetate], oxidative damage by carbonyl sulfhydryl radicals, and antioxidants such as SOD [superoxide dismutase] and GSH [glutathione]), and analysis of reepithelialization by assessing the wound healing rate. The topical application of A. indica extract in the excisional wounds of hyperglycemic rats accelerated wound healing through anti-inflammatory, antioxidant, angiogenic, and fibrillogenic effects. Thus, these results show that A. indica extract has the potential to be used as a therapeutic agent for treating excisional wounds in hyperglycemic.

Aims: To evaluate whether angiotensin-converting enzyme (ACE) inhibitor and angiotensin II receptor blocker (ARB) combination therapy is more nephroprotective than ACE inhibitor or ARB monotherapy in people with type 2 diabetes and overt nephropathy. Materials and methods: In this prospective, randomized, open, blind-endpoint phase III trial sponsored by the Italian Drug Agency, 103 consenting patients with type 2 diabetes, aged >40 years, with serum creatinine levels 159 to 309 μmol/L, spot morning urinary albumin–creatinine ratio > 1000 mg/g (or > 500 mg/g in those on ACE inhibitor or ARB therapy at inclusion) were stratified by centre and randomized to 4.5-year treatment with valsartan 320 mg/d (n = 36), benazepril 20 mg/d (n = 34) or halved doses of both medications (n = 33). The primary endpoint was end-stage renal disease (ESRD). Modified intention-to-treat analyses were performed. Results: Recruitment took place between June 2007 and February 2013 at 10 centres in Italy and one in Slovenia. A total of 77 participants completed the study and 26 were prematurely withdrawn. During a median (interquartile range) of 41 (18–54) months, 12 participants on benazepril (35.3%) and nine on combination therapy (27.3%) progressed to ESRD, versus five on valsartan (13.9%). Differences between benazepril (hazard ratio [HR] 3.59, 95% confidence interval [CI] 1.25–10.30; P = 0.018) or combination therapy (HR 3.28, 95% CI 1.07–10.0; P = 0.038) and valsartan were significant, even after adjustment for age, gender and baseline serum creatinine, systolic blood pressure and 24-hour proteinuria (HR 5.16, 95% CI 1.50–17.75, P = 0.009 and HR 4.75, 95% CI 1.01–22.39, P = 0.049, respectively). Adverse events were distributed similarly among the groups. Conclusions: In people with type 2 diabetes with nephropathy, valsartan (320 mg/d) safely postponed ESRD more effectively than benazepril (20 mg/d) or than halved doses of both medications.

Sequential covalent embedding of cobalt, osmium, and iron complexes of 4′-(pyridin-4-yl)-2,2′:6′,2′′-terpyridine to surface-enhanced supports pre-functionalized with a templating layer results in hetero-bimetallic (Os–Fe and Co–Fe) and hetero-trimetallic (Co–Os–Fe) monolayer materials. During sequential embedding, each subsequent metal complex deposits onto unoccupied sites of the templating layer leading to densely packed functional materials. Electrochemical and XPS results show that the packing density of Fe(II) species on the surface of the resulting materials is higher than those of pre-deposited Co(II) and Os(II) isostructural complexes. Moreover, according to Raman spectroscopy results, Fe-containing molecules experience steric hindrance when anchored to sequentially deposited Os– or Co–Os materials. The oxidation states of the metal centres in these materials can be successively electrochemically changed resulting in distinct colour-to-colour transitions. Importantly, monolayer materials formed on the conductive surface-enhanced indium-tin-oxide support can operate as entire working electrodes, which, in combination with a polymer gel electrolyte and an indium tin oxide/glass counter electrode, result in two-electrode electrochromic devices (ECDs). Upon operation these devices demonstrate some deterioration of the change in optical density (ΔOD), however, this deterioration is mostly due to gel electrolyte decomposition. The nature of the material allows easy replacement of the electrolyte and the counter electrode. Re-assembly of Co–Fe ECD using a new gel electrolyte and an ITO-glass counter electrode results in almost complete ΔOD recuperation.

Outer membrane vesicles (OMVs) of Neisseria meningitidis contain important antigens to trigger an immune response against meningococci and have been studied as vaccines compounds. The immune response to a vaccine may be affected by its constitution and route of administration. Therefore, Swiss mice were immunized by different routes with OMVs of N. meningitidis B with dimethyl dioctadecyl ammonium bromide in bilayer fragments (DDA-BF) or aluminum hydroxide (AH) as adjuvants. The adjuvants and different routes were compared regarding the immune responses by ELISA, western blot, delayed type hypersensitivity (DTH) and histopathologic analysis. The antigenic preparation generated humoral and cellular immune responses. In quantitative analyzes, in general, AH was superior to DDA-BF. However, analysis such as IgG avidity index, bactericidal activity and immunoblot, revealed no important differences regarding the adjuvant or route of immunization. Regarding the parameters tested, it was not possible to define a superiority between the adjuvants and routes of immunization proposed by this study.

The chemical composition of the essential oil obtained from the branches of Ocotea paranaensis was studied by gas chromatography/mass spectrometry (GC/MS). Twenty-seven compounds, comprising 94.82% of the total components, were identified. The oil showed relatively high concentration of sesquiterpenes (62.96%), oxygenated sesquiterpenes (33.33%), and diterpene (3.70%). Regarding the major compounds, Z-nerolidol (19.16%), germacrene D (12.92%) and α-bulnesene (8.47%) could be highlighted, which corresponded to 40.55% of the substances that were found. The essential oil from Ocotea paranaensis has phosphomolybdenum reducing action and is moderately toxic to the Artemia salina (LC50 = 147.91 μg/mL). It showed haemolytic potential and moderate activity against Staphylococcus aureus, (minimum inhibitory concentration MIC = 250 μg/mL) and Pseudomonas aeruginosa (MIC = 500 μg/mL). No satisfactory cytotoxicity results were observed in lineage H460 and HeLa.

Aims Alicyclobacillus acidoterrestris is a sporulating, acidophilic bacterial species which spoils acidic beverages such as fruit juices. This work aims to quantify the heat resistance of A. acidoterrestris spores and their recovery potential as a function of heating and recovery media pH. Methods and results The heat treatments were carried out with the strain of A. acidoterrestris Ad 746 in Bacillus acidoterrestris thermophilic medium. The pH of the heating medium from pH 7 to pH 2 nonsignificantly reduced the heat resistance. However, the pH levels of the recovery media strongly affected the apparent heat resistance of this strain. The maximum heat resistance was found when the pH was 4·70 and decreased when the pH decreased to pH 2·8, close to the minimum growth pH and when the recovery medium pH increased to pH 5·3. Conclusion The heating medium pH has a slight effect on the spore heat resistances of this acidophilic species. However, the pH of the recovery media strongly affected the apparent heat resistance of this strain. Significance and impact of the study The obtained parameters quantifying the heat resistance of A. acidoterrestris spores are tools to optimize the heat treatments and to control its development.

Shear cell tests have been conducted on twenty different lactose powders, most of which commercially available for oral or inhalation purposes, spanning a wide range of particle sizes, particle morphologies, production processes. The aims of the investigation were: i) to verify the reliability of the technique in evaluating and classifying the flowability of powders; ii) to understand the connection between the flowability of a powder and the morphological properties of its particles; iii) to find a general mathematical relationship able to predict the yield locus shape given the particle size, shape and consolidation state of a lactose powder. These aspects and their limitations are detailed in the manuscript together with other interesting findings on the stick-slip behavior observed in most of the lactose powders examined.

Highly concentrated sugar solutions are known to be effective antimicrobial agents. However, it is unknown whether this effect is solely the result of the collective osmotic effect imparted by a mixture of sugars or whether the type of carbohydrate used also has an individual chemical effect on bacterial responses, that is, inhibition/growth. In view of this, in this work, the antimicrobial properties of four sugars, namely, glucose, fructose, sucrose and maltose against three common food pathogens; Staphylococcus aureus, Escherichia coli and Salmonella enterica, were investigated using a turbidimetric approach. The results obtained indicate that the type of sugar used has a significant effect on the extent of bacterial inhibition which is not solely dependent on the water activity of the individual sugar solution. In addition, while it was shown that high sugar concentrations inhibit bacterial growth, very low concentrations show the opposite effect, that is, they stimulate bacterial growth, indicating that there is a threshold concentration upon which sugars cease to act as antimicrobial agents and become media instead. SIGNIFICANCE AND IMPACT OF THE STUDY: In this work, an analysis on the antimicrobial properties of glucose, fructose, sucrose and maltose in solution was conducted using a turbidimetric approach. Our findings indicate that while, as expected, all of these sugars exhibit significant antimicrobial effects at high concentrations, at low concentrations they appear to act as substrates for the bacteria which results in enhanced microbial growth instead of inhibition. In addition, the results obtained also suggest that the resultant osmotic stress imparted by the sugar solutions is not the only factor which determines their antimicrobial activity and that other chemical factors may be playing a significant role.

Resumo As pesquisas com fungos em busca de substâncias com potencial atividade biológica ganham cada vez mais destaque por serem fontes sustentáveis de novas moléculas. Diplodia pinea é um fungo fitopatogênico encontrado em espécies florestais, que tem apresentado poucos relatos de moléculas com atividade biológica e pode apresentar potencial biotecnológico promissor. Assim, o objetivo deste estudo foi detectar substâncias produzidas por esse fungo em meio líquido, sob condições de estresse. Um metabólito foi detectado na fração alcoólica e caracterizado por difração de raios-X como D-manitol. Esta é a primeira descrição deste monossacarídeo produzido por D. pinea. Abstract Research with fungi in search of substances with potential biological activity is gaining more prominence since they are sustainable sources of new molecules. Diplodia pinea is a phytopathogenic fungus found in forest species, which has shown few reports of molecules with biological activity and may present promising biotechnological potential. Thus, the objective of this study was to detect substances produced by this fungus in a liquid medium, under stressful conditions. A metabolite was detected in the alcoholic fraction characterized by X-ray diffraction as D-mannitol. This is the first description of this monosaccharide produced by D. pinea. Resumen La investigación con hongos en busca de sustancias con potencial actividad biológica está ganando más protagonismo ya que son fuentes sostenibles de nuevas moléculas. Diplodia pinea es un hongo fitopatógeno que se encuentra en especies forestales, que ha mostrado pocos informes de moléculas con actividad biológica y puede presentar un potencial biotecnológico prometedor. Así, el objetivo de este estudio fue detectar sustancias producidas por este hongo en un medio líquido, en condiciones de estrés. Se detectó un metabolito en la fracción alcohólica caracterizada por difracción de rayos X como D-manitol. Ésta es la primera descripción de este monosacárido producido por D. pinea Made available in DSpace on 2021-07-05T19:00:56Z (GMT). No. of bitstreams: 1 Celso-16424-Article-211106-1-10-20210621.pdf: 233954 bytes, checksum: 30751e54426cfbefcdf2d764b2ddfa00 (MD5) Previous issue date: 2021

Campania, due to its pedo-climatic conditions and to its large varietal heritage, is able to produce oils with high typicity, each different from the other. In this study a “minor” autochthonous cultivar of Campania “Oliva Bianca” was analyzed. In autumn 2020, on drupes from trees belonging to the Campania germplasm collection a varietal characterization through physical, chemical and chromatic parameters at the harvest was carried out. Phenolic compounds profile, fatty acids composition and volatile organic compounds have been investigated in the resulting oil. Quality indices, organoleptic and sensory qualities (panel test) were also determined on the oil. Drupe weight was 4.31 g, flesh/pit ratio was 3.68 and the accumulation of oil content at harvest in drupes was 18.63% FW. The drupes showed high anthocyanins content equal to 116.10 mg/kg. In the oil studied, the secoiridoids represented the 82.25% of total phenolic compounds, the concentration of oleic acid was 74.82% and the most present volatile compound was trans-2-hexenal (72.30%). High secoiridoid derivatives concentrations such as oleuropein (85.93 mg/kg) and ligstroside (122.43 mg/kg) aglycones were showed. This study showed a good content of qualitative and quantitative parameters of “Oliva Bianca” oil and drupe, that can have important beneficial effects on human health.

Introduction: To safely expand the donor pool, we introduced a strategy of biopsy-guided selection and allocation to single or dual transplantation of kidneys from donors >60 years old or with hypertension, diabetes, and/or proteinuria (older/marginal donors). Here, we evaluated the long-term performance of this approach in everyday clinical practice. Methods: In this single-center cohort study, we compared outcomes of 98 patients who received one or two biopsy-evaluated grafts from older/marginal donors (“recipients”) and 198 patients who received nonhistologically assessed single graft from ideal donors (“reference-recipients”) from October 2004 to December 2015 at the Bergamo Transplant Center (Italy). Results: Older/marginal donors and their recipients were 27.9 and 19.3 years older than ideal donors and their reference-recipients, respectively. KDPI/KDRI and donor serum creatinine were higher and cold ischemia time longer in the recipient group. During a median follow-up of 51.9 (interquartile range 23.1–88.6) months, 11.2% of recipients died, 7.1% lost their graft, and 16.3% had biopsy-proven acute rejection (BPAR) versus 3.5, 7.6, and 17.7%, respectively, of reference-recipients. Overall death-censored graft failure (rate ratio 0.78 [95% CI 0.33–2.08]), 5-year death-censored graft survival (94.3% [87.8–100.0] vs. 94.2% [90.5–98.0]), BPAR incidence (rate ratio 0.87 [0.49–1.62]), and yearly measured glomerular filtration rate decline (1.18 ± 3.27 vs. 0.68 ± 2.42 mL/min/1.73 m2, p = 0.37) were similar between recipients and reference-recipients, respectively. Conclusions: Biopsy-guided selection and allocation of kidneys from older/marginal donors can safely increase transplant activity in clinical practice without affecting long-term outcomes. This may help manage the growing gap between organ demand and supply without affecting long-term recipient and graft outcomes.

International audience; Previous studies have demonstrated that endocrine disruptors (EDs) can promote the transgenerational inheritance of disease susceptibility. Among the many existing EDs, 2,3,7,8-tetrachlordibenzo-p-dioxin (TCDD) affects reproductive health, including in humans, following direct occupational exposure or environmental disasters, for instance the Agent Orange sprayed during the Vietnam War. Conversely, few studies have focused on TCDD multigenerational and transgenerational effects on human reproductive health, despite the high amount of evidence in animal models of such effects on male and female reproductive health that mimic human reproductive system disorders. Importantly, these studies show that paternal ancestral TCDD exposure substantially contributes to pregnancy outcome and fetal health, although pregnancy outcome is considered tightly related to the woman’s health. In this work, we conducted a systematic review of the literature and a knowledge synthesis in order (i) to describe the findings obtained in rodent models concerning TCDD transgenerational effects on reproductive health and (ii) to discuss the epigenetic molecular alterations that might be involved in this process. As ancestral toxicant exposure cannot be changed in humans, identifying the crucial reproductive functions that are negatively affected by such exposure may help clinicians to preserve male and female fertility and to avoid adverse pregnancy outcomes.

Spent coffee ground (SCG) is a significant by-product generated by the coffee industry. It is considered a great source of bioactive molecules well-recognized for exerting biological properties. This study aimed to implement SCG in a baked foods, such as cookies (SCGc), to increase their bioactive potential. A comprehensive study of the polyphenolic fraction of the SCG and SCGc using a high-resolution mass spectrometry analysis was performed. Moreover, the polyphenol bioaccessibility and change in antioxidant activity during simulated gastrointestinal digestion (GiD) were assessed. Data showed that SCGc provided 780 mg of melanoidins, 16.2 mg of chlorogenic acid (CGA), 6.5 mg of caffeine, and 0.08 mg of phenolic acids per 100 g of sample. Moreover, the 5-caffeoylquinic acid was the most relevant CGA found in SCG (116.4 mg/100 g) and SCGc (8.2 mg/100 g) samples. The antioxidant activity evaluated through three spectrophotometric tests, and the total phenolic compounds of SCGc samples exhibited significantly higher values than the control samples. Furthermore, during simulated GiD, the highest bioaccessibility of SCGc polyphenols was observed after the colonic stage, suggesting their potential advantages for human health. Therefore, SCG with high content in bioactive molecules could represent an innovative ingredient intended to fortify baked food formulations.

Multiple myeloma (MM) is tightly dependent on inflammatory bone marrow microenvironment. IL-17 producing CD4+ T cells (Th17) sustain MM cells growth and osteoclasts-dependent bone damage. In turn, Th17 differentiation relies on inflammatory stimuli. Here, we investigated the role of miR-21 in Th17-mediated MM tumor growth and bone disease. We found that early inhibition of miR-21 in naive T cells (miR-21i-T cells) impaired Th17 differentiation in vitro and abrogated Th17-mediated MM cell proliferation and osteoclasts activity. We validated these findings in NOD/SCID-g-NULL mice, intratibially injected with miR-21i-T cells and MM cells. A Pairwise RNAseq and proteome/phosphoproteome analysis in Th17 cells demonstrated that miR-21 inhibition led to upregulation of STAT-1/-5a-5b, STAT-3 impairment and redirection of Th17 to Th1/Th2 like activated/polarized cells. Our findings disclose the role of miR-21 in pathogenic Th17 activity and open the avenue to the design of miR-21-targeting strategies to counteract microenvironment dependence of MM growth and bone disease.

Brassica L. microgreens are a fresh microscale vegetable crop of high antioxidant value and naturally dense in nutrients without the intervention of biofortification or genetic engineering. A climate chamber experiment on peat-based substrate was set up to test microgreens growth and accumulation of secondary metabolites in response to nutrient supplementation. Microgreens mineral content was analyzed through ion chromatography and total ascorbic acid through UV-Vis spectrophotometry, while carotenoids and phenolic acids were quantified by HPLC-DAD and UHPLC-HRMS, respectively. Brussels sprouts and cabbage yield was only reduced by 10%, while nitrate was reduced by 99% in the absence of nutrient supplementation. Rocket yield was prominently reduced by 47%, with a corresponding nitrate reduction of 118%. Brussels sprouts secondary metabolites were not improved by the absence of nutrient supplementation, whereas cabbage microgreens demonstrated a 30% increase in total ascorbic acid and a 12% increase in total anthocyanins. As for rocket, the absence of nutrient supplementation elicited an extensive increase in secondary metabolites, such as lutein (110%), β-carotene (30%), total ascorbic acid (58%) and total anthocyanins (20%), but caused a decrease in total phenolic acids. It is hereby demonstrated that growing microgreens on a commercial peat-based substrate without nutrient supplementation can be feasible for certain species. Moreover, it might elicit a species-dependent spike in bioactive secondary metabolites.

B-cell acute lymphoblastic leukaemia (B-ALL) reprograms the surrounding bone marrow (BM) stroma to create a leukaemia-supportive niche. To elucidate the contribution of immune cells to the leukaemic microenvironment, we investigated the involvement of monocyte/macrophage compartments, as well as several recruitment pathways in B-ALL development. Immunohistochemistry analyses showed that CD68-expressing macrophages were increased in leukaemic BM biopsies, compared to controls and predominantly expressed the M2-like markers CD163 and CD206. Furthermore, the "non-classical" CD14+ CD16++ monocyte subset, expressing high CX3CR1 levels, was significantly increased in B-ALL patients' peripheral blood. CX3CL1 was shown to be significantly upregulated in leukaemic BM plasma, thus providing an altered migratory pathway possibly guiding NC monocyte recruitment into the BM. Additionally, the monocyte/macrophage chemoattractant chemokine ligand 2 (CCL2) strongly increased in leukaemic BM plasma, possibly because of the interaction of leukaemic cells with mesenchymal stromal cells and vascular cells and due to a stimulatory effect of leukaemia-related inflammatory mediators. C5a, a macrophage chemoattractant and M2-polarizing factor, further appeared to be upregulated in the leukaemic BM, possibly as an effect of PTX3 decrease, that could unleash complement cascade activation. Overall, deregulated monocyte/macrophage compartments are part of the extensive BM microenvironment remodelling at B-ALL diagnosis and could represent valuable targets for novel treatments to be coupled with classical chemotherapy.

Hydrogen bonds are key interactions determining protein-ligand binding affinity and therefore fundamental to any biological process. Unfortunately, explicit structural information about hydrogen positions and thus H-bonds in protein-ligand complexes is extremely rare and similarly the important role of water during binding remains poorly understood. Here, we report on neutron structures of trypsin determined at very high resolutions ≤1.5 Å in uncomplexed and inhibited state complemented by X-ray and thermodynamic data and computer simulations. Our structures show the precise geometry of H-bonds between protein and the inhibitors N-amidinopiperidine and benzamidine along with the dynamics of the residual solvation pattern. Prior to binding, the ligand-free binding pocket is occupied by water molecules characterized by a paucity of H-bonds and high mobility resulting in an imperfect hydration of the critical residue Asp189. This phenomenon likely constitutes a key factor fueling ligand binding via water displacement and helps improving our current view on water influencing protein–ligand recognition., Trypsin is a serine protease. Here the authors present the high resolution X-ray and neutron diffraction structures of uncomplexed and inhibitor bound trypsin that provide insights into the geometry of H-bonds in the active site of the enzyme and molecular dynamics simulations reveal the kinetics of ligand binding induced desolvation.

Carbonic anhydrases (CAs, EC 4.2.1.1) are ubiquitous metalloenzymes, grouped into seven different classes, which catalyze the reaction of CO2 hydration to bicarbonate and protons. All of the fifteen human isoforms reported to date belong to the α-class and contain zinc as a cofactor. The structure of human Zn,Cu-CA II has been solved which contains a copper ion bound at its N-terminal, coordinated to His4 and His64. In the active site a dioxygen molecule is coordinated to the zinc ion. Since dioxygen is a rather unexpected CA ligand, molecular dynamics (MD) simulations were performed which suggested a superoxide character of the zinc bound O2.

Context Acetyl-l-carnitine (ALC), a mitochondrial carrier involved in lipid oxidation and glucose metabolism, decreased systolic blood pressure (SBP), and ameliorated insulin sensitivity in hypertensive nondiabetic subjects at high cardiovascular risk. Objective To assess the effects of ALC on SBP and glycemic and lipid control in patients with hypertension, type 2 diabetes mellitus (T2D), and dyslipidemia on background statin therapy. Design After 4-week run-in period and stratification according to previous statin therapy, patients were randomized to 6-month, double-blind treatment with ALC or placebo added-on simvastatin. Setting Five diabetology units and one clinical research center in Italy. Patients Two hundred twenty-nine patients with hypertension and dyslipidemic T2D >40 years with stable background antihypertensive, hypoglycemic, and statin therapy and serum creatinine, In a prospective randomized controlled trial in 229 patients with hypertension and T2D, 6-month oral acetyl-L-carnitine (1000 mg twice daily) or placebo treatment had similar effects on blood pressure, metabolic control, lipids, and GFR.

Drug-eluting balloon catheters (DEBc) coated with paclitaxel (PTX) have been associated with potential safety concerns. An efficacious but less toxic balloon coating may reduce these outcomes. We evaluated a novel DEBc, Epi-Solve, coated with metacept-3 (MCT-3), a member of the histone deacetylase inhibitor (HDACi) class of epigenetic agents, in a large animal model of neointimal hyperplasia (NIH). Plain balloon angioplasty (PABA) catheters were ultrasonically coated with MCT-3 to generate Epi-Solve DEBc. An ovine model of NIH formation was established utilising partial left common carotid artery (LCA) ligation. Twenty-eight days post neointima (NI) induction, PABA, Epi-Solve or PTX-coated DEBc were deployed at the site of induced NI formation. Twenty-eight days post-intervention, ligated vessels were evaluated for attenuation of NI formation, gene expression profiles and immunohistochemical analysis. Epi-Solve DEBc demonstrated attenuation of NIH over no intervention and a trend to inhibition of NIH over PABA. Gene expression analysis and immunohistochemical studies identified significant anti-proliferative and anti-inflammatory signatures and reduced vascular endothelial cell activation compared to PABA. Epi-Solve is a novel HDACi-coated DEBc which demonstrates significant anti-proliferative and anti-inflammatory signatures and reduced vascular endothelial cell activation compared to PABA in an ovine model and may afford endothelial protection.

Bixin is the main natural apocarotenoid extracted from the seeds of Bixa orellana, widely used as a cosmetic and textile colorant. Despite the description of several pharmacological properties of B. orellana extracts, little has been studied regarding the pharmacological properties of bixin. Then we aimed to investigate the potential anti-inflammatory and antinociceptive effect of bixin in preclinical models of inflammation and acute pain. The anti-inflammatory activity of bixin (15 or 30 mg/kg, orally) was determined using carrageenan-induced paw edema and the myeloperoxidase (MPO) activity in male Wistar rats. The antinociceptive effect of bixin was assessed in the formalin and hot plate tests in rats (at same doses) and in the acetic acid-induced writhing test in Swiss albino male mice (at doses of 27 or 53 mg/kg). General locomotor activity was evaluated in the open field test. Only the higher dose of bixin significantly decreased the carrageenan-induced paw edema and the MPO activity and increased the latency time in the hot plate. Both doses of bixin significantly reduced the number of flinches in both phases of the formalin test and the number of acetic acid-induced writhings without changing the locomotor performance in the open field test. This study validates the use of bixin as an anti-inflammatory trough mechanism related to the reduction of neutrophil migration. Furthermore, this is the first report showing the antinociceptive property of bixin, which does not appear to be related to the sedative effect. Further studies are necessary to characterize the mechanisms involved in these effects.

Extraction of scandium (Sc) from nickel processing residue using Cyanex 923 was studied. Cyanex 923 showed better selectivity for Sc compared with the other metallic components in the residue. Scandium was extracted by solvent extraction via a solvation mechanism. The aqueous-to-organic (A/O) phase ratio was set at 1:2. An increase in the scandium extraction rate was observed when increasing the concentration of H+. Cyanex 923 was shown to be more selective for Sc than Fe. Coextracted Fe was removed from the organic phase by 3 mol L−1 H2SO4 scrubbing with Sc loss of 1.3%. Stripping of scandium from the organic phase of 84.3% was achieved using 4% oxalic acid solution. The precipitated scandium oxalate was recovered by filtration and calcination. The obtained scandium oxide presented minimum purity of 99.0%.

Optical Solar Reflectors (OSRs) play a crucial role in thermal management of spacecraft as the outer-surface to the space environment. To achieve the thermal control, an ideal OSR should be able to reflect all the UV/Vis/NIR spectrum radiation to avoid heating up by the son and also emit blackbody radiation in infrared (IR) spectrum for thermal dissipation. Conventionally, OSRs are made of aluminium plated quartz tiles and therefore have serious issues for their heavy weight and fragile format, significantly adding the launch and assembly cost, respectively. Recently, we have proposed a meta-OSR solution based on thin film technology using Al-doped ZnO (AZO) [1] and the meta-OSRs can be fabricated on flexible substrates, making them light in weight and easy for surface mounting. In addition, we have also proposed a smart meta-OSR solution based on VO 2 , which has a tunable emissivity depending on the operational temperature [2] . In this work, we report a novel meta-OSR with a planar AZO metasurface unlike island-like metasurfaces formed by etch. The planar metasurface is fabricated through our patented plasma patterning technique [3] [4] , which can locally modulate carrier density in AZO.

In this paper a contribution to the set-up and optimization of the fabrication process of thermoplastic-based composites is made. Measurements are carried out of some parameters of interests, related to the regularity of texture. Some measuring techniques are used, all based on a vision system, but different with reference to the processing algorithms of the obtained images. Image processing methods based on artificial intelligence are also used. The analysis can allow to optimize the experimental set-up and the procedure in order to obtain a suitable level of uncertainty for an effective control of the production process.

Aberrant glycosylation has long been known to be associated with cancer, since it is involved in key mechanisms such as tumour onset, development and progression. This review will focus on protein glycosylation studies in cells, tissue, urine and serum in the context of prostate cancer. A dedicated section will cover the glycoforms of prostate specific antigen, the molecule that, despite some important limitations, is routinely tested for helping prostate cancer diagnosis. Our aim is to provide readers with an overview of mass spectrometry-based glycoproteomics of prostate cancer. From this perspective, the first part of this review will illustrate the main strategies for glycopeptide enrichment and mass spectrometric analysis. The molecular information obtained by glycoproteomic analysis performed by mass spectrometry has led to new insights into the mechanism linking aberrant glycosylation to cancer cell proliferation, migration and immunoescape.

This contribution is the result of our progressive engagement to develop and to apply a top-down liquid chromatography (LC) matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) (LC-MALDI-TOF) analysis for the histone post-translational modifications (PTMs) and variants characterization, mainly in order to provide comprehensive and fast results. The histone post-translational modifications and the differential expression of the histone variants play an essential role both in the DNA packaging mechanism in chromosomes and in the regulation of gene expression in different cellular processes, also in response to molecular agents of environmental origin. This epigenetic mechanism is widely studied in different field such as cellular differentiation, development and in the understanding of mechanisms underlying diseases. The characterization of histone PTMs has traditionally performed by antibodies-based assay, but immunological methods have significant limits, and today systems that use mass spectrometry are increasingly employed. We evaluated an in-source decay (ISD) analysis for the histone investigation on human lymphoblastoid cells, and by this approach, we were able to identify and quantify several PTMs such as the di-methylation in the lysine 20 and the acetylation in the lysine 16 in H4 and the mono-methylation, di-methylation and trimethylations at K9 of the histone H3.1. Moreover, we detected and quantified in the same H2B spectrum the prevalent H2B 1C/2E type but also the minor H2B 1D, 1M and 1B/1L/1N, 1O/2F, 1J/1K variants. In this work, we show that MALDI-ISD represents an excellent methodology to obtain global information on histone PTMs and variants from cells in culture, with rapidity and simplicity of execution. Finally, this is a useful approach to get label-free relative quantitative data of histone variants and PTMs.

Intending to highlight new fruits with nutraceutical potential, the present work reports the nutritional and antioxidant content of Syzygium cumini (L.) Skeels (S. cumini) and Syzygium malaccense (L.) Merr. & LM Perry (S. malaccense), and evaluates the anticancer potential against CP-H460 (lung carcinoma line) and its functionality over HEK-293 (healthy embryonic kidney line), two human cells. For this, the physical-chemical characterization of the lyophilized fruits was carried out, and the content of total phenolic compounds (Folin-Ciocalteau) and the antioxidant potential (DPPH, FRAP, and ORAC) was determined. For anticancer activity, aqueous extracts were prepared and evaluated using the MTT (3-[4,5-dimethyl-triazol-2-yl]-2,5-diphenyltetrazolium) assay for periods of 20 hours. These two species are rich in dietary fibers, mainly insoluble fibers, and are sources of natural compounds and antioxidants, which possibly explain the protective potential against cancer cells. Thus, it is expected, as two source fruits, results obtained (p

The aim of this study was to evaluate the biological and antimicrobial activities of commercial freeze-dried whey fermented by lactic acid bacteria in order to valorize this high polluting liquid waste of the dairy industry. Freeze-dried whey was fermented by different strains of Lactobacillus plantarum (CECT 220, 221, 748) at three different times of fermentation (24, 48, 72 h). Afterwards, the extract was purified on centricon amicon with a cut-off of 3 kDa to obtain a permeate consisting of small bioactive compounds reported in the literature to show greater bioactivity. The purified and diluted samples were subjected to the biological and antimicrobial tests for the evaluation of antioxidant, antihypertensive, iron binding, and antifungal activities and identification of phenolic compounds. The results highlighted a radical cation scavenging activity ranging from 1.415 to 2.083 mmol trolox equivalents TE per kg of dry weight, a percentage of iron binding capacity ranging between 23-55% and a percentage of ACE inhibitory activity ranging between 67-85%. The optimal biological activity was obtained from whey fermented by L. plantarum 220 for all the assays performed, except for the iron chelating activity. Furthermore, the antifungal analysis showed a good activity against the mycotoxigenic fungi belonging to Fusarium generum (F. moniliformis, F. graminearum and F. verticillioides), while a slight activity was obtained for Aspergillus and Penicillium generum. This antifungal activity could be correlated to the production of phenolic compounds during fermentation. The obtained results support the hypothesis of using whey as a functional ingredient to improve food preservation.

Background Autosomal dominant polycystic kidney disease (ADPKD) is the most frequent genetically determined renal disease. In affected patients, renal function may progressively decline up to end-stage renal disease (ESRD), and approximately 10% of those with ESRD are affected by ADPKD. The somatostatin analog octreotide long-acting release (octreotide-LAR) slows renal function deterioration in patients in early stages of the disease. We evaluated the renoprotective effect of octreotide-LAR in ADPKD patients at high risk of ESRD because of later-stage ADPKD. Methods and findings We did an internally funded, parallel-group, double-blind, placebo-controlled phase III trial to assess octreotide-LAR in adults with ADPKD with glomerular filtration rate (GFR) 15–40 ml/min/1.73 m2. Participants were randomized to receive 2 intramuscular injections of 20 mg octreotide-LAR (n = 51) or 0.9% sodium chloride solution (placebo; n = 49) every 28 days for 3 years. Central randomization was 1:1 using a computerized list stratified by center and presence or absence of diabetes or proteinuria. Co-primary short- and long-term outcomes were 1-year total kidney volume (TKV) (computed tomography scan) growth and 3-year GFR (iohexol plasma clearance) decline. Analyses were by modified intention-to-treat. Patients were recruited from 4 Italian nephrology units between October 11, 2011, and March 20, 2014, and followed up to April 14, 2017. Baseline characteristics were similar between groups. Compared to placebo, octreotide-LAR reduced median (95% CI) TKV growth from baseline by 96.8 (10.8 to 182.7) ml at 1 year (p = 0.027) and 422.6 (150.3 to 695.0) ml at 3 years (p = 0.002). Reduction in the median (95% CI) rate of GFR decline (0.56 [−0.63 to 1.75] ml/min/1.73 m2 per year) was not significant (p = 0.295). TKV analyses were adjusted for age, sex, and baseline TKV. Over a median (IQR) 36 (24 to 37) months of follow-up, 9 patients on octreotide-LAR and 21 patients on placebo progressed to a doubling of serum creatinine or ESRD (composite endpoint) (hazard ratio [HR] [95% CI] adjusted for age, sex, baseline serum creatinine, and baseline TKV: 0.307 [0.127 to 0.742], p = 0.009). One composite endpoint was prevented for every 4 treated patients. Among 63 patients with chronic kidney disease (CKD) stage 4, 3 on octreotide-LAR and 8 on placebo progressed to ESRD (adjusted HR [95% CI]: 0.121 [0.017 to 0.866], p = 0.036). Three patients on placebo had a serious renal cyst rupture/infection and 1 patient had a serious urinary tract infection/obstruction, versus 1 patient on octreotide-LAR with a serious renal cyst infection. The main study limitation was the small sample size. Conclusions In this study we observed that in later-stage ADPKD, octreotide-LAR slowed kidney growth and delayed progression to ESRD, in particular in CKD stage 4. Trial registration ClinicalTrials.gov NCT01377246; EudraCT: 2011-000138-12., In this double-blind, placebo-controlled, phase 3 randomised trial, Piero Ruggenenti and co-workers assessed the renoprotective effects of Octreotide-LAR in adults with ADPKD at high risk of ESRD, Author summary Why was this study done? Autosomal dominant polycystic kidney disease is the most frequent genetically determined renal disease and affects approximately 10% of patients on chronic dialysis therapy because of end-stage kidney disease. The disease is characterized by relentless growth of renal cysts, with consequent disruption of normal parenchyma and progressive reduction of kidney function up to terminal kidney failure. Previous studies found that octreotide long-acting release (octreotide-LAR), an analog of somatostatin, slowed cyst growth and progressive renal function loss in patients with normal or slightly impaired kidney function. What did the researchers do and find? In this randomized and placebo-controlled trial, we evaluated whether the renoprotective effect of octreotide-LAR could be extended to patients with severe renal disease, that is, with chronic kidney disease stage 3b or 4. One-hundred patients with estimated glomerular filtration rate 15–40 ml/min/1.73 m2 were randomized to receive two 20-mg intramuscular injections of octreotide-LAR (n = 51) or sodium chloride (placebo; n = 49) every 28 days for 3 years. We found that 3-year treatment with octreotide-LAR did not appreciably affect glomerular filtration rate decline compared to placebo, but significantly slowed cyst growth and progression to end-stage kidney failure, in particular in patients with more severe renal insufficiency (stage 4) to start with, and was safe and tolerated well. What do these findings mean? Our present data—combined with previous evidence of a protective effect against kidney and liver volume growth and renal function loss in patients with normal or moderately reduced kidney function—indicate that octreotide-LAR could be a novel disease-modifying therapy that benefits patients with autosomal dominant polycystic kidney disease, including those with later-stage disease. Octreotide-LAR is an expensive medication. The identification of a subgroup of patients with CKD stage 4, accounting for approximately 10% to 15% of patients with autosomal dominant polycystic kidney disease, who are at high risk of kidney failure and at the same time may benefit the most from treatment may help increase the cost-effectiveness of octreotide-LAR for the prevention of end-stage renal disease (and related costs of renal replacement therapy and its complications) in this population.

Hydrogen atoms play a key role in protein–ligand recognition. They determine the quality of established H-bonding networks and define the protonation of bound ligands. Structural visualization of H atoms by X-ray crystallography is rarely possible. We used neutron diffraction to determine the positions of the hydrogen atoms in the ligands aniline and 2-aminopyridine bound to the archetypical serine protease trypsin. The resulting structures show the best resolution so far achieved for proteins larger than 100 residues and allow an accurate description of the protonation states and interactions with nearby water molecules. Despite its low pKa of 4.6 and a large distance of 3.6 Å to the charged Asp189 at the bottom of the S1 pocket, the amino group of aniline becomes protonated, whereas in 2-aminopyridine, the pyridine nitrogen picks up the proton although its amino group is 1.6 Å closer to Asp189. Therefore, apart from charge–charge distances, tautomer stability is decisive for the resulting binding poses, an aspect that is pivotal for predicting correct binding.

Fungi are organisms capable of synthesizing metabolites of industrial interest and the standardization of biomass production for the extraction of these compounds has biotechnological applications. The objective of this work was to optimize the in vitro cultivation process for fungi isolated from Pinus sp., standardizing the best conditions for the production of biomass, contributing to its large scale production. Therefore, the conditions of in vitro cultivation of the fungi Botrytis cinerea, Rhizoctonia sp. and Suillus sp., were evaluated based on the maximum production of dry biomass (PBS), varying temperature, medium and cultivation time. The fungi were grown in glass flasks with liquid culture media, in a BOD chamber, without mechanical stirring. Potato-dextrose broth - PD broth (PD), Czapek - CZ broth (CZ) and Malt Extract - EM broth (EM) were evaluated at temperatures ranging from 8 to 32 ºC and incubation times from 7 to 35 days. PD broth showed better results for fungi B.cinerea and Rhizoctonia sp., when compared to CZ and EM broths, in PBS, while Suillus sp. showed better development in EM broth. The best growth temperature based on PBS was 12 ºC and 16 ºC, with 28 and 35 days of cultivation. Los hongos son organismos capaces de sintetizar metabolitos de interés industrial y la estandarización de la producción de biomasa para la extracción de estos compuestos tiene aplicaciones biotecnológicas. El objetivo de este trabajo fue optimizar el proceso de cultivo in vitro de hongos aislados de Pinus sp., Estandarizando las mejores condiciones para la producción de biomasa, contribuyendo a su producción a gran escala. Por tanto, las condiciones de cultivo in vitro de los hongos Botrytis cinerea, Rhizoctonia sp. y Suillus sp., se evaluaron con base en la producción máxima de biomasa seca (PBS), variación de temperatura, medio y tiempo de cultivo. Los hongos se cultivaron en frascos de vidrio con medio de cultivo líquido, en una cámara de DBO, sin agitación mecánica. Se evaluaron caldos de papa-dextrosa (PD), Czapek (CZ) y extracto de malta (EM) a temperaturas que oscilan entre 8 y 32 ºC y tiempos de incubación de 7 a 35 días. El caldo PD mostró mejores resultados para los hongos B. cinerea y Rhizoctonia sp., En comparación con los caldos CZ y EM, en PBS, mientras que Suillus sp. mostró un mejor desarrollo en caldo EM. La mejor temperatura de crecimiento basada en PBS fue de 12 ºC y 16 ºC, con 28 y 35 días de cultivo. Fungos são organismos capazes de sintetizar metabólitos de interesse industrial e a padronização da produção de biomassa para a extração desses compostos tem aplicações biotecnológicas. O objetivo deste trabalho foi otimizar o processo de cultivo in vitro para fungos isolados de Pinus sp., padronizando as melhores condições para a produção de biomassa, contribuindo para sua produção em larga escala. Portanto, as condições de cultivo in vitro dos fungos Botrytis cinerea, Rhizoctonia sp. e Suillus sp., foram avaliados com base na produção máxima de biomassa seca (PBS), variando temperatura, meio e tempo de cultivo. Os fungos foram cultivados em frascos de vidro com meios de cultura líquidos, em câmara BOD, sem agitação mecânica. Caldos de batata-dextrose (BD), Czapek (CZ) e extrato de malte (EM) foram avaliados em temperaturas variando de 8 a 32 ºC e tempos de incubação de 7 a 35 dias. O caldo BD apresentou melhores resultados para os fungos B. cinerea e Rhizoctonia sp., quando comparado com os caldos CZ e EM, na PBS, enquanto que Suillus sp. apresentou melhor desenvolvimento em caldo EM. A melhor temperatura de crescimento com base no PBS foi de 12 ºC e 16 ºC, com 28 e 35 dias de cultivo.

Industrial hemp (Cannabis sativa L. Family Cannabaceae) contains a vast number of bioactive relevant compounds, namely polyphenols including flavonoids, phenolic acids, phenol amides, and lignanamides, well known for their therapeutic properties. Nowadays, many polyphenols-containing products made of herbal extracts are marketed, claiming to exert health-promoting effects. In this context, industrial hemp inflorescence may represent an innovative source of bioactive compounds to be used in nutraceutical formulations. The aim of this work was to provide a comprehensive analysis of the polyphenolic fraction contained in polar extracts of four different commercial cultivars (Kompoti, Tiborszallasi, Antal, and Carmagnola Cs) of hemp inflorescences through spectrophotometric (TPC, DPPH tests) and spectrometry measurement (UHPLC-Q-Orbitrap HRMS). Results highlighted a high content of cannflavin A and B in inflorescence analyzed samples, which appear to be cannabis-specific, with a mean value of 61.8 and 84.5 mg/kg, meaning a ten-to-hundred times increase compared to other parts of the plant. Among flavonols, quercetin-3-glucoside reached up to 285.9 mg/kg in the Carmagnola CS cultivar. Catechin and epicatechin were the most representative flavanols, with a mean concentration of 53.3 and 66.2 mg/kg, respectively, for all cultivars. Total polyphenolic content in inflorescence samples was quantified in the range of 10.51 to 52.58 mg GAE/g and free radical-scavenging included in the range from 27.5 to 77.6 mmol trolox/kg. Therefore, C. sativa inflorescence could be considered as a potential novel source of polyphenols intended for nutraceutical formulations.

The health advantages of extra-virgin olive oil (EVOO) are ascribed mainly to the antioxidant ability of the phenolic compounds. Secoiridoids, hydroxytyrosol, tyrosol, phenolic acid, and flavones, are the main nutraceutical substances of EVOO. Applications of beneficial microbes and/or their metabolites impact the plant metabolome. In this study the effects of application of selected Trichoderma strains or their effectors (secondary metabolites) on the phenolic compounds content and antioxidant potential of the EVOOs have been evaluated. For this purpose, Trichoderma virens (strain GV41) and Trichoderma harzianum (strain T22), well-known biocontrol agents, and two their metabolites harzianic acid (HA) and 6-pentyl-&alpha, pyrone (6PP) were been used to treat plants of Olea europaea var. Leccino and var. Carolea. Then the nutraceutical potential of EVOO was evaluated. Total phenolic content was estimated by Folin&ndash, Ciocalteau&rsquo, s assay, metabolic profile by High-Resolution Mass spectroscopy (HRMS-Orbitrap), and antioxidant activity by DPPH and ABTS assays. Our results showed that in the cultivation of the olive tree, T22 and its metabolites improve the nutraceutical value of the EVOOs modulating the phenolic profile and improving antioxidants activity.

Food plays a central role in health, especially through consumption of plant-derived foods. Functional foods, supplements, and nutraceuticals are increasingly entering the market to respond to consumer demand for healthy products. They are foods, supplements, and ingredients which offer health benefits beyond the standard nutritional value. Some benefits are associated with phenolic compounds and phytochemicals with antioxidant properties. An olive p&acirc, t&eacute, (OP) was added with antioxidants derived from olive mill wastewater (OMWW) to obtain a functional product rich in phenolic compounds. The olive p&acirc, is produced from the ground olive pericarp, which shows an excellent natural antioxidant content. The OMWW is a waste product from oil processing, which is also rich in phenolic compounds. The result was a product rich in trans-resveratrol, OH tyrosol, and tyrosol in concentrations such as satisfying the European community&rsquo, s claims regarding the possible antioxidant action on plasma lipids with excellent shelf-life stability. The total phenolic content was assayed by a colorimetric method, the antioxidant activity by the ABTS [(2,2&prime, azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)] test, the phenolic profile by Q Exactive Orbitrap LC-MS/MS. The shelf-life stability was confirmed by yeast, molds, and total microbial count, pH, and water activity determinations, and the best pasteurization parameters were determined. The palatability was judged as excellent.

Milk is a nutritious food suitable for infants and adults, and it plays an important role in the human diet. However, it may also be a vehicle for food contaminants. In this report, we developed a method using ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-Q-Exactive Orbitrap HRMS; Thermo Fisher Scientific, Waltham, MA) for simultaneous identification of target pharmacologically active substances and mycotoxins in milk. We also used the Q-Orbitrap operating in full scan mode to identify other possible drugs and microbial metabolites that occurred in samples. Fifty-six commercially available milk samples from the Italian market were analyzed. Investigated analytes were extracted using a QuEChERS (quick, easy, cheap, effective, rugged, and safe) approach. Method detection and quantification limits and performance criteria set by European regulations were fulfilled. Pharmacologically active substances were detected in 49% of samples (range 0.007-4.53 ng/mL), including nontarget mycotoxins. Retrospective analysis allowed us to identify other antibiotics and pharmacologically active substances, as well as nonregulated fungal/bacterial metabolites at a relatively high incidence. From the obtained values, the need for continuous monitoring of contaminants in the milk production chain is clear. This is the first study to assess the presence of pharmacologically active substances, mycotoxins, and other microbial metabolites in Italian milk samples using the UHPLC-Q-Orbitrap HRMS system.

Alternanthera brasiliana (L.) Kuntze is recognized for its healing properties; however, its therapeutic effects remain unclear. Therefore, our study aimed to elucidate the wound healing activities of A. brasiliana using in vitro and in vivo assays. In vitro assays were used to evaluate the antibacterial, anti-inflammatory, and antioxidant effects of A. brasiliana extract. For the in vivo study, two dorsal excisions were established in Wistar rats using a punch (1.5 cm in diameter), which were topically treated daily with 2% carbopol gel (Ctrl group) or 20% hydroalcoholic plant extract with 2% carbopol gel (A. brasiliana–Ab group). After the 2nd, 7th, 14th, and 21st days, inflammation, oxidative damage, antioxidants, angiogenesis, tissue formation, and re-epithelialization were evaluated. In vitro, Ab reduced nitric oxide, anion superoxide, and pro-inflammatory cytokine production. In vivo, Ab presented lower levels of inflammatory infiltrate, although increased levels of IL-1β and TGF-β1 were observed. The plant extract controlled oxidative damage by antioxidants, which favored angiogenesis, collagenesis, and wound re-epithelialization. Thus, the topical application of the hydroalcoholic extract of 20% A. brasiliana was distinguished by its important anti-inflammatory and antioxidant activities both in vivo and in vitro. The plant extract also stimulated angiogenesis and tissue formation, accelerating total re-epithelization, which is promising for wound healing.

Essential oils are an important natural source of pesticides and can be suitable used in ectoparasite control. The goal of this paper was to investigate the chemical composition of the volatile oils obtained from leaves and stems of Philodendron meridionale Buturi & Sakur by gas chromatography/mass spectrometry and to evaluate their insecticidal effect against Ctenocephalides felis Bouché adult insects and their cytotoxicity on human Jurkat leukemic T-cells by 3-(4,5-dimethylthiazol-2-yl)-2,5diphenyltetrazoluim bromide (MTT) reduction method. The main compound was ent-kaur-16-ene at concentrations of 20.78 and 22.46% for volatile oils from leaves and stems, respectively. These volatile oils provided insecticidal effect against C. felis with an average knockdown time of 9.61 and 7.35 min for fumigant test performed with the leaves and stems samples, respectively. Considering the topical application, these P. meridionale volatile oils extracted from leaves and stems demonstrated average knockdown values of 14.70 and 20.0 min, respectively. A very low cytotoxic effect of 9.48 and 11.32% was observed for P. meridionale volatile oils from leaves and stems even at the highest tested concentration of 400 μg.mL-1, respectively. Therefore, it is possible to propose a biological selectivity to the volatile oils under study, which guarantees, at the same time, effect against this insect with no evident damage to human biological system. Los aceites esenciales son una importante fuente natural de pesticidas y pueden usarse de manera adecuada en el control de ectoparásitos. El objetivo de este trabajo fue investigar la composición química de los aceites volátiles obtenidos de hojas y tallos de Philodendron meridionale Buturi & Sakur por cromatografía de gases / espectrometría de masas y evaluar su efecto insecticida contra insectos adultos Ctenocephalides felis Bouché y su citotoxicidad em línea celular Jurkat - células-T leucêmicas humanos- por el método de reducción de bromuro de 3-(4,5-dimetiltiazol-2-il)-2,5difeniltetrazoluim (MTT). El compuesto principal fue ent-kaur-16-eno en concentraciones de 20,78 y 22,46% para aceites volátiles de hojas y tallos, respectivamente. Estos aceites volátiles proporcionaron un efecto insecticida contra C. felis con un tiempo de remoción promedio de 9,61 y 7,35 min para la prueba de fumigante realizada con las muestras de hojas y tallos, respectivamente. Considerando la aplicación tópica, estos aceites volátiles de P. meridionale extraídos de hojas y tallos mostraron valores promedio de caída de 14,70 y 20,0 min, respectivamente. Se observó un efecto citotóxico muy bajo de 9,48 y 11,32% para los aceites volátiles de P. meridionale de hojas y tallos, incluso a la concentración más alta probada de 400 μg.mL-1, respectivamente. Por tanto, es posible proponer una selectividad biológica a los aceites volátiles en estudio, lo que garantiza, al mismo tiempo, efecto contra este insecto sin daño evidente al sistema biológico humano. Os óleos essenciais são uma importante fonte natural de pesticidas e podem ser usados de forma adequada no controle de ectoparasitas. O objetivo deste trabalho foi investigar a composição química dos óleos voláteis obtidos de folhas e caules de Philodendron meridionale Buturi & Sakur por cromatografia gasosa / espectrometria de massa e avaliar seu efeito inseticida contra insetos adultos Ctenocephalides felis Bouché e sua citotoxicidade em linhagem celular Jurkat - células T leucêmicas humanas - pelo método de redução de brometo de 3- (4,5-dimetiltiazol-2-il) -2,5-difeniltetrazoluim (MTT). O principal composto majoritário foi o ent-kaur-16-eno em concentrações de 20,78 e 22,46% para óleos voláteis de folhas e caules, respectivamente. Esses óleos voláteis proporcionaram efeito inseticida contra C. felis com um tempo médio de remoção de 9,61 e 7,35 min para o teste de fumigação realizado com as amostras de folhas e caules, respectivamente. Considerando a aplicação tópica, esses óleos voláteis de P. meridionale extraídos de folhas e caules apresentaram valores médios de queda de 14,70 e 20,0 min, respectivamente. Um efeito citotóxico muito baixo de 9,48 e 11,32% foi observado para os óleos voláteis de P. meridionale de folhas e caules, mesmo na maior concentração testada de 400 μg.mL-1, respectivamente. Portanto, é possível propor uma seletividade biológica aos óleos voláteis em estudo, o que garante, ao mesmo tempo, um efeito contra este inseto sem danos evidentes ao sistema biológico humano.

In an effort to identify novel biopesticides, the present study aimed to assess the effects of Philodendron meridionale (Buturi & Sakur) stem and leaf ketonic and ethanolic extracts (SKE, SEE, LKE, and LEE, respectively) on the germination, growth, root respiration, and enzymatic activities of Lactuca sativa L. seeds, and to measure the associated saponins, phenolics, and flavonoids that may possess herbicidal, pharmaceutical, or pesticidal activities. The extracts were obtained using a modified Soxhlet apparatus and methanolic extracts of 0.1, 0.25, 0.5, 0.75 and 1.0 mg.mL-1 were tested, with water and methanol as a control. The presence of saponins and the concentration of phenolic compounds were determined. Allelopathic activity was evaluated in tests of germination, growth, cellular respiration and enzymatic activity. The presence of saponins and the concentration of phenolic compounds equivalent to 225.12 for LKE, 240.45 for LEE, 193.28 for SKE, and 265.14 for SEE (mg·g-1.gallic acid), and flavonoids 52.74 for LKE, 54.31 for LEE, 72.74 for SKE, and 67.21 for SEE (mg.g-1.quercetin) were determined. The allelopathy of the P. meridionale extracts against L. sativa was confirmed through negative effects on L. sativa seed germination, radical growth and morphology, dry mass, and the concentrations of α-amylase (EC 3.2.1.1), ascorbate peroxidase (EC 1.11.1.11), catalase (EC 1.11.1.6), and polyphenol oxidase (EC 1.10.3.1). It was likely that the allelopathic action of the P. meridionale extracts was related to its effects on the membrane permeability and oxidative stress of the treated L. sativa seeds. The P. meridionale extracts contained saponins, calcium oxalate crystals, and flavonoids, including phenolic compounds, which are known allelochemicals with herbicidal activities. En un esfuerzo por identificar nuevos bioplaguicidas, el presente estudio tuvo como objetivo evaluar los efectos de los extractos cetónicos y etanólicos de tallo y hoja de Philodendron meridionale (Buturi & Sakur) (SKE, SEE, LKE y LEE, respectivamente) sobre la germinación, el crecimiento y la raíz. respiración y actividades enzimáticas de las semillas de Lactuca sativa L., y medir las saponinas, fenólicos y flavonoides asociados que pueden poseer actividades herbicidas, farmacéuticas o pesticidas. Los extractos se obtuvieron utilizando un aparato Soxhlet modificado y se ensayaron extractos metanólicos de 0,1, 0,25, 0,5, 0,75 y 1,0 mg.mL-1, con agua y metanol como control. Se determinó la presencia de saponinas y la concentración de compuestos fenólicos. La actividad alelopática se evaluó en pruebas de germinación, crecimiento, respiración celular y actividad enzimática. La presencia de saponinas y la concentración de compuestos fenólicos equivalentes a 225,12 para LKE, 240,45 para LEE, 193,28 para SKE y 265,14 para SEE (mg.g-1.ácido gálico), y flavonoides 52,74 para LKE, 54,31 para LEE, 72,74 para SKE y 67,21 para SEE (mg.g-1.quercetina). La alelopatía de los extractos de P. meridionale contra L. sativa se confirmó através de los efectos negativos sobre la germinación de semillas de L. sativa, el crecimiento y morfología de radicales, la masa seca y las concentraciones de α-amilasa (EC 3.2.1.1), ascorbato peroxidasa (EC 1.11.1.11), catalasa (EC 1.11.1.6) y polifenol oxidasa (EC 1.10.3.1). Es probable que la acción alelopática de los extractos de P. meridionale estuviera relacionada con sus efectos sobre la permeabilidad de la membrana y el estrés oxidativo de las semillas de L. sativa tratadas. Los extractos de P. meridionale contenían saponinas, cristales de oxalato de calcio y flavonoides, incluidos compuestos fenólicos, que son aleloquímicos conocidos con actividad herbicida. Em um esforço para identificar novos biopesticidas, o presente estudo teve como objetivo avaliar os efeitos dos extratos cetônicos e etanólicos de caule e folha de Philodendron meridionale (Buturi & Sakur) (SKE, SEE, LKE e LEE, respectivamente) na germinação, crescimento, respiração radicular e atividades enzimáticas de sementes de Lactuca sativa L., e medir o índice de saponinas, fenólicos e flavonóides associados que podem possuir atividades herbicidas, farmacêuticas ou pesticidas. Os extratos foram obtidos em aparelho Soxhlet modificado. Foram testados os extratos metanólicos de 0,1; 0,25; 0,5; 0,75 e 1,0 mg.mL-1, tendo água e metanol como controle. Foram determinadas a presença de saponinas e a concentração de compostos fenólicos. A atividade alelopática foi avaliada em testes de germinação, crescimento, respiração celular e atividade enzimática. A presença de saponinas e a concentração de compostos fenólicos equivalente a 225,12 para LKE, 240,45 para LEE, 193,28 para SKE e 265,14 para SEE (mg.g-1.ácido gálico) e flavonóides 52,74 para LKE, 54,31 para LEE, 72,74 para SKE e 67,21 para SEE (mg.g-1.quercetina). A alelopatia dos extratos de P. meridionale contra L. sativa foi confirmada por meio de efeitos negativos na germinação de sementes de L. sativa, crescimento e morfologia do radical, massa seca e concentrações de α-amilase (EC 3.2.1.1), ascorbato peroxidase (EC 1.11.1.11), catalase (EC 1.11.1.6) e polifenol oxidase (EC 1.10.3.1). É provável que a ação alelopática dos extratos de P. meridionale esteja relacionada aos seus efeitos na permeabilidade da membrana e no estresse oxidativo das sementes tratadas de L. sativa. Os extratos de P. meridionale continham saponinas, cristais de oxalato de cálcio e flavonóides, incluindo compostos fenólicos, que são conhecidos aleloquímicos com atividades herbicidas.

Summary Infection by Mycoplasma pneumoniae has been identified as a preceding factor of Guillain–Barré–Stohl syndrome. The Guillain–Barré–Stohl syndrome is triggered by an immune reaction against the major glycolipids and it has been postulated that M. pneumoniae infection triggers this syndrome due to bacterial production of galactocerebroside. Here, we present an extensive comparison of 224 genome sequences from 104 Mycoplasma species to characterize the genetic determinants of galactocerebroside biosynthesis. Hidden Markov models were used to analyse glycosil transferases, leading to identification of a functional protein domain, termed M2000535 that appears in about a third of the studied genomes. This domain appears to be associated with a potential UDP‐glucose epimerase, which converts UDP‐glucose into UDP‐galactose, a main substrate for the biosynthesis of galactocerebroside. These findings clarify the pathogenic mechanisms underlining the triggering of Guillain–Barré–Stohl syndrome by M. pneumoniae infections., Mycoplasma pneumoniae is a preceding factor of Guillain‐Barré‐Stohl syndrome, triggered by an autoimmune reaction against a glycolipid named galactocerebroside, produced by the bacterium. In this work, we reconstruct the pathway and identify the functional protein domain responsible for the synthesis of galactocerebroside in Mycoplasmas.