Search

Failure to resolve inflammation underlies many prevalent pathologies. Recent insights have identified lipid mediators, typified by lipoxins (LXs), as drivers of inflammation resolution, suggesting potential therapeutic benefit. We report the asymmetric preparation of novel quinoxaline-containing synthetic-LXA4-mimetics (QNX-sLXms). Eight novel compounds were screened for their impact on inflammatory responses. Structure–activity relationship (SAR) studies showed that (R)-6 (also referred to as AT-02-CT) was the most efficacious and potent anti-inflammatory compound of those tested. (R)-6 significantly attenuated lipopolysaccharide (LPS)- and tumor-necrosis-factor-α (TNF-α)-induced NF-κB activity in monocytes and vascular smooth muscle cells. The molecular target of (R)-6 was investigated. (R)-6 activated the endogenous LX receptor formyl peptide receptor 2 (ALX/FPR2). The anti-inflammatory properties of (R)-6 were further investigated in vivo in murine models of acute inflammation. Consistent with in vitro observations, (R)-6 attenuated inflammatory responses. These results support the therapeutic potential of the lead QNX-sLXm (R)-6 in the context of novel inflammatory regulators.

Persistent cytopenia due to poor graft function (PoGF) is a relatively common complication which may affect up to 20% of patients undergoing allogeneic hematopoietic stem cell transplantation (HSCT). Treatment options for PoGF remain limited, and reinfusion of additional HSC is often the only way to rescue hematopoiesis. Here we describe a retrospective single-center experience with the thrombopoietin-mimetic agent eltrombopag for the treatment of PoGF. Thirteen patients have received eltrombopag for either PoGF (n = 12) or primary graft failure (n = 1). In the 12 PoGF patients eltrombopag was started at the median time of 79 days after HSCT, due to persistent thrombocytopenia, with concomitant anemia and neutropenia in 7 and 3 patients, respectively. The treatment was started at the dose of 50 mg per day, and eventually increased up to 150 mg in case of lack of response. Hematological response was seen in 7 patients, with 6 complete responses. Hematological responses were seen both in patients with evidence of immune-mediated pathophysiology, and with possible infectious/iatrogenic causes. In responding patients, eltrombopag was discontinued in 6/7 patients without further relapse. These results suggest that eltrombopag is safe and possibly effective in the setting of the treatment of PoGF, and pave the way for future prospective studies.

Glioblastoma represents the highest grade of brain tumors. Despite maximal resection surgery associated with radiotherapy and concomitant followed by adjuvant chemotherapy with temozolomide (TMZ), patients have a very poor prognosis due to the rapid recurrence and the acquisition of resistance to TMZ. Here, initially considering that TMZ is a prodrug whose activation is pH-dependent, we explored the contribution of glioblastoma cell metabolism to TMZ resistance. Using isogenic TMZ-sensitive and TMZ-resistant human glioblastoma cells, we report that the expression of O6-methylguanine DNA methyltransferase (MGMT), which is known to repair TMZ-induced DNA methylation, does not primarily account for TMZ resistance. Rather, fitter mitochondria in TMZ-resistant glioblastoma cells are a direct cause of chemoresistance that can be targeted by inhibiting oxidative phosphorylation and/or autophagy/mitophagy. Unexpectedly, we found that PARP inhibitor olaparib, but not talazoparib, is also a mitochondrial Complex I inhibitor. Hence, we propose that the anticancer activities of olaparib in glioblastoma and other cancer types combine DNA repair inhibition and impairment of cancer cell respiration.

Citrus aurantium L. dry extracts (CAde) improve adipogenesis in vitro. These effects are dependent from an early modulation of CCAAT/enhancer-binding protein beta (C/Ebp&beta, ) expression and cyclic Adenosine Monophosphate (cAMP) response element-binding protein (CREB) activation. C/Ebp&beta, and Creb are also targets of miR-155. This study investigated whether CAde regulates miR-155 expression in the early stages of adipogenesis and whether it ameliorates adipocyte differentiation of cells exposed to tumor necrosis factor-alpha (TNF&alpha, ). Adipogenic stimuli (AS) were performed in 3T3-L1 pre-adipocytes treated with CAde, TNF&alpha, or both. Gene and miRNA expression were determined by quantitative real-time PCR. Adipogenesis was evaluated by Oil-Red O staining. CAde treatment enhanced AS effects during the early adipogenesis phases by further down-regulating miR-155 expression and increasing both C/Ebp&beta, and Creb mRNA and protein levels. At variance, TNF&alpha, inhibited 3T3-L1 adipogenesis and abolished AS effects on miR-155, C/Ebp&beta, and Creb expression. However, in cells exposed to TNF&alpha, CAde improved adipocyte differentiation and restored the AS effects on miRNA and gene expression at early time points. In conclusion, this study identified miR-155 down-regulation as part of the mechanism through which CAde enhances adipogenesis of pre-adipocytes in vitro. Furthermore, it provides evidence of CAde efficacy against TNF&alpha, negative effects on adipogenesis.

Controlling unmodified serotonin levels in brain synapses is a primary objective when treating major depressive disorder - a disease that afflicts ~20% of the world's population. Roughly 60% of patients respond poorly to first-line treatments and thus new therapeutic strategies are sought. Toward this end, we have constructed isoform-specific inhibitors of the human cytosolic sulfotransferase 1A3 (SULT1A3) - the isoform responsible for sulfonating ~80% of the serotonin in extracellular brain fluid. The inhibitor design includes a core ring structure, which anchors the inhibitor into a SULT1A3-specific binding pocket located outside the active site, and a sidechain crafted to act as a latch to inhibit turnover by fastening down the SULT1A3 active-site cap. The inhibitors are allosteric, they bind with nanomolar affinity and are highly specific for the 1A3 isoform. The cap-stabilizing effects of the latch can be accurately calculated and are predicted to extend throughout the cap and into the surrounding protein. A free energy correlation demonstrates that the percent inhibition at saturating inhibitor varies linearly with cap stabilization - the correlation is linear because the rate-limiting step of the catalytic cycle, nucleotide release, scales linearly with the fraction of enzyme in the cap-open form. Inhibitor efficacy in cultured cells was studied using a human mammary epithelial cell line that expresses SULT1A3 at levels comparable to those found in neurons. The inhibitors perform similarly in ex vivo and in vitro studies; consequently, SULT1A3 turnover can now be potently suppressed in an isoform-specific manner in human cells.

To evaluate whether treatment with 100,000 IU/month (equivalent to 3200 IU/day) of cholecalciferol and 1 g/day of dietary calcium supplementation in HIV patients following different cART regimens yields normal levels of vitamin D3 and PTH as well as whether changes in bone mineral density are clinically significant.Consecutive HIV patients following different cART regimens received 100,000 IU/month (equivalent to 3200 IU/day) of cholecalciferol and 1 g/day of dietary calcium supplementation. The participants underwent BMD assessment via dual energy X-ray absorptiometry of the spine and hip at baseline (T0) and after 24 months (T1). Levels of 25(OH) vitamin D3 and parathyroid hormone (PTH) were assessed at T0 and T1. Quantitative variables were assessed with a paired t-test, independent t-test or analysis of variance, as appropriate. A chi-squared analysis was used to assess the association between qualitative variables. A p-value0.05 was considered significant. Patients were divided into three groups depending on the cART regimen.A total of 79 patients were included (40 males, 51% and 39 females, 49%), with a mean age of 46.6 (SD ±11.2) years, a baseline CD4 count of 649 cells/µl and a mean 25 hydroxycholecalciferol (25(OH) D3) value of 25 + 10 ng/ml. After 24 months, the 25(OH) D3 increased to 40 + 11 ng/ml. The initial BMDs at T0 were estimated as 0.919 (±0.27) and 0.867 (±0.14) g/cmThe proposed protocol of cholecalciferol and dietary calcium supplementation is safe and valid for correcting vitamin D abnormalities in almost all patients as well as reducing PTH levels in a high percentage of patients; however, it is not sufficient for normalization, particularly in patients exposed to tenofovir or efavirenz. At the spine, no significant BMD change was found in any of the therapy groups. At the hip, our data confirm a modest negative effect on bone mass caused by tenofovir and efavirenz.

Glucocorticoids are utilized for their anti-inflammatory properties in the skeletal muscle and arthritis. However, the major drawback of use of glucocorticoids is that it leads to senescence and toxicity. Therefore, based on the idea that decreasing particle size allows for increased surface area and bioavailability of the drug, in the present study, we hypothesized that nanodelivery of dexamethasone will offer increased efficacy and decreased toxicity. The dexamethasone-loaded poly(lactic-co-glycolic acid) (PLGA) nanoparticles were prepared using nanoprecipitation method. The morphological characteristics of the nanoparticles were studied under scanning electron microscope. The particle size of nanoparticles was 217.5 ± 19.99 nm with polydispersity index of 0.14 ± 0.07. The nanoparticles encapsulation efficiency was 34.57% ± 1.99% with in vitro drug release profile exhibiting a sustained release pattern over 10 days. We identified improved skeletal muscle myoblast performance with improved closure of the wound along with increased cell viability at 10 nmol/L nano-dexamethasone-PLGA. However, dexamethasone solution (1 μmol/L) was injurious to cells because the migration efficiency was decreased. In addition, the use of dexamethasone nanoparticles decreased lipopolysaccharide-induced lactate dehydrogenase release compared with dexamethasone solution. Taken together, the present study clearly demonstrates that delivery of PLGA-dexamethasone nanoparticles to the skeletal muscle cells is beneficial for treating inflammation and skeletal muscle function.

Lysosomal storage disorders (LSDs) are inherited diseases characterized by lysosomal dysfunction and often showing a neurodegenerative course. There is no cure to treat the central nervous system in LSDs. Moreover, the mechanisms driving neuronal degeneration in these pathological conditions remain largely unknown. By studying mouse models of LSDs, we found that neurodegeneration develops progressively with profound alterations in presynaptic structure and function. In these models, impaired lysosomal activity causes massive perikaryal accumulation of insoluble α‐synuclein and increased proteasomal degradation of cysteine string protein α (CSPα). As a result, the availability of both α‐synuclein and CSPα at nerve terminals strongly decreases, thus inhibiting soluble NSF attachment receptor (SNARE) complex assembly and synaptic vesicle recycling. Aberrant presynaptic SNARE phenotype is recapitulated in mice with genetic ablation of one allele of both CSPα and α‐synuclein. The overexpression of CSPα in the brain of a mouse model of mucopolysaccharidosis type IIIA, a severe form of LSD, efficiently re‐established SNARE complex assembly, thereby ameliorating presynaptic function, attenuating neurodegenerative signs, and prolonging survival. Our data show that neurodegenerative processes associated with lysosomal dysfunction may be presynaptically initiated by a concomitant reduction in α‐synuclein and CSPα levels at nerve terminals. They also demonstrate that neurodegeneration in LSDs can be slowed down by re‐establishing presynaptic functions, thus identifying synapse maintenance as a novel potentially druggable target for brain treatment in LSDs.

Glioma Cancer Stem-Like Cells (GSCs) are a small subset of CD133+ cells with self-renewal properties and capable of initiating new tumors contributing to Glioma progression, maintenance, hierarchy, and complexity. GSCs are highly resistant to chemo and radiotherapy. These cells are believed to be responsible for tumor relapses and patients' fatal outcome after developing a recurrent Glioblastoma (GBM) or High Grade Glioma (HGG). GSCs are cells under replicative stress with high demands on NAD+ supply to repair DNA, maintain self-renewal capacity and to induce tumor plasticity. NAD+ feeds Poly-ADP polymerases (PARP) and NAD+-dependent deacetylases (SIRTUINS) contributing to GSC phenotype. This energetic core axis is mainly controlled by the rate-limiting enzyme nicotinamide phosphoribosyltransferase (NAMPT), an important oncogene contributing to tumor dedifferentiation. Targeting GSCs depicts a new frontier in Glioma therapy; hence NAMPT could represent a key regulator for GSCs maintenance. Its inhibition may attenuate GSCs properties by decreasing NAD+ supply, consequently contributing to a better outcome together with current therapies for Glioma control., AC lab was supported by grants from the Spanish Ministry of Economy and Competitivity, Plan Estatal de I+D+I 2013-2016, ISCIII (Fis: PI15/00045), and CIBER de Cáncer (CB16/12/00275), Co-funded by FEDER from Regional Development European Funds (European Union), Consejeria de Ciencia e Innovacion (CTS-1848), and Consejeria de Salud of the Junta de Andalucia (PI-0096-2014). Especial thanks to the AECC Foundation for supporting this work.

Mantle cell lymphoma (MCL) accounts for approximately 3–6% of all Non-Hodgkin Lymphomas (NHL). Patients’ median age at diagnosis is 68 years [1] with male predominance and median survival of 4–5 ye...

The objectives of our study were to evaluate free amino acid (FAA) concentrations in the serum of patients affected by fibromyalgia syndrome (FMS) and to determine the relationships between FAA levels and FMS clinical parameters. Thus, serum amino acid concentrations were quantified (HPLC analysis) in 23 females with fibromyalgia (according to the American College of Rheumatology classification criteria) and 20 healthy females. The results showed significantly higher serum concentrations of aspartate, cysteine, glutamate, glycine, isoleucine, leucine, methionine, ornithine, phenylalanine, sarcosine, serine, taurine, tyrosine and valine in FMS patients vs. healthy controls. Patients with higher Fibromyalgia Impact Questionnaire (FIQ) scores showed increased levels of alanine, glutamine, isoleucine, leucine, phenylalanine, proline and valine. In conclusion, our results indicate an imbalance in some FAAs in FMS patients. Increased Glu is particularly interesting, as it could explain the deficit in monoaminergic transmission involved in pain.

Objective Recent studies have suggested a possible role of high levels of plasma lysophosphocholines (lysoPCs) in fibromyalgia syndrome (FMS). The aim of this study was to evaluate the content of plasma phospholipases (e.g., Platelet Activating Factor Acetyl Hydrolase [PAF-AH], secretory Phospholipase A2 [sPLA2], Total Antioxidant Capacity [TAOC] and 2,7,8-trimethyl-2-(2-carboxyethyl)-6-hydroxy chroman [γ-CEHC]) in FMS patients and their association with clinical status and quality of life. Methods Thirty-six females meeting the 2011 American College of Rheumatology criteria for the classification of FMS and thirty-four healthy females were enrolled for the study. Plasma enzyme levels were quantified using commercial enzyme-linked-immunosorbent-assay (ELISA). In order to assess the disease severity and the functional status of patients, the Fibromyalgia Impact Questionnarie (FIQ) was used. Results Higher levels of sPLA2 and lower PAF-AH and γ-CEHC were observed in the plasma of FMS patients compared to the controls. A decrease in PAF-AH and TAOC levels were found in severe FMS (S-FMS) compared to mild/slight (MS-FMS) forms. Conclusion The results of the study indicate a possible involvement of phospholipases and γ-CEHC in fibromyalgia syndrome.

We demonstrate that ultrasound field calibration and imaging are achievable in a vertical resonator using digital holography, showing that it is a flexible tool to assist the diffusion of acoustophoresis microfluidic devices.

Fundamental challenges of targeting specific brain regions for treatment using pharmacotherapeutic nanoparticle (NP) carriers include circumventing the blood-brain-barrier (BBB) and tracking delivery. Angiopep-2 (AP2) has been shown to facilitate the transport of large macromolecules and synthetic nanoparticles across the BBB. Thus, conjugation of AP2 to an MS2 bacteriophage based NP should also permit transport across the BBB. We have fabricated and tested a novel MS2 capsid-based NP conjugated to the ligand AP2. The reaction efficiency was determined to be over 70%, with up to two angiopep-2 conjugated per MS2 capsid protein. When linked with a porphyrin ring, manganese (Mn2+) remained stable within MS2 and was MRI detectable. Nanoparticles were introduced intracerebroventricularly or systemically. Systemic delivery yielded dose dependent, non-toxic accumulation of NPs in the midbrain. Design of a multifunctional MRI compatible NP platform provides a significant step forward for the diagnosis and treatment of intractable brain conditions, such as tinnitus.

In this work the profile of primary metabolites in potato (Solanum tuberosum L., cultivar Agria) grown under organic vs traditional farming system was studied, and the influence of heat processing (for producing potato powder) on nutritionally important compounds such as essential amino acids, proteins and carbohydrates was evaluated. The potato powder was obtained by means of a drum dryer using different process parameters (temperature, time and product thickness) to minimise thermal damage on the final product. The effects of these different processes were evaluated by analysing also some other quality indicators, as final moisture, colour of the powder, and rheological properties of the rehydrated puree. Moreover, because of its potential cancerogenity and genotoxicity, acrylamide content was determined in the potato powder obtained and compared with that of its main precursors (asparagine and reducing sugars). The steam pressure of 0.2 MPa and 2 r.p.m. allowed to obtain potato powder with a lower acrylamide contents and still a very low water content that caused no microbiological safety problems. This powder, both from conventional and organic farming, when rehydrated, resulted in purees with minimal nutritional losses and colour and consistency very similar to the fresh ones. © 2011 Elsevier Ltd. All rights reserved. In this work the profile of primary metabolites in potato (Solanum tuberosum L, cultivar Agria) grown under organic vs traditional farming system was studied, and the influence of heat processing (for producing potato powder) on nutritionally important compounds such as essential amino acids, proteins and carbohydrates was evaluated. The potato powder was obtained by means of a drum dryer using different process parameters (temperature, time and product thickness) to minimise thermal damage on the final product. The effects of these different processes were evaluated by analysing also some other quality indicators, as final moisture, colour of the powder, and theological properties of the rehydrated puree. Moreover, because of its potential cancerogenity and genotoxicity, acrylamide content was determined in the potato powder obtained and compared with that of its main precursors (asparagine and reducing sugars). The steam pressure of 0.2 MPa and 2 r.p.m. allowed to obtain potato powder with a lower acrylamide contents and still a very low water content that caused no microbiological safety problems. This powder, both from conventional and organic farming, when rehydrated, resulted in purees with minimal nutritional losses and colour and consistency very similar to the fresh ones. (C) 2011 Elsevier Ltd. All rights reserved.

European Geosciences Union General Assembly 2016, EGU Division Energy, Resources & the Environment (ERE) / Edited by Maria Ask, Viktor Bruckman, Chris Juhlin, Thomas Kempka and Michael Kühn European Geosciences Union General Assembly 2016, EGU Division Energy, Resources & the Environment (ERE), Vienna, Austria, 16 Apr 2016 - 22 Apr 2016; Amsterdam [u.a.] : Elsevier, Energy procedia, 97, 302-309 (2016). doi:10.1016/j.egypro.2016.10.005, Published by Elsevier, Amsterdam [u.a.]

Aspherical and freeform optics are applied to reduce geometrical aberrations as well as to reduce the required number of components, the size and the weight of the system. To measure these optical components with nanometre level uncertainty is a challenge. The NANOMEFOS machine was developed to provide suitable metrology (high accuracy, universal, non-contact, large measurement volume and short measurement time) for use during manufacturing of these surfaces. This paper describes the design, realization and testing of this machine. In particular it describes the design and testing of the air-bearing motion system with parallel stage configuration, and the separate metrology system with Silicon Carbide metrology frame and an interferometry system for direct displacement measurement of the optical probe. Preliminary validation measurements demonstrate the nanometer level repeatability for freeform surface measurement. © 2011 Elsevier Inc.

The feasibility of extracting lignans from flaxseed meal using pressurized low polarity water (PLPW) in a fixed bed extraction cell was evaluated. Optimization experiments were performed using surface response methodology for flow rate, bed depth and solvent to solid ratio (S/S). Maximum yield of lignan was obtained at flow rate of 0.6–2 mL/min, bed depth to internal diameter ratio (L/ID) of 20–25, and S/S ratio of 77–150 mL/g. The effect of flow rate and the mass transfer mechanisms governing the extraction of lignans from flaxseed meal was also evaluated. A diffusion model and a two site kinetic model were used to describe the extraction of lignans from flaxseed meal by PLPW. The two models gave similar quality of data fits. The diffusion model yielded diffusivities ranging from 2 × 10 −13 to 9 × 10 −13 m 2 s −1 for extraction of lignans at 180 °C, pH 9 and a 1:1.5 meal to co-packing material ratio.

We report adhesion, growth, and differentiation of mouse neural cells on ultra-thin films of an organic semiconductor, pentacene. We demonstrate that i) pentacene is structurally and morphologically stable upon prolonged contact with water, physiological buffer, and cell culture medium; ii) neural stem cells adhere to pentacene and remain viable on it for at least 15 days; iii) densely interconnected neural networks and glial cells develop on the pentacene surface after several days. This implies that adhesion proteins secreted by the cells find suitable adsorption loci to anchor the cells. Pentacene is also a suitable substrate for casting thin layers of cell adhesion molecules, such as laminin and poly-L-lysine. Our results show that pentacene, albeit being an aromatic molecule, allows neurons to adhere to and grow on it, which is possibly due to its tightly packed solid state structure. This structure remains unaltered upon exposure to water and interfacial force exerted by the cells. The integration of living cells into organic semiconductors is an important step towards the development of bio-organic electronic transducers of cellular signals from neural networks.

This study examined the application of pressurized low polarity water (PLPW) extraction of lignans, proteins and carbohydrates from defatted flaxseed meal. Key processing conditions included temperature (130, 160, 190 °C), solvent pH (4, 6.5 and 9), solvent to solid ratio (90, 150 and 210 mL/g) and introduction of co-packing material (0 and 3 g glass beads). The addition of 3 g glass beads increased the yields for all target compounds. The maximum yield of lignans (21 mg/g meal) was obtained at 170 °C with solvent to solid ratio of 100 mL/g meal at pH 9. Optimal conditions for protein extraction were pH 9, solvent to solid ratio of 210 mL/g meal and 160 °C. Total carbohydrates recovery was maximized at 215 mg/g meal (50% recovery) at pH 4 and 150 °C with solvent to solid ratio of 210 mL/g meal. The increase of temperature accelerated extraction, thus reducing solvent volume and time to reach equilibrium. For the extraction of proteins and carbohydrates, however, a temperature of 130–160 °C is recommended, as proteins and carbohydrates are vulnerable to thermal degradation.

The application of pressurized low polarity water (PLPW) extraction to the production of flaxseed lignans-rich products has been studied, and the key geometric and process conditions, including temperature, flow rate, and total volume have been determined and optimized. Maximum amounts of lignans and other flaxseed bioactive, including proteins were extracted at 160 °C and 5.2 MPa. However, on a dry weight basis the most concentrated extracts in terms of lignans and other phenolic compounds were obtained at 140 °C and 5.2 MPa. A flow rate of 0.5 mL/min was optimal for the extraction of lignans from flaxseed (Linum usitatissimum L.) and a total volume of 30–40 mL/g of seed was required to maximize the recovery. Higher flow rates increased the rate of the extraction but required larger water volumes. Bed depth to ID ratios of 5–18 resulted in faster extraction and maximum recovery (90–95%) at water to seed ratios of 30–50 mL/g. Larger depth to ID ratios (15–18) would allow the use of lower solvent to solid ratios (14–20 mL/g) and would still result in yields of 84–90%.

Fish is a good source of nutrients for humans but can pose a risk to human health because of the possible presence of some xenobiotics such as heavy metals and persistent organic contaminants. Constant monitoring is needed to minimize health risks and ensure product quality and consumer safety. The aim of the present study was to use atomic absorption spectrometry to determine the concentrations of some heavy metals (Hg, Pb, and Cd) in tuna packaged in different kinds of packages (cans or glass) in various countries (Italy and elsewhere). Concentrations of Cd and Hg were within the limits set by European Commission Regulation (EC) No 1881/2006 and in many samples were below the detection limit. Pb concentrations exceeded European limits in 9.8% of the analyzed samples. These results are reassuring in terms of food safety but highlighted the need to constantly monitor the concentrations of heavy metals in fish products that could endanger consumer health.

Alcohol abuse is a substantial and growing health problem in Western societies. In the last years in vivo and in vitro studies have suggested that males and females display a different alcohol drinking behaviour, with swingeing differences not only in the propensity for alcohol use but also in the metabolic and behavioural consequences. In this study we investigated, in adult female rats, ethanol self-administration and preference pattern using a 3-bottle paradigm with water, 10% ethanol solution, and white wine (10%, v/v), along a four-week period. The influence of alcohol free-access on explorative behaviour in the open field (OF), and on spatial learning and reference memory in the Morris water maze (MWM) were also evaluated. Our results indicate that: (i) female rats show a higher preference for alcohol, in the first two weeks of the paradigm, displaying a higher consumption of 10% ethanol solution than white wine; in the last two weeks, they reduce their alcoholic preference, drinking the same moderate amounts of the two alcoholic beverages; (ii) at the fourth week of the free-access paradigm rats show a lower explorative behaviour in the open field and a worsening in spatial memory retention in the Morris water maze. In conclusion our data suggest that, despite the ability to self-regulate alcohol intake, female rats suffer from relevant impairments in spatial memory retention and cognitive flexibility, displaying a sexually dimorphic modification in the adaptive strategies.

The growth of Lactobacillus plantarum WCFS1 and of its ΔccpA ery mutant, WCFS1-2, was compared in batch fermentations in a complex medium at controlled pH (6.5) and temperature (30 °C) with or without aeration, in order to evaluate the effect of ccpA inactivation and aeration on growth, metabolism and stress resistance. Inactivation of ccpA and, to a lesser extent, aeration, significantly affected growth, expression of proteins related to pyruvate metabolism and stress, and tolerance to heat, oxidative and cold/starvation stresses. The specific growth rate of the mutant was ca. 60% of that of the wild type strain. Inactivation of ccpA and aerobic growth significantly affected yield and production of lactic and acetic acid. Stationary phase cells were more stress tolerant than exponential phase cells with little or no effect of inactivation of ccpA or aeration. On the other hand, for exponential phase cells inactivation of ccpA impaired both heat stress and cold/starvation stress, but increased oxidative stress tolerance. For both strains, aerobically grown cells were more tolerant of stresses. Evidence for entry in a viable but non-culturable status upon prolonged exposure to cold and starvation was found. Preliminary results of a differential proteomic study further confirmed the role of ccpA in the regulation of carbohydrate catabolism and class I stress response genes and allow to gain further insight on the role of this pleiotropic regulator in metabolism and stress. This is the first study in which the impact of aerobic growth on stress tolerance of L. plantarum is evaluated. Although aerobic cultivation in batch fermentations does not improve growth it does improve stress tolerance, and may have significant technological relevance for the preservation of starter and probiotic cultures.

Lactobacillus plantarum is a facultative heterofermentative lactic acid bacterium widely used in the production of most fermented food due to its ability to thrive in several environmental niches, including the human gut. In order to cope with different growth conditions, it has developed complex molecular response mechanisms, characterized by the induction of a large set of proteins mainly regulated by HrcA and CtsR repressors as well as by global regulators such as carbon catabolite control protein A (CcpA).In this study, the role of CcpA in the regulation of growth under anaerobiosis and aerobiosis, and the adaptation to aeration in L. plantarum WCFS1 were comprehensively investigated by differential proteomics. The inactivation of ccpA, in both growth conditions, significantly changed the expression level of 76 proteins, mainly associated with carbohydrate and energy metabolism, membrane transport, nucleotide metabolism, protein biosynthesis and folding. The role of CcpA as pleiotropic regulator was particularly evident at the shift from homolactic fermentation to mixed fermentation. Proteomic results also indicated that the mutant strain was more responsive to aerobic growth condition. © 2012 Elsevier B.V.

Anthocyanins are the largest group of water-soluble pigments in the plant kingdom. They are responsible for most of the red, blue, and purple colors of fruits, vegetables, flowers, and other plant tissues or products. The analysis of anthocyanins is complex as a result of their ability to undergo structural transformations and complexation reactions. In addition, they are difficult to measure independently of other flavonoids, as they have similar structural and reactivity characteristics. Anthocyanins are generally extracted with weakly acidified alcohol-based solvents, followed by concentration (under vacuum), and purification of the pigments. Paper and/or thin-layer chromatography and UV-Vis spectroscopy have traditionally been used for the identification of anthocyanins. Capillary zone electrophoresis, a hybrid of chromatography and electrophoresis, is gaining popularity for the analysis of anthocyanins; however, liquid chromatography (LC) has become the standard method for identification and separation in most laboratories and may be used for both preparative and quantitative analysis. LC with mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy are possibly the most powerful methods for the structural elucidation of anthocyanins available, to date. At present, the most satisfactory method for mixture analysis is the multistep method of separation, isolation, and quantification by LC with peak identification by MS and high-field NMR.

The HOOO− anion, the conjugate base of dihydrogen trioxide, HOOOH, is suggested as a key intermediate in ozonation reactions relevant to atmospheric chemistry, water purification, preparative organic chemistry, etc. Whereas recent theoretical calculations characterize HOOO− as a long-lived metastable species, so far the anion proved experimentally elusive, defying attempts at its direct detection even in solution, where it should be stabilized by solvation. We report the preparation and the proof-positive detection of HOOO− as an isolated species with a lifetime >0.8 μs. The preparation proved particularly difficult, since the only viable approach is the indirect route involving the preliminary formation of HOOO upon neutralization of HOOO+, followed by electron attachment to the radical, utilizing neutralization–reionization ( + NR − ) mass spectrometry. Direct two-electron reduction of the HOOO+ cation to the HOOO− anion by the vertical process typical of charge reversal (CR) experiments proved inefficient. This observation, and application of neutral and ion decomposition difference (NIDD) mass spectrometry, pioneered by Schwarz and coworkers [Int. J. Mass Spectrom. Ion Processes 172 (1998) 181] suggest that the geometries of HOOO+ and HOOO− are significantly different, whereas that of the HOOO radical is structurally intermediate between them, which accounts for the diversity of the Franck–Condon factors characterizing the mutual vertical transitions of the three species. This inference is consistent with the available optimized geometries of HOOO and HOOO+, computed at various levels of theory, and with that of the anion recently reported by Plesnicar et al. [J. Am. Chem. Soc. 124 (2002) 8462].

Extraction of anthocyanins from black currants using aqueous ethanol was optimized for yield and antioxidant activity. The process variable having the most effect on the extraction was the solvent to solid ratio, which increased phenolic extraction in the whole range from 0 to 19 L/kg. Total phenolics increased with ethanol concentration up to a maximum at about 60% and then decreased with further increase in solvent concentration irrespective of the solvent to solid ratio. Temperature only affected the extraction of anthocyanins. Increasing the temperature beyond 30 to 35 °C resulted in degradation of anthocyanins and reduction of yields. Variation in extract composition was not sufficiently large to affect antioxidant activity.

Chronic ethanol consumption is able to modify emotional behaviour and cognition in humans. In particular, the effects exerted by alcohol may depend on doses, time and modalities of administration. In this study we investigated, in adult male rats, ethanol self-administration and preference patterns using a 3-bottle choice paradigm with water, 10% ethanol solution, and white wine (10%, v/v), along a four-week period. The influence of alcohol free-access on novelty-induced explorative behaviour in the open field, and on spatial learning and reference memory in the Morris water maze was also evaluated. Our results indicate that: (i) rats show a higher preference for alcohol, in the first two weeks of the paradigm, displaying a higher consumption of 10% ethanol solution than white wine; in the last two weeks, they reduce their alcoholic preference, drinking the same moderate amounts of the two alcoholic beverages; (ii) at the fourth week of the free-access paradigm rats show a high explorative behaviour in the central squares of the open field and an improvement in spatial information processing in the new-place learning task of the Morris water maze. In conclusion our data suggest that, interestingly, rats exposed to the free-access paradigm were able to self-regulate their alcoholic intake, and indicated that a moderate alcohol consumption was able to induce an increase in behavioural reactivity and an enhancement in spatial learning flexibility.

BACKGROUND AND METHODS Ethanol (EtOH) activates hypothalamic-pituitary-adrenal (HPA) axis, resulting in adrenocorticotropin hormone, glucocorticoid release, and in modifications of the response of the axis to other stressors. The initial site of EtOH action within the HPA system seems to be the hypothalamus. Thus, to determine the mechanisms responsible for these effects, we investigated: (i) whether EtOH was able to release corticotrophic releasing hormone (CRH) from incubated hypothalamic explants; (ii) whether acetaldehyde (ACD), its first metabolite formed in the brain by catalase activity, might play a role in EtOH activity. To this aim, rat hypothalamic explants were incubated with: (i) medium containing EtOH at 32.6 x 10(3) microM; (ii) different concentration of ACD (1, 3, 10, and 30 microM); (iii) EtOH plus 3amino-1,2,4-triazole (3AT, 32 x 10(3) microM) an inhibitor of cerebral catalase; (iv) ACD plus D-penicillamine (DP, 50.3 x 10(3) microM) an ACD-trapping agent. CRH levels were evaluated by a radioimmunoassay. RESULTS Incubation with EtOH induced a 7-fold increase in CRH secretion, with respect to basal levels; ACD was able to stimulate CRH release in a dose-dependent manner; the inhibition of cerebral catalase by 3AT blocked EtOH-induced CRH outflow; the inactivation of ACD by DP reverted the ACD-stimulating effect on CRH secretion. CONCLUSIONS These data show that both EtOH and acetaldehyde are able to increase hypothalamic CRH release from the rat hypothalamus and that acetaldehyde itself appears to be the mediator of EtOH activity.

Background: The cancer chaperone, Heat Shock Protein 90 (HSP90), is known for its ability to regulate the stability of different oncogenic proteins. Thus, its overexpression has been related to unfavorable prognosis in some types of tumors. EGFR and EML4-ALK, two of the most important drivers in non-small cell lung cancer (NSCLC), are HSP90 clients and extremely depend on it. As a consequence, this chaperone is especially important in NSCLC hence HSP90 inhibition shows a lot of possibilities to future treatments in this tumor type. Nevertheless, to obtain a successful clinical development, will be essential supporting evidence of inhibitory efficacy in several molecularly defined subgroups of NSCLC. Methods: NSCLC cell lines carrying different gene mutations, whose direct (HCC827: EGFR mutated and H3122: EML4-ALK rearrangement) and indirect (A549: KRAS mutated) relationship with HSP90 has been reported, were used. In these cell lines, along with H1781 (EGFR, KRAS, ALK wild type) as control, the activity of the chaperone studied was interrupted. To this end, pharmacological inhibition of HSP90 was achieved through geldanamycin and resorcinol derivates. First, western blotting was carried out to confirm the effect of this inhibition. Later, to identify a proteomic profile associated with HSP90 inhibition, two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) of selected cell lines were performed. Results: The expression of the oncogenic HSP90 client proteins studied was decreased by the inhibitors in the NSCLC cell lines. The oncoproteins drivers EGFR and EML4-ALK showed a strong dependence on HSP90 observed through a high sensitivity of the cell lines HCC827 and H3122 to the inhibition. Therefore, these cell lines were selected with the purpose of identify a proteomic signature linked to HSP90 inhibition by 2D-PAGE. The untreated EGFR positive cell line presented 104 protein spots significantly up-regulated, whereas 80 spots were down-regulated compared to inhibited cell line. Meanwhile, the cell line harboring the EML4-ALK translocation showed 16 spots up-regulated and 5 down-regulated in the untreated versus inhibited cell line. In addition HSP70 overexpression, compared to unprocessed cell lines, was detected after treatments. This feedback, previously reported, confirmed HSP90 inhibition in the two cell lines studied. Conclusions: The evidence of treatment response, in the cell lines studied, was showed through oncogenic client proteins reduction as well as HSP70 induction. The proteomic profiles identified, of HSP90 inhibited and untreated tumor cells, revealed several deregulated pathways involved in the tumorigenesis. Citation Format: Angela Marrugal, Irene Ferrer, Maria Dolores Pastor, Alvaro Quintanal, Antonio Lucena-Cacace, Amancio Carnero, Luis Paz-Ares, Sonia Molina-Pinelo. HSP90 inhibition leads to a differential proteomics profile in non small cell lung cancer cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2207. doi:10.1158/1538-7445.AM2017-2207

Artificial bioreactors are desirable for in vitro biochemical studies and as protocells. A key challenge is maintaining a favourable internal environment while allowing substrate entry and product departure. We show that semipermeable, size-controlled bioreactors with aqueous, macromolecularly crowded interiors can be assembled by liposome stabilization of an all-aqueous emulsion. Dextran-rich aqueous droplets are dispersed in a continuous polyethylene glycol (PEG)-rich aqueous phase, with coalescence inhibited by adsorbed ~130-nm diameter liposomes. Fluorescence recovery after photobleaching and dynamic light scattering data indicate that the liposomes, which are PEGylated and negatively charged, remain intact at the interface for extended time. Inter-droplet repulsion provides electrostatic stabilization of the emulsion, with droplet coalescence prevented even for submonolayer interfacial coatings. RNA and DNA can enter and exit aqueous droplets by diffusion, with final concentrations dictated by partitioning. The capacity to serve as microscale bioreactors is established by demonstrating a ribozyme cleavage reaction within the liposome-coated droplets.

Background/Aims: we have investigated whether cultured cardiomyocytes of the cell line HL-1 have the ability to perform regulatory volume responses both in hypotonic and hypertonic conditions. Furthermore, we characterized those regulatory responses and studied the effects of bumetanide and DIDS in volume regulation of HL-1 cells. Methods: we used a light scattering system to measure the transient volume changes of HL-1 cells when subjected to osmotic challenge. Results: We found that HL-1 cells correct for their volume excess by undergoing regulatory volume decrease (RVD), and also respond to hypertonic stress with a regulatory volume increase (RVI). Rate of RVD was 0.08 ± 0.04 intensity/min, and rate of RVI was 0.09 ± 0.01 intensity/min. Volume recovery was 83.68 ± 5.73 % for RVD and 92.3 ± 2.3 % for RVI. Bumetanide 50 µM inhibited volume recovery, from 92.3 ± 2.3 % (control) to 24.6 ± 8.8 % and reduced the rate of RVI from 0.070 ± 0.020 intensity/min (control) to 0.010 ± 0.005 intensity/min. 50 µM DIDS reduced volume recovery to 42.93 ± 7.7 % and rate of RVI, to 0.03 ± 0.01 intensity/min. Conclusions: these results suggest that bumetanide- and DIDS-sensitive mechanisms are involved in the RVI of HL-1 cells, which points to the involvement of the Na + / K + /2Cl - cotransporter and Cl - /bicarbonate exchanger in RVI, respectively.

The reactivity of XeF + ions towards methanol was investigated in the gas phase by a combination of mass spectrometric techniques and theoretical methods. The charged reagent, generated from the chemical ionization of xenon difluoride, undergoes both fluorine and xenon transfer to methanol, following dissociation of primary (CH 3 OH)XeF + electrostatic adducts. Structural analysis performed by low-energy collisionally activated dissociation and reactivity probing techniques, and the results of calculations carried out up to the B3LYP/CCSD(T) level of theory, concur in suggesting the formation of two different primary adducts of O-Xe-F and O-F-Xe connectivity that evolve into CH 3 O(H)Xe + and CH 3 O(H)F + , upon loss of F and Xe, respectively. Evidence was obtained for the structure of the fluorinated species, identified as O-protonated methyl hypofluorite. This finding allowed the first experimental evaluation of the proton affinity of CH 3 OF, which falls in the range 648.0-680.5 kJ mol -1 , not inconsistent with the 644.8 kJ mol -1 value calculated at the CCSD(T) level and 298 K. The proton affinity of methyl hypoxenite, CH 3 OXe, was theoretically calculated to amount to 747.7 kJ mol -1 .

Reverse gyrase is a peculiar DNA topoisomerase, specific of thermophilic microorganisms, which induces positive supercoiling into DNA molecules in an ATP-dependent reaction. It is a modular enzyme and comprises an N-terminal helicase-like module fused to a C-terminal topoisomerase IA-like domain. The exact molecular mechanism of this unique reaction is not understood, and a fundamental mechanistic question is how its distinct steps are coordinated. We studied the cross-talk between the components of this molecular motor and probed communication between the DNA-binding sites and the different activities (DNA relaxation, ATP hydrolysis and positive supercoiling). We show that the isolated ATPase and topoisomerase domains of reverse gyrase form specific physical interactions, retain their own DNA binding and enzymatic activities, and when combined cooperate to achieve the unique ATP-dependent positive supercoiling activity. Our results indicate a mutual effect of both domains on all individual steps of the reaction. The C-terminal domain shows ATP-independent topoisomerase activity, which is repressed by the N-terminal domain in the full-length enzyme; experiments with the isolated domains showed that the C-terminal domain has stimulatory influence on the ATPase activity of the N-terminal domain. In addition, the two domains showed a striking reciprocal thermostabilization effect.

Gaseous (FCN)H+ ions were obtained by protonation of cyanuric fluoride by strong Bronsted acids such as H3+ (D3+) and CnH5+ (n = 1, 2), and their structure and reactivity were examined by FT-ICR mass spectrometry and computational methods at the G2 level of theory. The results show that FCNH+ is considerably more stable than the HFCN+ protomer, the gap amounting to 360 kJ mol-1 at the G2 level. The gas-phase basicity (GB) of FCN was estimated by “bracketing” experiments by measuring the efficiency of H+ transfer from FCNH+ to reference bases. Utilizing a general correlation between the rate and the ΔG° change of the proton-transfer we obtain GB(FCN) = 668.4 ± 10 kJ mol-1, to be compared with the G2 computed GB of 651.8 ± 8 kJ mol-1, both values being referred to N protonation. The reactions of FCNH+ with isolated (C2H4) and conjugated (C6H6) π systems were also investigated. The ion behaves as a FC+ = NH electrophile promoting aromatic cyanation, e.g., yielding C6H5CNH+ from benzene. Consideration is give...

The positive ion chemistry of peroxynitric acid ( 1 ) was investigated in the gas phase by mass-analyzed ion kinetic, collisionally activated dissociation, and Fourier transform-ion cyclotron resonance mass spectrometric techniques and theoretical methods up to the B3LYP/6-311++g(3 df ,2 pd ) and G2, i.e. QCISD(T)/6-311+g(3 df ,2 pd ), levels. The ion–neutral complex HOOH–NO 2 + ( 1a ) is the only detectable protomer in CI experiments involving the protonation of 1 by H 3 O + , and can also be obtained from the reaction of NO 2 + with H 2 O 2 . 1a behaves as a protonating and nitrating agent toward gaseous nucleophiles. The experimental proton affinity of 1 is estimated to be 176 ± 3 kcal mol −1 , in excellent agreement with the 175 ± 2 kcal mol −1 G2 PA. The theoretical results show that 1a is more stable than the HOONO 2 H + ( 1b ) and the H 2 OONO 2 + ( 1c ) protomers by 13 and 16 kcal mol −1 , respectively, at the B3LYP level of theory, and account for the exclusive formation of 1a in the CI experiments. The experimental and B3LYP theoretical binding energy of NO 2 + to H 2 O 2 amounts to 18 ± 2 kcal mol −1 .

Recent mass spectrometric studies of simple inorganic species (both charged and neutral) of main-group elements are reviewed, focusing attention on radicals and ions of interest to the chemistry of the atmosphere and its pollution. The examples illustrated concern the detection of HO 3 , hydrogen trioxide, the O 2 /O 3 isotope exchange and its charged intermediate O 5 + , the reactions promoted by ionization of ozone/halocarbon mixtures in atmospheric gases, the ion chemistry of NO x oxides, and that of elemental chlorine and chlorine fluoride. Among the results of specific interest to gas-phase ion chemistry, the examples illustrated concern the intracluster ligand-switching reactions in ternary NO + complexes, the NO 2 + reactivity towards ethylene and acetylene, the gas-phase basicity of Cl 2 , the formation and characterization of Cl 2 X + ions (X = Cl, F) and of [H 3 C-Cl-Cl] + , a new isomer of protonated dichloromethane.

A phenomenological theory of salt-induced Hofmeister phenomena is presented, based on a relation between protein solubility in salt solutions and protein-water interfacial tension. As a generalization of previous treatments, it implies that both kosmotropic salting out and chaotropic salting in are manifested via salt-induced changes of the hydrophobic/hydrophilic properties of protein-water interfaces. The theory is applied to describe the salt-dependent free energy profiles of proteins as a function of their water-exposed surface area. On this basis, three classes of protein conformations have been distinguished, and their existence experimentally demonstrated using the examples of bacteriorhodopsin and myoglobin. The experimental results support the ability of the new formalism to account for the diverse manifestations of salt effects on protein conformation, dynamics, and stability, and to resolve the puzzle of chaotropes stabilizing certain proteins (and other anomalies). It is also shown that the relation between interfacial tension and protein structural stability is straightforwardly linked to protein conformational fluctuations, providing a keystone for the microscopic interpretation of Hofmeister effects. Implications of the results concerning the use of Hofmeister effects in the experimental study of protein function are discussed.

A mechanistic link has been found between the gas-phase preparation of ethylenexenonium ions and the reactions of unsaturated compounds with XeF+ salts, utilized in solution as stereoselective fluorinating agents. Complex C2H4XeF+ (2) is the charged intermediate in the formation process of C2H4Xe+ (1) from XeF+ and ethylene. This was evidenced through FT-ICR and TQ mass spectrometry, complemented by DFT calculations at the BLYP/DZVP//CCSD(T) level of theory.

Gaseous [HN2O5]+ ions formed upon addition of NO2+ to nitric acid have been studied by mass spectrometric and computational methods. The results from MIKE, CAD and FT-ICR spectrometry and calculations at the B3LYP 6-311++G(3df, 3dp)//6-311G(d,p) level of theory show that the most stable adduct formed is an electrostatic HNO3·NO2+ complex where NO2+ is coordinated to the nitro group of nitric acid. Consequently, the nitro group is the energetically preferred protonation site of N2O5, whose experimental proton affinity (PA) amounts to 189.8 ± 2 kcal mol-1, vs theoretically computed values ranging from 182 to 188 kcal mol-1. Addition of NO2+ to XNO2 molecules (X = CH3O, C2H5O and NH2) also yields electrostatic complexes where the nitronium ion is coordinated to the NO2 group. The most stable [HN2O4]+ ion from the addition of NO+ to HNO3 is also identified as a cluster characterized by coordination of the nitrosonium ion to the nitro group, whose almost thermoneutral isomerization into a cluster where a nitro...

Fibromyalgia Syndrome (FMS) is a chronic disease characterized by widespread pain, and difficult to diagnose and treat. We analyzed the plasma metabolic profile of patients with FMS by using a metabolomics approach combining Liquid Chromatography-Quadrupole-Time Of Flight/Mass Spectrometry (LC-Q-TOF/MS) with multivariate statistical analysis, aiming to discriminate patients and controls. LC-Q-TOF/MS analysis of plasma (FMS patients: n = 22 and controls: n = 21) identified many lipid compounds, mainly lysophosphocholines (lysoPCs), phosphocholines and ceramides. Multivariate statistical analysis was performed to identify the discriminating metabolites. A protein docking and molecular dynamic (MD) study was then performed, using the most discriminating lysoPCs, to validate the binding to Platelet Activating Factor (1-alkyl-2-acetyl-sn-glycero-3-phosphocholine, PAF) Receptor (PAFr). Discriminating metabolites between FMS patients and controls were identified as 1-tetradecanoyl-sn-glycero-3-phosphocholine [PC(14∶0/0∶0)] and 1-hexadecanoyl-sn-glycero-3-phosphocholine [PC(16∶0/0∶0)]. MD and docking indicate that the ligands investigated have similar potentialities to activate the PAFr receptor. The application of a metabolomic approach discriminated FMS patients from controls, with an over-representation of PC(14∶0/0∶0) and PC(16∶0/0∶0) compounds in the metabolic profiles. These results and the modeling of metabolite-PAFr interaction, allowed us to hypothesize that lipids oxidative fragmentation might generate lysoPCs in abundance, that in turn will act as PAF-like bioactivators. Overall results suggest disease biomarkers and potential therapeutical targets for FMS.

Kinase suppressor of Ras (KSR) is an evolutionarily conserved component of Ras-dependent signaling pathways. Here, we find that murine KSR (mKSR1) translocates from the cytoplasm to the plasma membrane in the presence of activated Ras. At the membrane, mKSR1 modulates Ras signaling by enhancing Raf-1 activity in a kinase-independent manner. The activation of Raf-1 is mediated by the mKSR1 cysteine-rich CA3 domain and involves a detergent labile cofactor that is not ceramide. These findings reveal another point of regulation for Ras-mediated signal transduction and further define a noncatalytic role for mKSR1 in the multistep process of Raf-1 activation.

Summary: While a variety of immunologic factors are likely to be involved in the initial acute phlogistic events in IgA nephropathy (IgAN), progression to renal failure occurs through a relatively silent course histologically related to prevalent sclerotic changes. In this phase of IgAN, haemodynamic mechanisms are likely to play the most important role. Among the others, angiotensin II and endothelin 1 are thought to be critical mediators. Angiotensin II promotes mesangial cell contraction, modulates membrane permselectivity and induces glomerular hypertension. Evidence for a local angiotensin II hyperactivity in IgAN (especially in patients at definite risk of progression) has been previously provided by our group. Endothelin 1 is mitogenetic for mesangial cells and increases matrix production. Moreover, it exerts local vasoconstrictor effects and induces mesangial cell contraction, both resulting in glomerular afterload. Again, plasma and urinary endothelin 1 are elevated in IgAN. Moreover, those patients with increased risk of progression show an increased ratio between urinary endothelin 1 and cyclic guanosine 3′,5′ monophosphate (GMP), a local messanger with counterbalancing effects. Several data of clinical benefits of angiotensin-converting enzyme inhibitors in IgAN have been reported. Less is known about the intrinsic mechanism of the antiproteinuric action, either reduction of filtration fraction or changes in glomerular permselectivity. The clinical usefulness of factors counterbalancing endothelin 1 effects, such as nitric oxide donors, remains to be established. A sample of eight IgAN patients with poor prognostic indicators was enrolled to examine the renal haemodynamic effects of angiotensin receptor antagonism, angiotensin-converting enzyme inhibition and administration of an exogenous source of nitric oxide. Study periods (7 days each) were randomized and spaced out by one week wash-out. Data showed that angiotensin receptor antagonism and angiotensin-converting enzyme inhibition were equally effective in modifying renal haemodynamics, but only the latter condition significantly reduced albuminuria, possibly indicating a prevalent action on membrane permselectivity in the short time period of the study. The antiproteinuric effects of nitric oxide donors were strictly confined to patients with increased urinary endothelin 1/cyclic GMP ratio. Finally, each treatment condition was found to be associated with an increased nitric oxide production, which appeared to be an additional factor in modulation of filtration fraction changes.

In this research compositional changes of tinplate-canned tomato purées, with or without the addition of essential onion oil were investigated. The study was focused on the analyses of carbohydrates and carboxylic acids in two groups of canned samples (with or without nitrates) to determine whether their chemical composition was affected with storage time. The measurements were performed by high performance liquid chromatography, during six months of storage. The contents of glucose, fructose and two major organic acids, citric and malic, were found in the concentration range 1.77–1.97%, 1.86–2.09%, 0.60–0.75% and 0.23–0.30%, respectively, in all canned samples. Compared to carbohydrates and organic acids, amino acids were found in minor quantities, among them, as most abundant ones were glutamic acid, arginine, aspartic and γ-amino butyric acids. The results show that contents of carbohydrates and carboxylic acids are significantly affected by the change of storage time in majority of analyzed samples. The results also indicated that the influence of essential onion oil on composition of canned tomato purée is within the range of changes due to storage time measured for all other types of cans. Therefore the addition of essential onion oil as natural efficient corrosion inhibitor, as it was found in our previous work, can be recommended for canned tomato purée.

To evaluate serum purine metabolite concentrations in patients affected by fibromyalgia syndrome (FMS) and the relationships between their levels and FM clinical parameters.Serum purine levels were quantified using LC/UV-vis in 22 fibromyalgic females (according to the American College of Rheumatology classification criteria) and 22 healthy females.Significantly higher serum inosine, hypoxanthine and xanthine levels (p0.001) and significantly lower serum adenosine (p0.05) were detected in the FMS patients vs healthy controls. Our data show a negative correlation between adenosine and Fibromyalgia Impact Questionnaire (FIQ).Study results suggest that purines, in particular adenosine and inosine, may be involved in pain transmission in fibromyalgia.

On etudie l'alkylation du biphenyle et des methyl biphenyles par (CH 3 ) 2 F + et (CH 3 ) 2 Cl +

The structure, stability, interconversion, and unimolecular decomposition processes of gaseous protonated nitrosyl fluoride, ONF, have been investigated by mass-analyzed ion kinetic energy (MIKE) and collisionally activated dissociation (CAD) mass spectrometry, as well as ab initio GAUSSIAN-1 calculations. Positive evidence has been obtained for the existence of two distinct, not easily interconvertible isomers, HO-NF + , 1, and ON-FH + , 3. These two species are distinguishable by both CAD and MIKE spectrometry, whose results are consistent with the theoretical description of the system. The fluorine-protonated isomer 3 is the global minimum on the surface and is more stable than the oxygen-protonated isomer 1 by 70.1 kcal mol -1