Morphoanatomy of the stem in Cyperaceae

Cyperaceae are usually perennial, with underground stems mainly rhizomatous, however, other stem types may also occur, such as corms and tubers. The underground stems of five Cyperaceae species were examined. Cyperus rotundus and Fuirena umbellata have plagiotropic rhizomes, while C. esculentus, C. odoratus, Hypolytrum schraderianum and Bulbostylis paradoxa have orthotropic rhizomes. Corms occur in C. rotundus and C. esculentus, and stolons in C. esculentus. The primary body originates from the activity of the apical meristem and later, from the primary thickening meristem (PTM). Secondary growth results from secondary thickening meristem (STM) activity, and occurs in rhizomes of H. schraderianum, B. paradoxa, C. odotarus and F. umbellata. The procambium and the PTM give rise to collateral bundles in H. schraderianum, and amphivasal bundles in the remaining species. The STM gives rise to the vascular system with the associated phloem and xylem. According to our results, the concept of stem type in Cyperaceae depends on external morphology, function, life phase, activity of the thickening meristems and the relative amount of parenchyma.

Gladiolus development in response to bulb treatment with different concentrations of humic acids

Gladiolus is an ornamental species produced for cut flowers and propagated by corms. The early flowering and increase in the number of flower buds, besides the production of commercial corms are constant challenges to be addressed in the crop improvement. Commercial production of ornamentals is technologically accelerated by means of growth regulators. Among them, the auxins stand out for their key role in the adventitious rooting and cell elongation. Alternatively, the humic substances present in the organic matter also have biostimulating effect, which is very similar to the auxinic effect. Therefore, this work aimed to study the growth and development of gladiolus in response to application of different concentrations of humic acids (HA) isolated from vermicompost. Corms were soaked for 24 hours in solutions containing 0, 10, 20, 30 and 40 mmol L-1 of C from HA. The corms were planted in 10-dm³ plastic bags filled with substrate and kept in a greenhouse. Growth of shoots and roots was evaluated. The results showed that the use of HA accelerates growth, and anticipates and increases flowering of Gladiolus.

Interacções de proteínas do plasma com potenciais fármacos: estudos 3D a nível molecular

Dissertação para obtenção do Grau de Mestre em Biotecnologia

Carbon monoxide, autophagy and cytoprotection in response to cerebral ischemia

Dissertação para obtenção do Grau de Mestre em Genética Molecular e Biomedicina

Complexos Organometálicos: caracterização preliminar de compostos de vanádio e de moléculas libertadoras de CO como potenciais fármacos

Vários complexos metálicos têm sido explorados visando uma melhoria de atuais tratamentos das mais variadas doenças. Os compostos de vanádio apresentam uma capacidade de mimetizar a ação da insulina, insurgindo-se assim como uma aposta no tratamento da doença Diabetes mellitus. O monóxido de carbono (CO), outrora visto como apenas um agente tóxico, atualmente é explorado pelas suas capacidades anti-inflamatórias, tendo-se desenvolvido moléculas libertadoras de CO, os CORMs, de modo a tirar partido dos seus efeitos biológicos. Com vista a determinar a formação de aductos entre os complexos organometálicos de vanádio e diferentes proteínas e caracterizar o tipo de interação estabelecida recorreu-se à Cristalografia de raios-X. A estrutura cristalográfica do aducto VOIV(picolinato)2–tripsina bovina foi obtida a 1,09 Å de resolução, mostrando o complexo VOIV(pic)2 no centro ativo da enzima, coordenado à cadeia lateral da serina catalítica (Ser195), numa geometria octaédrica. Ensaios cinéticos preliminares realizados com o complexo VO(pic)2 foram efetuados para determinar potenciais efeitos inibitórios na atividade das enzimas lisozima da clara do ovo (HEWL) e tripsina. O potencial terapêutico dos CORMs associado à Nanotecnologia foi explorado para o [Ru(CO)3Cl(glicinato)] (CORM-3), visando desenvolver nanopartículas de ouro (AuNPs) funcionalizadas com o composto e diferentes proteínas. A toxicidade celular direta do CORM-3, bem como alguns potenciais efeitos biológicos de CORM-3–albumina de soro humano (BSA) e CORM-3–HEWL foram também avaliados, recorrendo a ensaios de viabilidade celular , riscados celulares, e RT-qPCR em várias linhas celulares tumorais e linha primária de fibroblastos humanos. Foi possível iniciar o desenvolvimento de nanosistemas de AuNPs@PEG@BSA e AuNPs@PEG@HEWL, seguindo-se futuramente a conjugação com o CORM-3. Não foi possível determinar o efeito anti-inflamatório do CORM-3 e seus conjugados através da análise dos genes de resposta por RT-qPCR em tempo real, no entanto determinou-se que o composto CORM-3 não possui toxicidade em linhas celulares tumorais, e que aparenta fomentar a regeneração celular em fibroblastos e células tumorais NINA, sendo este efeito potenciado aquando conjugação com BSA.

First report of black rot of Colocasia esculenta caused by Ceratocystis fimbriata in Brazil

Ceratocystis fimbriata was found sporulating in gray to black discolored areas on edible corms of Colocasia esculenta found in supermarkets in the states of São Paulo, Rio de Janeiro, Bahia, Rondônia and the Distrito Federal. In most cases the corms were grown in the state of São Paulo. The black rot appeared to occur post-harvest. Sequences of rDNA indicated that the Colocasia sp. isolates belong to the Latin American clade of the C. fimbriata complex, but the isolates were more aggressive than isolates from Ficus carica and Mangifera indica, in pseudopetioles of C. esculenta.

Diversity, local uses and availability of medicinal plants and wild yams in the Mahafaly region of south-western Madagascar

Short summary: This study was undertaken to assess the diversity of plant resources utilized by the local population in south-western Madagascar, the social, ecological and biophysical conditions that drive their uses and availability, and possible alternative strategies for their sustainable use in the region. The study region, ‘Mahafaly region’, located in south-western Madagascar, is one of the country’s most economically, educationally and climatically disadvantaged regions. With an arid steppe climate, the agricultural production is limited by low water availability and a low level of soil nutrients and soil organic carbon. The region comprises the recently extended Tsimanampetsotsa National Park, with numerous sacred and communities forests, which are threatened by slash and burn agriculture and overexploitation of forests resources. The present study analyzed the availability of wild yams and medicinal plants, and their importance for the livelihood of the local population in this region. An ethnobotanical survey was conducted recording the diversity, local knowledge and use of wild yams and medicinal plants utilized by the local communities in five villages in the Mahafaly region. 250 households were randomly selected followed by semi-structured interviews on the socio-economic characteristics of the households. Data allowed us to characterize sociocultural and socioeconomic factors that determine the local use of wild yams and medicinal plants, and to identify their role in the livelihoods of local people. Species-environment relationships and the current spatial distribution of the wild yams were investigated and predicted using ordination methods and a niche based habitat modelling approach. Species response curves along edaphic gradients allowed us to understand the species requirements on habitat conditions. We thus investigated various alternative methods to enhance the wild yam regeneration for their local conservation and their sustainable use in the Mahafaly region. Altogether, six species of wild yams and a total of 214 medicinal plants species from 68 families and 163 genera were identified in the study region. Results of the cluster and discriminant analysis indicated a clear pattern on resource, resulted in two groups of household and characterized by differences in livestock numbers, off-farm activities, agricultural land and harvests. A generalized linear model highlighted that economic factors significantly affect the collection intensity of wild yams, while the use of medicinal plants depends to a higher degree on socio-cultural factors. The gradient analysis on the distribution of the wild yam species revealed a clear pattern for species habitats. Species models based on NPMR (Nonparametric Multiplicative Regression analysis) indicated the importance of vegetation structure, human interventions, and soil characteristics to determine wild yam species distribution. The prediction of the current availability of wild yam resources showed that abundant wild yam resources are scarce and face high harvest intensity. Experiments on yams cultivation revealed that germination of seeds was enhanced by using pre-germination treatments before planting, vegetative regeneration performed better with the upper part of the tubers (corms) rather than the sets of tubers. In-situ regeneration was possible for the upper parts of the wild tubers but the success depended significantly on the type of soil. The use of manure (10-20 t ha¹) increased the yield of the D. alata and D. alatipes by 40%. We thus suggest the promotion of other cultivated varieties of D. alata found regions neighbouring as the Mahafaly Plateau.

Tetrachlorocarbonyliridates: Water-Soluble Carbon Monoxide Releasing Molecules Rate-Modulated by the Sixth Ligand

A new family of compounds is presented as potential carbon monoxide releasing molecules (CORMs). These compounds, based on tetrachlorocarbonyliridate(III) derivatives, were synthesized and fully characterized by X-ray diffraction, electrospray mass spectrometry, IR. NMR, and density functional theory calculations. The rate of CO release was studied via the myoglobin assay. The results showed that the rate depends on the nature of the sixth ligand, trans to CO, and that a significant modulation on the release rate can be produced by changing the ligand. The reported compounds are soluble in aqueous media, and the rates of CO release are comparable with those for known CORMs, releasing CO at a rate of 0.03-0.58 mu M min(-1) in a 10 mu M solution of myoglobin and 10 mu M of the complexes.

Tetrachlorocarbonyliridates: Water-Soluble Carbon Monoxide Releasing Molecules Rate-Modulated by the Sixth Ligand

A new family of compounds is presented as potential carbon monoxide releasing molecules (CORMs). These compounds, based on tetrachlorocarbonyliridate(III) derivatives, were synthesized and fully characterized by X-ray diffraction, electrospray mass spectrometry, IR. NMR, and density functional theory calculations. The rate of CO release was studied via the myoglobin assay. The results showed that the rate depends on the nature of the sixth ligand, trans to CO, and that a significant modulation on the release rate can be produced by changing the ligand. The reported compounds are soluble in aqueous media, and the rates of CO release are comparable with those for known CORMs, releasing CO at a rate of 0.03-0.58 mu M min(-1) in a 10 mu M solution of myoglobin and 10 mu M of the complexes.

First report of black rot of Colocasia esculenta caused by Ceratocystis fimbriata in Brazil

Ceratocystis fimbriata foi encontrado em tubérculos de inhame (Colocasia esculenta), apresentando lesões escuras, pouco profundas, contendo estruturas de reprodução do fungo, cuja coloração variava do cinza ao negro. As amostras foram coletadas em supermercados, quitandas e varejões nos Estados de São Paulo, Rio de Janeiro, Bahia, Rondônia e Distrito Federal que, na maioria dos casos, comercializavam inhame produzido no Estado de São Paulo. Os sintomas de podridão negra indicam se tratar de uma doença de pós-colheita. Seqüências de rDNA indicam que os isolados de Colocasia sp. pertencem ao clado da America Latina do complexo C. fimbriata, embora esses isolados sejam mais agressivos em pseudo-pecíolos de C. esculenta do que os isolados de Ficus carica e Mangifera indica.

Mechanical Properties of Plant Underground Storage Organs and Implications for Dietary Models of Early Hominins

The diet of early human ancestors has received renewed theoretical interest since the discovery of elevated d13C values in the enamel of Australopithecus africanus and Paranthropus robustus. As a result, the hominin diet is hypothesized to have included C4 grass or the tissues of animals which themselves consumed C4 grass. On mechanical grounds, such a diet is incompatible with the dental morphology and dental microwear of early hominins. Most inferences, particularly for Paranthropus, favor a diet of hard or mechanically resistant foods. This discrepancy has invigorated the longstanding hypothesis that hominins consumed plant underground storage organs (USOs). Plant USOs are attractive candidate foods because many bulbous grasses and cormous sedges use C4 photosynthesis. Yet mechanical data for USOs—or any putative hominin food—are scarcely known. To fill this empirical void we measured the mechanical properties of USOs from 98 plant species from across sub-Saharan Africa. We found that rhizomes were the most resistant to deformation and fracture, followed by tubers, corms, and bulbs. An important result of this study is that corms exhibited low toughness values (mean = 265.0 J m-2) and relatively high Young’s modulus values (mean = 4.9 MPa). This combination of properties fits many descriptions of the hominin diet as consisting of hard-brittle objects. When compared to corms, bulbs are tougher (mean = 325.0 J m-2) and less stiff (mean = 2.5 MPa). Again, this combination of traits resembles dietary inferences, especially for Australopithecus, which is predicted to have consumed soft-tough foods. Lastly, we observed the roasting behavior of Hadza hunter-gatherers and measured the effects of roasting on the toughness on undomesticated tubers. Our results support assumptions that roasting lessens the work of mastication, and, by inference, the cost of digestion. Together these findings provide the first mechanical basis for discussing the adaptive advantages of roasting tubers and the plausibility of USOs in the diet of early hominins.

Development of advanced analytical methodologies for Alzheimer’s disease drug discovery

Advanced analytical methodologies were developed to characterize new potential active MTDLs on isolated targets involved in the first stages of Alzheimer’s disease (AD). In addition, the methods investigated drug-protein bindings and evaluated protein-protein interactions involved in the neurodegeneration. A high-throughput luminescent assay allowed the study of the first in class GSK-3β/ HDAC dual inhibitors towards the enzyme GSK-3β. The method was able to identify an innovative disease-modifying agent with an activity in the micromolar range both on GSK-3β, HDAC1 and HDAC6. Then, the same assay reliably and quickly selected true positive hit compounds among natural Amaryllidaceae alkaloids tested against GSK-3β. Hence, given the central role of the amyloid pathway in the multifactorial nature of AD, a multi-methodological approach based on mass spectrometry (MS), circular dichroism spectroscopy (CD) and ThT assay was applied to characterize the potential interaction of CO releasing molecules (CORMs) with Aβ1-42 peptide. The comprehensive method provided reliable information on the different steps of the fibrillation process and regarding CORMs mechanism of action. Therefore, the optimal CORM-3/Aβ1−42 ratio in terms of inhibitory effect was identified by mass spectrometry. CD analysis confirmed the stabilizing effect of CORM-3 on the Aβ1−42 peptide soluble form and the ThT Fluorescent Analysis ensured that the entire fibrillation process was delayed. Then the amyloid aggregation process was studied in view of a possible correlation with AD lipid brain alterations. Therefore, SH-SY5Y cells were treated with increasing concentration of Aß1-42 at different times and the samples were analysed by a RP-UHPLC system coupled with a high-resolution quadrupole TOF mass spectrometer in comprehensive data-independent SWATH acquisition mode. Each lipid class profiling in SH-SY5Y cells treated with Aß1-42 was compared to the one obtained from the untreated. The approach underlined some peculiar lipid alterations, suitable as biomarkers, that might be correlated to Aß1-42 different aggregation species.