Metabolite Profiling of 96% Ethanol Extract from Marsilea crenata Presl. Leaves Using UPLC-QToF-MS/MS and Anti-Neuroinflammatory Predicition Activity with Molecular Docking

Phytoestrogen is a group of compounds that can replace the estrogen function in the body. One of its roles was as anti-neuroinflammatory by inhibiting the microglia M1 polarity activation. Marsilea crenata Presl. is a plant that suspected to contain phytoestrogens. The aim of this research was to determine the metabolite profile of 96% ethanol extract of M. crenata using UPLC-QToF-MS/MS, and prediction the anti-neuroinflammatory activity of compounds with molecular docking. The 100 ppm of 96% ethanol extract in DCM and methanol were injected 5 µl each into the UPLC-QToF-MS/MS, and then analyzed by Masslynx 4.1 software to determine the compounds. The result of metabolite profiling shows a total 59 compounds in both DCM and methanol. Molecular docking was done with Autodock 4.2.6. After being analyzed, there are 3 compounds that are predicted to have activities similar to 17?-estradiol, they are prochlorperazine, 12-Aminododecanoic acid, and 1-methyl-2-[(4-methylpiperazin-1-yl)methyl]benzimidaol-5-amine hydrochloride. The results showed that the three compounds were predicted to be phytoestrogen compounds from M. crenata leaves, which have potential as anti-neuroinflammatory

Metabolite profiling of 96% ethanol extract from Marsilea crenata presl. leaves using UPLC-QToF-MS/MS and anti-neuroinflammatory predicition activity with molecular docking

Phytoestrogen is a group of compounds that can replace the estrogen function in the body. One of its roles was as anti-neuroinflammatory by inhibiting the microglia M1 polarity activation. Marsilea crenata Presl. is a plant that suspected to contain phytoestrogens. The aim of this research was to determine the metabolite profile of 96% ethanol extract of M. crenata using UPLC-QToF-MS/MS, and prediction the anti-neuroinflammatory activity of compounds with molecular docking. The 100 ppm of 96% ethanol extract in DCM and methanol were injected 5 µl each into the UPLC-QToF-MS/MS, and then analyzed by Masslynx 4.1 software to determine the compounds. The result of metabolite profiling shows a total 59 compounds in both DCM and methanol. Molecular docking was done with Autodock 4.2.6. After being analyzed, there are 3 compounds that are predicted to have activities similar to 17β-estradiol, they are prochlorperazine, 12-Aminododecanoic acid, and 1-methyl-2-[(4-methylpiperazin-1-yl)methyl]benzimidaol-5-amine hydrochloride. The results showed that the three compounds were predicted to be phytoestrogen compounds from M. crenata leaves, which have potential as anti-neuroinflammatory

Paederia foetida Linn Leaves-Derived Extract Showed Antioxidant and Cytotoxic Properties Against Breast Carcinoma Cell

Paederia foetida Linn. is a rich source of active compounds, yet its antioxidant and cytotoxic properties remain poorly studied. This current research aimed to investigate the chemical profiles of a crude extract derived from P. foetida leaves, its fractionated substances, and their antioxidant and cytotoxic properties on MCF-7 cells, a breast carcinoma cell. Six fractionated substances have been separated in thin layer chromatography (TLC) plate. Five fractions showed antioxidant properties, as indicated by the formation of the yellowish band on the TLC plate after spraying with the DPPH reagent. Furthermore, crude extract of P. foetida leaves and its fractions are also characterized by antioxidant properties against 2, 2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radicals. F1 fraction exhibited the most potent antioxidant activity with IC50 values of 10.94±2.67 and 50.04±0.48 µg/ml, respectively, toward DPPH and ABTS radicals. Crude extract and all six fractions also varied in their cytotoxic properties against MCF-7 cells, with IC50 values ranging from 410.24±0.30 to 831.57±6.91 µg/ml. F3 fractions showed the strongest cytotoxicity (IC50 value: 410.24±0.30 µg/ml). Methanolic extract and active plant leaf fractions contained flavonoid and phenolic compounds, as measured by aluminum chloride and Folin-Ciocalteu reagent, respectively. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis showed that phenol, flavonoid, alkaloid, limonoid, and steroid were the major compound found in the crude extract and fractions. In conclusion, the present study proved that crude extract of P. foetida leaves and its fractionated substances were potential as antioxidant and cytotoxic agents

Invetigation into the Mechanism of Action and Effects of Triticum Aestivum (Wheat) Grass

Wheat, (Triticum species) a cereal grass of the Gramineae (Poaceae) family, is the world's largest edible grain cereal-grass crop. Wheat has been a food crop for mankind since the beginning of agriculture. For over fifty years, researchers have known that the cereal plant, at this young green stage, is many times richer in the levels of vitamins, minerals and proteins as compared to seed kernel, or grain products of the mature cereal plant. The young germinated plant is a factory of enzyme and growth activity. In the early stages of growth they store large amounts of vitamins and proteins in the young blades. After jointing stage, the nutritional level in the leaves drops rapidly while the fiber content increases rapidly. Agriculturally, important species of Triticum include - Triticum aestivum, Triticum durum and Triticum dicoccum. Wheatgrass has been traditionally used, since ancient times, to treat various diseases and disorders. Presently, there are number of wheatgrass suppliers, in almost all cities of India, supply fresh wheatgrass, on daily basis to their regular customers by home-delivery system for various ailments and as a health tonic. Dr. Ann Wigmore, U. S. A. founder director of the Hippocrates Health Institute, Boston, U.S.A. was one of the proponents of the ‘‘Wheatgrass Therapy'. Dr. Wigmore claimed that wheatgrass is a safe and effective treatment for ailments such as high blood pressure, some cancers, obesity, diabetes, gastritis, ulcers, anemia, asthma and eczema. Scientific reports on nutritional analysis of wheatgrass have been published frequently in various journals. These reports and the chemical analyses undertaken reveal that wheatgrass is rich in chlorophyll, minerals like magnesium, selenium, zinc, chromium, antioxidants like beta-carotene (provitamin A), vitamin E, vitamin C, antianemic factors like vitamin B12, iron, folic acid, pyridoxine and many other minerals, amino acids and enzymes, which have significant nutritious and medicinal value

Étude des effets photosensibilisés de nouvelles molécules de synthèse : applications potentielles en photochimiothérapie antitumoral cutanée

Tese de doutoramento (co-tutela), Medicina (Dermatologia e Venereologia), Université Picardie Jules Verne, Universidade de Lisboa, 2011La thérapie photodynamique (PDT) cutanée est utilisée pour traiter les kératoses actiniques, la maladie de Bowen et les carcinomes basocellulaires. Malgré des résultats prometteurs, elle manque d'efficacité pour les lésions profondes et l'amélioration des photosensibilisateurs est d'actualité. Nous avons étudié les propriétés photophysiques et photosensibilisatrices de porphyrines tricationiques substituées. Elles sont fluorescentes et leur état excité triplet a une grande durée de vie et un rendement quantique élevé à l'exception du conjugué poly-S-lysine. L'oxygène singulet est formé quantitativement depuis ce triplet. L’étude chez des kératinocytes cutanés humains hyperproliférants de l'incorporation et de la photocytotoxicité montrent que 3 d'entre eux soutiennent favorablement la comparaison avec la protoporphyrine IX induite par l'acide δ-aminolévulinique (traitement actuel en PDT cutanée). Ils se localisent dans des vésicules d'endo- et pinocytose mais pas dans le noyau ni les mitochondries. La structure de leur chaîne substituée conditionne l'incorporation, la localisation et donc la phototoxicité. Les mécanismes de mort cellulaire étudiés avec le dérivé le plus efficace montrent l'absence de fixation de l'annexine V, d'activation des caspases, de condensation de la chromatine et donc d'apoptose. L'apparition de vésicules LC3-positives dans ces cellules transfectées par GFP-LC3 et la dégradation de p62 démontrent l’implication de l’autophagie dans la photocytotoxicité avec une régulation négative par JNK. En conclusion, ce sont des photosensibilisateurs intéressants nécessitant des études complémentaires dans des modèles cellulaires et animaux

53rd National Meeting of the Italian Society of Biochemistryand Molecular Biology (SIB)andNational Meeting of Chemistry of Biological Systems – Italian Chemical Society (SCI - Section CSB)

The 53rd National Congress of the Italian Society of Biochemistry and Molecular Biology (SIB), which will be held in Riccione from 23 to 26 September, is characterised by the elevated scientific level and interdisciplinary interest of the numerous sessions in which it is organised. The Scientific Programme comprises three joint Symposia of the SIB and the Chemistry of Biological Systems section of the Italian Chemistry Society (SCI) on Molecular Systems Biology, Chemistry of Nucleic Acids, Protein and Drug Structure, and Environmental Biotechnology. These Symposia address groundbreaking arguments, making the joint interest of the two societies particularly fascinating; the joint organisation of these events in fact signals the shared intention to proceed along the path of scientific exchange. The topics of the other sessions have been chosen by the Scientific Committee on the basis of their scientific relevance and topicality, with particular attention paid to the selection of the speakers. The SIB sessions will range from Signal Transduction and Biomolecular Targets, Protein Misfolding and its Relationship with Disease, Emerging Techniques in Biochemistry, Gene Silencing, Redox Signalling and Oxidative Stress, Lipids in Cell Communication and Signal Transduction, Mitochondrial Function and Dysfunction

Targeted Photodynamic Therapy of cancer using photoimmunoconjugates based on pyropheophorbide a derivatives

Photodynamic therapy (PDT) utilises light, oxygen and organic macrocycles, called photosensitisers, to produce reactive oxygen species that can kill malignant cells. Conventional PDT is associated with side effects that have stifled its advance and widespread use. These include low tumour selectivity, slow blood clearance and poor formulation. We proposed that an antibody fragment could be used to carry the photosensitiser to the target cells, significantly overcoming these limitations. Pyropheophorbide-a (PPa) was synthetically modified to enhance its water solubility obtaining two compounds, PS1 and PS4 each more water soluble than PPa. The use of Sonogashira coupling and short polyethylene glycol chains gave PS1, whereas the use of Suzuki coupling and a single positive charge gave PS4. The singlet oxygen quantum yields of these were improved compared to PPa with that of PS4 being 1.5 times higher than PPa. The in vitro characterisation of PPa, PS1 and PS4 using cytotoxicity assays did not correlate with their photophysical characterisation. PS4 was significantly less potent than PPa and PS1 on SKOV3 and KB human cancer cell lines. Confocal microscopy aided further characterisation using stains for intracellular organelles. PS1 was found to localise primarily in the ER and Golgi apparatus, similarly to PPa, while PS4 was found to localise mainly in the lysosomes. PS1 was conjugated to C6.5(-k), an anti-HER2 single chain variable fragment (scFv) using lysine coupling, to obtain a photoimmunoconjugate that was characterised in vitro and subsequently in vivo. In vitro characterisation showed increased potency and specificity but non-specific cell death attributed to the non-covalently bound photosensitiser was observed. However, in vivo therapy studies showed that the C6.5(-k)-PS1 photoimmunoconjugate could be used to cure SKOV3 subcutaneous tumours in nude mice, validating the use of targeted PDT as a successful targeted therapy with the potential to lower the effective drug dose and minimise side effects

Characterisation of duramycin as an anti-cancer agent

Duramycin is a relatively small (~2kDa) tetracyclic peptide that has a defined three-dimensional (3D) structure, this structure forms a stable binding pocket which specifically recognises the plasma membrane phospholipid phosphatidylethanolamine (PE). PE usually resides on the inner membrane layer though can become exposed on the outer membrane during physiological processes such as apoptotic cell death, cytokinesis and coagulation. Expression of cell surface PE has been observed on cancer cell lines, tumour endothelium and tumour-derived microparticles and observed in higher abundance in a variety of tumour xenografts compared to their normal tissue counterparts. There is a need in medical oncology for the development of novel, effective therapies and anti-cancer agents. A form of therapy that has gained interest is targeted anti-cancer therapy (TAT) which aims to bring about selective damage to tumour cells while limiting effect on surrounding normal tissue by utilising agents that recognise structures specific to tumours. Therefore, as duramycin had the potential to bind to PE exposed on cancer cell surfaces, it was theorised that it could be a promising targeted anti-cancer agent. Thus the aim of this project was to characterise duramycin’s anti-cancer properties and enhance its specificity to tumour cells. To achieve this a duramycin-porphyrin conjugate was developed as a novel photosensitiser and cancer cell lines were treated with photodynamic therapy (PDT). Duramycin was able to detect cell surface PE expression on cancer cell lines representing different cancer types including lymphoma, multiple myeloma and leukaemia and ovarian, pancreatic, breast and colon cancer. Duramycin was shown to have cytotoxic and anti-proliferative effects on cancer cell lines (ovarian and pancreatic), in both two-dimensional and 3D cell cultures, and a cytotoxic dose-dependent effect on a normal human endothelial cell line. Duramycin had been reported in the literature to have an effect on plasma membrane integrity and on a multitude of ion transport systems through the formation of membrane pores. Thus duramycin’s effect on cancer cell membranes was investigated along with a focus on whether the cytotoxic effect observed in the cancer cell lines was due to a form of calcium ion (Ca²⁺) overload. It was shown that duramycin treatment resulted in a concentration-dependent rise in intracellular Ca²⁺ concentration in cancer cell lines. The duramycin-porphyrin conjugate plus PDT light irradiation reduced cell proliferation of ovarian and pancreatic cancer cell lines in a dose-dependent manner and had a significantly enhanced effect over unconjugated duramycin and the free porphyrin. A preliminary investigation into the effect of duramycin-PDT treatment on tumour xenografts on the chorioallantoic membrane (CAM) of fertilised chicken eggs was undertaken with promising results. In summary, duramycin was shown to be an effective anti-cancer agent and was applicable to the PDT treatment of cancer in vitro and in a low level-in vivo tumour model and thus merits further studies

Non-covalent interactions in organotin(IV) derivatives of 5,7-ditertbutyl- and 5,7-diphenyl-1,2,4-triazolo[1,5-a]pyrimidine as recognition motifs in crystalline self- assembly and their in vitro antistaphylococcal activity

Non-covalent interactions are known to play a key role in biological compounds due to their stabilization of the tertiary and quaternary structure of proteins [1]. Ligands similar to purine rings, such as triazolo pyrimidine ones, are very versatile in their interactions with metals and can act as model systems for natural bio-inorganic compounds [2]. A considerable series (twelve novel compounds are reported) of 5,7-ditertbutyl-1,2,4-triazolo[1,5-a]pyrimidine (dbtp) and 5,7-diphenyl- 1,2,4-triazolo[1,5-a]pyrimidine (dptp) were synthesized and investigated by FT-IR and 119Sn M\uf6ssbauer in the solid state and by 1H and 13C NMR spectroscopy, in solution [3]. The X-ray crystal and molecular structures of Et2SnCl2(dbtp)2 and Ph2SnCl2(EtOH)2(dptp)2 were described, in this latter pyrimidine molecules are not directly bound to the metal center but strictly H-bonded, through N(3), to the -OH group of the ethanol moieties. The network of hydrogen bonding and aromatic interactions involving pyrimidine and phenyl rings in both complexes drives their self-assembly. Noncovalent interactions involving aromatic rings are key processes in both chemical and biological recognition, contributing to overall complex stability and forming recognition motifs. It is noteworthy that in Ph2SnCl2(EtOH)2(dptp)2 \u3c0\u2013\u3c0 stacking interactions between pairs of antiparallel triazolopyrimidine rings mimick basepair interactions physiologically occurring in DNA (Fig.1). M\uf6ssbauer spectra suggest for Et2SnCl2(dbtp)2 a distorted octahedral structure, with C-Sn-C bond angles lower than 180\ub0. The estimated angle for Et2SnCl2(dbtp)2 is virtually identical to that determined by X-ray diffraction. Ph2SnCl2(EtOH)2(dptp)2 is characterized by an essentially linear C-Sn-C fragment according to the X-ray all-trans structure. The compounds were screened for their in vitro antibacterial activity on a group of reference staphylococcal strains susceptible or resistant to methicillin and against two reference Gramnegative pathogens [4] . We tested the biological activity of all the specimen against a group of staphylococcal reference strains (S. aureus ATCC 25923, S. aureus ATCC 29213, methicillin resistant S. aureus 43866 and S. epidermidis RP62A) along with Gram-negative pathogens (P. aeruginosa ATCC9027 and E. coli ATCC25922). Ph2SnCl2(EtOH)2(dptp)2 showed good antibacterial activity with a MIC value of 5 \u3bcg mL-1 against S. aureus ATCC29213 and also resulted active against methicillin resistant S. epidermidis RP62A

53rd National Meeting of the Italian Society of Biochemistry and Molecular Biology (SIB) and National Meeting of Chemistry of Biological Systems – Italian Chemical Society (SCI - Section CSB)

Il 53° Congresso Nazionale della Società Italiana di Biochimica e Biologia Molecolare che si tiene a Riccione dal 23 al 26 Settembre si distingue per l'alto livello scientifico e l'interesse interdisciplinare delle numerose sessioni nelle quali è strutturato. Il Programma scientifico vede tre Simposi congiunti della SIB con la Sezione della Chimica dei Sistemi Biologici della Società Italiana di Chimica (SCI) su Molecular Systems Biology, Chemistry of Nucleic Acids, Protein and Drug Structure, Environmental Biotechnology. Questi Simposi, riguardano argomenti di avanguardia per i quali fa piacere l'interesse condiviso delle due Società, che per la prima volta organizzano dei Simposi congiunti a significare l'intento di procedere insieme negli scambi scientifici. Gli argomenti delle altre sessioni sono stati scelti dal comitato scientifico in base alla loro rilevanza e attualità scientifica, con particolare cura nella individuazione dei relatori. Le sessioni SIB spazieranno da Signal Transduction and Biomolecular Targets, Protein Misfolding and its Relationship with Diseases, Emerging Techniques in Biochemistry, Gene Silencing, Redox Signalling and Oxidative Stress, Lipids in Cell Communication and Signal Transduction, Mitochondrial Function and Dysfunction