Adenosine receptors as promising targets for the management of ocular diseases

The ocular drug discovery arena has undergone a significant improvement in the last few years culminating in the FDA approvals of 8 new drugs. However, despite a large number of drugs, generics, and combination products available, it remains an urgent need to find breakthrough strategies and therapies for tackling ocular diseases. Targeting the adenosinergic system may represent an innovative strategy for discovering new ocular therapeutics. This review focused on the recent advance in the field and described the numerous nucleoside and non-nucleoside modulators of the four adenosine receptors (ARs) used as potential tools or clinical drug candidates

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

CHEMICAL COMPOSITION OF TEN ESSENTIAL OILS FROM CALOPHYLLUM INOPHYLLUM LINN AND THEIR TOXICITY AGAINST ARTEMIA SALINA

Essential oils from ten different parts of Calophyllum inophyllum were analyzed by Gas Chromatography-Mass Spectrometry (GC-MS) to study their chemical compositions. The yields were between 0.219 and 0.506 %. A total of 102 compounds were identified in the ten C. inophyllum essential oils, which are mostly monoterpenes, sesquiterpenes and their oxygenated derivatives. The numbers and percentages of identified compounds varied in the different parts of the plant: leaf (71, 54.94%), leaf stalk (22, 79.55%), flower (25, 51.24%), seed (25, 89.39%), seed-coat (69, 73.80%) fruit-pulp (15, 46.10%), stem wood (55, 59.40%), stem bark (9, 69.38%) root wood (51, 58.73%), and root bark (24, 74.66%). High content of cymene, terpinene, and limonene in the oils may be responsible for the vast ethno-medicinal applications of the plant. Toxicity experiments show that the oils were fairly toxic. Each part after 24 hours of exposure against Artemia salina gave the following LC50 values in μg/mL: leaf (68.8740 μg/mL), leaf-stalk (102.5692 μg/mL), flower (114.4410 μg/mL), seed (132.2324 μg/mL), seed coat (137.1206 μg/mL), fruit-pulp (135.0350 μg/mL) stem wood (126.1410 μg/mL), stem bark (149.7237 μg/mL), root wood (110.6539 μg/mL) and root bark (110.6539 μg/mL). The chemical compositions and toxicity levels of these ten Calophyllum inophyllum essential oils are reported for the first time in literature

Inhibition of HIV-1 replication in macrophages by red blood cell-mediated delivery of a heterodinucleotide of azidothymidine and 9-(R)-2-(phosphono methoxypropyl)adenine.

Monocyte-derived macrophages (M/M) are considered important in vivo reservoirs for different kinds of viruses, including HIV. Hence, therapeutic strategies are urgently needed to protect these cells from virus infection or to control viral replication. In this paper, we report the synthesis, target delivery and in vitro efficacy of a new heterodinucleotide (AZTpPMPA), able to inhibit HIV-1 production in human macrophages. AZTpPMPA consists of two established anti-HIV drugs [zidovudine (AZT) and tenofovir (PMPA)] chemically coupled together by a phosphate bridge. This drug is not able to prevent p24 production when administered for 18 h to M/M experimentally infected with HIV-1 Bal (inhibition 27%), but can almost completely suppress virus production when given encapsulated into autologous erythrocytes (inhibition of p24 production 97%). AZTpPMPA is slowly converted to PMPA, AZT monophosphate and AZT (36 h half-life at 37°C) by cell-resident enzymes. Thus AZTpPMPA should be considered a new prodrug of AZT and PMPA that is able to provide stechiometric amounts of both nucleoside analogues to macrophage cells and to overcome the low phosphorylating activity of M/M for AZT and the modest permeability of PMPA

5'-C-ethyl-tetrazolyl-N 6-substituted adenosine and 2-chloro-adenosine derivatives as highly potent dual acting A1 adenosine receptor agonists and A3 adenosine receptor antagonists

A series of N(6)-substituted-5'-C-(2-ethyl-2H-tetrazol-5-yl)-adenosine and 2-chloro-adenosine derivatives was synthesized as novel, highly potent dual acting hA1AR agonists and hA3AR antagonists, potentially useful in the treatment of glaucoma and other diseases. The best affinity and selectivity profiles were achieved by N(6)-substitution with a 2-fluoro-4-chloro-phenyl- or a methyl- group. Through an in silico receptor-driven approach, the molecular bases of the hA1- and hA3AR recognition and activation of this series of 5'-C-ethyl-tetrazolyl derivatives were explained

Studying GGDEF Domain in the Act: Minimize Conformational Frustration to Prevent Artefacts

GGDEF-containing proteins respond to different environmental cues to finely modulate cyclic diguanylate (c-di-GMP) levels in time and space, making the allosteric control a distinctive trait of the corresponding proteins. The diguanylate cyclase mechanism is emblematic of this control: two GGDEF domains, each binding one GTP molecule, must dimerize to enter catalysis and yield c-di-GMP. The need for dimerization makes the GGDEF domain an ideal conformational switch in multidomain proteins. A re-evaluation of the kinetic profile of previously characterized GGDEF domains indicated that they are also able to convert GTP to GMP: this unexpected reactivity occurs when conformational issues hamper the cyclase activity. These results create new questions regarding the characterization and engineering of these proteins for in solution or structural studies

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

Synthesis and potency of novel uracil nucleotides and derivatives as P2Y2 and P2Y6 receptor agonists

The phosphate, uracil, and ribose moieties of uracil nucleotides were varied structurally for evaluation of agonist activity at the human P2Y2, P2Y4, and P2Y6 receptors. The 2-thio modification, found previously to enhance P2Y2 receptor potency, could be combined with other favorable modifications to produce novel molecules that exhibit high potencies and receptor selectivities. Phosphonomethylene bridges introduced for stability in analogues of UDP, UTP and uracil dinucleotides markedly reduced potency. Truncation of dinucleotide agonists of the P2Y2 receptor, in the form of Up4-sugars, indicated that a terminal uracil ring is not essential for moderate potency at this receptor and that specific SAR patterns are observed at this distal end of the molecule. Key compounds reported in this study include: 9, α,β-methylene-UDP, a P2Y6 receptor agonist; 30, Up4-phenyl ester and 34, Up4-[1]glucose, selective P2Y2 receptor agonists; 43, the 2-thio analogue of INS37217 (P1-(uridine 5′)-P4- (2′-deoxycytidine 5′) tetraphosphate), a potent and selective P2Y2 receptor agonist

Discovery of multitargeting chemical entities for the treatment of Parkinson's disease

Since half of the world's older population lives in Asia, neurodegenerative diseases including Parkinson‘s disease (PD) become a serious problem that should be investigated in greater depth in this region. Indeed, there is a huge effort towards the identification of new, effective and safe drugs for the successful therapy of PD: previous approaches focused on highly selective compounds targeting individual proteins (i.e., ―one compound-one target‖ approach). Subsequently, poly-pharmacology has gained much interest, since the heterodimerization of A2A adenosine receptors and D2 dopamine receptors in the brain was established. Hence, recent anti-parkinson therapies combine A2A antagonists and D2 selective agonists through a ―cocktail drugs-multiple targets‖ approach. Although a few multi-component therapies have entered clinical trials, these methodologies are often associated with side effects, due to the complicated interaction among drugs and different pharmacokinetic profiles. In view of these limitations, we have brought together key leaders with diversified expertise, in order to identify New Chemical Entities that are able to modulate the activity of both D2 and A2A receptors simultaneously (i.e., ―one compound-multiple targets‖ approach). The advantages associated with this strategy encompass avoidance of drug-drug interactions, clear pharmacokinetic and pharmacodynamic profiles, more predictable metabolic instabilities and more likely lower side effects. Indeed, none of the proposed treatments for PD uses a single molecule to trigger the desired therapeutic effects. This project includes molecular modeling, organic syntheses, crystallography, genetic studies as well as in vitro and in vivo pharmacological evaluations, through a multi-institutional initiative involving the National University of Singapore, the Nanyang Technological University, the Duke-NUS Medical School, the Drug Development Unit and a few European academic partners. This team effort will result not only in the development of more effective drug candidates or better models of disease, but it will enable the translation of these new methodologies into clinically relevant medicine paradigms