Oviposition inhibitory activity of the Mexican sunflower Tithonia diversifolia (Asteraceae) polar extracts against the two-spotted spider mite Tetranychus urticae (Tetranychidae)

The Mexican sunflower (Tithonia diversifolia, Asteraceae) is an invasive shrub of agricultural and non-agricultural lands in tropical countries. Besides extensive utilizations in the traditional medicine, mainly to treat malaria, the plant is believed to have a great potential in agriculture of developing countries as a green biomass to produce fertilizer, fodder and biopesticides. The plant is known to produce tagitinins, which are sesquiterpene lactones with a bitter taste endowed with toxicity against several insects such as mosquitoes, aphids, and beetles. Here, we evaluated the potential of T. diversifolia against the two-spotted spider mite Tetranychus urticae (Tetranychidae), which is one of the most economically important arthropod pests worldwide. The leaf methanolic extract and its ethyl acetate fraction were tested for acute and chronic toxicity and for oviposition inhibitory effects. The chemical composition of the extracts was analyzed by HPLC-MSn and NMR. The main constituents were flavonoid derivatives, phenylpropanoids and sesquiterpene lactones. Among the latter, tagitinin C and tagitinin A were the major compounds. In acute toxicity assays, mortality did not exceed 50% even for the highest tested dose of 150 \u3bcg cm-3. However, in chronic toxicity assays, on day 5 from application, the methanolic extract LD50 was 41.3 \u3bcg cm-3 while LD90 was 98.7 \u3bcg cm-3. Furthermore, both T. diversifolia extracts inhibited oviposition in T. urticae. The ethyl acetate extract was the most active oviposition inhibitor, with an ED50 value of 44.3 \u3bcg cm-3 and an ED90 of 121.5 \u3bcg cm-3. Overall, the good yield rate of the extract and the high crop yield highlighted good prospects of using the extract from this plant for the development of oviposition inhibitors against mite

Identification of tagitinin C from Tithonia diversifolia as antitrypanosomal compound using bioactivity-guided fractionation

Tithonia diversifolia (Asteraceae), is used as traditional medicine in tropical countries for the treatment of various diseases, including malaria. Although numerous studies have assessed the antimalarial properties, nothing is known about the effect of T. diversifolia extracts on trypanosomiasis. In this study extracts of T. diversifolia aerial parts were evaluated for their bioactivity against Trypanosoma brucei. The activity was studied against bloodstream forms of T. brucei (TC221), as well as against mammalian cells (BALB/3T3 mouse fibroblasts), as a counter-screen for toxicity. Both methanolic and aqueous extracts showed significant effects with IC50 values of 1.1 and 2.2\u3bcg/mL against T. brucei (TC221) and 5.2 and 3.7\u3bcg/mL against BALB/3T3 cells, respectively. A bioassay-guided fractionation on the methanolic extract yielded in identification of active fractions (F8 and F9) with IC50 values of 0.41 and 0.43\u3bcg/mL, respectively, against T. brucei (TC221) and 1.4 and 1.5\u3bcg/mL, respectively, against BALB/3T3 cells,. The phytochemical composition of the extracts and the purified fractions were investigated using HPLC-ESI-MS/MS and 1D and 2D NMR spectra showing the presence of sesquiterpene lactones that in turn were subjected to the isolation procedure. Tagitinin A and C were rather active but the latter presented a very strong inhibition on T. brucei (TC221) with an IC50 value of 0.0042\u3bcg/mL. This activity was 4.5 times better than that of the reference drug suramin. The results of this study shed light on the antitrypanosomal effects of T. diversifolia extracts and highlighted tagitinin C as one of the possible responsible for this effect. Further structure activity relationships studies on tagitinins are needed to consider this sesquiterpenes as lead compounds for the development of new antitrypanosomal drugs

Tyttöjen Tila : kehittämishanke Loviisassa

Tytti Eriksson. Tyttöjen tila- kehittämishanke Loviisassa. Diakonia-ammattikorkeakoulu. Diak, Helsinki, syksy 2016. sivuja 52 liitteitä 5. Sosiaalialan koulutusohjelma, Diakonia ja kristillinen kasvatus, sosionomi, (YAMK) Opinnäytetyön tavoite oli selvittää loviisalaisten yläasteikäisten tyttöjen toiveita ja osallisuutta Loviisaan perustettavan Tyttöjen Tilan toimintaan. Lisäksi työn tarkoituksena oli suunnitella toimintaa ja hakea rahoitusta toiminnan pyörittämiseen ja avata tila Loviisassa. Opinnäytetyö on kehittämishanke, joka toteutettiin kaksiosaisena, kyselytutkimuksena sekä kehittävänä toimintana, joka päättyi tyttöjen tilan perustamiseen. Opinnäytetyön tutkimuksellinen osuus toteutettiin Loviisan kaupungin tilauksesta kyse-lytutkimuksena. Loviisa on kaksikielinen kunta, jossa suomenkielisten osuus väestöstä on 55 % ja ruotsinkielisten 42 %. Kysely tehtiin kaikille yläkouluikäisille loviisalaisty-töille ja se toteutettiin kaksikielisenä. Kyselyt tehtiin keväällä 2016 ja tila avattiin loka-kuussa 2016. Tutkimus oli määrällinen kyselytutkimus ja kehittämishanke. Lisäksi sa-maan aikaan opinnäytetyön tekijä suunnitteli tyttöryhmän kanssa tyttöjen omaa huonetta Loviisan keskustan Forumin nuorisotilaan. Tekijä haki myös rahoitusta toimintaan kol-mella erillisellä rahoitushakemuksella. Tyttötyö ja sukupuolisensitiivinen työ ovat olleet tärkeimpiä suuntaviivoja opinnäyte-työssä. Myös nuorten osallisuus ja nuorten oman äänen kuuluminen ovat vaikuttaneet opinnäytetyöhön ja ovat siinä esillä. Toimintaa halutaan jatkaa yhdessä tyttöjen kanssa ja kehittää sitä heidän toiveidensa mukaisesti. Opinnäytetyön valmistumisvaiheessa yhdestä rahoitushakemuksista on saatu rahaa toi-mintaan. Lisäksi International Soroptimistit Loviisa- naisjärjestö oli tukemassa toimintaa. Tulevaisuudessa kuntasektorilla tulee olemaan tarve hakea rahoitusta toimintaan erilaisista hankkeista ja projektirahoituksista, jotta voidaan turvata laadukkaan ja ennal-taehkäisevän nuorisotyön jatkuvuus kuntien taloudellisen tilanteen heikentyessä. Itse uskon, että työ tulee tulevaisuudessa olemaan enemmän projektiluonteista ja ostopalve-lut tulevat lisääntymään, joten rahoituskanavien ja erilaisten projektien tuntemus tulee jatkossa olemaan myös kuntatyöntekijöiden osaamisaluetta. Opinnäytetyö oli tarpeellinen työn kehittämisen kannalta ja antoi suuntaviivaa toiminnan jatkosta. Tilan avaamiseen lokakuussa osallistui 120 tyttöä. Asiasanat: nuorten osallisuus, sukupuolisensitiivinen työ, tyttötyö, Loviisan kaupunki  Eriksson, Tytti. Girls´ place – development project in Loviisa. Autum 2016.52p.,5 ap-pendices Language Finnish. Diaconia University of Applied Sciences. Degree pro-gramme in Social Services. Diaconia, Christian Education and Youth Work. Degree: Master of Social Services. The purpose of this thesis was to understand the gender-sensitive hopes and aspirations of teenage girls participating in to-be-established Girls Club activities. In addition, the aim was to advise the planning of those activities and to seek funding for opening and running a girls´ club in Loviisa, Finland. This thesis is a development project, which was carried out in two parts, as a survey and as a development activity, which led to the establishment of the girls´ club and its services. In accordance with the town of Loviisa´s research guidance, the research was conducted as a survey. Loviisa is a bilingual municipality consisting of population of 55% Finnish speakers and 42% Swedish speakers. The sample included high-school aged girls in Loviisa and the surveys were conducted bi-lingually. The research was qualitative. In addition, the author planned together with the interviewees the facilities in the town-centre Forum Youth Club. The author also sought funding with three independent appli-cations. The survey took place in the Spring 2016 and the facilities opened in October 2016. Working with the girls and addressing the gender-sensitive issues has been a significant driver for this thesis. The actual participation and the desire to hear the ‘true voice’ of the youth have had a significant impact on this work, as can be seen in the findings. The aspiration is to develop this work, and to do so according to the wishes of teenage girls in Loviisa. One of the original funding applications out of the three has been successful so far. In addition, the International Soroptimist Loviisa - a women’s institution – has been sup-porting this project. It is essential that the municipality- level sector continues to find and to secure funding from a variety of projects, to ensure the continuity of high quality and preemptive youth support work, especially under currently weakening municipality- level economy. The author firmly believes that future work will be more project-based and that the outsourcing of services will increase. Therefore the council and youth workers will need ongoing training about the appropriate funding sources and venues. This thesis is necessary for developing the provision of youth services and to provide pointers for the future services and activities. The first night opening, in October 2016, of the Girls´ Club in Loviisa attracted 120 girls. Keywords: youth participation, youth work, youth support, gender sensitivity, working with teenage girls, Loviisa

Evaluation of the in vitro trypanocidal activity of Vernonia amygdalina leaf extracts and isolated compounds

Human African trypanosomiasis (HAT), also known as African sleeping sickness, is a neglected disease caused by two subspecies of the protozoan parasite Trypanosoma brucei, Trypanosoma brucei gambiense and Trypanosoma brucei rhodesiense. The parasites are introduced into the mammalian host by the bite of an infected tsetse fly and HAT progresses in two stages: the hemo-lymphatic or early stage and the meningoencephalitic or late stage on which the parasites enter the central nervous system (CNS) (1). HAT threatens primarily rural populations and is fatal unless treated. To date, there are five drugs approved, although none of them are satisfactory, due to treatment failures and toxicity, and the parenteral administration that is inappropriate in settings with poor medical infrastructure. Therefore, there is an urgent need to improve HAT treatment by enhancing the oral administration and the discovery and development of cost-effective new drugs. Drug discovery efforts are nowadays directed towards natural products and medicinal plants represent a validated source for discovery of new lead compounds and standardized herbal medicines against trypanosomiasis (2). Vernonia amygdalina def., generally called “bitter leaf”, is a small shrub that can grow as tall as 5 m in height, and is member of the Asteraceae (or Compositae) family. The species is indigenous to tropical Africa and is found wild or cultivated all over sub-Saharan Africa. Due to its abundant availability in sub-Saharan countries, V. amygdalina has become commonly used in the traditional medicine by local ethnic groups. The leaves of V. amygdalina are used in phyto-medicine to treat fever, hiccups, kidney disease and stomach discomfort, as well as anthelmintic and for treatment and prevention of malaria (3). On the basis of the above-illustrated therapeutic properties, the V. amygdalina leaf extracts and isolated compounds have been selected as valid candidates for investigation as potential inhibitors of Trypanosoma brucei. The results of this study will be discussed. (1) Jacobs, R. T.; Nare, B.; Phillips, M. A. State of the Art in African Trypanosome Drug Discovery. Curr. Top. Med. Chem. 2011, 11, 1255−1274. (2) Scotti, L.; Mendonça, F.J.; da Silva, M.S.; Scotti, M.T. Enzymatic Targets in Trypanosoma brucei. Curr. Protein Pept. Sci. 2016, 17, 243-259. (3) Abay, S.M; Lucantoni, L.; Dahiya, N.; Esposito, F.; Lupidi, G.; Taglialatela-Scafati, O.; Bramucci, M.; Quassinti, L.; Habluetzel, A. Plasmodium transmission blocking activities of Vernonia amygdalina extracts and isolated compounds. Malar J. 2015, 14, 288-306

SYNTHESIS AND PHARMACOLOGICAL CHARACTERIZATION OF RIBOSE-MODIFIED ADENOSINE DERIVATIVES AS P1 RECEPTOR LIGANDS

Adenosine, the natural ligand of P1 receptors, is implicated in the control of many physiological and pathological conditions such as inflammation, pain, cardiovascular and central nervous system (CNS) diseases.1 P1 receptors belong to the large family of GPCR receptors and are divided in four subtypes: A1, A2A, A2B and A3 adenosine receptors (ARs). Even though a large number of P1 ligands have been synthesized and characterized byin vitro and in vivo pharmacological studies, only very few of them are commercially available. Modifications at the ribose moiety and substitution at the N6-position of adenosine, lead to adenosine derivatives endowed with increased potency at A1 or A3AR. Our previous SAR studies showed that the replacement of OH at 5’-position of the ribose moiety of N6-substituted adenosine derivatives by a chlorine improved A1AR potency and selectivity versus A3AR, with 5′-chloro-5′-deoxy-N6-(±)-(endo- norborn-2-yl)-adenosine (5′Cl5′d-(±)-ENBA) as one of the most potent and selective A1AR agonists,2 while a 5’-C-ethyl-tetrazolyl moiety maintained the A1AR potency, but restored high A3AR affinity, leading to very potent dual A1AR and A3AR ligands.3 Interestingly, both modifications at 5’-position of adenosine derivatives brought to human A3AR antagonism. In order to further explore the structural determinants of this class of P1 ligands, a new series of ribose- modified N6-substituted adenosine derivatives was synthesized and their pharmacological profile was assayed. The results of this study will be discussed.References 1. Jacobson KA, Muller CE, Neuropharmacology 2015, doi: 10.1016/J.neuropharm.2015.12.001. 2. (a) Franchetti, P.; Cappellacci, L.; Vita, P.; Petrelli, R.; Lavecchia, A.; Kachler, S.; Klotz, K.-N.; Marabese, I.; Luongo, L.; Maione, S.; Grifantini, M. J. Med. Chem. 2009, 52, 2393−2406. (b) Luongo, L.; Petrelli, R.; Gatta, L.; Giordano, C.; Guida, F.; Vita, P.; Franchetti, P.; Grifantini, M.; De Novellis, V.; Cappellacci, L.; Maione, S. Molecules 2012, 17, 13712−13726. (c) Luongo, L.; Guida, F.; Imperatore, R.; Napolitano, F.; Gatta, L.; Cristino, L.; Giordano, C.; Siniscalco, D.; Di Marzo, V.; Bellini, G.; Petrelli, R.; Cappellacci, L.; Usiello, A.; de Novellis, V.; Rossi, F.; Maione, S. Glia 2014, 62, 122−132. 3. Petrelli, R.; Torquati, I.; Kachler, S.; Luongo, L.; Maione, S.; Franchetti, P.; Grifantini, M.; Novellino, E.; Lavecchia, A.; Klotz, K.-N-; Cappellacci, L. J. Med. Chem. 2015, 58, 2560-2566

NOVEL 5'-C-ETHYL-TETRAZOLYL-N6-SUBSTITUTED-ADENOSINE DERIVATIVES: SYNTHESIS AND BIOLOGICAL EVALUATION

Adenosine is an endogenous purine ribonucleoside implicated in the control of the function of many tissues and organs. It exerts a protective action throughout all organs of the body and plays an important role not only in the pathophysiological processes, but also in the modulation of normal processes. Adenosine exerts its effects by interacting with specific G-protein coupled receptors, classified in 4 subtypes: A1, A2A, A2B and A3 (1). A1 adenosine receptor (A1AR) is widely distributed throughout the brain but also in the heart, aorta, liver kidney, bladder and eye. A1AR agonists have shown neuro- and cardioprotective effects, but their clinical use is hampered by severe cardiovascular side effects. A3AR are expressed in multiple organs and at low levels in the CNS. A3AR agonists are in clinical trials for the treatment of cancer and inflammatory diseases, and recent preclinical studies reported their analgesic effects in chronic neuropathic pain (2). Our previous work showed that potent dual A1AR agonists and A3AR antagonists have been obtained combining a 5'-C-ethyl-tetrazolyl moiety and an appropriate N6-substitution in adenosine derivatives (3). A dual A1 agonist and A3 antagonist might be useful in the treatment of glaucoma and other diseases, and might have advantages respect to the combination of two drugs. In order to study the influence of N6-substitution on affinity and selectivity at ARs, a novel series of 5'-C-2-ethyl-tetrazolyl-N6-substituted adenosine derivatives were synthesized and assayed at all human adenosine receptor subtypes. The results of this study will be discussed. (1) Fredholm, B.B.; Ijzerman, A.P.; Jacobson, K:A., Linden, J.; Muller, C.E. International Union of Basic and Clinical Pharmacology. LXXXI. Nomenclature and calssification of adenosine receptors - an update. Pharmacol. Rev. 2011, 63, 1-34. (2) Little, J.W.; Ford, A.; Symons-Liguori, A.M.; Chen, Z.; Janes, K.; Doyle, T.; Xie, J.; Luongo,, L.; Tosh, D.K.; Maione, S.; Bannister, K.; Dickenson, A.; Vandrah, T.W.; Porreca, F.; Jacobson, K.A.; Salvemini, D. Endogenous adenosine A3 receptor activation selectively alleviates persistent pain states. Brain 2015, 138, 28-35 (3) Petrelli, R.; Torquati, I.; Kachler, S.; Luongo, L.; Maione, S.; Franchetti, P.; Grifantini, M.; Novellino, E.; Lavecchia, A.; Klotz, K.-N-; Cappellacci, L. 5′‑C‑Ethyl-tetrazolyl‑N6‑Substituted Adenosine and 2‑Chloroadenosine Derivatives as Highly Potent Dual Acting A1 Adenosine Receptor Agonists and A3 Adenosine Receptor Antagonists. J. Med. Chem. 2015, 58, 2560-2566

An overlooked horticultural crop, Smyrnium olusatrum, as a potential source of compounds effective against African trypanosomiasis

Among natural products, sesquiterpenes have shown promising inhibitory effects against bloodstream forms of Trypanosoma brucei, the protozoan parasite causing human African trypanosomiasis (HAT). Smyrnium olusatrum (Apiaceae), also known as Alexanders or wild celery, is a neglected horticultural crop characterized by oxygenated sesquiterpenes containing a furan ring. In the present work we explored the potential of its essential oils obtained from different organs and the main oxygenated sesquiterpenes, namely isofuranodiene, germacrone and β-acetoxyfuranoeudesm-4(15)-ene, as inhibitors of Trypanosoma brucei. All essential oils effectively inhibited the growth of parasite showing IC50 values of 1.9-4.0μg/ml. Among the main essential oil constituents, isofuranodiene exhibited a significant and selective inhibitory activity against T. brucei (IC50 of 0.6μg/ml, SI=30), with β-acetoxyfuranoeudesm-4(15)-ene giving a moderate potentiating effect. These results shed light on the possible application of isofuranodiene as an antiprotozoal agent to be included in combination treatments aimed not only at curing patients but also at preventing the diffusion of HAT

Identification of Tagitinin C from Tithonia diversifolia as antitrypanosomal compound using bioactivity-guided fractionation

Tithonia diversifolia (Asteraceae), is used as traditional medicine in tropical countries for the treatment of various diseases, including malaria. Although numerous studies have assessed the antimalarial properties, nothing is known about the effect of T. diversifolia extracts on trypanosomiasis. In this study extracts of T. diversifolia aerial parts were evaluated for their bioactivity against Trypanosoma brucei. The activity was studied against bloodstream forms of T. brucei (TC221), as well as against mammalian cells (BALB/3T3 mouse fibroblasts), as a counter-screen for toxicity. Both methanolic and aqueous extracts showed significant effects with IC50 values of 1.1 and 2.2 μg/mL against T. brucei (TC221) and 5.2 and 3.7 μg/mL against BALB/3T3 cells, respectively. A bioassay-guided fractionation on the methanolic extract yielded in identification of active fractions (F8 and F9) with IC50 values of 0.41 and 0.43 μg/mL, respectively, against T. brucei (TC221) and 1.4 and 1.5 μg/mL, respectively, against BALB/3T3 cells,. The phytochemical composition of the extracts and the purified fractions were investigated using HPLC-ESI-MS/MS and 1D and 2D NMR spectra showing the presence of sesquiterpene lactones that in turn were subjected to the isolation procedure. Tagitinin A and C were rather active but the latter presented a very strong inhibition on T. brucei (TC221) with an IC50 value of 0.0042 μg/mL. This activity was 4.5 times better than that of the reference drug suramin. The results of this study shed light on the antitrypanosomal effects of T. diversifolia extracts and highlighted tagitinin C as one of the possible responsible for this effect. Further structure activity relationships studies on tagitinins are needed to consider this sesquiterpenes as lead compounds for the development of new antitrypanosomal drugs

Oviposition inhibitory activity of the Mexican sunflower Tithonia diversifolia (Asteraceae) polar extracts against the two-spotted spider mite Tetranychus urticae (Tetranychidae)

The Mexican sunflower (Tithonia diversifolia, Asteraceae) is an invasive shrub of agricultural and non-agricultural lands in tropical countries. Besides extensive utilizations in the traditional medicine, mainly to treat malaria, the plant is believed to have a great potential in agriculture of developing countries as a green biomass to produce fertilizer, fodder and biopesticides. The plant is known to produce tagitinins, which are sesquiterpene lactones with a bitter taste endowed with toxicity against several insects such as mosquitoes, aphids, and beetles. Here, we evaluated the potential of T. diversifolia against the two-spotted spider mite Tetranychus urticae (Tetranychidae), which is one of the most economically important arthropod pests worldwide. The leaf methanolic extract and its ethyl acetate fraction were tested for acute and chronic toxicity and for oviposition inhibitory effects. The chemical composition of the extracts was analyzed by HPLC-MSn and NMR. The main constituents were flavonoid derivatives, phenylpropanoids and sesquiterpene lactones. Among the latter, tagitinin C and tagitinin A were the major compounds. In acute toxicity assays, mortality did not exceed 50% even for the highest tested dose of 150 μg cm-3. However, in chronic toxicity assays, on day 5 from application, the methanolic extract LD50 was 41.3 μg cm-3 while LD90 was 98.7 μg cm-3. Furthermore, both T. diversifolia extracts inhibited oviposition in T. urticae. The ethyl acetate extract was the most active oviposition inhibitor, with an ED50 value of 44.3 μg cm-3 and an ED90 of 121.5 μg cm-3. Overall, the good yield rate of the extract and the high crop yield highlighted good prospects of using the extract from this plant for the development of oviposition inhibitors against mites