Antioxidant and anti-inflammatory potential of Lavandula spp.

Lavenders have been used in folk medicine as disinfectant, expectorant, analgesic, anti-spasmodic, laxative and stimulant. Some species are endemic of Iberian Peninsula and often found in the Portuguese montado, including Lavandula stoechas subsp. luisieri and Lavandula pedunculata. The investigation of the antioxidant and anti-inflammatory potential of the Portuguese montado flora is very poor and restricted to a few botanical families, and their biological activities are mainly attributed to the essential oils. So, it is crucial to know the properties of Lavandula EOs, contributing for animal health and the valorisation of Portuguese montado flora. The aim of this study was to evaluate chemical composition, antioxidant properties and screening anti-inflammatory potential of EOs and extracts of Lavandula stoechas L. subsps. luisieri Rozeira, Lavandula pedunculata (Mill.) Cav. subsp. pedunculata and Lavandula viridis L’Hér, wild grown in the south of Portugal.

Seasonal variation, Chemical composition and antioxidante and antimicrobial proprieties of Calamintha nepeta

The aim of this study is to evaluate the chemical composition and the antioxidant and antimicrobial activities of EOs of C. nepeta collected in two different seasons, spring (leaves) and autumn (leaves and flowers) and to understand the relationship between seasonality composition and these biological activities. EOs were extracted by hydrodistillation of aerial parts of the plants wild grown in Évora (Alentejo) and their chemical composition was evaluated by GC-FID and GC-MS. Antioxidant activity was determined by β -carotene/linoleic acid system, total reducing power assay and DPPH radical methods [1]. Antimicrobial activity was assessed against Gram-negative and Gram-positive clinical isolates and food spoilage fungi [2,3].

Síntese de biguanidas e avaliação da sua atividade biológica em linhas de células tumorais

As biguanidas são um grupo de compostos com diversas atividades biológicas. Recentemente, esta família de compostos tem sido estudada não só pela sua atividade hipoglicemiante, mas também pela sua atividade anti-proliferativa. Um dos objetivos deste estudo foi a síntese de biguanidas, com cadeias laterais com diferentes estruturas e grupos funcionais. O trabalho desenvolvido permitiu a síntese de diversas biguanidas, tendo sido isolados quatro compostos. Outro dos objetivos deste estudo foi avaliar a atividade anti-proliferativa de biguanidas, na linha celular MDST8. Para esse efeito, foi desenvolvido inicialmente um método de quantificação celular com base na atividade ATPásica, testado nas linhas celulares MDST8, MCF7 e BRIN-BD11, tendo sido utilizado como referência a quantificação pelo método das desidrogenases. Os compostos estudados com melhor atividade anti-proliferativa apresentaram IC50 da ordem de 2,5 – 2,9 x10-3 M. Estes valores foram observados em biguanidas cujos grupos substituintes possuíam cadeias hidrocarbonadas cíclicas, alifáticas ou aromáticas p-substituídas, na sua estrutura; Abstract: Synthesis of biguanides and evaluation of their biologic activity in tumor cell lines Biguanides are a group of compounds which have diverse biological activities. Recently, this family of compounds has been studied not only for its hypoglycemic activity, but also for its anti-proliferative activity. One purpose of this study was the synthesis of biguanides, with side chains with different structures and functional groups. The work led to the synthesis of several biguanides, with the isolation of four compounds. Another objective of this study was the evaluation of the anti-proliferative activity of biguanides, in the cell line MDST8. To this aim, it was initially developed a cell quantification method based on the ATPase activity, tested in MDST8, MCF7 and BRIN-BD11 cell lines, with the dehydrogenases method used as reference. The studied compounds with better anti-proliferative activity had IC50 in the range from 2.5 to 2.9 x10-3 M for biguanides whose substituent groups had cyclic hydrocarbon, aliphatic or p-substituted aromatic chains in their structure.

Dedifferentiation of Leaf Cells and Growth Pattern of Calluses of Capsicum annuumcv. Etna.

Background:In vitrocell suspension cultivation systems have been largely reported assafe and standardized methods for production of secondary metabolites with medicinaland agricultural interest.Capsicum annuumis one of the most widely grown vegetablein the world and its biological activities have been demonstrated against insects, fungi,bacteria and other groups of organisms. The determination of procedures for thededifferentiation of cells into callus cells and the subsequent study of the callus growthpattern are necessary for the establishment of cellsuspensions and also to subsidizestudies regarding the bioactivity of its secondarymetabolites. To date, no study hasdescribed the development of protocols for callus induction inC. annuumL. cv. Etna. Objective:The objective of this study was to establish a protocol for dedifferentiationof leaf cells of the cultivarC. annuumcv. Etna and to determine the growth pattern ofthe calluses with a focus on the deceleration phase, when the callus cells must besubcultured into a liquid medium in order to establish cell suspension cultivationsaiming at the production of secondary metabolites.Results:The treatment that resultedin the highest %CI, ACCC and callus weight was thecombination of 4.52 μ M 2,4-D +0.44 μ M BA. The calluses produced were friable andwhitish and their growth patternfollowed a sigmoid shape. The deceleration phase started on the 23rdday of cultivation.Conclusion:Callus induction in leaf explants ofC. annuumcv. Etnacan be achieved inMS medium supplemented with 4.52 μ M 2,4-D + 0.44 μ MBA, which results in highcellular proliferation; in order to start a cell suspension culture, callus cells on the 23rdday of culture should be used.

Design and synthesis of (pro)electrophilic compounds for investigating the multifactorial nature of neurodegenerative diseases: focus on inflammation-driven events

Neuroinflammation constitutes a major player in the etiopathology of neurodegenerative diseases (NDDs), by orchestrating several neurotoxic pathways which in concert lead to neurodegeneration. A positive feedback loop occurs between inflammation, microglia activation and misfolding processes that, alongside excitotoxicity and oxidative events, represent crucial features of this intricate scenario. The multi-layered nature of NDDs requires a deepen investigation on how these vicious cycles work. This could further help in the search for effective treatments. Electrophiles are critically involved in the modulation of a variety of neuroprotective responses. Thus, we envisioned their peculiar ability to switch on/off biological activities as a powerful tool for investigating the neurotoxic scenario driven by inflammation in NDDs. In particular, in this thesis project, we wanted to dissect at a molecular level the functional role of (pro)electrophilic moieties of previously synthesized thioesters of variously substituted trans-cinnamic acids, to identify crucial features which could interfere with amyloid aggregation as well as modulate Nrf2 and/or NF-κB activation. To this aim, we first synthesized new compounds to identify bioactive cores which could specifically modulate the intended target. Then, we systematically modified their structure to reach additional pathogenic pathways which could in tandem contribute to the inflammatory process. In particular, following the investigation of the mechanistic underpinnings involving the catechol feature in amyloid binding through the synthesis of new dihydroxyl derivatives, we incorporated the identified antiaggregating nucleus into constrained frames which could contrast neuroinflammation also through the modulation of CB2Rs. In parallel, Nrf2 and/or NF-κB antinflammatory structural requirements were combined with the neuroprotective cores of pioglitazone, an antidiabetic drug endowed with MAO-B inhibitory properties, and memantine, which notably contrasts excitotoxicity. By acting as Swiss army knives, the new set of molecules emerge as promising tools to deepen our insights into the complex scenario regulating NDDs.

In Vitro Biological Screening Of The Anticholinesterase And Antiproliferative Activities Of Medicinal Plants Belonging To Annonaceae.

The aim of this research was to investigate the antiproliferative and anticholinesterase activities of 11 extracts from 5 Annonaceae species in vitro. Antiproliferative activity was assessed using 10 human cancer cell lines. Thin-layer chromatography and a microplate assay were used to screen the extracts for acetylcholinesterase (AchE) inhibitors using Ellman's reagent. The chemical compositions of the active extracts were investigated using high performance liquid chromatography. Eleven extracts obtained from five Annonaceae plant species were active and were particularly effective against the UA251, NCI-470 lung, HT-29, NCI/ADR, and K-562 cell lines with growth inhibition (GI50) values of 0.04-0.06, 0.02-0.50, 0.01-0.12, 0.10-0.27, and 0.02-0.04 µg/mL, respectively. In addition, the Annona crassiflora and A. coriacea seed extracts were the most active among the tested extracts and the most effective against the tumor cell lines, with GI50 values below 8.90 µg/mL. The A. cacans extract displayed the lowest activity. Based on the microplate assay, the percent AchE inhibition of the extracts ranged from 12 to 52%, and the A. coriacea seed extract resulted in the greatest inhibition (52%). Caffeic acid, sinapic acid, and rutin were present at higher concentrations in the A. crassiflora seed samples. The A. coriacea seeds contained ferulic and sinapic acid. Overall, the results indicated that A. crassiflora and A. coriacea extracts have antiproliferative and anticholinesterase properties, which opens up new possibilities for alternative pharmacotherapy drugs.

Monotherapy and combined therapy of new potential antitumor compounds: antiproliferative activities and biological targets

Review article Martins, P., Marques, M., Coito, L., Pombeiro, A.J.L., Baptista, P.V., Fernandes, A.R. 2014. Organometallic Compounds in Cancer Therapy: Past Lessons and Future Directions. Anti-cancer Agents in Medicinal Chemistry 14. PMID: 25173559

In vitro biological screening of the anticholinesterase and antiproliferative activities of medicinal plants belonging to Annonaceae

The aim of this research was to investigate the antiproliferative and anticholinesterase activities of 11 extracts from 5 Annonaceae species in vitro. Antiproliferative activity was assessed using 10 human cancer cell lines. Thin-layer chromatography and a microplate assay were used to screen the extracts for acetylcholinesterase (AchE) inhibitors using Ellman's reagent. The chemical compositions of the active extracts were investigated using high performance liquid chromatography. Eleven extracts obtained from five Annonaceae plant species were active and were particularly effective against the UA251, NCI-470 lung, HT-29, NCI/ADR, and K-562 cell lines with growth inhibition (GI50) values of 0.04-0.06, 0.02-0.50, 0.01-0.12, 0.10-0.27, and 0.02-0.04 µg/mL, respectively. In addition, the Annona crassiflora and A. coriacea seed extracts were the most active among the tested extracts and the most effective against the tumor cell lines, with GI50 values below 8.90 µg/mL. The A. cacans extract displayed the lowest activity. Based on the microplate assay, the percent AchE inhibition of the extracts ranged from 12 to 52%, and the A. coriacea seed extract resulted in the greatest inhibition (52%). Caffeic acid, sinapic acid, and rutin were present at higher concentrations in the A. crassiflora seed samples. The A. coriacea seeds contained ferulic and sinapic acid. Overall, the results indicated that A. crassiflora and A. coriacea extracts have antiproliferative and anticholinesterase properties, which opens up new possibilities for alternative pharmacotherapy drugs.

Directory of field activities of the Bureau of Biological Survey, 1939.

Includes index.

Directory of field activities of the Bureau of biological survey.

Issued June 1929; Revised October, 1932.

A Comparison Between Characterization And Biological Properties Of Brazilian Fresh And Aged Propolis.

As propolis is a highly valued bee product, we aimed to verify the quality of aged propolis, investigating their phenolic and flavonoid composition, levels of toxic metals, radical scavenging and antimicrobial activities. Samples of fresh and aged propolis of six different beekeepers, from the same geographical location, were investigated in terms of their phenolic and flavonoid composition and levels of Pb, Cd, and Cr, as well as radical scavenging and antimicrobial activities. The two groups of propolis had similar qualitative composition by HPLC-PDA and ESI(-)-MS. Fresh propolis and aged propolis show no differences when average values of extraction yield, flavonoids, EC50, or MIC were compared and both types of propolis showed good antimicrobial activity at low concentrations. Only levels of phenolic compounds were higher in fresh propolis. The propolis samples considered in this study, aged or fresh, had similar qualitative composition, although they were collected in different periods. Samples only differed in their levels of total phenolic content. Moreover, aged propolis conserves significant radical scavenging and antimicrobial properties. We suggest that aged propolis should not be discarded but explored for alternative applications.

A new approach for in vitro bioassay to measure tannin biological effects based on a gas production technique

The aim of this paper was to study a method based on gas production technique to measure the biological effects of tannins on rumen fermentation. Six feeds were used as fermentation substrates in a semi-automated gas method: feed A - aroeira (Astronium urundeuva); feed B - jurema preta (Mimosa hostilis), feed C - sorghum grains (Sorghum bicolor); feed D - Tifton-85 (Cynodon sp.); and two others prepared mixing 450 g sorghum leaves, 450 g concentrate (maize and soybean meal) and 100 g either of acacia (Acacia mearnsii) tannin extract (feed E) or quebracho (Schinopsis lorentzii) tannin extract (feed F) per kg (w:w). Three assays were carried out to standardize the bioassay for tannins. The first assay compared two binding agents (polyethylene glycol - PEG - and polyvinyl polypirrolidone - PVPP) to attenuate the tannin effects. The complex formed by PEG and tannins showed to be more stable than PVPP and tannins. Then, in the second assay, PEG was used as binding agent, and this assay was done to evaluate levels of PEG (0, 500, 750, 1000 and 1250 mg/g DM) to minimize the tannin effect. All the tested levels of PEG produced a response to evaluate tannin effects but the best response was for dose of 1000 mg/g DM. Using this dose of PEG, the final assay was carried out to test three compounds (tannic acid, quebracho extract and acacia extract) to establish a curve of biological equivalent effect of tannins. For this, five levels of each compound were added to I g of a standard feed (Lucerne hay). The equivalent effect showed not to be directly related to the chemical analysis for tannins. It was shown that different sources of tannins had different activities or reactivities. The curves of biological equivalence can provide information about tannin reactivity and its use seems to be important as an additional factor for chemical analysis. (C) 2007 Elsevier B.V. All rights reserved.

3`3-Ditrifluoromethyldiphenyl diselenide: A new organoselenium compound with interesting antigenotoxic and antimutagenic activities

The trace element selenium (Se), once known only for its potential toxicity, is now a well-established essential micronutrient for mammals. The organoselenium compound diphenyl diselenide (DPDS) has shown interesting antioxidant and neuroprotective activities. On the other hand, this compound has also presented pro-oxidant and mutagenic effects. The compound 3`3-ditrifluoromethyldiphenyl diselenide (DFDD), a structural analog of diphenyl diselenide, has proven antipsychotic activity in mice. Nevertheless, as opposed to DPDS, little is known on the biological and toxicological properties of DFDD. In the present study, we report the genotoxic effects of the organoselenium compound DFDD on Salmonella typhimurium, Saccharomyces cerevisiae and Chinese hamster lung fibroblasts (V79 cells). DFDD protective effects against hydrogen peroxide (H(2)O(2))-induced DNA damage in vitro are demonstrated. DFDD did not cause mutagenic effects on S. typhimurium or S. cerevisiae strains; however, it induced DNA damage in V79 cells at doses higher than 25 mu M, as detected by comet assay. DFDD protected S. typhimurium and S. cerevisiae against H(2)O(2)-induced mutagenicity, and, at doses lower than 12.5 mu M, prevented H(2)O(2)-induced genotoxicity in V79 cells. The in vitro assays demonstrated that DFDD mimics catalase activity better than DPDS, but neither presents Superoxide dismutase action. The products of the reactions of DFDD or DPDS with H(2)O(2) were different. as determined by electrospray mass spectrometry analysis (ESI-MS). These results suggest that DFDD is not mutagenic for bacteria or yeast; however, it may induce weak genotoxic effects on mammalian cells. In addition, DFDD has a protective effect against H(2)O(2)-induced damage probably by mimicking catalase activity, and the distinct products of the reaction DFDD with H(2)O(2) probably have a fundamental role in the protective effects of DFDD. (C) 2009 Elsevier B.V. All rights reserved.

Mercury Exposure Increases Circulating Net Matrix Metalloproteinase (MMP)-2 and MMP-9 Activities

Mercury (Hg) exposure causes health problems that may result from increased oxidative stress and matrix metalloproteinase (MMP) levels. We investigated whether there is an association between the circulating levels of MMP-2, MMP-9, their endogenous inhibitors (the tissue inhibitors of metalloproteinases; TIMPs) and the circulating Hg levels in 159 subjects environmentally exposed to Hg. Blood and plasma Hg were determined by inductively coupled plasma-mass spectrometry (ICP-MS). MMP and TIMP concentrations were measured in plasma samples by gelatin zymography and ELISA respectively. Thiobarbituric acid-reactive species (TBARS) were measured in plasma to assess oxidative stress. Selenium (Se) levels were determined by ICP-MS because it is an antioxidant. The relations between bioindicators of Hg and the metalloproteinases levels were examined using multivariate regression models. While we found no relation between blood or plasma Hg and MMP-9, plasma Hg levels were negatively associated with TIMP-1 and TIMP-2 levels, and thereby with increasing MMP-9/TIMP-1 and MMP-2/TIMP-2 ratios, thus indicating a positive association between plasma Hg and circulating net MMP-9 and MMP-2 activities. These findings provide a new insight into the possible biological mechanisms of Hg toxicity, particularly in cardiovascular diseases.