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Funding Acknowledgements Type of funding sources: Foundation. Main funding source(s): Velux Stiftung Background Ageing of cardiomyocytes (CM) involves structural and functional adverse remodelling that finally could result in heart failure (HF) insurgence, which incidence rise along with age (1). Current medical therapies for HF may not always be tolerated in elder patients(2). Having shown that cardiac progenitor cells (CPCs) secrete nanoparticles named exosomes (EXO) enriched of cardioprotective factors(3,4), we are exploring EXO’s capacity to ameliorate senescence-derived modification into CMs. However, human models of in vitro cardiac aging are currently missing(5). Aim This study exploits CMs derived from human induced pluripotent stem cells (hiPSCs) as an in vitro model for cardiac senescence, that will be used as platform to characterize mechanisms involved in cardiac ageing and to test protective effect of CPC-derived EXO. Methods Patient-derived CPCs were reprogrammed into hiPSCs and subsequently expanded and differentiated into cardiomyocytes (hiPSC-CMs). Senescence-like phenotype was induced by short exposure (3 hours) to doxorubicin (DOX) at sub-lethal concentration (0.2 µM), followed by washing and medium change. Following DOX exposure, cells were exposed to EXO, derived from the purification of conditioned culture media of CPCs using an ultracentrifugation-based isolation method and quantified and sized using a NTA counter. Senescence induction was highlighted by protein and gene expression analysis and senescence-associated b-galactosidase (SA-β-gal) assay.Electrical activity of hiPSC-CMs was evaluated recording extracellular field potentials through multi-microelectrode arrays (MEA) and by single cell patch clamp. Metabolic features were analysed with western blot, real time RT-PCR and specific biochemical assays. Results DOX treatment in hiPSC-CMs induced senescence, as confirmed by activation of p21 and p16 pathways and increasing of SA-β-gal staining as compared to untreated cells (CTR). Biochemical and gene expression analysis revealed an increased ROS production and a reduction in mitochondrial potential, which drives a strong decrease in the ATP/AMP ratios. Real Time PCR analysis reveal an increased transcription of molecules related to the senescence associated secretory phenotype in DOX-CMs. Moreover, DOX-CMs showed impaired Ca++ handling, prolonged multicellular QTc and single cell APD, with increased APD variability and delayed afterdepolarizations (DADs) incidence in comparison to CTR. EXO treatment mitigated the senescent phenotype induced by DOX, as shown by a decreased ROS induction, higher mitochondrial potential which drives a restored ATP/AMP ratio. Furthermore, DOX-induced QTc prolongation was prevented by EXO treatment. Conclusion Our hiPSC-CMs based cellular model recapitulates the phenotype of aged CMs in terms of senescence markers, electrical and metabolic proprieties. CPC-derived EXOs limit age-related modifications, highlighting the cardioprotective role of small molecules released by EXO.

The astrophysical community is currently focusing its efforts in the development of a new generation of gamma-ray telescopes to detect low-energy photons in the MeV-GeV energy range, operating both in the Compton and pair conversion regimes. The reconstruction of the incident photons energy and direction is not straightforward, as the range of secondary particles produced by photon interactions is usually short. We propose a detector consisting of a tracker system based on scintillating fibers and of a Compton-pair imaging calorimeter made of CsI(Na) crystals coupled to wavelength shifting (WLS) fibers read out by Silicon Photomultiplier (SiPM) arrays. We have developed a dedicated simulation code to study the performance of this detector. The simulation takes into account the optical photon production and propagation inside the fibers and is used to optimize the fiber geometrical and optical properties and the design of the readout system.

Satellite experiments for gamma-ray and cosmic-ray detection employ plastic scintillators to discriminate charged from neutral particles in order to correctly identify gamma-rays and charged nuclei. The High Energy Cosmic Radiation Detection (HERD) facility will be among these experiments, to be installed onboard the future Chinese Space Station (CSS), to detect cosmic-rays and gamma-rays up to TeV energies. The plastic scintillator detector (PSD) will consist of scintillator tiles or bars coupled to Silicon Photomultipliers (SiPMs). To discriminate gamma-rays from charged particles and measure the ion charge up to iron nuclei a wide dynamic range is required, from few tens up to thousands of photoelectrons. We have equipped a plastic scintillator tile prototype with SiPMs produced by Hamamatsu and AdvanSiD and coupled their analog signals to the DT5550W board based on the CITIROC ASIC, produced by CAEN SpA. The CITIROC ASIC allows both the formation of a fast trigger with a configurable threshold and the digitization of analog waveforms after a preamplification and shaping stage along two paths with different gain settings. The performance of our prototype will be shown.

Plastic scintillators are widely used for anti-coincidence systems and for the identification of charged cosmic-ray nuclei in satellite experiments. For this reason, a plastic scintillator detector (PSD) should have a high detection efficiency for charged cosmic rays and a very good capability of measuring charges. We implemented a full and customizable simulation tool to investigate the performance of a PSD coupled to Silicon Photomultipliers. The overall performance of the detector is studied by tracking optical photons produced inside the scintillator. The simulation will be used for the design of a PSD for future space experiments, such as HERD, AMEGO, e-Astrogam. In this work we investigated in detail the effect of Birks’ saturation in the discrimination of charged ions up to iron nuclei. We will show the comparison between simulations and measurements conducted on prototype scintillator tiles.

Ospemifene is a selective estrogen-receptor modulator approved for treating menopause-related moderate to severe dyspareunia and vaginal dryness, symptoms of vulvovaginal atrophy (VVA), in the United States, and for treating menopause-related, symptomatic VVA in women not appropriate for local estrogen therapy in Europe. This review summarizes the effects of ospemifene on bone, including bone biomarker data from a phase 3 vaginal dryness study. Early-phase studies of postmenopausal women showed that ospemifene dose-dependently decreased bone turnover markers versus placebo, similar to raloxifene. A 12-week, phase 3 study of ospemifene 60 mg/day in postmenopausal women showed improvements in all VVA parameters and significantly greater decreases in seven of nine bone biomarkers versus placebo. Lower bone resorption markers with ospemifene were observed regardless of time since menopause (≤5 years or5 years) or baseline bone mineral density (BMD) (normal [

Dantrolene is an orphan drug representing the sole therapeutic treatment for malignant hyperthermia, a life-threatening pathology affecting 0.2 in every 10,000 people in the EU. [...]

The Pan-Asian REVIVE survey aimed to examine women's experiences with genitourinary syndrome of menopause (GSM) and their interactions with health-care professionals (HCPs).Self-completed surveys were administered face-to-face to 5992 women (aged 45-75 years) in Indonesia, Malaysia, Singapore, Taiwan, and Thailand.Of 638 postmenopausal women with GSM symptoms, only 35% were aware of the GSM condition, most of whom first heard of GSM through their physician (32%). The most common symptoms were vaginal dryness (57%) and irritation (43%). GSM had the greatest impact on sexual enjoyment (65%) and intimacy (61%). Only 25% had discussed their GSM symptoms with a HCP, and such discussions were mostly patient-initiated (64%) rather than HCP-initiated (24%). Only 21% had been clinically diagnosed with GSM and only 24% had ever used treatment for their symptoms. Three-quarters of those who had used treatment for GSM had discussed their symptoms with a HCP compared to only 9% of those who were treatment-naïve.GSM is underdiagnosed and undertreated in Asia. As discussion of GSM with HCPs appears to be a factor influencing women's awareness and treatment status, a more active role by HCPs to facilitate early discussions on GSM and its treatment options is needed.

Management of plant parasitic nematodes with nematode predators, parasites or antagonists is an eco-friendly approach that may avoid the problems arisen by the use of toxic chemicals. Fungi belonging to Trichoderma spp. are well known in literature for their role in control of plant parasitic nematodes. Root-knot nematodes (RKNs), Meloidogyne spp., are obligate parasites that cause the formation of familiar galls on the roots of many cultivated plants. The interaction between the M. incognita motile second stage juveniles (J2s) and the isolate ITEM 908 of Trichoderma harzianum was examined in its effect on the nematode infestation level of susceptible tomato plants. To gain insight into the mechanisms by which ITEM 908 interacts with nematode-infected tomato plants, the expression patterns of the genes PR1 (marker of Salycilic Acid-depending resistance signalling pathway) and JERF3 (marker of the Jasmonic Acid/Ethylene-depending resistance signalling pathway) were detected over time in: i) untreated roots; ii) roots pre-treated with the fungus; iii) roots inoculated with the nematode; iv) pre-treated and inoculated roots. Infestation parameters were checked in untreated plants and plants treated with the fungus to test the effect of the fungus on nematode infestation level and to compare this effect with the expression of the genes PR1 and JERF3, involved in induced resistance.

Fusapyrone (FP), an antifungal natural compd., was tested against the three main ochratoxigenic species of the Aspergillus section Nigri. The MICs at 24 h were 6.0, 11.6, and 9.9 g/mL for Aspergillus carbonarius, Aspergillus tubingensis, and Aspergillus niger, resp. Strong inhibition of growth and morphol. changes were still obsd. at half the MIC after 7 days. The application of a 100 g/mL FP soln. in a lab. assay on artificially inoculated grapes resulted in a significant redn. (up to 6 orders of magnitude) of A. carbonarius CFU counts. Dramatic redns. of the ochratoxin A (OTA) content, compared to the content of the pos. control (av. amt. of OTA, 112.5 ng/g of grape; three expts.), were obtained with the application of either 100 or 50 g/mL of FP (0.6 or 5.1 ng/g of grape, resp.).

A large series of coumarin derivatives (71 compounds) were tested for their monoamine oxidase A and B (MAO-A and MAO-B) inhibitory activity. Most of the compounds acted preferentially on MAO-B with IC(50) values in the micromolar to low-nanomolar range; high inhibitory activities toward MAO-A were also measured for sulfonic acid esters. The most active compound was 7-[(3, 4-difluorobenzyl)oxy]-3,4-dimethylcoumarin, with an IC(50) value toward MAO-B of 1.14 nM. A QSAR study of 7-X-benzyloxy meta-substituted 3,4-dimethylcoumarin derivatives acting on MAO-B yielded good statistical results (q(2)() = 0.72, r(2)() = 0.86), revealing the importance of lipophilic interactions in modulating the inhibition and excluding any dependence on electronic properties. CoMFA was performed on two data sets of MAO-A and MAO-B inhibitors. The GOLPE procedure, with variable selection criteria, was applied to improve the predictivity of the models and to facilitate the graphical interpretation of results.

Mitochondrial respiratory failure secondary to complex I inhibition may contribute to the neurodegenerative process underlying nigral cell death in Parkinson's disease (PD). Isoquinoline derivatives structurally related to 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) or 1-methyl-4-phenylpyridinium (MPP+) may be inhibitors of complex I, and have been implicated in the cause of PD as endogenous neurotoxins. To determine the potency and structural requirements of isoquinoline derivatives to inhibit mitochondrial function, we examined the effects of 22 neutral and quaternary compounds from three classes of isoquinoline derivatives (11 isoquinolines, 2 dihydroisoquinolines, and 9 1,2,3,4-tetrahydroisoquinolines) and MPP+ on the enzymes of the respiratory chain in mitochondrial fragments from rat forebrain. With the exception of norsalsolinol and N,n-propylisoquinolinium, all compounds inhibited complex I in a time-independent, but concentration-dependent manner, with IC50s ranging from 0.36-22 mM. Several isoquinoline derivatives were more potent inhibitors of complex I than 1-methyl-4-phenylpyridinium ion (MPP+) (IC50 = 4.1 mM), the most active being N-methyl-6-methoxy-1,2,3,4-tetrahydroisoquinoline (IC50 = 0.36 mM) and 6-methoxy-1,2,3,4-tetrahydroisoquinoline (IC50 = 0.38 mM). 1,2,3,4-Tetrahydroisoquinoline was the least potent complex I inhibitor (IC50 approximately 22 mM). At 10 mM, only isoquinoline (23.1%), 6,7-dimethoxyisoquinoline (89.6%), and N-methylsalsolinol (34.8%) inhibited (P < 0.05) complex II-III, but none of the isoquinoline derivatives inhibited complex IV. There were no clear structure-activity relationships among the three classes of isoquinoline derivatives studied, but lipophilicity appears to be important for complex I inhibition. The effects of isoquinoline derivatives on mitochondrial function are similar to those of MPTP/MPP+, so respiratory inhibition may underlie their reported neurotoxicity.

The 1-octanol/water partition coefficients of a number of toxic and nontoxic analogues of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) were determined using centrifugal partition chromatography (CPC), a novel and effective technique for measuring lipophilicity, and found to be highly correlated with values calculated by a fragmental method. Some conformational properties of these compounds were also assessed by molecular mechanics calculations and 1H-NMR spectroscopy. A quantitative structure-metabolism relationship (QSMR) study of MPTP and analogues based on literature data was undertaken in order to determine the key features eliciting MAO-A and MAO-B reactivity and selectivity and influencing toxication. Multiple regression analysis (MRA) and comparative molecular field analysis (CoMFA) showed that MAO-B activity is nonlinearly (parabolically or bilinearly) correlated to the lipophilicity of MPTP analogues and influenced negatively by steric effects exerted by bulky substituents in the ortho position. With regard to MAO-A activity, while lipophilicity was shown to play no relevant role, electrostatic and steric fields led to a 3D-QSAR model with an acceptable predictive value (cross-validated r2 = 0.571). The results of this study bring evidence at a quantitative level that the MAO-B and MAO-A catalytic sites differ in their hydrophobic, steric, and stereoelectronic requirements.

A number of isomers and analogs of the neurotoxin 1-methyl-4-phenylpyridinium (MPP+) were examined for their lipophilic behavior. Their partition coefficients in the 1-octanol/H-2O system were measured by centrifugal partition chromatography (CPC), which, being much faster and markedly more precise than the classical shakeflask method, proved a very promising alternative for assessing lipophilicity. A smaller-than-expected lipophilicity was shown by the ortho-isomer of MPP+ (M2PP+) and is explained in terms of electronic effects and rigidity, as revealed by UV and NMR spectroscopy, and conformational analysis. Implications of lipophilicity in modulating the in vivo dopaminergic neurotoxicity of the examined compounds are also discussed

In recent years, many transmission companies have established sets of performance measures for their customer delivery systems. Such measures are used for various purposes such as investment, comparison of performance, etc. When it comes to transmission stations, there are no performance measures in place at the present time that reflect the performance of a station as a whole. In addition, outages to station-related equipment may have different consequences. A station related-outage can affect customers directly in case of a load station or the transfer capability between two locations in the system in case of a transmission station. The asset manager of a transmission system needs to use any available data on asset performance, conditions, utilization and available analysis tools in driving business decisions. He or she may need to know, for example, how stations of different sizes with the same voltage level are compared from the different point of views such as equipment performance, station utilization, station security and personnel safety. He or she may need to identify the worst performing stations (outliers) so that fund may be allocated appropriately. Also, performance measures can help quantify benefits of investments over time. This paper proposes some new quantitative performance measures for transmission and load stations. The proposed measures covers a variety of station related performance aspects such as reliability, utilization, security and safety to personnel. The new measures was used to determine relative station performance, ranking of stations, transmission station component unavailability for the entire transmission network and performance trends. The new performance measures was illustrated by examples.

Based on the results obtained with different automated computational approaches as applied to the study of eleven high-affinity agonists of the neuronal nicotine acetylcholine receptor (nAChR), belonging to different chemical classes, new relevant features were detected which complement the existing pharmacophores. Convergent results from DISCO (Distance Comparison), QXP (Quick Explore), Catalyst/HipHop, and MIPSIM (Molecular Interaction Potential Similarity) allowed us to identify and locate, in a well defined spatial arrangement, three geometrically independent key structural features: (i) a positively charged nitrogen atom for ionic or hydrogen bond interactions, (ii) a lone pair of the pyridine nitrogen or a specific lone pair of a carbonyl oxygen, as a hydrogen bond acceptor, and (iii) a centre of a hydrophobic area generally occupied by aliphatic cycles. The pharmacophore presented herein, along with predictive 2D and 3D QSAR models recently developed in our group, could represent valuable computational tools for the design of new nAChR agonists having therapeutical potential.

A set of 17 coumarin and 2 chromone derivatives with known inhibitory activity toward monoamine oxidase (MAO) A and B were tested as acetylcholinesterase (AChE) inhibitors. All compounds inhibited AChE with values in the micromolar range (3-100 microM). A kinetic study showed that most compounds acted as noncompetitive AChE inhibitors. This finding may be of interest in the context of Alzheimer's disease because recent observations suggest that MAO and AChE inhibition might decrease beta-amyloid deposition.

The acid-base properties of pyridine-2(1H)-one derivatives, analogues of the cardiotonic agent milrinone, were studied by capillary zone electrophoresis (CZE). Electrophoretic mobility and pH data were fitted to equilibrium expressions and apparent dissociation constants (pKa) calculated by non-linear regression. Compared with the ultraviolet (UV) spectrophotometric method and potentiometric titrations, the CZE technique showed advantages, such as rapidity and applicability to compounds that are sparingly soluble in water. Based on the pKa values, intramolecular electronic interactions were assessed. The lipophilicity of a number of derivatives was also examined, by determining their n-octanol/water distribution coeAcients over a wide pH range, and found to be significantly aAected by 2-pyridone/2-hydroxypyridine tautomerism. As revealed by a comparison between experi- mental and calculated log P values, electron withdrawing substituents, especially at the C(6) position of 2-pyridone, favour the less polar hydroxypyridine tautomers both in water and octanol. Our results indicate that the positive inotropism of milrinone-related compounds could be explained taking ionization and tautomerism into account. # 2000 Elsevier Science Ltd. All rights reserved.

A simple, very sensitive and rapid HPLC method was developed for the simultaneous quantitative analysis of both fusapyrone (FP) and deoxyfusapyrone (DFP), the two antifungal 3-substituted-4-hydroxy-6-alkyl-2-pyrones isolated from rice culture of Fusarium semitectum, in crude extracts. Such method was optimized on C-18 reverse phase column, using the isolated metabolites as standards, with a sequence of linear elution steps with a MeOH-H(2)O mixture and using an ultraviolet detector fixed at 285 nm, where both alpha-pyrones showed a characteristic absorption maximum. This method was used to quantify the bioactive metabolites in crude organic extracts from two F. semitectum strains. The recovery of FP and DFP was measured in a crude extract from a poor metabolite producer F. semitectum strain. The recovery values ranged from 84% to 99% for FP and from 99% to 101% for DFP, indicating that the method was close to quantitative recovery. Furthermore, an efficient medium pressure column chromatography and TLC combined method was developed for the isolation and purification of FP and DFP from fungal culture extracts.

Molecular lipophilicity can be expressed by log P or more conveniently by log kw, determined by the so-called "shake-flask" (SF) technique or by reversed-phase high-performance liquid chromatography (RP-HPLC), respectively. In the present study, the lipophilicity of a large set of solutes was measured by RP-HPLC on a silanol-deactivated octylsilane (OS) reversed phase, namely Inertsil OS (IN). IN lipophilicity parameters were found to be fairly well correlated with log P. A Linear Solvation Energy Relationship (LSER) study revealed that cavity term (modeled by intrinsic molar volume, V1) and hydrogen-bond acceptor capacity (modeled by the solvatochromic parameter beta) of solutes are the most important factors affecting retention on Inertsil OS. The comparison of LSER model for retention on Inertsil with those obtained for two other silanol-deactivated OS stationary phases, namely Deltabond OS and RP Select B, shows that alpha (H-bond donor activity of solutes) can be used as a parameter able to discriminate the behaviour of different stationary phases. QSAR studies indicated that lipophilicity parameters determined on silanol-deactivated OS phases have potential in medicinal chemistry applications.

Title compounds 1-36 were synthesized by reacting hydrazine with aldol adducts obtained from ninhydrin and methyl or alpha methylene ketones and aldehydes. Some of them showed a low, but significant benzodiazepine receptor affinity whose variation was interpreted through a structure-activity relationships study based on qualitative correlations and Comparative Molecular Field Analysis (CoMFA). Some indeno-pyridazine derivatives were found to possess an interesting anticonvulsant activity which however does not seem simply related to the benzodiazepine receptor modulation.

Fifteen strains of Trichoderma belonging to 12 different species for potential use in biological control were examined for their ability to produce polypeptide antiobitics (peptaibols). Gas chromatography was used to detect the hydrolysis products of peptaibols after esterification with n-propanol and derivatisation with heptafluorobutyric anhydride. In particular, amino acids (alpha-aminoisobutyric acid, and isovalin) and amino alcohols (phenylalaninol, leucinol, and valinol) were used as markers for the detection of different peptaibols. alpha-Aminoisobutyric acid, an amino acid characterising all peptaibols, was detected in all cultures examined, indicating that the production of peptaibols is a common feature of the various Trichoderma species. Different amino acid/amino alcohol combinations suggested specific peptaibol production. On this basis, the peptaibol paracelsin was confirmed by HPTLC in 11 out of 15 Trichoderma strains cultures. Valinol was not found in any culture extract, indicating the lack of trichotoxins production. Culture extracts of the 6 tested Trichoderma strains were extremely toxic to Artemia salina larvae; for 5 of them this toxicity could be attributed to paracelsin (calculated LD50 = 2.2 microM).

A large homologous series of quaternary ammonium derivatives of perhydropyrrolo[3,4-c]pyridine 5 was synthesized and tested for in vitro antibacterial activity against different Gram-positive and Gram-negative bacteria. Compounds 5 proved to be always more potent than benzalkonium chloride, taken as reference. Antibacterial activity, expressed as log 1/MIC, was found linearly related to lipophilicity up to C13-C14 homologs, where a break in the linear relationship was observed.

Lipophilicity is a molecular property encoding a wealth of structural information. Solvatochromic analysis of partition coefficients allow their factorization into a steric term accounting for hydrophobic and dispersive solute/solvent interactions, and polar terms accounting for induction and orientation forces and for hydrogen bonds. Two lipophilicity-related parameters express H-bonding capacity, namely Δlog P and δ. Some examples have been published which document their relation with skin and blood-brain barrier permeation. A QSAR study of carbonic anhydrase inhibitors shows that these parameters may also be related to pharmacodynamic responses.

The lipophilic character of two large series of substituted benzenesulphonamides (BzSA) and 4-aminodiphenylsulphones (4-ADS) has been assessed by two chromatographic methods, i.e. reversed-phase HPLC using a relatively novel octadecylpolyvinyl packing and centrifugal counter-current chromatography (CPC). The octadecylpolyvinyl stationary phase proved an interesting alternative to the more common octadecylsilane type stationary phase for obtaining retention parameters correlated to partition coefficients (i.e. log P). The CPC method, being far less time-consuming and markedly more precise than the classical shake-flask method, offers a promising alternative for measuring partition coefficients. The parameter Δlog Poct-hep, i.e. log Poctanol minus log Pheptane, was also determined for both congeneric series and was indicative of a similar H-bonding capacity for the SO2NH2 and 4-NH2-C6H4-SO2 groups. QSAR analyses of carbonic anhydrase inhibition by BzSA and antimycobacterial activity of 4-ADS show the capacity of the new lipophilicity parameters to express the hydrophobic component of the drug-enzyme interactions and to reveal a possible role of H-bond donor capacity in governing the antimycobacterial activity of 4-ADS.

A simple and efficient method is described for the direct determination of Isoniazide (INH) and Pyrazinamide (PZAm) in human plasma, based on reversed-phase HPLC. A number of other antitubercular drugs such as p-Aminosalycylic acid (PAS), Streptomycin (STM) and Ethambutol (ETH) and the major metabolites of the two investigated drugs, namely N-Monoacetylisoniazide (AcINH) and Pyrazinoic acid (PZAc), were shown not to interfere with the assay method. A potential application of the method to routine clinical monitoring of antitubercular drugs is discussed.

Several N,N'-bis-alkyl-perhydropyrrolo[3,4-c]pyridines 4 and the corresponding bis-quaternary derivatives 5 have been prepared through standard methods starting from the bicyclic dilactam 2. Compounds 4 and 5 were evaluated for their inhibitory properties against acetylcholine (ACh), 1,1-dimethylphenyl piperazinium iodide (DMPP), 5-hydroxytryptamine (5-HT) and histamine (H1), and the guinea pig isolated ileum. Pharmacological data indicated that the strongest anticholinergic activity was shown by the dibenzyl amine 4 g, which among the tested molecules presented also a rather potent effect at the ganglia level, resulting about three times more potent than hexamethonium as ganglionic blocking agent. The preliminary SAR coming from the analysis of the pharmacological results are presented and discussed.