NEW APPROACHES IN THE DISCOVERY OF NATURAL PRODUCT-BASED AGROCHEMICALS

Currently, the increase of food demand of a steadily growing human population is not balanced by a global agricultural supply. As a consequence, crop protection plays a crucial role in maximising crop productivity and preventing losses caused by biological and not biological agents. Biological agents, collectively named as pests, are responsible for quantitative losses ranging from 20 % to 40 %, and qualitative losses, thus reducing the value of crops. For these reasons, new methods to control pests must be developed.The discovery of new agrochemicals is necessary to face the problem of the emergence of resistances and the increasing of stringent regulatory standards, which are leading to the ban of many products no longer considered acceptable. All these factors are changing the research of new pesticides that is becoming a more expensive and time-consuming process. Thus, new methods to improve and speed up the discovery of new agrochemicals must be developed. In this context, the aim of this Ph.D. thesis was to explore new approaches to the synthesis of agrochemicals. Our attention was focused on the naturally derived compound-based molecules, due to the proved value of these compounds in development of agrochemicals and more in general of biologically active compounds. The first part of this Ph.D. thesis is focused on the development of dual-target fungicides obtained by combination of the naturally derived strobilurin fungicides with succinate dehydrogenase inhibitors. The development of dual-target molecules is a well-known approach in pharmaceutic research, while it is still underexplored in the field of agrochemicals. The research work was organized following a cycle of design, synthesis, in vitro tests, and evaluation of the mechanism of action. The dual-target compounds were designed on the basis of the reported structure-activity relationship for both the classes of fungicides. The designed compounds were synthetized, and the antifungal activity was screened in vitro by measurement of the mycelium growth inhibition. Finally, the mechanism of action was evaluated in silico by docking studies and in vitro in a cell free system. The second part of this dissertation was focused on the development of versatile synthetic approaches in the synthesis of biologically active natural compounds. SmI2 chemistry was employed in light of the remarkable synthetic power of this reagent. First, a SmI2-mediated Reformatsky reaction was developed as a complementary approach to Evans aldol reaction for the synthesis of precursors of cembranoids. Then, a SmI2-mediated pinacol coupling was investigated as a key step for the synthesis of the natural herbicide auscalitoxin aglycone

Samarium iodide-promoted asymmetric Reformatsky reaction of 3-(2-Haloacyl)-2-oxazolidinones with enals

3-(2-Haloacyl)-2-oxazolidinones were shown to react with enals in an asymmetric SmI2-promoted Reformatsky reaction to give stereochemically well-defined 3-hydroxy-4-alkenyl- and 3-hydroxy-2-methyl-4-alkenyl imides. Chirality transfer of the Evans (S)-oxazolidinone unit via a Zimmerman-Traxler-like transition state resulted in Reformatsky products with a relative syn-configuration. The absolute configuration of compounds obtained is opposite to the corresponding products obtained via aldol addition of boron enolates to enals using the same Evans oxazolidinones

Modeling RR Tel through the Evolution of the Spectra

We investigate the evolution of RR Tel after the outburst by fitting the emission spectra in two epochs. The first one (1978) is characterized by large fluctuations in the light curve and the second one (1993) by the slow fading trend. In the frame of a colliding wind model two shocks are present: the reverse shock propagates in the direction of the white dwarf and the other one expands towards or beyond the giant. The results of our modeling show that in 1993 the expanding shock has overcome the system and is propagating in the nearby ISM. The large fluctuations observed in the 1978 light curve result from line intensity rather than from continuum variation. These variations are explained by fragmentation of matter at the time of head-on collision of the winds from the two stars. A high velocity (500 km/s) wind component is revealed from the fit of the SED of the continuum in the X-ray range in 1978, but is quite unobservable in the line profiles. The geometrical thickness of the emitting clumps is the critical parameter which can explain the short time scale variabilities of the spectrum and the trend of slow line intensity decrease.Comment: 26 pages, LaTeX (including 5 Tables) + 6 PostScript figures. To appear in "The Astrophysical Journal

Prenatal smoking, alcohol and caffeine exposure and offspring externalizing disorders:a systematic review and meta-analysis

BACKGROUND AND AIMS: Several studies have indicated an association between maternal prenatal substance use and offspring externalizing disorders; however, it is uncertain whether this relationship is causal. We conducted a systematic review to determine: (1) if the literature supports a causal role of maternal prenatal substance use on offspring externalizing disorders diagnosis and (2) whether these associations differ across externalizing disorders. METHODS: We searched Web of Science, Embase, PsycINFO and Medline databases. Risk of bias assessment was conducted using the Newcastle-Ottawa Scale (NOS), and where possible meta-analysis was conducted for studies classed as low risk of bias. We included studies of any design that examined prenatal smoking, alcohol or caffeine use. Studies in non-English language, fetal alcohol syndrome and comorbid autism spectrum disorders were excluded. Participants in the included studies were mothers and their offspring. Measurements included prenatal smoking, alcohol or caffeine use as an exposure, and diagnosis of attention-deficit hyperactivity disorder (ADHD), conduct disorder (CD) and oppositional defiant disorder (ODD) in offspring as an outcome. RESULTS: We included 63 studies, 46 of which investigated smoking and ADHD. All studies were narratively synthesized, and seven studies on smoking and ADHD were meta-analysed. The largest meta-analysis based on genetically sensitive design included 1 011 546 participants and did not find evidence for an association [odds ratio (OR)  = 0.90, 95% confidence interval (CI) = 0.83-1.11; OR   = 1.04, 95% CI = 0.79-1.36). Studies on alcohol exposure in all the outcomes reported inconsistent findings and no strong conclusions on causality can be made. Studies on caffeine exposure were mainly limited to ADHD and these studies do not support a causal effect. CONCLUSIONS: There appears to be no clear evidence to support a causal relationship between maternal prenatal smoking and offspring attention-deficit hyperactivity disorder. Findings with alcohol and caffeine exposures and conduct disorder and oppositional-defiant disorder need more research, using more genetically sensitive designs

Maternal and child genetic liability for smoking and caffeine consumption and child mental health:An intergenerational genetic risk score analysis in the ALSPAC cohort

Background and aims: Previous studies suggest an association between maternal tobacco and caffeine consumption during and outside of pregnancy and offspring mental health. We aimed to separate effects of the maternal environment (intrauterine or postnatal) from pleiotropic genetic effects. Design: Secondary analysis of a longitudinal study. We (i) validated smoking and caffeine genetic risk scores (GRS) derived from published genome-wide association study (GWAS) for use during pregnancy, (ii) compared estimated effects of maternal and offspring GRS on childhood mental health outcomes and (iii) tested associations between maternal and offspring GRS on their respective outcomes. Setting: We used data from a longitudinal birth cohort study from England, the Avon Longitudinal Study of Parents and Children (ALSPAC). Participants: Our sample included 7921 mothers and 7964 offspring. Measurements: Mental health and non-mental health phenotypes were derived from questionnaires and clinical assessments: 79 maternal phenotypes assessed during and outside of pregnancy and 71 offspring phenotypes assessed in childhood (<10years) and adolescence (11–18years). Findings: The maternal smoking and caffeine GRS were associated with maternal smoking and caffeine consumption during pregnancy (2nd trimester: Psmoking=3.0 × 10−7, Pcaffeine=3.28 × 10−5). Both the maternal and offspring smoking GRS showed evidence of association with reduced childhood anxiety symptoms (βmaternal=−0.033; βoffspring=−0.031) and increased conduct disorder symptoms (βmaternal=0.024; βoffspring=0.030), after correcting for multiple testing. Finally, the maternal and offspring smoking GRS were associated with phenotypes related to sensation seeking behaviours in mothers and adolescence (e.g. increased symptoms of externalising disorders, extraversion and monotony avoidance). The caffeine GRS showed weaker evidence for associations with mental health outcomes. Conclusions: We did not find strong evidence that maternal smoking and caffeine genetic risk scores have a causal effect on offspring mental health outcomes. Our results confirm that the smoking genetic risk scores also captures liability for sensation seeking personality traits

Characterization of alcohol polygenic risk scores in the context of mental health outcomes:Within-individual and intergenerational analyses in the Avon Longitudinal Study of Parents and Children

Background: Heavy alcohol consumption often co-occurs with mental health problems; this could be due to confounding, shared biological mechanisms, or causal effects. Polygenic risk scores (PRS) for alcohol use can be used to explore this association at critical life stages. Design: We characterized a PRS reliably associated with patterns of adult alcohol consumption by 1) validating whether it predicts own alcohol use at different life-stages (pregnancy, adolescence) of interest for mental health impact. Additionally, we explored associations of alcohol PRS on mental health phenotypes 2) within-individuals (using own alcohol PRS on own phenotypes) and 3) intergenerationally (using maternal alcohol PRS on offspring phenotypes). We used data from the Avon Longitudinal Study of Parents and Children (ALSPAC) (n = 960–7841). Additional substance abuse behaviors and mental health/behavioral outcomes were investigated (alcohol phenotypes n = 22; health phenotypes n = 91). Findings: Maternal alcohol PRS was associated with consumption during pregnancy (strongest signal: alcohol frequency at 18 weeks’ gestation: β = 0.041, 95%CI = 0.0.02–0.06), p = 1.01 × 10−5, adjusted R2 = 1.6 %), offspring alcohol PRS did not predict offspring alcohol consumption. We found evidence for an association of maternal alcohol PRS with own perinatal depression (OR = 1.10, 95% CI = 1.02 to 1.18, p = 0.022) and decreased offspring intellectual ability (β=-0.209, 95% CI -0.38 to -0.04, p= 0.016). Conclusions: These alcohol PRS are a valid proxy for maternal alcohol use in pregnancy. Offspring alcohol PRS was not associated with drinking in adolescence. Consistently with results from different study designs, we found evidence that maternal alcohol PRS are associated with both prenatal depression and decreased offspring intellectual ability

Prenatal smoking, alcohol and caffeine exposure and maternal-reported attention deficit hyperactivity disorder symptoms in childhood:triangulation of evidence using negative control and polygenic risk score analyses

Background and aims Studies have indicated that maternal prenatal substance use may be associated with offspring attention deficit hyperactivity disorder (ADHD) via intrauterine effects. We measured associations between prenatal smoking, alcohol and caffeine consumption with childhood ADHD symptoms accounting for shared familial factors. Design First, we used a negative control design comparing maternal and paternal substance use. Three models were used for negative control analyses: unadjusted (without confounders), adjusted (including confounders) and mutually adjusted (including confounders and partner's substance use). The results were meta-analysed across the cohorts. Secondly, we used polygenic risk scores (PRS) as proxies for exposures. Maternal PRS for smoking, alcohol and coffee consumption were regressed against ADHD symptoms. We triangulated the results across the two approaches to infer causality. Setting We used data from three longitudinal pregnancy cohorts: Avon Longitudinal Study of Parents and Children (ALSPAC) in the United Kingdom, Generation R study (GenR) in the Netherlands and Norwegian Mother, Father and Child Cohort study (MoBa) in Norway. Participants Phenotype data available for children were: NALSPAC = 5455–7751; NGENR = 1537–3119; NMOBA = 28 053–42 206. Genotype data available for mothers was: NALSPAC = 7074; NMOBA = 14 583. Measurements A measure of offspring ADHD symptoms at age 7–8 years was derived by dichotomizing scores from questionnaires and parental self-reported prenatal substance use was measured at the second pregnancy trimester. Findings The pooled estimate for maternal prenatal substance use showed an association with total ADHD symptoms [odds ratio (OR)SMOKING = 1.11, 95% confidence interval (CI) = 1.00–1.23; ORALCOHOL = 1.27, 95% CI = 1.08–1.49; ORCAFFEINE = 1.05, 95% CI = 1.00–1.11], while not for fathers (ORSMOKING = 1.03, 95% CI = 0.95–1.13; ORALCOHOL = 0.83, 95% CI = 0.47–1.48; ORCAFFEINE = 1.02, 95% CI = 0.97–1.07). However, maternal associations did not persist in sensitivity analyses (substance use before pregnancy, adjustment for maternal ADHD symptoms in MoBa). The PRS analyses were inconclusive for an association in ALSPAC or MoBa. Conclusions There appears to be no causal intrauterine effect of maternal prenatal substance use on offspring attention deficit hyperactivity disorder symptoms

Arachidonic acid-evoked Ca^{2+} signals promote nitric oxide release and proliferation in human endothelial colony forming cells

Arachidonic acid (AA) stimulates endothelial cell (EC) proliferation through an increase in intracellular Ca^{2+} concentration ([Ca^{2+}]_{i}), that, in turn, promotes nitric oxide (NO) release. AA-evoked Ca^{2+} signals are mainly mediated by Transient Receptor Potential Vanilloid 4 (TRPV4) channels. Circulating endothelial colony forming cells (ECFCs) represent the only established precursors of ECs. In the present study, we, therefore, sought to elucidate whether AA promotes human ECFC (hECFC) proliferation through an increase in [Ca^{2+}]_{i} and the following activation of the endothelial NO synthase (eNOS). AA induced a dose-dependent [Ca^{2+}]_{i} raise that was mimicked by its non-metabolizable analogue eicosatetraynoic acid. AA-evoked Ca^{2+} signals required both intracellular Ca^{2+} release and external Ca^{2+} inflow. AA-induced Ca^{2+} release was mediated by inositol-1,4,5-trisphosphate receptors from the endoplasmic reticulum and by two pore channel 1 from the acidic stores of the endolysosomal system. AA-evoked Ca^{2+} entry was, in turn, mediated by TRPV4, while it did not involve store-operated Ca^{2+} entry. Moreover, AA caused an increase in NO levels which was blocked by preventing the concomitant increase in [Ca^{2+}]_{i} and by inhibiting eNOS activity with NG-nitro-l-arginine methyl ester (l-NAME). Finally, AA per se did not stimulate hECFC growth, but potentiated growth factors-induced hECFC proliferation in a Ca^{2+} - and NO-dependent manner. Therefore, AA-evoked Ca^{2+} signals emerge as an additional target to prevent cancer vascularisation, which may be sustained by ECFC recruitment

Maternal and offspring genetic risk score analyses of fetal alcohol exposure and attention-deficit hyperactivity disorder risk in offspring

Background: Studies investigating the effects of prenatal alcohol exposure on childhood attention-deficit hyperactivity disorder (ADHD) symptoms using conventional observational designs have reported inconsistent findings, which may be affected by unmeasured confounding and maternal and fetal ability to metabolize alcohol. We used genetic variants from the alcohol metabolizing genes, alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH), as proxies for fetal alcohol exposure to investigate their association with risk of offspring ADHD symptoms around age 7–8 years. Methods: We used data from 3 longitudinal pregnancy cohorts: Avon Longitudinal Study of Parents and Children (ALSPAC), Generation R study (GenR), and the Norwegian Mother, Father and Child Cohort study (MoBa). Genetic risk scores (GRS) for alcohol use and metabolism using 36 single nucleotide polymorphisms (SNPs) from ADH and ALDH genes were calculated for mothers (NALSPAC = 8196; NMOBA = 13,614), fathers (NMOBA = 13,935), and offspring (NALSPAC=8,237; NMOBA=14,112; NGENR=2,661). Associations between maternal GRS and offspring risk of ADHD symptoms were tested in the full sample to avoid collider bias. Offspring GRS analyses were stratified by maternal drinking status. Results: The pooled estimate in maternal GRS analyses adjusted for offspring GRS in ALSPAC and MoBa was OR = 0.99, 95%CI 0.97–1.02. The pooled estimate in offspring GRS analyses stratified by maternal drinking status across all the cohorts was as follows: ORDRINKING = 0.98, 95% CI 0.94–1.02; ORNO DRINKING = 0.99, 95% CI 0.97–1.02. These findings remained similar after accounting for maternal genotype data in ALSPAC and maternal and paternal genotype data in MoBa. Conclusions: We did not find evidence for a causal effect of fetal alcohol exposure on risk of ADHD symptoms in offspring. The results may be affected by limited power to detect small effects and outcome assessment.publishedVersio