<p>Primenom tečne hromatografije na obrnutim fazama<br />ispitano je retenciono pona&scaron;anje 14 novosintetisanih<br />derivata s-triazina na tankom sloju RP C-18 i silika gelu<br />impregniranim parafinskim uljem. Kao pokretne faze<br />kori&scaron;ćene su dvokomponentne sme&scaron;e vode i organskih<br />rastvarača. Izračunate su R_M^0&nbsp;vrednosti i ispitana je<br />korelacija sa različitim deskriptorima i prediktorima<br />biolo&scaron;ke aktivnosti.</p> / <p>Retentional behaviour of 14newly syntetised derivates&nbsp;of s-triazine on RP C-18 and paraffin oil-impregnated&nbsp;silica gel suport had been investigated using thin-layer&nbsp;<br />chromatography. As a mobilephases were used two-component water based mobile &nbsp;phases. Calculated R_M^0&nbsp;values were corelated to various molecular descriptors&nbsp;<br />as well as biological activity indicators.</p>

A matéria orgânica do solo (MOS) é composta de uma mistura complexa e heterogênea de compostos orgânicos, sendo as substâncias húmicas (SH) a fração mais recalcitrante da MOS. As SH têm grande influência sobre as propriedades químicas e biológicas do solo, contribuindo como fonte de energia para microrganismos e também contribuindo com a disponibilidade de nutrientes. Além disso, as SH podem contribuir de forma expressiva para o desenvolvimento e produção das plantas cultivadas. Neste sentido, o objetivo deste trabalho foi avaliar a atividade e diversidade da comunidade bacteriana do solo, a disponibilidade de nutrientes e a produtividade da cultura de trigo após aplicação de doses crescentes de SH. Dois experimentos foram montados e avaliados, sendo um ensaio de incubação com um solo arenoso e outro um experimento de casa de vegetação com plantas de trigo em dois solos tropicais distintos (textura arenosa e argilosa). Houve um estímulo à atividade microbiana em ambos os experimentos, sendo este estímulo transitório, o que provavelmente ocorreu devido a um efeito priming provocado pela adição de SH. No entanto não houve alteração na estrutura da comunidade bacteriana pela adição de SH, o que pode ser devido ao fato de ser um material recalcitrante e já presente no solo. Também houve aumento na disponibilidade de fósforo, potássio, cálcio e nitrogênio, bem como da CTC, tanto em solo arenoso quanto argiloso, expressando o efeito das SH na melhoria das propriedades químicas de solos tropicais. Além disso, um efeito significativo foi visto em parâmetros relacionados à produtividade do trigo. Houve aumento de massa seca de raízes e parte aérea, além de aumento no peso e número de grãos, o que pode se refletir em aumento considerável de produtividade. Esses resultados são importantes por mostrarem que mesmo a fração mais recalcitrante da MOS pode influenciar positivamente aspectos químicos e biológicos do solo, contribuindo com a melhoria do sistema de produção e também com maiores produtividades vegetais. / The soil organic matter (SOM) is composed of a complex and heterogeneous mixture of organic compounds, in which the humic substances (HS) are considered to be the most recalcitrant fraction. HS have great influence on chemical and biological soil properties, contributing as energy source for microorganisms and as nutrient reservoir for plants. Furthermore, HS can contribute significantly to the development and production of crop plants. Therefore, the objective of this study was to evaluate the activity and diversity of the soil bacterial community, nutrient availability and productivity of a wheat crop after application of increasing doses of HS. For this purpose, two experiments were performed and evaluated, one is an incubation essay with a sandy soil and the other a greenhouse experiment with wheat plants using two distinct tropical soils as substrates (sandy and clayey). Microbial activity was stimulated in both experiments, and this transient stimulus may have been due to a priming effect caused by the addition of HS. However, there was no change in the bacterial community structure which may be due to the fact that HS is a recalcitrant material and already present in the soil. Nevertheless, we found an increase in the availability of phosphorus, potassium, calcium and nitrogen, as well as in the CEC in both, sandy and clayey soils, showing that HS do improve chemical properties of tropical soils. Furthermore, we found a significant effect on wheat productivity related parameters. There was an increase of dry weight of roots and shoots, as well as increased weight and number of grains, which may be reflected in a significant increase in productivity. These results are important because they show that even the most recalcitrant SOM fraction can positively influence soil chemical and biological aspects, contributing to the improvement of the production system and also to higher plant productivity.

Este trabalho apresenta uma análise comparativa da atividade biológica das células imobilizadas em um reator anaeróbio, tratando esgoto sanitário. Os materiais utilizados como suporte para o desenvolvimento do biofilme foram: um polimérico poroso (espuma de poliuretano), um polímero menos poroso (PVC), uma cerâmica porosa (cerâmica especial, desenvolvida pelo DEMA-UFSCar) e uma cerâmica menos porosa (tijolo refratário). A concepção do reator anaeróbio possibilitou o desenvolvimento do biofilme de forma igual, assim como a retirada desse suporte mantendo íntegra a biomassa aderida. Alguns suportes e o lodo retirados do reator anaeróbio foram colocados em reatores diferenciais para determinação dos parâmetros cinéticos. Esses parâmetros foram utilizados para avaliar a atividade biológica, tanto do biofilme aderido aos suportes quanto do lodo. As células imobilizadas nos materiais poliméricos apresentaram maior atividade biológica. Não foi verificada nenhuma diferença significativa na remoção dos constituintes da matéria orgânica (proteínas, carboidratos e lipídeos) entre as células imobilizadas. As análises microscópicas não constataram diferenças na predominância de algum gênero nos suportes ou lodo. O reator foi operado durante 183 dias, com remoção média de DQO de 50,5%, a taxa de carregamento orgânico média foi de 0,193 kg DQO.m3.dia-1. / This work presents a comparative analysis of the biological activity of immobilized biomass in a fixed-bed anaerobic reactor treating domestic sewage. The matrices used for biofilm growth were a porous polymer (polyurethane foam), a polymer with low porosity (PVC), a porous ceramic (special ceramic) and a ceramic with low porosity (refractory brick). The conception of the fixed-bed anaerobic reactor warranty the structure and integrity of the biofilm when samples of support material must be taken. Moreover, the conditions for biofilm growth were similar for all the materials used. After start-up period, some supports were taken from the fixed-bed reactor and transferred to differential reactors, where kinetic studies were carried out. Kinetic parameters were estimated for the different supports and for granulated sludge and used to evaluate the biological activity in a comparative way. The cells adhered to the polymeric supports presented higher activities and the highest activity was obtained when polyurethane foam was used. No differences were verified for organic matter consumption (as proteins, carbohydrates and lipids) when different matrices were used for cell immobilization. Microscopic analysis indicated that the morphological types in the biofilms were similar in ali support materiais. The fixed-bed reactor was operated for 183 days with mean COD removal efficiency of 50.5%, subjected to a organic loading rate of 0.193 Kg COD.m3.dia-1.

O estudo fitoquímico dos extratos de plantas adultas e plântulas da espécie Piper reticulatum, coletada na Floresta Nacional de Carajás, levou ao isolamento de quatro amidas, incluindo diidrowisanidina, descrita previamente nesta espécie, wisanidina, (2E,4E)-N-isobutileicosa-2,4-dienamida, (3E,5E,14E)-N-pirrolidileicosa-3,5,14-trienamida (inédita em função da posição das insaturações da cadeia alquílica), uma lignana (siringaresinol) e uma nitrila (benzoato de cianobenzila), além de terem sido propostas as estruturas de oito amidas contendo longa cadeia alquílica, com base nos dados de cromatografia gasosa e líquida acoplada a espectrometria de massas. Os extratos e produtos naturais isolados foram avaliados quanto aos seus potenciais antifúngicos frente aos fitopatógenos Cladosporium cladosporioides e C. sphaerospermum e atividade inibidora da enzima acetilcolinesterase, sendo constatada forte atividade para as amidas em ambos ensaios, além de forte atividade para a nitrila contra C. sphaerospermum. Além disso, os extratos de plantas adultas e plântulas de P. reticulatum foram analisados através de EM (ESI+) e os dados analisados por PCA (análise dos componentes principais) e através de HCA (análise por agrupamento hierárquico) juntamente com a espécie P. amalago, filogeneticamente relacionada com P. reticulatum, P. crassinervium, P. solmsianum, P. fuligineum, P. gaudichaudianum, P. aduncum e P. tuberculatum, afim de uma distinção ou classificação de grupos seguindo parâmetros metabólicos. / The phytochemical study of extracts from adult plants and seedlings of the species Piper reticulatum, collected in the Carajás National Forest, afforded the isolation of four amides, including dihydrowisanidine, previously described from this species, wisanidine, (2E,4E)-N-2-isobutyleicosa-2,4-dienamide, (3E,5E,14E)-N-3,5,14-pyrrolidyleicosa-trienamide (novel based on the unsaturations position at the alkyl chain), a lignan (syringaresinol) and a nitrile (cianobenzyl benzoate) and, additionaly eigh amides containing long alkyl chain had their structures proposed based on gas and liquid chromatography coupled to mass spectrometry data. The extracts and the isolated natural products were evaluated for their potential antifungal activity against the phytopathogens Cladosporium cladosporioides and C. sphaerospermum and acetylcholinesterase inhibitory activity, being found strong activity for amides for both two tests, and strong activity for nitrile against C. sphaerospermum. In addition, extracts of adult plants and seedlings of P. reticulatum were analyzed by MS (ESI +) and by PCA (principal component analysis) and HCA (hierarchical cluster analysis) together with the species P. amalago, phylogenetically related to P. reticulatum, P. crassinervium, P. solmsianum, P. fuligineum, P. gaudichaudianum, P. aduncum and P. tuberculatum in order to distinguish or classify them according to the metabolic profile.

4 ABSTRACT Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Botany Candidate: Viera Bodoríková Supervisor: doc. Ing. Lucie Cahlíková, Ph.D. Title of Diploma thesis: Synthesis of haemanthamine derivatives and their biological activity Haemanthamine, an isoquinoline Amarillidaceae alkaloid, exhibits a wide and important range of biological activities, including antitumor, antiviral, antioxidant, antimalarial and anticonvulsant. Biological activity of haemanthamine relatives closely with its structure. By modifying the different parts of the molecule, we can identify some structure-activity relationships. With this aim, the thirteen semisynthetic analogues of alkaloid haemathamine were prepared and purified using analytic and preparative TLC methods. The obtained substances were then subjected to structural analysis, specifically, there were used MS, HRMS, 1D and 2D NMR spectroscopic techniques. Prepared compounds were tested on its possibility to inhibit human erythrocytic acetylcholinesterase (HuAChE) and human serum butyrylcholinesterase (HuBuChE). The most promising biological activities have been shown by aromatic esters labelled as LC- 70 (IC50 HuAChE = 0,12 ± 0,01 µM) and LC-73 (IC50 HuAChE =0,17 ± 0,01 µM). The cytotoxic activity of prepared compounds has been...

Šípková, P.: Biological activity of secondary plants metabolites X. Alkaloids of Vinca minor L. Diploma thesis, Charles University, Faculty of Pharmacy in Hradec Králové, Department of Pharmaceutical Botany, Hradec Králové, 2019, 66 s. Key words: Apocynaceae, Vinca minor, alkaloids, isolation, biological activity, screening Alzheimer's disease is a progressive neurodegenerative disease. The number of affected patients is constantly increasing. This disease cannot be treated casually, therefore discovering and testing new substances that could potentially be used in a treatment is very important. The Vinca minor L. fraction after column chromatography was separated by flash chromatography. Isolation of the individual alkaloids was performed by preparative TLC. Based on NMR and MS analyses and comparison with literature, alkaloids were identified as vincarubine and (-)-vinoxine. Modified Ellman's method was used to test cholinesterase inhibitory activity of isolated alkaloids. Acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) are enzymes, that play a very important role in the pathofysiology of Alzheimer's diasease. (-)-Vinoxine showed relatively high activity against BuChE (IC50 = 24,61 ± 1,71 µM), inhibitory activity against AChE was insignificant (IC50 > 1000µM). Vincarubin did not...

Perennial plants from the Amaryllidaceae family are generally known for their beauty but also like herbs which contain wide range of alkaloids. To these days more than 500 alkaloids have been isolated. Amaryllidaceae alkaloids (AmA) are derivatived from aminoacid tyrosine and divided into nine basic groups. Biological activity of these substances icludes antitumor, antibacterial, antifungal, antiviral, antimalarial activity and some of them are used for treatment Alzheimer's disease (AD). Narcissus cv. PROFESSOR EINSTEIN have been choosen thanks to previous research of summary extract. Twelve alkaloids have been detected by GC-MS and ten of them have been identified (e.g.: lykoramine, pluviine, haemanthamine, pancracine, homolycorine). Due to this diversity of alkaloids and the fact that summary extract has relatively high inhibitory activity (IC50 = 49,99 ± 5,38 μg/mL) against HuBuChE Narcissus have been appropriate for isolation of the alkaloids and for further study of their biological activity. Summary ethanolic extract for gain pure compounds was prepared from 34.3 kg fresh bulbs. Separation was initiated by column chromatography and extract was divided into almost 500 fractions. Some of them were put together owing to TLC analysis and finally 27 subfractions were formed. Subfraction Nr. 26...

[Formulae and special characters can only be approximated here. Please see the pdf version of the Abstract for an accurate reproduction.] The kinetics for outer-sphere electron transfer between a series of cobalt(II) poly-aza cage ligand complexes and the iron(III) sarcophagine-type hexa-aza cage complex, [Fe(sar)]3+ (sar = 3,6,10,13,16,19-hexaazabicyclo[6.6.6]icosane), in aqueous solution have been investigated and the Marcus correlation is used to deduce the electron self-exchange rate constant for the [Fe(sar)]3+/2+ couple from these cross-reactions. The deduced electron self-exchange rate constant is in relatively good agreement with the experimentally determined rate constant (k ex calc = 4 ´ 10 5 M -1 s -1 ; k ex obs = 8 ´ 10 5 M -1 s -1 ). The successful application of the Marcus correlation to the electron transfer reactions of the Fe cage complex is consistent with the trend for the Co, Mn, Ni and Ru cage complexes which all follow the pattern of outer-sphere electron transfer reactivity expected from the Marcus-Hush formalism. A comparison of predictions based on the Marcus correlation with the experimentally determined kinetics of an extended series of cross reactions involving cobalt cage complexes with low-spin-high-spin cobalt(III)/(II) couples shows that electron transfer reactions involving large spin changes at the metal centre are not necessarily anomalous in the context of the adiabatic Marcus-Hush formalism. The results of this study also show that for suitable systems, the Marcus correlation can be used to reliably calculate the rates of outer-sphere electron transfer cross-reactions, with reaction free-energy changes spanning the range -6 to -41 kJ mol -1 and many different combinations of initial electronic configurations. Together, these results provide a coherent and internally consistent set of experimental data in support of the Marcus-Hush formalism for outer-sphere electron transfer. The results with the caged metal-ion systems also highlight the special nature of the mechanism of electron transfer in reactions of metal-aqua ions. ¶ A new range of symmetrically disubstituted hexa-aza sarcophagine-type cage ligand complexes are prepared in this study by the base-catalysed co-condensation of formaldehyde and a-methylene aliphatic aldehydes with cobalt(III) tris(1,2-diamine) precursors in acetonitrile solution. Encapsulation reactions based on the condensation of the weak carbon di-acids propanal and decanal with formaldehyde and the cobalt(III) tris(1,2-diamine) precursors, [Co(en)3 ] 3+ (en = 1,2-ethanediamine) and D-lel3 -[Co((R, R)-chxn)3 ] 3+ (chxn = 1,2-cyclohexanediamine), yield unsaturated cobalt(III) cage complexes with an endo-cyclic imine function in each cap. The Co III -coordinated endo-cyclic imine units of the cage ligands are reactive electrophiles that are readily reduced by the BH4 - ion to give the corresponding symmetrically di-substituted hexaamine macrobicyclic cage ligands. The nitromethane carbanion is also shown to add at the endo-cyclic imine function to yield a novel nitromethylated cage ligand complex. The latter reaction introduces a new method for the regioselective functionalisation of cage ligands at sites removed from the more commonly substituted bridgehead positions. The capping of cobalt(III) tris(1,2-diamine)-type complexes with weak CH-acids developed in this study introduces a new and more direct route to symmetrically di-substituted cage ligand complexes. ¶ A new range of cobalt(III) surfactant cage complexes, with linear octyl, dodecyl and hexadecyl hydrocarbon chains built directly into the bridgehead structure of the cage ligand, have been prepared by the base catalysed co-condensation of formaldehyde and long chain aliphatic aldehydes with the tripodal cobalt(III) hexaamine complex, [Co(sen)]3+ (sen = 4,4',4''-ethylidynetris(3-azabutan-1-amine)), in acetonitrile solution. Chiral surfactant cage complexes are obtained by capping reactions beginning with the optically pure L-[Co(sen)]3+ precursor complex. The cobalt(III) cage complexes with octyl to hexadecyl substituents are surface active and reduce the surface tension of water to levels approaching those of organic solvents. The dodecyl substituted cage complex forms micelles in aqueous solution when the concentration of cage complex is > 1 ´ 10 -3 mol dm -3 at 25 °C. The cobalt(III) cage head-group of these surfactants undergoes an electrochemically reversible one-electron reduction to the corresponding cobalt(II) cage complex. The reduction potential of the surfactant head group can be tuned to more positive potentials by replacing the bridgehead hydrocarbon chain substituent with an ether linked hydrocarbon chain. The cobalt(III) surfactant-cage complexes are biologically active and are lethal to the tapeworm Hymenolepis diminuta, and the vaginal parasites, Trichomonas vaginalis and Tritrichomonas foetus. The surfactant cage complexes also cause lysis in red-blood cell membranes at concentrations as low 10 -5 mol dm -3 . Their biological activity is linked to the high head-group charge (3+) and size which cause distortions in biological membranes when the membrane is treated with these molecules. The combination of the chemically reversible outer-sphere redox properties of the cobalt cage head-groups and the chirality of the head group introduces a new and possibly unique series of chiral surfactant coordination complexes which are also redox active. ¶ The chiral carboxylic-acid ionophore, lasalocid A, has been used to promote the selective supramolecular transport and extraction of cobalt(III) hexa-aza cage cations and related tripodal cobalt(III) complexes. The conjugate base anion of lasalocid A forms stoichiometric outer-sphere complexes with the cobalt(III) cage and tripod complexes. These outer-sphere complexes are highly lipophilic and partition strongly from water into a chloroform phase. The extraction of the dissymmetric cobalt(III) complexes by the chiral polyether anion is enantioselective for many systems and results in the partial resolution of initially racemic complexes in the aqueous phase. A strong structural preference was demonstrated by the ionophore for symmetrically disubstituted cobalt(III) hexa-aza cage cations with a D-absolute configuration of the ligand about the metal-ion and an R configuration of the coordinated secondary amine N-H groups. The lasalocid A anion was also shown to promote the transport of the complexes, intact, across a chloroform bulk-liquid membrane against an NH4 + concentration gradient. The transport of the cobalt(III) complexes was also enantioselective and resulted in partial resolution of the initially racemic aqueous phase. The most efficiently transported enantiomer of each complex was also the most efficiently extracted isomer in all systems examined, consistent with a transport process limited by interfacial diffusion. The magnitude of the enantiomer separation obtained in some systems was sufficient to indicate that lasalocid A mediated extraction and transport may become a practical method for the resolution of particular types of kinetically-inert chiral metal-amine complexes.

Heparin binding (HB) proteins mediates a wide range of important cellular processes, which makes this class of proteins biopharmaceutically important. Engineering HB proteins could bring many advantages, but it necessitates cost effective and efficient purification methodologies compared to the currently available methods. One of the most important classes of heparin binding protein is the fibroblast growth factors (FGFs) and its receptors (FGFRs). In this study, we report an efficient off-column purification of FGF-1 from soluble fractions and purification of the D2 domain of FGFR from insoluble inclusion bodies, using a weak amberlite cation (IRC) exchanger. This approach is an alternative to conventional affinity column chromatography, which exhibit several disadvantages, including time-consuming experimental procedures and regeneration and results in high cost for production of recombinant proteins. Authenticity of the purified proteins was verified by SDS-PAGE and MALDI mass spectrum analysis. Results of the heparin binding chromatography and steady state fluorescence experiments showed that the FGF-1 and the D2 are in a native biologically active conformation. The findings of this study will not only aid an in-depth investigation of this class of proteins but will also provide avenues for inexpensive and efficient purification of other important biological macromolecules.

Pyridazines are heterocyclic compounds with an N-N bond in their ring structure. Heterocyclic aromatic compounds are of great interest as a result of their novel properties and commercial applications. Our current research is focused on the potential role of pyridazines in next generation electronic devices that utilize organics as the semiconducting material. The synthesis of 5, 6-fused ring pyridazines beginning from fulvenes (Scheme 1) is described herein. These fused heterocycles will serve as synthetic models and building blocks for potential organic or organometallic conducting polymers. Our goal was to modify the route of Snyder et al. previously reported for the direct synthesis of pyridazine 2. This required improved synthesis of fulvene 1 and higher yields of 5. Additionally, a thorough analysis of the x-ray data was obtained to better understand the 3D aspects of this compound (pyridazine 2).This route was quite general and features an efficient and convenient synthesis. Single crystal X-ray analysis confirms the molecular structure of pyridazine 2. Full synthesis and characterization of newly formed pyridazine 2 and Fulvene 1 are reported.