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Made available in DSpace on 2016-12-08T16:44:36Z (GMT). No. of bitstreams: 1 PGPV10MA058.pdf: 1288592 bytes, checksum: a78d5ad5cb453cad5bba3781ca902dfd (MD5) Previous issue date: 2010-12-15 / High land fields in the Coxilha Rica region of Santa Catarina state have a worthy biodiversity due to low anthropic interference and the singular predominant ecosystem. Traditional extensive cattle system practiced during several generations allowed to keep some reminiscent forestry. The ecosystems that had low human interference may keep high biodiversity with several species, having therapeutic, aromatic, nutritional, and condiment properties. It is very important to study the native flora in such region once biodiversity is threatened by water powering (PCH`s), re-forestation, and agriculture large plantations. Native plant species that are endemically in such region may reduce gradually its occurrence or even became extinct. Culture and customs in this region have been strongly influenced by indigenous, slaves, and gauchos knowledge. The use of plants as medicines in the local communities of Coxilha Rica is a part of its tradition, but it has been decreases with time. Ethno-knowledge may help to maintain the communities integrated with local genetic resources. To recognize and prospect native genetic resources would be essential to rescue dignity and to empowering people for local sustainable development, which may improve live quality. Mostly of plant species that are characteristic successional species in the high land fields were not yet studied for their potential economic exploration, even they are already useed for local communities. Medicinal plants consisted of the most large group of plant that are currently used by the communities living in the high land fields and it gives the spatial name of medicina campeira . One of the most frequently used species is Poiretia spp., that is indicated for stomach pain. The presence of volatile essential oil gives the potential to be used also as cosmetic. The objective of this work was to do assessment of medicinal plants used by farmers living in Coxilha Rica region of Santa Catarina state. Studies were carried out in the follow communities: Cajurú, Morrinhos, Pinheiro Seco, São Jorge, Faxinal, Km 14, Raposo, Escurinho, Pelotinhas, Vigia, Santo Cristo, Borel, and Rincão do Perigo. Semi-structured questionnaire was applied to each visited farmer. Informations included social-cultural aspects, indicated treatment, medicinal plant occurrence, habitat, processing, and plant storage methods. The medicinal species of Poiretia latifolia was characterized by taking data of its natural habitat and the associated plant community. Multiplication systems of Poiretia latifolia were studied in the laboratory. Results showed that the farmers living in the Coxilha Rica region used frequently plant for human treatment. It was reported 121 plant species as medicinal and 50 of them to be native occurrence in the South Brazil. The most used plants include Achyrocline satureioides, Maytenus ilicifolia, Chionolaena latifolia, Persea pirifolia, Baccharis trimera, Malva sylvestris, Mentha pulegium, Poiretia latifolia, and Trichocline macrocephala. The most frequent treatments with plants were for stomach pain, hunting, and colt. Poiretia latifolia occurred in field areas, shallower soil covering rocks and stones. It is dispersed in all communities of Coxilha Rica region. It may be conclude that Poiretia latifolia is a frequent genetic resorce presente in the Planalto Serrano Catarinense and it has a valuable local knowledge associated with it. This makes the species P. latifolia a potential plant species for domestication and used in economic cropping. For domestication purposes of Poiretia latifolia it is necessary further studies about multiplication and ecology. There are several native plant species occurring in high land fields of the Coxilha Rica region that would be multiplied to cultivate as medicinal plant once local farmers had been used from long time / Os campos nativos de altitude da região da Coxilha Rica, SC, possuem biodiversidade peculiar pouco alterada, mantendo características naturais do ecossistema Campos. Isto se deve, sobretudo, pela pecuária extensiva tradicionalmente explorada há várias gerações associada a manutenção de remanescentes florestais,. Os ecossistemas pouco modificados apresentam alta riqueza de recursos genéticos e podem ser portadores de genótipos que contêm propriedades singulares para usos terapêuticos, aromáticos, condimentares, nutricionais, entre outros. A caracterização de espécies da flora silvestre dessa região é primordial para a manutenção e conservação da biodiversidade, ameaçada pela ação antrópica tais como agricultura extensiva, plantio de maciços florestais e construção de pequenas centrais hidrelétricas (PCH s). Com isto, várias espécies vegetais de ocorrência endêmica poderão reduzir gradativamente suas populações ou até mesmo desaparecerem. A região da Coxilha Rica é fortemente influenciada por uma cultura que preserva saberes tradicionais herdados de indígenas, escravos e tropeiros. O uso de plantas no tratamento de doenças na chamada medicina campeira faz parte da cultura popular e é uma alternativa de medicação de baixo custo. Porém, este conhecimento popular está sendo perdido ao longo do tempo. Estudos etnobotânicos poderiam auxiliar na perpetuação dos processos culturais integrados ao uso dos recursos genéticos locais. O reconhecimento e a prospecção destes germoplasmas silvestres, aliados a valorização da cultura tradicional local são essenciais para atingir a eqüidade social, resgatando a auto-estima, a dignidade e qualidade de vida do homem que ai reside. A maioria das espécies de plantas sucessionais nos campos naturais não tem ainda potencial de exploração econômica conhecido, embora já fazem parte do uso na cultura local. As plantas medicinais compõem um dos grupos de maior riqueza, tem sido utilizadas pelos moradores da região dos Campos Naturais de Altitude, originando o termo local de medicina campeira . Entre as espécies conhecidas e mais utilizadas pelas famílias locais, cita-se a erva-de-touro, empregada para tratar problemas estomacais e com alto potencial farmacológico e cosmético devido a sua composição com óleos voláteis aromáticos peculiares. O objetivo deste trabalho foi realizar levantamento de espécies utilizadas na medicina campeira por agricultores e pecuaristas da região da Coxilha Rica, SC. Foram realizadas visitas a agricultores/pecuaristas da região da Coxilha Rica, abrangendo as comunidades de Cajurú, Morrinhos, Pinheiro Seco, São Jorge, Faxinal, Km 14, Raposo, Escurinho, Pelotinhas, Vigia, Santo Cristo, Borel e Rincão do Perigo. Para cada visita foi aplicado questionário semi-estruturado abordando aspectos sócio-culturais do informante e informações sobre as plantas utilizadas medicinalmente, local de ocorrência destas plantas, formas de armazenagem, manipulação e uso. A erva-de-touro (Poiretia latifolia) teve caracterizado seu hábitat natural de ocorrência e a comunidade vegetal associada. Formas de multiplicação da erva-de-touro. foram estudadas em laboratório. Dados levantados permitiram verificar que moradores da região da Coxilha Rica usam com frequência plantas medicinais.Foram relatadas 121 espécies de plantas, sendo 50 de ocorrência natural da região Sul do Brasil. As plantas mais usadas foram a macela (Achyrocline satureioides), espinheira-santa (Maytenus ilicifolia), arnica (Chionolaena latifolia), pau-andrade (Persea pirifolia), carqueja (Baccharis trimera), malva (Malva sylvestris), poejo (Mentha pulegium), erva-de-touro (Poiretia latifolia) e cravo-do-campo (Trichocline macrocephala). Os usos mais comuns relatados foram para problemas digestivos, ferimentos e gripe. A erva-de-touro (P. latifolia) ocorreu predominantemente em áreas de campo e em solos pedregosos e é naturalmente dispersa em toda região da Coxilha Rica. Pode-se concluir que a erva-de-touro é um recurso genético do Planalto Serrano Catarinense, dispõe de considerável conhecimento associado à população local e tem potencial de exploração econômica. Para a domesticação da espécie e inclusão em sistemas de produção agrícola são necessários maiores estudos sobre a ecologia da espécie e formas de multiplicação. Por outro lado, outras espécies nativas são igualmente de alto valor medicinal conforme apontado pelos moradores da Coxilha Rica

The ham mite, Tyrophagus putrescentiae (Schrank) (Sarcoptiformes: Acaridae) is the predominant pest of dry cured hams during aging in the processing facilities. Methyl bromide is currently the only known fumigant that is effective at controlling ham mite infestations in aging houses. However, methyl bromide is being phased out of all industries and will be depleted in the near future. The research objectives were to 1) evaluate dry cured hams that have been treated with previously developed food grade coatings for sensory differences, and 2) to develop and determine the efficacy of ham nets incorporated with food grade coatings on controlling mite infestations and sensory properties. Food grade coating combinations of 1) propylene glycol (PG), xanthan gum, and water or 2) PG, propylene glycol alginates, carrageenan and water were dipped and sprayed on whole hams in commercial facilities in the summers of 2014 and 2015 (composition patent pending). The lowest concentration of propylene glycol needed to control mites in laboratory studies was 15% with xanthan gum and 7.5% with propylene glycol alginate and carrageenan. Sensory difference from control tests with trained panelists indicated that there were slight to moderate differences detected in some of the treated hams in comparison to untreated control hams (P < 0.05) when hams were dipped with coatings. However, there were no differences (P > 0.05) detected between the treated hams and the control hams when hams were only sprayed rather than dipped with these coatings. Polyester/cotton blend or cotton nets were infused with various food grade coatings and evaluated on the bench top by inoculating 20 adult mites onto one inch ham cubes for their efficacy at controlling mite infestations. Live adults and mobile immature stages were counted after 14 d of incubation (23 ± 2 °C and 70 ± 5% RH). Mite infestation tests demonstrated that coatings and coating-treated nets were effective at controlling mite growth. Therefore, food grade coatings can be applied to dry cured hams and also can be infused into nets as a potential means to control mite growth in ham processing facilities.

Acrocomia é do gênero pertencente à família Arecacea, nativa de florestas tropicais. A polpa do fruto macaúba tem usos alimentícios, como sucos, geleias, sorvetes que possuem substâncias bioativas, “ditas funcionais”, como antioxidantes, fibras e minerais imunomodularodes como o Zinco. Sabe-se que fibras e compostos antioxidantes têm ação estimulante para o tratamento de inflamações. O presente trabalho teve como objetivo investigar a atividade antibacteriana e anti-inflamatória dos extratos e das frações do óleo do mesocarpo da Acrocomia aculeataobtidas por extração a frio e as frações por cromatografia de coluna. Ascepas bacterianas foram tratadas com os extratos à uma concentração de 5mg/ml para a avaliação da atividade bacteriana e para controle positivo foi utilizado o antimicrobiano convencional Imipenen. Na avaliação da atividade antibacteriana foi observado que nenhum dos extratos testados mostraram atividade frente às bactérias Grampositivas e Gram-negativas. Quanto a avaliação da atividade de mediadores da inflamação, as células foram tratadas com os bioativos oleoso do mesocarpo da macaúba e estimuladas com LPS. Após 24 horas o sobrenadante celular foi coletado para avaliação da produção de óxido nítrico (NO•) pela reação de Griess. Analisandoos efeitos sobre a produção de mediadores inflamatórios, foi observado no primeiro ensaio que as frações hexânica (FHex) e metanólica (FMeOH)na concentração de 25 e 50 μg/ml inibiram a produção de NO•, em células macrófagos murino J774 estimulados por LPS. Em ensaios posteriores nas mesmas condições não demonstrou nenhum tipo de atividade, sugerindo a realização de estudos futuros na obtenção do extrato, ou no fracionamento em cromatografia de coluna e até mesmo adaptações nos métodos usados neste trabalho. Este estudo pode fornecer uma base para investigações futuras sobre o papel terapêutico do óleo do mesocarpo no tratamento da inflamação. / Acromonia is a gender belonging to the Arecaceae family and native from the tropical forests. Food is one of the many subjects related to the fruit pulp from macauba, for example: juices, jellies, ice-creams which possess bio actives substances, called as antioxidant, fibers and immune modulators minerals like the Zinc; it is known that fibers and antioxidant compounds have stimulating action to inflammations treatments. This study had the purpose to investigate the antibacterial activity and anti-inflammatory from the extracts and fractions of the Acrocomia aculeata mesocarp oil achieved by extraction of fractions from the spine chromatography. The bacterial vines were treated with the extracts concentration of 5mg/ml to evaluate the bacterial activity e to the positive control was used the conventional antimicrobial Imipenen. In the evaluation of the antibacterial activity was observed that none of the tested extractions showed activity upon the Gram-positives and Gram-negatives bacteria’s. As for the activity evaluation of inflammation mediators, the cells were treated with oil bio actives from macauba’s mesocarp and stimulated with LPS. After 24 hours the cellular supernatant was collected to evaluation of nitric oxide production (NO°) for reaction of Griess. Analyzing the effects under the production of inflammatory mediators, was observed that in the first test that the hexanica fractions (FHex) and metalonica (FmeOH) in concertation of 25 and 50 mg/ml inhibited the NO° production, on macrophages murine J774 cells stimulated by LPS. On further tests in the same conditions doesn’t showed any kind of activity, suggesting for future studies futures observations on obtain the extract or on fractioning on spine chromatography or even for the same adaptations on methods used in this study. This work can provide a base to future investigation about the therapeutic paper of mesocarp oil on inflammation treatment.

The thesis entitled “Enantioselective synthesis of bio-active bicyclic acetals, cyclic ethers and lactones” demonstrates the utility of chiral pool tartaric acid as the source in the synthesis of natural products. The results are discussed in three chapters; 1) Enantioselective synthesis of bio-active bicyclic acetals, 2) Enantioselective synthesis of bio-active cyclic ethers and 3) Enantioselective synthesis of bio-active lactones. A brief introduction is provided in each chapter to keep the present work in proper perspective. Compounds (in bold) and references (in superscripts) are sequentially numbered differently for each chapter and references are given as foot notes. Experimental procedures are given differently for each chapter and placed at the end of chapter. Scanned 1H and 13C NMR spectras are given with description of signals. Chapter 1 describes the enantioselective synthesis of bicyclic acetal containing insect pheromones. First part of this chapter deals with the enantiodivergent synthesis of both enantiomers of hydroxy-exo-brevicomin and 2-hydroxy-exo-brevicomin starting from a single chiral compound, bis-Weinreb amide derived from L-(+)-tartaric acid. Controlled addition of Grignard reagent to bis-Weinreb amide followed by diastereoselective reduction of the resultant ketone was employed as the key step for the enantiodivergent synthesis of hydroxy-exo-brevicomin and 2-hydroxy-exo-brevicomin. In the second part, enantioselective synthesis of exo-brevicomin, iso-exo-brevicomin and formal synthesis of frontalin comprising similar framework is demonstrated, utilizing á -benzyloxy aldehydes derived from L-(+)-tartaric acid as chiral building block. Second Chapter describes the enantioselective synthesis of bio-active cyclic ethers, disparlure, centrolobine and isolaurepan. Employing á-benzyloxy aldehydes derived from L-(+)-tartaric acid as the chiral building block, synthesis of both enantiomers of insect pheromone disparlure is achieved involving the diastereoselective addition of allyltributyl tin to the á-benzyloxy aldehyde and cross metathesis of the resultant homoallylic alcohol with 4-methyl-1-pentene. Formal synthesis of centrolobine and isolaurepan are accomplished. Pivotal step involved in the synthesis of centrolobine is iron(III) mediated cyclization of 1,5-diol derived from L-(+)-tartaric acid, while Lewis acid mediated reductive cyclization of the hydroxy ketone derived from á-benzyloxy aldehyde is the key step in the synthesis of isolaurepan. Third chapter in the thesis deals with the enantioselective synthesis of bio-active lactones muricatacin, 6-acetoxy-5-hexadecanolide and boronolide. Utilizing á-benzyloxy aldehyde as the building block, synthesis of five and six membered lactones, muricatacin and 6-acetoxy-5-hexadecanolide were accomplished via the diastereoselective addition of 3-butenylmagnesium bromide and allyltributyl tin to á-benzyloxy aldehyde, respectively. Stereoselective formal synthesis of boronolide was described, starting from D-(–)-tartaric acid. Key reaction sequence includes the elaboration of ã-hydroxy amide obtained by a combination of controlled Grignard addition and diastereoselective reduction from bis- Weinreb amide derived from D-(–)-tartaric acid.

Bumblebees are important pollinators, commercially used in large-scale plant growing in greenhouses. Their males produce marking pheromones for mating, which attract young bumblebee queens. These pheromones are often a complicated mixture of chemicals, which is produced in the male labial gland, and the mixture itself is specific for each bumblebee species. The regulation of bumblebee sexual pheromone biosynthesis is largely unknown, and this Master's thesis is focused on the analysis of the mechanisms which lead to the regulation of the Bombus terrestris male pheromone's fat-acid and terpenes biosynthesis, specifically on stereospecific enzymatic reduction of double bond of farnesol. This thesis studies the influence of potential neurohormones on a specific enzymatic mixture, which is involved in the pheromone biosynthesis. Methods used in this project include biochemical, analytic and molecular-biology methods.

The thesis entitled “Total synthesis of bio-active macrolide natural products and sulphonamide based ligands in asymmetric catalysis” is divided into two chapters. First chapter of the thesis describes the total synthesis of bio-active macrolide natural products cladospolide A 1, seimatopolide A 2 and synthetic studies towards aetheramides A 3 and B 4 (Figure 1). Figure 1: Bio-active macrolide natural products. Section A of chapter 1 describes the enantiospecific total synthesis of cladospolide A (ent-1). Cladospolide A was isolated from three different sources such as culture filtrate of cladosporium fulvam FI-113, Fungus cladosporium tenuissimum and Fermentation broath of cladosporium sp. FT-0012. Cladospolide A is shown to inhibit the root growth of lettuce seedlings. Enantiospecific total synthesis of cladospolide A ent-1 was accomplished in 9% overall yield in 11 linear steps using D-ribose as a chiral pool precursor. Key reactions in the present approach include olefin cross metathesis and Yamaguchi macrolactonization reactions (Scheme 1). Scheme 1: Total synthesis of cladospolide A (ent-1). Section B of chapter 1 describes the use of furan as a surrogate for the E-but-2-ene-1, 4-dione unit in the total synthesis of seimatopolide A 2. Seimatopolide A 2 was isolated by Heip and co-workers from the fungus Seimatosporium discosioides in 2012 and is shown to activate the γ-subtype peroxysome proliferator-activated receptors (PPAR-γ), which is a pivotal process in the type-2 diabetes. Total synthesis of ent-seimatopolide A was accomplished in 7.8% overall yield in 14 linear steps from furfural. Nagao acetate aldol and Shiina macrolactonization reactions were employed as key reactions for the synthesis of ent-seimatopolide A (ent-2) (Scheme 2). Scheme 2: Stereoselective total synthesis of seimatopolide A (ent-2). In section C of Chapter 1, studies towards the synthesis of aetheramides A 3 and B 4 are described. Aetheramides A 3 and B 4 are isolated by Müller’s group in 2012 from the novel myxobacterial genus “Aetherobacter”. Aetheramides are cyclic depsipeptides, which are shown to inhibit the HIV-I infection with IC50 values of ∼0.015 μM and cytostatic activity against human colon carcinoma (HCT-116) cells with IC50 values of 0.11 μM. Stereochemistry at two chiral centers present in the molecules is unassigned. The first approach (Scheme 3) relied on macrolactonization as the key step while the second approach (Scheme 4) relied on RCM to accomplish the macrolactonization. The required precursors were synthesized from elaboration of chiral furyl carbinol, while synthesis of the RCM precursor was accomplished employing the aldol reaction. Scheme 3: Macrolactonization strategy for synthesis of 3 from chiral furyl carbinol. Scheme 4: RCM strategy for synthesis of 3 from chiral furyl carbinol. The successful synthesis of the macrolactone core of aetheramide A 1 is accomplished by employing the ring closing metathesis reaction to construct the C18-C19 bond. RCM precursor has been synthesized by the amidation of the amine derived from R-mandelic acid, while the acid fragment is synthesized from allyl trityl ether (Scheme 5). Scheme 5: RCM strategy for synthesis of 3 from R-mandelic acid. Second chapter of the thesis describes the synthesis and application of novel sulfinamide ligands in asymmetric catalysis. In section A of chapter 2, chiral 2-pyridylsulfinamides are shown to be effective catalysts in the alkylation of aryl and alkyl aldehydes with diethylzinc providing the corresponding alcohols in excellent enantioselectivity. It was found that the chirality present at sulfur in the ligand is pivotal for the asymmetric induction (Scheme 6). Scheme 6: Asymmetric alkylation of benzaldehyde with some of the 2-Pyridyl sulfinamide catalysts. Second section of chapter 2 describes the synthesis and application of C2-symmetric bis-sulfinamides in Rh (I) catalyzed conjugate addition of PhB(OH)2 to enones. Chirality present at sulphur in sulfonamide as well as symmetry present in the ligand plays crucial role in the outcome of the reaction (Scheme 7). Scheme 7: Asymmetric arylation of enones using C2-symmetric bis-sulfinamide/olefin ligands. The thesis entitled “Total synthesis of bio-active macrolide natural products and sulphonamide based ligands in asymmetric catalysis” is divided into two chapters. First chapter of the thesis describes the total synthesis of bio-active macrolide natural products cladospolide A 1, seimatopolide A 2 and synthetic studies towards aetheramides A 3 and B 4 (Figure 1). Figure 1: Bio-active macrolide natural products. Section A of chapter 1 describes the enantiospecific total synthesis of cladospolide A (ent-1). Cladospolide A was isolated from three different sources such as culture filtrate of cladosporium fulvam FI-113, Fungus cladosporium tenuissimum and Fermentation broath of cladosporium sp. FT-0012. Cladospolide A is shown to inhibit the root growth of lettuce seedlings. Enantiospecific total synthesis of cladospolide A ent-1 was accomplished in 9% overall yield in 11 linear steps using D-ribose as a chiral pool precursor. Key reactions in the present approach include olefin cross metathesis and Yamaguchi macrolactonization reactions (Scheme 1). Scheme 1: Total synthesis of cladospolide A (ent-1). Section B of chapter 1 describes the use of furan as a surrogate for the E-but-2-ene-1, 4-dione unit in the total synthesis of seimatopolide A 2. Seimatopolide A 2 was isolated by Heip and co-workers from the fungus Seimatosporium discosioides in 2012 and is shown to activate the γ-subtype peroxysome proliferator-activated receptors (PPAR-γ), which is a pivotal process in the type-2 diabetes. Total synthesis of ent-seimatopolide A was accomplished in 7.8% overall yield in 14 linear steps from furfural. Nagao acetate aldol and Shiina macrolactonization reactions were employed as key reactions for the synthesis of ent-seimatopolide A (ent-2) (Scheme 2). Scheme 2: Stereoselective total synthesis of seimatopolide A (ent-2). In section C of Chapter 1, studies towards the synthesis of aetheramides A 3 and B 4 are described. Aetheramides A 3 and B 4 are isolated by Müller’s group in 2012 from the novel myxobacterial genus “Aetherobacter”. Aetheramides are cyclic depsipeptides, which are shown to inhibit the HIV-I infection with IC50 values of ∼0.015 μM and cytostatic activity against human colon carcinoma (HCT-116) cells with IC50 values of 0.11 μM. Stereochemistry at two chiral centers present in the molecules is unassigned. The first approach (Scheme 3) relied on macrolactonization as the key step while the second approach (Scheme 4) relied on RCM to accomplish the macrolactonization. The required precursors were synthesized from elaboration of chiral furyl carbinol, while synthesis of the RCM precursor was accomplished employing the aldol reaction. Scheme 3: Macrolactonization strategy for synthesis of 3 from chiral furyl carbinol. Scheme 4: RCM strategy for synthesis of 3 from chiral furyl carbinol. The successful synthesis of the macrolactone core of aetheramide A 1 is accomplished by employing the ring closing metathesis reaction to construct the C18-C19 bond. RCM precursor has been synthesized by the amidation of the amine derived from R-mandelic acid, while the acid fragment is synthesized from allyl trityl ether (Scheme 5). Scheme 5: RCM strategy for synthesis of 3 from R-mandelic acid. Second chapter of the thesis describes the synthesis and application of novel sulfinamide ligands in asymmetric catalysis. In section A of chapter 2, chiral 2-pyridylsulfinamides are shown to be effective catalysts in the alkylation of aryl and alkyl aldehydes with diethylzinc providing the corresponding alcohols in excellent enantioselectivity. It was found that the chirality present at sulfur in the ligand is pivotal for the asymmetric induction (Scheme 6). Scheme 6: Asymmetric alkylation of benzaldehyde with some of the 2-Pyridyl sulfinamide catalysts. Second section of chapter 2 describes the synthesis and application of C2-symmetric bis-sulfinamides in Rh (I) catalyzed conjugate addition of PhB(OH)2 to enones. Chirality present at sulphur in sulfonamide as well as symmetry present in the ligand plays crucial role in the outcome of the reaction (Scheme 7). Scheme 7: Asymmetric arylation of enones using C2-symmetric bis-sulfinamide/olefin ligands.

Total synthesis of natural products is of contemporary interest in organic synthesis. One of the useful ways to synthesize the natural products is to originate from inexpensive chiral pool compounds abundantly available in nature. In this context, our research group is actively involved in the use of tartaric acid as the four carbon four hydroxy building block in the synthesis of a number of natural products of therapeutic importance. Our strategy relies on the utility of γ-hydroxy amides derived from tartaric acid involving a controlled addition of Grignard reagents and stereoselective reduction. We were successful in application o f this useful building block for the synthesis of a variety of natural products possessing varied functional groups (Chart-1). derived from tartaric acid in the synthesis of oxylipins such as pinellic acid and diyne containing natural products. Chapter 1 of the thesis describes the total synthesis of (+) pinellic acid 6 and (Z,8S,9S,10R)-8,9,10-trihydroxyoctadec-6-enoic acid 10. Key strategy in the synthesis of pinellic acid is elaboration of the aldehyde 3, derived from the γ-hydroxy amide 2 via Horner-Emmons-Wadsworth reaction to yield the α,β-unsaturated ketone 4. Stereoselective reduction of the ketone with (R)-BINAL-H produced the alcohol with requisite stereochemistry which was further extended to pinellic acid 6 (Scheme 1). Wittig homologation of the aldehyde 8 derived from γ-hydroxy amide 7 is the key step for the synthesis of the (Z,8S,9S,10R)-8,9,10-trihydroxyoctadec-6-enoic acid 10. Second chapter of the thesis deals with total synthesis of diyne containing natural products. In the first part of this chapter enantioselective synthesis of possible diastereomers of heptadeca1-ene-4,6-diyne-3,8,9,10-tetrol, a structure proposed for the natural product isolated from Hydrocotyle leucocephala, is accomplished. The alkyne precursors 13 and 14 were synthesized from the α-hydroxy ester 12 derived from γ-hydroxy amide 11 while the alkyne 17 is synthesized from the masked tetrol 16 derived from lactol 15 which was obtained from D-ribose. yne to assemble the diyne unit which was further elaborated to heptadeca-1-ene-4,6-diyne3,8,9,10-tetrol (Scheme 3). It was found that the NMR spectral data of the putative structure assigned for the natural product did not match with any of the diasteromers that were synthesized. This establishes that the structure proposed for the natural product is wrong and requires revision. OH OH OH 18 OH OH 19 OH OH 20 OH OH Scheme 3: Synthesis of diastereomers of heptadeca-1-ene-4,6-diyne-3,8,9,10-tetrol. [Part of this work is published: Prasad, K. R.; Swain, B. J. Org. Chem. (in press)] Second part of this chapter deals with the synthesis of panaxytriol 26 and panaxydiol 28. Key reaction in the synthesis of panaxytriol and panaxdiol is the coupling of bromoalkynes 25 and 27 with 3-silyloxy pent-1-en-4-yne and further elaboration to the triol and diol. The required alkynes were synthesized from the primary alcohol 24 which was obtained from the γ-hydroxy amide 11 involving a series of simple synthetic operations. (Scheme 4). (For structural formula pl see the abstract file)

Ideally, every person in the world should have access to a safe and clean water supply; if not all sources of water are clean and safe, at the very least, an effective method to detect water contamination should be readily available. An effective detection method should not only be sensitive, rapid, robust, and affordable, but, ideally, it should also be equipment-free and easy to transport and deliver to the end-users. The main goal of this project is to develop a variety of bits and pieces of bioassay systems, with a particular focus on paper-based bioactive devices in order to provide portable and ready-to-use biosensors which can be useable by anyone anywhere around the world without requiring formal training. According to the World Health Organization (WHO), 76,000 people each year die in India alone because of pesticide poisoning. Long term exposure to organophosphate pesticides is known to have adverse effects on neurological function and can lead to Alzheimer's Disease, Attention Deficit Hyperactivity Disorder (ADHD), and reduced Intelligence Quotient (IQ). The likelihood of long term exposure to pesticides is heightened in developing countries, so a reliable and inexpensive pesticide sensor is a much-needed device in the developing world. To address this need, this project reports on the development of a fully-automated bioactive paper-based sensor for the detection of organophosphate pesticides. In the proposed biosensor, two innovations were implemented to achieve a full-automated format for the pesticide sensor: (I) First is a PUMP ON A PAPER (Jahanshahi-Anbuhi et al., LOC, 2012) that increases the flow rate of fluids within paper-based microfluidic analytical devices and sequentially brings two separate liquid streams to the enzyme test zone on the paper sensor, and (II) the second innovation is a PIPETTE ON A PAPER (Jahanshahi-Anbuhi et al., LOC, 2014) that involved the creation of a pullulan (a natural non-ionic polysaccharide) temporary bridge-system to transfer a known amount of solution to the sensing zone that, gives the enzyme zone a chance to dry and accept the substrate solution from the slow channel after a fixed period of time. This proposed format results in a simplified assay that detects the presence of pesticides automatically without any further manipulation from the user. However, the shelf life of this assay kit is challenging due to instability of both enzyme (AChE) and substrate (IDA) at room temperature. AChE loses its enzymatic activity when stored at room temperature and IDA becomes oxidized quickly. This problem is not unique to these two bio reagents, however; almost all bioassays which use bio-reagents (such as enzymes and small-molecular substrates) are unstable to varying degrees and require special shipping and storage. The instability of these molecules can arise from either thermal denaturation or chemical modification, such as oxidation or hydrolysis. Because of these issues, they often have to be shipped on dry ice with special packaging, which is costly. The cost of maintaining a cold chain for distributing bio-reagents accounts for up to 80% of the cost. Aside from the cost, these reagents also have to be stored in bulk in refrigerators or freezers to minimize the loss of activity, but they must be thawed and aliquoted for their intended tests. Repeated freezing and thawing can result in a significant loss of activity, which often leads to less reliable test results. These issues make running such assays in resource-limited settings a significant challenge. There is, therefore, an urgent need for an assay system with stable reagents that is easy to use, simple to read, inexpensive, and that includes a method for the long-term stabilization of enzymes and other unstable reagents in pre-measured quantities. To overcome to all these issues, pullulan is utilized for the development of pill-based-biosensors. Pullulan dissolves quickly in aqueous solutions and shows very high oxygen barrier properties in its film form. Considering the unique properties of pullulan, it is hypothesized that pullulan may be suitable for producing assay pills with encapsulated enzymes or other unstable molecules and may provide a simplified platform for carrying out bioassays in resource-limited settings. The application of these pill-based-biosensors is shown via the entrapment of AChE and IDA for the creation of an assay kit that can detect organophosphate pesticides (Jahanshahi-Anbuhi et al., Angew. Chem., 2014). Moreover, this thesis reports on the stabilization of highly unstable firefly luciferase for the detection of microorganisms and, more particularly, ATP. Through the use of pullulan, this thesis demonstrates that both the enzyme and the substrate can be protected, immobilized, and stabilized at room temperature, instead of the existing storage methods, which require temperatures <-20˚C. This innovation allows for a more convenient method of shipping the bioassay kits around the world without any extra care. Furthermore, pullulan-based films are utilized for the development of a method for controlled multidirectional flow within paper-based biosensors. This method provides the possibility of trapping labile and volatile reagents and stabilizing them by forming thin films with pullulan. The trapped reagents within pullulan films can be strategically stacked and assembled on a paper strip in different directions. Furthermore, should the need arise, these reagents can be released and delivered sequentially or simultaneously in both vertical and lateral directions through the paper. The application of this method is shown for: (I) creation of "ready-to-use" assay kit for the detection of Escherichia coli (E. Coli). This assay kit has the step of cell lysing and proceeds automatically to the step in which enzymes react. The second application (II) shows the trapping of Simon’s reagents, which is widely used for methamphetamine detection. Overall, these unique fabrication techniques can be widely used for the preparation of highly stable, ready-to-use, and user-friendly biosensors. We are currently working on the detection of other contaminants such as heavy metals, and we are starting on vaccine stabilization and delivery, which would have a tremendous impact for society. / Dissertation / Doctor of Engineering (DEng)